Derivatization preferences for compounds containing active hydrogens.
\r\n\tIn the last decades, particular attention to this field has been paid to the coastal erosion problem all over the world. Indeed, the deployment of artificial reservoirs, modification of the runoff characteristics of internal areas, sand extraction from rivers, and harbor siltation, caused a decrease of sediment input on the coastal environments, and, therefore, a generalized deficit in the sediment budget. Often, dredging activities are required to collect sediment finalized to “soft” techniques to restore beaches or to move the sand trapped in the harbor (clean or contaminated).
\r\n\tMoreover, the coastal protections induced hydrodynamics and morphodynamics modifications inducing sometimes strong variations to the sediment transport regime.
\r\n\tHistorically, all these aspects are related to specific research areas ranging from engineering, geology, geomorphology, biology, etc, but it is difficult to find a comprehensive overview of these topics.
\r\n\r\n\tThis book is intended to collect original works and review concerning numerical and experimental investigation, theoretical works, methodological approaches, and any other technique that allow giving the actual state-of-the-art in the field of sediment transport.
",isbn:"978-1-80355-868-4",printIsbn:"978-1-80355-867-7",pdfIsbn:"978-1-80355-869-1",doi:null,price:0,priceEur:0,priceUsd:0,slug:null,numberOfPages:0,isOpenForSubmission:!1,isSalesforceBook:!1,isNomenclature:!1,hash:"e7b1c1592e32fe87af399022616ad0f8",bookSignature:"Dr. Davide Pasquali",publishedDate:null,coverURL:"https://cdn.intechopen.com/books/images_new/11136.jpg",keywords:"Longshore Sediment Transport, Sediment Budget, Morphodynamics, Hydrodynamics, Sediment Transport, Sedimentation, Mathematical Modelling, Erosion and Deposition, Dredging, Harbor Siltation, Contaminated Sediment, Water Quality",numberOfDownloads:65,numberOfWosCitations:0,numberOfCrossrefCitations:0,numberOfDimensionsCitations:0,numberOfTotalCitations:0,isAvailableForWebshopOrdering:!0,dateEndFirstStepPublish:"November 4th 2021",dateEndSecondStepPublish:"February 23rd 2022",dateEndThirdStepPublish:"April 24th 2022",dateEndFourthStepPublish:"July 13th 2022",dateEndFifthStepPublish:"September 11th 2022",dateConfirmationOfParticipation:null,remainingDaysToSecondStep:"4 months",secondStepPassed:!0,areRegistrationsClosed:!0,currentStepOfPublishingProcess:4,editedByType:null,kuFlag:!1,biosketch:"Davide Pasquali is currently a Research Fellow in the Department of Civil, Construction-Architectural, and Environmental Engineering (DICEAA) at the University of L’Aquila. 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One is the continuous demand for more sensitive and accurate analytical methods. The other is the desire for simpler methods that require as little as possible human intervention. One of the various procedures to make the analytical methods more sensitive and accurate is the use of specific chemical changes (e.g., derivatization) applied on the analytes or even on the whole sample. However, these changes frequently involve more human intervention than the direct use of advanced instrumentation. For this reason, the methods involving chemical changes such as derivatizations are not necessarily the first choice when selecting an analytical method. Nevertheless, in many cases, the benefits of derivatization are more important than the disadvantage of requiring human intervention, and for this reason, derivatization is still frequently used in the analytical practice. Also, modern GC, GC/MS (or GC/MS/MS) instrumentation may offer autosampling with the capability of adding reagents to the sample, as well as stirring, heating, and injecting the sample at specific time intervals in the GC system. This type of instrumentation may reduce significantly the human handling involved in derivatization.
\nVarious chemical changes can be performed on an analyte in order to make it suitable for a specific method of analysis. The most common is derivatization, but other chemical changes can be utilized, for example, pyrolytic decomposition and, in the case of polymers, polymer fragmentation using reagents. The choice depends on the nature of the analyte, the sample matrix, the intended changes in the analyte properties, and the analytical method to be used.
\nThe addition of a reagent on a sample may produce a chemical reaction only with the analytes without affecting the matrix. However, it is also possible that some matrix components are derivatized unintentionally. Usually, it is preferable to have only the analytes derivatized since in this way a better separation from the matrix is expected. Some derivatizations are used in the sample cleanup or concentration process. Also, the derivatization process may be combined with simultaneous extraction and concentration of the sample or may be followed by a second preparation step before the chromatographic analysis. More frequently, the derivatization is done to change the analyte properties for the core analytical procedure (GC, GC/MS, etc.).
\nDerivatization can be applied before the core chromatographic process or after it. Precolumn derivatization takes place before the separation and postcolumn derivatization after it. In GC precolumn derivatization is much more common and most derivatizations are performed “offline.” There are however derivatizations that can be done “online,” for example, in the injection port of the GC such as some methylations using tetramethyl ammonium hydroxide (TMAH). Postcolumn derivatizations are performed only for enhancing the detectability of the analytes. Typically, they must be done “online” and should be completed in the specific time frame needed by the analyte to reach the detector.
\nA wide variety of derivatization reagents and procedures are described in the literature, with the reagents carrying specific moieties that provide a desired property to the analytes, as well as with specific reactive groups that permit the reaction with the analyte. Multiple step derivatizations as well as derivatizations followed by a second one are known.
\nDerivatization is not always the first step in sample preparation. Sample preparation typically includes other operations, besides derivatization. Some of these steps are more complex such as sample cleanup or concentration and others more simple such as pH adjustments, addition of proton acceptors or donors, change of the medium (from one solvent to another), and addition of catalysts to enhance the derivatization, and these may be necessary for a successful derivatization.
\nAlthough derivatization is performed in order to make possible or to improve the results of a chemical analysis, there are also some disadvantages of using derivatization. Besides the potential need of more manpower for the analysis, the addition of more operations applied on the sample (including the analytes) can be a source of additional errors. In particular the involvement of a chemical reaction that may not be perfectly controlled can bring significant errors in the analytical results. To minimize the potential errors when using derivatization, specific aspects of the derivatization must be considered in its choice, such as the efficiency of the chemical reaction used in the derivatization, the stability of the derivatized analytes, the availability of reagents and necessary equipment, and the time necessary for performing the analysis. For a given analyte or group of analytes, the reaction with the derivatization reagent must be complete or at least close to complete, must take place in a length of time that is not prohibitive, and must have very little loss of the analyte with formation of artifacts or decomposition products. Only when such criteria are satisfied can a specific chosen derivatization be applied successfully.
\nThe application of derivatization in chromatography is the subject of many studies. Numerous derivatizations have been reported in journals (e.g.,
For GC analysis, the effect of derivatization can be beneficial in a variety of circumstances. Some of the most common uses of derivatization for improving the GC separation are the following:
(a) Derivatization that replaces active (polar) hydrogen atoms in the analyte to decrease its boiling point. The active hydrogens in a chemical compound typically enhance the capability to form hydrogen bonds and increase the compound polarity. For this reason, many compounds containing active (polar) hydrogens are not volatile, the volatility being necessary for using GC or GC/MS as a core analytical method. Derivatization can be used to replace active hydrogens from an analyte Y-H (or Y
In reaction (1), the reagent R-X contains an “active” group X and a group R that carries a desired property (e.g., lack of polarity for GC). Group R in the reagent can be a low molecular mass fragment such as CH3 or C2H5, a short-chain fluorinated alkyl in alkylation reactions, Si(CH3)3 or other silyl groups in silylations, COCH3 or short-chain fluorinated acyl groups in acylations, etc. An example of a chromatogram resulting from the GC/MS analysis of a silylated tobacco sample is given in Figure 1. Tobacco contains many hydroxy acids such as malic, trihydroxybutanoic, citric, quinic, glucuronic, and chlorogenic. Also, it contains monosaccharides (e.g., glucose, fructose), disaccharides (e.g., sucrose), and even trisaccharides. None of these compounds are volatile, having numerous active hydrogens. The replacement of these hydrogens with Si(CH3)3 by silylation renders these compounds volatile, and they can be analyzed by GC/MS as seen in Figure 1.
(b) Derivatization for enhancing the separation. Specific moieties added to an analyte may be necessary for enhancing the separation. This is frequently practiced for general GC separations and is also very useful for the separation of chiral molecules (see Section 4). The derivatized analytes may have significantly different properties from each other, for example, regarding polarity and implicitly in their boiling point, allowing separations that are difficult to achieve otherwise. Also, derivatization may generate more significant differences between the analytes and the matrix components.
(c) Derivatization that replaces active hydrogens in the analyte to improve the behavior of the analyte in the chromatographic separation. The chromatographic column (e.g., a capillary column coated with a bonded stationary phase) may display additional capability to interact with polar molecules, besides the intended interactions due to its bonded phase. This may come, for example, from the silica wall of the column. Secondary interactions taking place with only a portion of the molecules of the analyte generate peak tailing. This is exemplified in Figure 2 which shows a hypothetical case of two different types of interaction between the column and a specific molecular species.
(d) Derivatization for the improvement of stability of a compound. This stability may refer to thermal stability, a property which overlaps to a certain extent to what was described at point (a). However, even some volatile compounds may be further thermally stabilized by derivatization. Also, chemical stability can be enhanced by protecting specific groups in the analyte using derivatization. For example, thiols can be protected using derivatization against oxidation by the traces of oxygen in the heated injection port of the GC.
GC/MS chromatogram of a silylated tobacco sample, with separation on a DB-5 MS column from Agilent (Agilent Technologies Inc., Wilmington, DE, USA) (Note: an internal standard I.S. was added to the sample).
Peak tailing due to multiple retention mechanisms.
The choice of the appropriate derivatization is not always a simple task. The replacement of a hydrogen atom with a group of atoms may increase the molecular weight of the derivatized analyte. In such cases, it must be verified that the increase in the molecular weight by derivatization brings no or only a small increase in the boiling point of the analyte. Most of the time, low molecular weight substituents such as CH3 or Si(CH3)3 are preferable for GC analysis to the active hydrogens for achieving the previously described goals. Large substituents may increase the boiling point too much and make the compound not acceptable for GC analysis.
\nBesides replacement of active hydrogens, other derivatization reactions can be utilized. For example, condensation reactions may decrease the boiling point and improve the thermal stability of an analyte. However, the generation of new active hydrogens must be avoided in condensation reactions or must be followed by a second derivatization.
\nThe compounds with structures that are mirror images to each other are indicated as enantiomers, and their molecules are not superimposable, having the property called chirality. Chirality is commonly caused by the existence in the molecule of at least one tetrahedral carbon atom substituted with groups that are different. However, chiral molecules may be generated with a phosphorus or a sulfur chiral atom. Not only chiral centers (such as an asymmetric carbon) generate enantiomers, but a chiral axis or a chiral plane can lead to enantiomers. The chirality in an enantiomer is specified using the symbols R and S based on specific rules. For the assignment of a symbol R or S to a chiral carbon, the substituents are arranged in a sequence a > b > c > d. For the four atoms directly attached to the asymmetric carbon, a higher atomic number outranks the lower, and a higher atomic mass outranks the lower mass. For the same atoms directly attached to the asymmetric carbon, the priorities are assigned at the first point of difference. After the sequence is established, the molecule is oriented in space with the group “d” of the lowest priority behind the asymmetric carbon. When viewed along the C─d bond (from C) and the three substituents a, b, and c are oriented clockwise, the compound contains an R asymmetric carbon, and it contains an S asymmetric carbon for counterclockwise arrangement.
\nMore than one asymmetric carbon can be present in a molecule, as in the case of carbohydrates. The stereoisomers generated by more than one asymmetric carbon can be mirror image one to the other (enantiomers) or may have different steric arrangements being diastereoisomers. These types of molecules are schematically shown in Figure 3.
\nCompounds with two chiral centers.
The (S,S)- and the (R,R)-compounds from Figure 3 are enantiomers, while the (S,R)-compound is a diastereoisomer to both (S,S)- and to (R,R)-compounds (it is an enantiomer to the (R,S)-compound). The gas chromatographic separation of enantiomers can be done only using chromatographic columns having chiral stationary phases. The derivatization of enantiomers with non-chiral reagents generates molecules that remain enantiomers. This type of derivatization may improve the chromatographic separation from other molecules, but the derivatized compounds of remaining enantiomers cannot be separated except on chiral stationary phases. Sometimes, better separation can be obtained even between the enantiomers (on chiral chromatographic columns) after derivatization. One such example is the separation of (R)- and (S)-nornicotine derivatized with isobutyl chloroformate on a chiral Rt-BDEXsm column with separation improved compared to that of underivatized enantiomers [6]. The derivatization reaction is indicated below:
\nDiastereoisomers can be separated on chromatographic columns with non-chiral stationary phases which offer a much wider possibility to select the column. For this reason, an alternative procedure toward the separation of enantiomers is using derivatization with chiral reagents. This type of derivatization generates diastereoisomers which can be separated on non-chiral stationary phases.
\nA discussion on the separation of enantiomers on chiral phases without derivatization is beyond the purpose of this chapter. Numerous publications are dedicated to this subject, including papers published in general chromatography journals or in dedicated journals (e.g.,
The separation after derivatization with a pure enantiomer reagent is based on formation of diastereoisomers that can be separated on regular stationary phases. Depending on the nature of the analyte and of the derivatization, different separation techniques can be applied. A variety of common columns are used for such GC separations. The choice of the column is again dependent on the analyte and the derivatization procedure. For example, α-substituted organic acids such as α-chloropropionic, α-bromocaproic, etc. can be derivatized with a specific enantiomer of an amino acid ester (e.g., ethyl 2-aminopropanoate) in the presence of a peptide coupling reagent (benzotriazol-1-yl-oxy-tris(dimethylamino)-phosphonium hexafluorophosphate or BOP) in a reaction of the type:
The derivatized acids that are now diastereoisomers (R,S) and (S,S) can be separated on a common capillary column (e.g., a DB-1701 column from Agilent). Another example of derivatization with a chiral reagent is that of methamphetamines with (R)-menthyl chloroformate. This derivatization allows the separation of over-the-counter (R)-methamphetamine from the illicit (S)-methamphetamine. The reaction of the (R)-enantiomer is indicated below [8]:
The separation of the (R,R) and (S,R) derivatives was possible on a non-chiral column for a GC/MS analysis.
\nGas chromatography (not coupled with mass spectrometry, GC/MS being separately presented) used as an analytical technique can involve various detectors. The variety of such detectors is rather large, and several types include the following: thermal conductivity detector (TCD), flame ionization detector (FID), nitrogen-phosphorus detector (NPD), electron capture detector (ECD), flame photometric detector (FPD), photoionization detector (PID), electrolytic conductivity (Hall), sulfur chemiluminescence, nitrogen chemiluminescence, aroyl luminescence detector (ALD), atomic emission detector (AED), helium ionization detector (HID), vacuum ultraviolet (VUV) absorbance, infrared Doppler (IRD) absorption, FID with catalytic conversion of all analytes in CH4 (e.g., Polyarc system [9]), etc. The derivatization with the purpose of improving detectability in GC is determined by the type of detector utilized. Most derivatizations are performed precolumn, even if they are applied only with the purpose of improving detection. However, it is important that the derivatization for improving detection does not deteriorate the separation. Preferably, both the detection and the chromatographic separation are improved by the same derivatization. Some specific postcolumn reactions applied to the analytes are part of certain types of detectors such as chemiluminescence detectors, atomic emission detectors (AED), and FID with catalytic conversion into CH4. Some of these chemical changes in the analytes are not necessarily classified as derivatization reactions.
\nNo specific derivatization is usually recommended to improve sensitivity when using nonselective detectors such as TCD and FID. However, in some cases when the detector is not sensitive to a specific analyte, such as formaldehyde or heavily halogenated compounds, derivatization can be used to enhance detection.
\nIn case of NPD detector, derivatization with nitrogenous compounds can be done, which should give a higher sensitivity. However, this type of derivatization is not very common. An adverse result occurs for the NPD detectors when silylation is performed on the sample. Besides a possible reduction in the NPD response on silylated compounds containing nitrogen, a drastic decrease in the lifetime of the detector may occur, probably due to the excess of silylating reagent that commonly is injected with a derivatized sample and affects the alkali active element of the NPD.
\nThe response of the photoionization detector (PID) depends on the ionization potential of the analyte, and compounds with higher ionization potential are not sensitive in PID, while those with lower ionization potential may have excellent sensitivity, as low as 10−12 mg of sample. A derivatization resulting in lowering the ionization potential of the analyte may be beneficial for PID detection. However, derivatization for enhancing PID response is not frequently used.
\nSome detectors such as electron capture detectors (ECD) may benefit very much from certain derivatization types. ECD (as well as negative chemical ionization mass spectrometry or NCI-MS) can be extremely sensitive, but they are selective to compounds that are able to form more stable negative ions. ECD, for example, can have sensitivity as low as 10−13 mg of analyte in the detector compared to the best sensitivity of FID that can be 10−8 to 10−11 mg of analyte. The efficiency of the process seems to be related to the ease of attaching an electron on the molecule. In ECD this process can be written as follows:
With some exceptions, ECD response can be correlated with the electron affinity of the analyte [4]. In general, the halogen substituents increase the sensitivity in ECD in the order I > Br > Cl > F. Multiple substitutions seem to have a cumulative effect. Besides halogens, nitro groups seem to have an effect similar to chlorine groups. For aromatic compounds, the substituents affect the sensitivity of the ECD according to their electron withdrawing capability. Strong electron withdrawing groups such as NO2 increase the sensitivity of the detection, while electron donating groups reduce it.
\nA variety of substitution groups containing electronegative elements (halogens) or nitro groups can be attached to an analyte. The procedure to attach these groups is in most cases the typical substitution of an active hydrogen in the analyte Y-H with a group R from a reagent R-X that has the appropriate active X group. Some groups used for enhancing ECD (as well as NCI-MS) sensitivity following an alkylation or aryl derivatization reaction are shown in Figure 4, and several substitution groups introduced by acylation, chloroformylation, or sulfonation used for the same purpose are shown in Figure 5. Besides alkylation or aryl derivatization, other derivatization techniques used to replace an active hydrogen are applied to introduce into a molecule as a substituent containing halogens or nitro groups enhancing significantly the detectability of the derivatized analytes by ECD (as well as NCI-MS). Silylation, for example, can be used for this purpose when silyl groups used for derivatization contain halogens. Several silyl groups containing halogens that can be attached to an analyte by silylation with special reagents are given in Figure 6 [4].
\nSubstitution groups used in alkylation and aryl derivatization for enhancing ECD (and NCI-MS) detectability (the masses are considered only for the most abundant isotope.).
Substitution groups used in acylation chloroformation and sulfonation for enhancing ECD (and NCI-MS) detectability.
Substitution groups used in silylation for enhancing ECD (and NCI-MS) detectability.
The most powerful tool used for compound identification purposes is very likely mass spectrometry (spectroscopy). This technique is capable to provide information from very low amounts of material such as that eluting from a chromatographic column and can be easily coupled with a gas chromatograph. Most analyses performed with MS detection (GC/MS or GC/MS/MS) are using EI+ ionization mode with electron impact at 70 eV. The electrons interact with the molecule A to eject an additional electron leaving a positively charged species (with an odd number of electrons) of the type A▪+. The ions also receive energy during electron impact and the excess of energy induces fragmentation. For most molecules, this process can be written as follows:
The fragments Bi+ are commonly but not always with an even number of electrons. The formation of molecular ions takes place with a range of internal energies, and more than one fragmentation path is possible for a given molecule. Also, the fragments can suffer further fragmentations. In general, the most abundant fragment ion results from the fragmentations that form the most stable products (ion and neutral radical). The abundance of a fragment ion is affected by its stability. For this reason, the intensity of the response of a mass spectrometric detector can be very different for different molecular species, and the prediction of this intensity is difficult. As a result, the improvements in the sensitivity in EI + −type mass spectrometry (in GC/MS using EI+ ionization) are not usually sought (but not impossible) through derivatization.
\nDerivatization for enhancing sensitivity is, however, frequently applied in NCI-MS. In this technique, the electrons interact with the molecules of the CI gas which is lowering their energy but without forming ions. The ionization of analyte molecules takes place by interaction with the low-energy electrons or with already formed negative ions by electron capture, dissociative electron capture, ion pair formation, or ion molecule reaction. The ionization process with the formation of negative ions is efficient only for molecules with positive electron affinities. For this reason, the sensitivity in NCI-MS is highly dependent on the electron affinity of the analyte, similarly to the sensitivity in ECD. For enhancing the electron affinity, the derivatization with reagents containing, for example, fluorinated moieties (indicated in Figures 4, 5, 6) is practiced. The sensitivity of the analytical methods where such derivatization is applicable can have very good sensitivity. For example, derivatization with heptafluorobutyric anhydride of aromatic amines that are present at low trace level in cigarette smoke leads to limit of detection (LOD) values as low as 0.05 ng/cig. for compounds such as 4-aminobiphenyl [10, 11].
\nThe fragmentation pattern generated by EI+ ionization mode that generates a specific mass spectrum of a molecule is very likely the most utilized technique for the identification of the molecular species. For this identification, large libraries of mass spectra are available, and computer algorithms are used for automatic searches. The identification of compounds using mass spectroscopy is not a simple process even with the capabilities offered by the electronic searches in the mass spectral libraries. This is particularly true for analysis of complex mixtures or when the analyzed compound is present in traces. Some compounds do not have a very characteristic mass spectrum, or during the chromatographic process, the separation is not achieved, and it is difficult to make an identification due to the spectra overlapping. Also, numerous compounds may have a mass spectrum that matches more than one compound (with a good quality fit). In such cases, a derivatization with the purpose of obtaining a compound that forms more informative fragments in the mass spectrum can be very useful.
\nThe fragments from derivatized compounds can be used for the identification of unknown compounds using library searches and even when the mass spectrum is not available in the libraries. As an example, the derivatization by silylation allowed the identification of a new pentacyclic triterpenoid present in several bioactive botanicals [12]. An unidentified compound with MW = 456.7 was detected by LC/MS/MS in a rosemary extract. The structure of the compound was elucidated after silylation of the plant material based on the comparison of mass spectrum of the unidentified compound with that of silylated betulinic acid. The new compound was identified as (3β)-3-hydroxy-lupa-18,20(29)-dien-28-oic acid (or betul-18-en-oic acid). The mass spectra of the two acids are shown in Figure 7.
\nMass spectrum of silylated betulinic acid and that of silylated betul-18-en-oic acid.
The two mass units difference between different fragments from the mass spectra of the two compounds allowed the identification of the new compound structure. Neither free betulinic acid nor betul-18-en-oic acid are volatile, such that the use of GC/MS for identification was possible only after derivatization.
\nAnother special procedure that may be utilized for compound identification based on mass spectra is the use of two parallel derivatizations, one of them being done with an isotope-labeled reagent. Common labeling isotopes are 2H (deuterium, d), 13C, 15N, etc. One such isotopic labeling can be done, for example, using silylation with d18-N,O-bis(trimethylsilyl)-trifluoroacetamide (d18-BSTFA). Derivatization of an aliquot of sample with regular BSTFA and another with d18-BSTFA provides a pairing chromatogram with peaks at retention times that have only small differences from the first but with spectra differing by a number of units. The comparison of the spectra for corresponding peaks (based on retention time) of a given compound allows the calculation of the number of silyl groups attached to that compound. In addition, the fragmentation in the spectra can be better interpreted allowing easier compound identification.
\nDerivatization in GC/MS analysis may have multiple other utilizations and benefits. For example, quantitative analysis frequently utilizes isotopically labeled internal standards. In an analysis with multiple analytes, addition of an isotopically labeled internal standard for each analyte may become a complex process. When a derivatization is involved in the analysis, this can be done with a non-labeled reagent for the analytes in the sample, while the internal standards are obtained by derivatization of standards with the same reagent but isotopically labeled. Such technique has been proven to be very successful, for example, in the analysis of multiple amino acids (but using an LC/MS/MS procedure [13]).
\nDerivatizations as chemical reactions can be classified as follows: (1) reactions with formation of alkyl or aryl derivatives, (2) silylation reactions, (3) reactions with formation of acyl derivatives, (4) reactions of addition to carbon-hetero multiple bonds, (5) reactions with formation of cyclic compounds, and (6) other reactions specific to a certain analysis. The selection of the derivatization reaction is typically done based on the desired property to be brought to the analyte and its possible reactivity. For this reason, the reagent is selected to have moieties that add the desired property to the analyte and also to have the capability to react with the specific functional group of the analyte. The matrix of the sample also has a role in the choice of a specific derivatization procedure. Initial matrix of the sample is not always suitable for derivatization, and in some cases preliminary sample preparation is necessary to change this matrix. The change can be as simple as drying the initial sample but can also be rather complex [14]. Table 1 gives a simplified view of preferences for the choice of a derivatization reagent for compounds containing active hydrogens [14].
\nDerivatization preferences for compounds containing active hydrogens.
Besides functionalities with active hydrogens, other functionalities can also be derivatized. Compounds containing carbonyls can be derivatized, for example, using condensation reactions. Some analytes may contain multiple functional groups such as the amino acids. Specific derivatization reactions can be selected for such cases.
\nThe formation of alkyl or aryl derivatives is applied to replace the active hydrogens from an analyte with an alkyl (R) or aryl (Ar) group. The replacement can be done in functionalities such as OH, COOH, SH, NH, or CONH. For example, the derivatization with short-chain alkyl bromides or iodides has numerous analytical applications for compounds such as steroids, amino acids, catecholamines, sulfonamides, phenols, barbiturates, organic acids, and mono- and oligosaccharides. A large number of reagents R-X are known, and in a simplified approach, it can be considered that R is carrying a specific property and X a specific reactivity, although the reactivity of a reagent is influenced by both R and X components of the molecule. The type of moiety R and that of reactive group X are guiding the selection process of selecting a reagent for a specific derivatization.
\nIn most alkylation reactions, the analyte acts as a nucleophile (Y
Various reagents and conditions were utilized in the derivatizations for analytical purposes. As reagents R-X for alkylations, one of the most commonly used are the alkyl halides, especially alkyl iodides and alkyl bromides. Because some of the derivatizations can be slow and inefficient depending on the analyte and on the reagent, the reaction rate becomes an important parameter for the analytical applicability. The reaction with an alkyl halide for the preparation of methyl or ethyl substituents, for example, is frequently performed either with a specific methylation reagent, in the presence of a catalyst, or in some instances using a particular solvent. The enhancement of the alkylation efficiency can be achieved using several other procedures. For example, for the analytical alkylation of carboxylic acids, specific cryptands such as crown ethers can be used to solvate the alkali metal portion of an organic acid salts, allowing the anion to be freer and increasing the rate of nucleophilic substitution. One other approach for enhancing the alkylation efficiency is the use of phase transfer alkylation. This approach is based on the formation of a compound easily extractable in an organic phase and on the displacement of the equilibrium in the direction of the formation of the desired product.
\nOne different way of enhancing the alkylation efficiency is the use of different alkylating reagents besides short-chain alkyl bromides or iodides. One example of a halide that is particularly reactive is pentafluorobenzyl bromide. This reagent can be used for the derivatization of a variety of compounds containing active hydrogens. Another reactive halide is 2-bromoacetophenone (phenacyl bromide). This reagent is used mainly for the alkylation of compounds containing more acidic hydrogens such as carboxylic acids. Another example of methylation using a special reagent R-X is applied on carbohydrates [15]. This methylation uses methylsulfinylmethanide anion. The reagent is prepared from dry DMSO and NaH or KH in a reaction as follows:
A polyol or a monosaccharide dissolved in DMSO is easily methylated with methylsulfinyl-methanide anion.
\nOther alkylating reagents are known (different X in R-X), also reacting in a nucleophilic substitution. For example, dimethyl sulfate can be used for alkylations. Alkylfluoromethyl-sulfonates are even more reactive than sulfates, and the reaction may take place with the active hydrogen even from alcohols or amines as follows:
Even tertiary amines, such as pyridine, also react with this type of reagent forming quaternary ammonium salts. The alkylation with alkylfluorosulfonates can be catalyzed as other alkylation reactions for increasing the reaction rate. A catalyst that can be used in this reaction is Hg(CN)2.
\nDiazomethane is another common alkylating (methylating) reagent. The alkylation using diazomethane is assumed to take place as follows:
Diazomethane is a gaseous unstable substance, which cannot be stored for long periods of time. It is usually prepared in small quantities and used immediately with or without an intermediate step of dissolution in ether. The preparation can be done from different N-nitroso-N-alkyl compounds in a reaction with a base. A common preparation uses N-nitroso-N-alkyl-p-toluenesulfonamide (Diazald). Methylation with diazomethane may require addition of a Lewis acid catalyst such as BF3. The methylation of partly acetylated sugars and amino sugars using diazomethane and BF3 in ether leads to the methylation of the free OH groups without the migration or substitution of the existent acyl groups.
\nA common alkylation of acidic analytes such as carboxylic acids, phenols, and thiols is performed using another type of alkylating reagent, namely, N,N-dimethylformamide dialkyl acetals. N,N-Dimethylformamide dimethyl acetal (Methyl-8®) is commonly used for methylations. For a compound containing a COOH group, the reaction with this reagent takes place as follows:
The compounds with acidic hydrogens can also be alkylated (methylated) using trimethyl orthoacetate, alkyl-p-tolyltriazenes (R─NH─N═N─C6H4─CH3), and O-alkyl isoureas are also used for the formation of analytes containing acidic hydrogens, imino esters, etc.
\nAlcohols can also act as alkylating reagents in particular when the analyte contains a more acidic hydrogen. Catalyst such as HCl, BF3, CF3 COOH or a cation exchange resin in H+ form is also frequently added to facilitate the reaction. The addition of HCl can be made as a water solution or as gaseous HCl that does not bring additional water to the reaction medium. The formation of alkyl or aryl derivatives of acids is a particularly important reaction known as esterification. Derivatization by esterification has been used with acids as the analyte and the alcohol as the reagent and also with the alcohol as the analyte and the acid the reagent. The esterification can be viewed either as the acid alkylation or as the acylation of the alcohol (see also the esterification mechanism). This reaction is typically catalyzed by strong acids and can be written as follows:
The mechanism of ester formation can be summarized by the following series of reactions:
The esterification efficiency can be improved by removing the water formed in this reaction. This can be done using a chemical reagent or distillation when the compounds of interest boil above 100°C. Among the materials able to eliminate water are desiccants such as anhydrous MgSO4, molecular sieves, or substances that react with water such as CaC2, (CH3)2C(OCH3)2 (2,2-dimethoxypropane), and even an appropriately chosen acid anhydride that reacts faster with water than with the reacting alcohol. The derivatization also may be performed in the presence of SOCl2 (thionyl chloride), which reacts with the water assisting in its removal, and when present in excess, may react with the alcohols forming alkyl chlorides or with the acids forming acyl chlorides. Chloride is a better leaving group in a nucleophilic alkylation reaction, and the efficiency of alkylation increases. Acids also can be esterified using a mixture of an alcohol and an acyl halide.
\nOne procedure for the formation of esters with less active organic acids applies the addition of dicyclohexylcarbodiimide (DCCI) in the derivatization process, to facilitate esterification. The reaction can be performed by adding to the acids that need to be analyzed the appropriate alcohol and DCCI usually in a solvent such as pyridine. Dicyclohexylurea, which is formed in the reaction, is not soluble in pyridine and can be separated. Besides DCCI, other carbodiimides can be used in the reaction of acids and alcohols. Among these are carbonyldiimidazole (CDI), 6-chloro-1-p-chlorobenzensulfonyloxybenzotriazole (CCBBT), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (EDAC), etc. Also, 2-chloro-1-methylpyridinium iodide, 2,4,6-triisopropylbenzenesulfonyl chloride, trialkyloxonium fluoroborate, etc. can be used to facilitate esterification.
\nTransesterification is another technique applicable for obtaining certain alkyl derivatives of acids (or acyl derivatives of alcohols). The reaction can be written as follows:
Transesterification can be catalyzed by acids (or Lewis acids) such as HCl, BF3, and H2SO4 or by bases such as CH3OK, CH3ONa, or C4H9ONa. The basic catalysts are commonly used for the methanolysis of triglycerides, followed by the analysis of the fatty acid methyl esters using GC or GC/MS [16].
\nA special alkylation can be achieved online during the heating in the injection port of a gas chromatograph using tertraalkylammonium hydroxides or alkylarylammonium hydroxides. Tetramethylammonium hydroxide (TMAH) is the most common reagents of this type. The reaction takes place as follows (Δ indicates heating):
Numerous other reactive compounds may be used for replacing active hydrogens in specific compounds. For example, epoxides, aziridines, and episulfides react easily with compounds with active hydrogens. Formation of a second group containing an active hydrogen may preclude the use of such reagents for analytical purposes.
\nBesides the desired derivatives, certain unexpected compounds that can be considered artifacts for the particular analysis can also be formed in alkylation reactions. The artifacts may be formed from unexpected interactions of the reagent with the analyte or may be a result of undesired effects of the catalysts or medium used for derivatization. In some cases, the control of the alkylation process may be difficult. Longer or shorter reaction times or intervals between derivatization and analysis may lead to errors, even when an internal standard is used for quantitation.
\nOne common case of artifact formation occurs during the reaction with compounds containing O-acyl or N-acyl groups, such as previously acylated carbohydrates, glycolipids, or glycoproteins, in particular when the reaction is done with short-chain alkyl bromides or iodides. When the OH groups of different sugars or NH2 groups of amino sugars were already protected with acyl groups, it was noted that, depending on the catalyst and the chosen medium, these acyl groups can be replaced by alkyl groups, or they may migrate from one position (such as C1) to other positions.
\nOxidation is another common side reaction when using Ag2O as a catalyst. The oxidation effect of Ag2O can be seen on free sugars as well as when attempting to permethylate peptides. Sulfhydryl groups are particularly sensitive to oxidation with Ag2O as a catalyst. The use of methylsulfinyl carbanion as a methylating reagent may also produce undesired side reactions with certain esters generating methylsulfinylketones. Also, strong alkylating reagents may produce undesired artifacts by unexpected alkylations.
\nThe derivatization with the purpose of obtaining aryl derivatives is similar in many respects to the alkylation reaction. The reaction takes place with compounds containing active hydrogens. Simple aryl halides are generally resistant to be attacked by nucleophiles and do not react similar to alkyl halides. This low reactivity can be significantly increased by changes in the structure of aryl halide or in the reaction conditions. The nucleophilic displacement can become very rapid when the aryl halide is substituted with electron attracting groups such as NO2.
\nSilylation is the chemical reaction of replacing a reactive hydrogen atom in OH, COOH, SH, NH, CONH, POH, SOH, or enolisable carbonyl with a silyl group, most frequently with trimethylsilyl (TMS). A large number of analytical methods involve silylation applied to alcohols including carbohydrates [17], phenols [18], amines, sterols [19], etc. The purpose of silylation in chromatography is mainly to reduce the polarity of the analyte, increase its stability, and improve the GC behavior. The differences in the mass spectra of the silylated compounds as compared to the initial analyte may also be an advantage for detectability. However, the mass spectra of many silylated compounds may not be available in common mass spectral libraries. Also, the silylated compounds plus the commonly present excess of silylating reagent may deteriorate some types of stationary phases such as that of Carbowax (polyethylene glycol)-type columns, and for this reason, their separation cannot be done on such columns.
\nSilylation can be performed on specific analytes or directly on complex samples such as a plant material (see, e.g., [12]). The silylating agent and the solvent can play the double role of extractant and silylating reagent. Many publications describe the use of silylation reactions for analytical purposes (e.g., [1, 5, 20]). The reaction of an analyte Y
The molecular weight for TMS is 73.047 calculated considering in the elemental composition of only the masses of the most abundant isotope. Numerous reagents have been synthesized to be used in silylations. Various aprotic solvents can be used as medium for silylation. The analysis can be focused on one analyte or on a mixture of analytes. The main factors contributing to the increase of the efficiency and the rate of the silylation reaction are the silyl donor ability of the reagent and the ease of silylation of different functional groups in the analyte. The solvent (or mixture of solvents) used as a medium and the compounds present or added in the silylation medium may also play a role for silylation efficiency. The reagent excess is sometimes important for displacing the equilibrium in the desired direction, and usually an excess up to ten times larger than stoichiometrically needed is used for silylation. Temperature also increases reaction rate, as expected, and heating of the sample with the reagents at temperatures around 70°C for 15 to 30 min is common. Some reagents used for trimethylsilylation are shown in Figure 8 [14].
\nSome reagents used for trimethylsilylation.
The approximate order of the increasing silyl donor ability for the reagents shown in Figure 8 is HMDS < TMCS < MSA < TMSA < TMSDEA < TMSDMA < MSTFA < BSA < BSTFA < TMSI. This order may be different on particular substrates where other reagents or reagent mixtures may be more reactive.
\nSilylation reagents can be used pure or in mixtures of two or even three reagents. The reagent mixtures may provide a more efficient silylation for specific compounds. For example, silylation of 3,4-dimethoxyphenylethylamine with BSA leads to the substitution of only one active hydrogen in the NH2 group, while the silylation with BSA in the presence of 5% TMCS produces silylation of both hydrogens in the NH2 [21]. A common silylating mixture is BSTFA with 1% TMCS.
\nOne of the determining factors regarding the silylation efficiency is the nature of the molecule Y
Several functional groups that can be silylated (listed in the approximate order of decreasing ease of silylation).
In general, the silylation of OH and COOH groups takes place with better results than that of NH2, CONH, or NH groups. Excellent results are obtained, for example, for the analysis of phenols after silylation [19]. A chromatogram of a solution containing a mixture of phenols at concentrations between 2.0 and 2.5 μg/mL in DMF, derivatized with BSTFA, separated on a BPX-5 chromatographic column (SGE Anal. Sci.), followed by MS analysis in single-ion monitoring (SIM) mode is shown in Figure 9. Details regarding the analyzed phenols are given in Table 3.
\nChromatogram of a set of phenol standards in DMF with the concentrations between 2.0 and 2.5 μg/mL derivatized with BSTFA, separated on a BPX-5 chromatographic column followed by MS analysis.
No. | \nCompound | \nRet. time | \nm/z | \nAbrrev. | \nNo. | \nCompound | \nRet. time | \nm/z | \nAbrrev. | \n
---|---|---|---|---|---|---|---|---|---|
(1) | \nPhenol | \n6.88 | \n166 | \nPh | \n(14) | \n3,4-Dimethylphenol | \n12.32 | \n194 | \n3,4-diMePh | \n
(2) | \no-Cresol | \n8.57 | \n180 | \no-Cr | \n(15) | \n3-Methoxyphenol | \n13.17 | \n196 | \n3-MeOPh | \n
(3) | \nm-Cresol | \n8.76 | \n180 | \nm-Cr | \n(16) | \n4-Methoxyphenol | \n13.47 | \n196 | \n4-MeOPh | \n
(4) | \np-Cresol | \n9.08 | \n180 | \np-Cr | \n(17) | \nCatechol | \n13.88 | \n254 | \nCa | \n
(5) | \n2-Ethylphenol | \n10.28 | \n194 | \n2-EtPh | \n(18) | \nResorcinol | \n16.05 | \n254 | \nRe | \n
(6) | \n2,5-Dimethylphenol | \n10.70 | \n194 | \n2,5-diMePh | \n(19) | \n4-Methylcatechol | \n16.27 | \n268 | \n4-MeCa | \n
(7) | \n3,5-Dimethylphenol | \n11.07 | \n194 | \n3,5-diMePh | \n(20) | \nHydroquinone | \n16.73 | \n254 | \nHy | \n
(8) | \n2,4-Dimethylphenol | \n11.20 | \n194 | \n2,4 diMePh | \n(21) | \n3-Methylcatechol | \n16.71 | \n268 | \n3-MeCa | \n
(9) | \n2-Methoxyphenol | \n11.28 | \n196 | \n2-MeOPh | \n(22) | \n5-Methylresorcinol | \n18.19 | \n268 | \n5-MeCa | \n
(10) | \n4-Ethylphenol | \n11.59 | \n194 | \n4-EtPh | \n(23) | \n2-Methylresorcinol | \n18.66 | \n268 | \n2-MeRe | \n
(11) | \n4-Chlorophenol | \n11.71 | \n185 | \n4-ClPh | \n(24) | \n4-Ethylresorcinol | \n19.90 | \n282 | \n4-EtRe | \n
(12) | \n2,6-Dimethylphenol | \n11.79 | \n194 | \n2,6-diMePh | \n(25) | \n2,5-Dimethylresorcinol | \n20.18 | \n282 | \n2,5-diMeRe | \n
(13) | \n2,3-Dimethylphenol | \n12.02 | \n194 | \n2,3-dimePh | \n\n | \n | \n | \n | \n |
Details regarding the analyzed phenols with the chromatogram shown in Figure 9.
Besides organic active hydrogens, several inorganic compounds with active hydrogens can also react with silylating reagents. Among these are H2O, H2O2, and strong inorganic acids. Also, some salts of the acids may be silylated. The reaction of silylating reagents with water imposes that water should be at the low level in the matrix or the solution of the analytes. The reaction with water takes place as follows:
In many solvents used as medium for derivatization, the trimethylsilanol formed in the reaction with water is separated as a distinct layer of solvent. The formation of two layers impedes a proper sampling of the derivatized material in the GC/MS instrument. In addition to that, the presence of an excess of water suppresses the derivatization of other compounds. The silylation is not recommended on samples with a water content higher than about 10%.
\nThe silylation reaction is commonly performed in a solvent that does not have active hydrogens. The most commonly used solvents as a medium for silylation are dimethylformamide (DMF), pyridine, and acetonitrile. The main role of the solvent is to dissolve the analyte and the reagents. The by-product HX of silylation shown in reaction (17) can be an acid, a base, or a neutral compound. As examples, for TMCS the by-product is HCl, for HMDS the by-product is NH3, for BSTFA the by-product is N-TMS-trifluoroacetamide, and for TMSI the by-product is imidazole. When the silylation reagent generates an acid as a by-product of the reaction, this may interfere with the silylation. For this reason, silylation can be promoted by any acid acceptor used as solvent or present in the solvent. Among such solvents are pyridine, triethylamine, and to a lower extent DMF. They can be used as both solvents and acid acceptors. Mixtures of solvents are commonly used for both enhancing solubility and promoting silylation. For example, formamide in the presence of pyridine may react with an acidic by-product generating CO and an ammonium salt. The addition of basic compounds to the silylation reaction may also influence the efficiency of the silylation. Also, some compounds may act as catalysts for silylation.
\nAlthough the TMS derivatives are by far the most commonly used in the derivatization for analytical purposes, other substituents in the silyl group can be used as reagents. Several such groups are indicated in Figure 10. The groups can be present in a variety of reagents connected to leaving groups “X-” such as Cl-, imidazolyl, F3C-(CO)-N(CH3)-, etc. For example, a common reagent containing
Examples of silyl groups different from TMS used in silylation reagents.
The use of different groups than TMS may serve different purposes. For example, a fluorinated or brominated group may enhance significantly the detection sensitivity when using ECD or NCI-MS. Also, the stability toward hydrolysis of compounds silylated with different groups than TMS may be higher, and such silylation can be advantageous. This is, for example, the case of
As an example, silylation of amino acids with MTBSTFA is commonly used [22, 23], and it is preferred to the silylation generating TMS derivatives. The chromatogram of a set of amino acid standards with the concentration of 0.05 μmol/mL derivatized with MTBSTFA and separated on a DB-5MS chromatographic column (from Agilent) followed by MS analysis is shown in Figure 11. Details regarding the analyzed amino acids are given in Table 4.
\nChromatogram of a set of amino acid standards with the concentration of 0.05 μmol/mL derivatized with MTBSTFA separated on a DB-5MS chromatographic column.
Peak No. | \nAmino acid | \nAbbrev. | \nMW | \nFormula + x TBDMS | \nMW + x TBDMS | \nCharact. ion | \nRet. time | \n
---|---|---|---|---|---|---|---|
(1) | \nα-Alanine | \nα-Ala | \n89.09 | \nC15H35NO2Si2 | \n317 | \n260 | \n31.69 | \n
(2) | \nGlycine | \nGly | \n75.07 | \nC14H33NO2Si2 | \n303 | \n246 | \n32.63 | \n
(3) | \nSarcosine | \nSar | \n89.09 | \nC15H35NO2Si2 | \n317 | \n260 | \n33.85 | \n
(4) | \nα-Amino-n-butyric acid | \nα-ABu | \n103.10 | \nC16H37NO2Si2 | \n331 | \n274 | \n34.36 | \n
(5) | \nβ-Alanine | \nβ-Ala | \n89.09 | \nC15H35NO2Si2 | \n317 | \n260 | \n35.58 | \n
(6) | \nUrea | \n\n | 60.06 | \nC13H32N2OSi2 | \n288 | \n231 | \n36.01 | \n
(7) | \nβ-Aminoisobutyric acid | \nβ-ABu | \n103.10 | \nC16H37NO2Si2 | \n331 | \n274 | \n36.11 | \n
(8) | \nValine | \nVal | \n117.15 | \nC17H39NO2Si2 | \n345 | \n186 | \n36.15 | \n
(9) | \nLeucine | \nLeu | \n131.17 | \nC18H41NO2Si2 | \n359 | \n200 | \n37.71 | \n
(10) | \nNorleucine | \n\n | 131.17 | \nC18H41NO2Si2 | \n359 | \n200 | \n38.8 | \n
(11) | \nIsoleucine | \niLeu | \n131.17 | \nC18H41NO2Si2 | \n359 | \n200 | \n38.8 | \n
(12) | \nγ-Aminobutyric acid | \nγ-ABu | \n103.10 | \nC16H37NO2Si2 | \n331 | \n274 | \n39.79 | \n
(13) | \nProline | \nPro | \n115.13 | \nC17H37NO2Si2 | \n343 | \n184 | \n39.87 | \n
(14) | \n2-Phenylglycine | \nPhGly | \n151.17 | \nC20H37NO2Si2 | \n379 | \n220 | \n46.16 | \n
(15) | \n5-Oxoproline | \noPro | \n129.13 | \nC17H35NO3Si2 | \n357 | \n300 | \n46.18 | \n
(16) | \nMethionine | \nMet | \n149.20 | \nC17H39NO2SSi2 | \n377 | \n320 | \n46.68 | \n
(17) | \nSerine | \nSer | \n105.09 | \nC21H49NO3Si3 | \n447 | \n390 | \n47.52 | \n
(18) | \nThreonine | \nThr | \n119.12 | \nC22H51NO3Si3 | \n461 | \n404 | \n48.43 | \n
(19) | \nPhenylalanine | \nPhe | \n165.19 | \nC21H39NO2Si2 | \n393 | \n336 | \n50.35 | \n
(20) | \nAspartic acid | \nAsp | \n133.10 | \nC22H49NO4Si3 | \n475 | \n418 | \n52.47 | \n
(21) | \nHydroxyproline | \nHyPro | \n131.13 | \nC23H51NO3Si3 | \n473 | \n314 | \n53.23 | \n
(22) | \n3-Methyl-L-histidine | \n3MeHys | \n169.20 | \nC19H39N3O2Si2 | \n397 | \n340 | \n55.15 | \n
(23) | \nGlutamic acid | \nGlu | \n147.13 | \nC23H51NO4Si3 | \n489 | \n432 | \n55.53 | \n
(24) | \nOrnithine | \nOrn | \n132.20 | \nC23H54N2O2Si3 | \n474 | \n286 | \n55.64 | \n
(25) | \n1-Methyl-L-histidine | \n1MeHys | \n169.20 | \nC19H39N3O2Si2 | \n397 | \n302 | \n57.03 | \n
(26) | \nLysine | \nLys | \n146.19 | \nC24H56N2O2Si3 | \n488 | \n300 | \n58.02 | \n
(27) | \nα-Aminoadipic acid | \n\n | 161.20 | \nC24H53NO4Si3 | \n503 | \n446 | \n58.06 | \n
(28) | \nHistidine | \nHys | \n155.16 | \nC24H51N3O2Si3 | \n497 | \n440 | \n62.29 | \n
(29) | \nTyrosine | \nTyr | \n181.19 | \nC27H53NO3Si3 | \n523 | \n302 | \n63.29 | \n
(30) | \nArginine | \nArg | \n174.20 | \nC24H56N4O2Si3 | \n516 | \n144 | \n64.26 | \n
(31) | \nTryptophan | \nTrp | \n204.22 | \nC29H54N2O2Si3 | \n546 | \n244 | \n67.98 | \n
(32) | \nCystine | \nCys | \n240.30 | \nC28H64N2O4S2Si4 | \n668 | \n348 | \n72.65 | \n
(33) | \nHomocystine | \nhCys | \n268.30 | \nC32H72N2O4S2Si4 | \n724 | \n362 | \n76.59 | \n
Details regarding the analyzed amino acids with the chromatogram shown in Figure 11.
In most situations, silylation generates only the desired derivatives. However, there are cases when the expected silylated compound is not formed, and either the silylation is not complete, or some compounds such as aldehydes, ketones, or esters with no obvious active hydrogen generate silylated compounds. Incomplete silylation is usually the result of inappropriate reaction conditions. However, when compounds with multiple functionalities are silylated, it is possible to generate a variety of derivatized compounds, regardless of the intention to obtain fully silylated or partly silylated compounds.
\nIn some cases, artifacts are formed due to the modification of the analyte under the influence of the reagents during derivatization. For example, when the silylation is done in basic or acidic conditions, the analytes that are sensitive to acidic or basic media may suffer unexpected transformations. The most frequent artifacts with compounds not containing obvious active hydrogens occur with aldehydes. Some aldehydes are able to undergo two types of chemical reactions with formation of OH groups, namely, enolization and acetal formation in the presence of water. The OH groups formed in this manner react with different silylating reagents and give the corresponding silylated products. Although the enolization or the acetal formation is negligible for the initial aldehyde, the reactions may be significantly displaced toward the formation of the silylated compounds of the enol or of the acetal. Artifacts can also be generated when the reaction is allowed to continue for an extended period of time. Other uncommon reactions with a specific silylation reagent and analyte may occur. An example of an uncommon reaction is the ring opening of flavanones.
\nThe formation of acyl derivatives is applied for replacing the active hydrogens from an analyte in functionalities such as OH, SH, NH [11, 24], CONH, etc. The acylation is also used for reducing polarity and improving the behavior of the analytes in the chromatographic column. Acylation may confer a better volatility of the analytes, although not as marked as for silylation or methylation. Only the derivatization with acetyl groups or with fluorinated acyl groups (not heavier than heptafluorobutyryl) improves volatility, while other heavier acyl groups are not suitable for this purpose. Acetylation, for example, can be used for compounds such as monosaccharides and amino acids to allow GC analysis. The detectability improvement on the other hand is a very common purpose for acylation. Acylation with fluorinated compounds is frequently used for enhancing detectability in GC with ECD or NCI-MS detection. Other uses of acylation include the enhancement of separation of chiral compounds, etc.
\nMost acylation reactions are nucleophilic substitutions where the analyte is a nucleophile (Y
Some common acyl groups present in acylation reagents are indicated in Table 5.
\nSome common groups present in acylating reagents used in derivatizations for GC analysis [14].
As shown in Table 5, the acyl groups in the reagent can be attached to various “X” groups. One such group is OH and among the acylating reagents are some free acids. When nucleophile is an alcohol, the reaction is known as esterification and has been discussed in Section 7. The acylation with acids can be applied besides alcohols to certain thiols, phenols, amines, etc. and can be written as follows:
The reaction can be displaced toward the formation of the acyl derivatives by eliminating the water using compounds such as anhydrous MgSO4, molecular sieve, or substances that react with water such as CaC2, or (CH3)2C(OCH3)2. Dicyclohexylcarbodiimide (DCCI) also is used for modifying the yield of the desired product. The reaction with reagents containing a carboxylic acid reactive group also can be done in the presence of 2,4,6-trichlorobenzoyl chloride or with various sulfonyl chlorides such as 2,4,6-triisopropyl-benzenesulfonyl chloride or 2,4,6-trimethyl-benzenesulfonyl chloride. The reaction of amines with acids can be displaced toward the formation of the amides using a peptide coupling reagent such as benzotriazol-1-yl-oxy-tris(dimethyl-amino)-phosphonium hexafluorophosphate (BOP), diethyl cyanophosphonate, O-benzotriazol-1-yl-N,N,N′,N′-bis(tetramethylene)uronium hexafluorophosphate, 2,2′-dipyridyl disulfide + triphenylphosphine, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (EDAC), etc.
\nCommon acylating reagents are acyl halides such as chlorides or bromides, which are reactive compounds suitable for acylation. The reaction of an acyl chloride with an amine, for example, takes place as follows:
Since the reactivity of amides is lower than that of amines, the second hydrogen in the amine is more difficult to replace. Also, steric hindrance may negatively influence the reaction. The generation of a strong acid such as HCl is a disadvantage in the reaction with acyl halides, and usually the acid should be removed either by adding basic compounds such as Na2CO3 or MgCO3 or using pyridine as the reaction medium. The high reactivity of acyl halides is used for the acylation of compounds with less reactive hydrogens. Certain carbonyl cyanides react similarly to acyl chlorides.
\nThe disadvantage of generating a strong inorganic acid in the acylation with acyl halides also can be avoided by having, instead of the acyl halide, an anhydride. The reaction of Y
The acid resulting together with the acylated compound is not a strong acid such as HCl. The anhydrides of trifluoroacetic acid (TFA), pentafluoropropionic anhydride (PFPA), and heptafluorobutyric (HFBA) acids are commonly used for derivatization of alcohols, phenols, amines, etc., with the purpose of enhancing detectability (by ECD or NCI-MS) and also for improving the chromatographic behavior (higher volatility, better thermal stability, better separation). The volatility of fluorinated compounds allows the GC applications. The reactivity of the perfluorinated anhydrides increases in the order HFBA < PFPA < TFA. However, the differences are not significant. Once formed, the heptafluorobutyrates are more stable to hydrolysis than the trifluoroacetates. An inert solvent such as CH2Cl2, ether, ethyl acetate, acetone, tetrahydrofuran or in CH3CN, etc. can be used as a medium for the reaction with perfluoroanhydrides. For the neutralization of the acids formed during derivatization, the basic compounds such as triethylamine, pyridine, or even solid NaHCO3 can be utilized.
\nIn order to avoid the formation of water or of a strong acid in the acylation reaction, certain amides such as N-methyl-bis(trifluoroacetamide), bis(trifluoroacetamide), or 2,2,2-trifluoro-N-methyl-N-(2,2,2-trifluoroacetyl)acetamide (MBTFA) can be used as reagents. Acylation of amines takes place at room temperature. Solvents such as CH3CN, pyridine, DMSO, or THF can be used as a reaction medium:
One other procedure successfully applied to obtain acyl derivatives is the use of acyl imidazoles as reagents. This class of compounds reacts with analytes containing alcohol, primary and secondary amino groups, or thiols. The reaction generates as a by-product imidazole:
Succinimidyl esters also can be used for acylation purposes. Amines and the amino group in amino acids also can be acylated using urethane-protected α-amino acid-N-carboxyanhydrides or oxycarbonyl-amino acid-N-carboxyanhydrides. Alkylketenes and their dimers may be used for acylation.
\nA special type of acylation is that using chloroformates. Carbonic acid, O═C(OH)2, can form amides, esters, halides, etc., due to the presence of two OH groups bonded to the CO group. Carbonic acid ester halides, also called chloroformates or chloroformate esters, with the formula R─O─C(═O)─X, where R is an alkyl or aryl group and X is F, Cl, Br, or I, can react with various compounds containing active hydrogens, such as acids [25], amines, alcohols, thiols, and amino acids. Amino acids, for example, in the presence of an alcohol in water form carbamate esters (urethanes) reacting as follows [26]:
The formation in reaction (25) of the alcohol Ra–OH may lead to traces of a resulting compound with both substituted radicals being Ra. For this reason it is typically recommended to perform the reaction in the presence of an alcohol having the same radical as the chloroformate reagent (Ra = Rb). Chloroformates containing in the alkyl or aryl group halogen substituents are particularly reactive. Even tertiary amines can react with specific chloroformates, such as pentafluorobenzoyl chloroformate or with trichloroethyl chloroformate, by displacing an alkyl group connected to the nitrogen atom and forming the carbamate ester.
\nSimilar in many respects to that of acyl derivatives R–CO–X are the reactions of sulfonyl derivatives R–SO2–X. Sulfonyl halides are in general less reactive than halides of carboxylic acids. The reaction of a sulfonyl derivative may take place with alcohols, phenols, amines, etc. The reactivity toward the sulfonyl sulfur is RNH2 > CH3COOR > H2O > ROH.
\nHigh reactivity toward active hydrogens in alcohols, amines, etc. can also be achieved using reagents with other functionalities. These functionalities include isocyanates, isothiocyanates, carbonyl azides, etc. These reactions can be seen as a replacement of an active hydrogen with a CO-R group or CS-R group as it occurs in other acylations.
\nA variety of other derivatization reactions are reported in the literature (see, e.g., [1]) and used for GC and GC/MS analyses. Among these are the addition to hetero multiple bonds in functional groups such as C═O, C═S, C═N, or C☰N. Many such reactions are additions to multiple bonds. Such reactions are, for example, the additions to the C═O groups in aldehydes and ketones. Reagents containing active hydrogens in groups such as NH2, OH, H2N-NH-, etc. can react, for example, with aldehydes and ketones. Alcohols, for example, form hemiacetals or acetals with aldehydes and ketals with ketones, and although most of such compounds are not stable enough to be suitable for derivatization, cyclic acetals and ketals may be stable and used for analytical purposes. A common reaction of carbonyl compounds is with amines. The initial addition reaction usually continues with water elimination forming a substituted imine or a Schiff base. Similar to the reaction of amines is the reaction with substituted hydroxylamines or hydrazines. A typical reaction of derivatization of carbonyl compounds is that using dinitrophenylhydrazine (DNPH). The derivatized compound can be analyzed either by LC [27] or by GC/MS [28]. The reaction takes place as follows:
The groups Ra and Rb can be H or alkyl or various other substituents.
\nAnother reagent that can be used for ketone derivatization is N-aminopiperidine in the presence of catalytic amounts of acetic acid. The resulting substituted hydrazone can be used in GC analysis:
β-Diketones may react differently with hydrazines generating pyrazole derivatives as shown below:
Several other classes of compounds similar to hydrazines react with the carbonyl compounds. Among these are hydrazones (NH2─N═CR2), hydrazides (NH2NH-COR), and semicarbazide (NH2NH-CONH2). Hydroxylamines also react with carbonyl compounds forming oximes. Hydroxylamine itself, hydroxylamine hydrochloride (STOX® reagent), or derivatives such as H2N-OSO3H in a solvent like pyridine can be used in this reaction:
When the reaction is performed with hydroxylamine, the generated oxime contains an active hydrogen. This can be further derivatized, for example, by silylation in a reaction with a common silylation reagent.
\nFor derivatization purposes other reagents can be used, such as substituted hydroxylamines like methoxyamine hydrochloride NH2OCH3•HCl (MOX® reagent) and O-(pentafluorobenzyl)-hydroxylamine hydrochloride (FLOROX® reagent). The reaction of a ketone or aldehyde with FLOROX is shown below:
The oximes existing in
The transformation of the oximes into nitriles generates one single compound from the two (syn- and anti-) isomers and can be used to simplify the chromatograms of sugars derivatized as oximes.
\nAlcohols, amines, and thiols also can react at other hetero multiple bonds leading to analytical applications. This addition may occur at the isocyanates (─N═C═O), ─C═O group in an amide, at a nitrile, at CS2, or at other groups. One example is the addition under special conditions of alcohols to dimethylformamide. The resulting acetals are very reactive and are used themselves as reagents, as shown previously for N,N-dimethylformamide dimethyl acetal (see reaction 12). Another example is the reaction of CS2 with alcohols in the presence of a base, leading to the formation of xanthates. Amines also react with CS2, and the formed isothiocyanate can be analyzed using GC analysis. The reaction takes place as follows:
Formation of new cycles from noncyclic compounds or replacement of old cycles with new ones that are more stable or have a desired property is also exploited in sample processing using derivatization. Epoxides, for example, can be formed in the reaction of a compound with a carbon–carbon double bond and a peroxy acid. Among the peroxy acids more frequently used for the formation of epoxides are peracetic, performic, perbenzoic, trifluoroperacetic, and 3,5-dinitroperoxybenzoic acids. However, in this reaction a mixture of enantiomers is formed, as shown below for a
The separation of the epoxides may be easier to achieve than that of olefins, and this type of derivatization has been utilized, for example, for better separation of various
Another reaction with formation of new cycles is that of amino acids with phenyl isothiocyanate leading to a thiohydantoin derivative:
This reaction has been successfully used for the analysis of amino acids in proteins [29, 30].
A variety of aromatic cycles can be formed in reactions involving bifunctional compounds. Addition reactions to hetero multiple bonds in bifunctional molecules frequently lead to cyclic compounds. For example, formaldehyde can react with tryptophan or tryptamine generating a β-carboline derivative as follows:
The new compound can be analyzed by GC, usually after further derivatization by silylation of the carboxyl group.
\nA typical reaction leading to pyrazoles is the reaction of hydrazines with diketones such as 2,4-pentandione (acetylacetone). For example, the reaction between hydrazine or methylhydrazine and acetylacetone takes place as follows:
Activated carbonyl groups such as those in hexafluoroacetone are known to react with difunctional compounds. The reaction may take place with an amino acid as follows:
Amino acids can react with an activated anhydride such as trifluoroacetic anhydride (TFAA):
The reaction takes place by heating the amino acids with an excess of TFAA. The reaction mixture is then dissolved in ethyl acetate and analyzed by GC.
\nNumerous other types of derivatization reactions were used for making the analytes suitable for GC and GC/MS analyses. These include formation of various cyclic types of compounds such as azines, siliconides, boronates, etc., that are thermally stable and do not have polar hydrogens such that GC or GC/MS analysis is possible. In addition to reagents that add specific moieties to the analytes, oxidation and reduction were sometimes used for the analyte modification (see, e.g., [4]).
\nSolid-phase reagents are polymeric materials with specific groups that are reactive and can be transferred to the analyte molecule producing derivatization. For an analyte of the form Y
Solid-phase reagents must work analogously to the corresponding small-molecule reagents containing the group R (a tag). Reagents that are insoluble in certain solvents at high concentrations can often provide a high ratio of analyte/substrate in a polymeric microenvironment that yields a high kinetic rate for the heterogeneous reaction.
\nA variety of materials can be used as solid support, such as specifically bound reagents on a silica support (used, e.g., for online derivatization in HPLC analysis), ion exchange resins, as well as other supports [31]. One example of solid-phase support that can produce derivatization is trifluoroacetyl nylon 6,6. This solid-phase reagent can be obtained from poly(hexamethylene adipamide) (nylon 6,6) and trifluoroacetyl anhydride. This solid-phase reagent can be used in amine derivatization in a reaction as follows:
This derivatization of the amine is done by mixing the solid-phase reagent with a solution of amine solution in CH3CN. Following derivatization, the solid-phase reagent is separated by centrifugation, and the solution is concentrated by evaporating part of the solvent and analyzed by GC (an amine internal standard must be used in this procedure). However, some such derivatizations require a long time of interaction between the solid-phase reagent and the analytes and found only limited applications.
\n(Another) alternative of derivatization of specific analytes is using the reaction between the reagent and the analyte both adsorbed on a solid support. This type of derivatization has been used, for example, in connection with a solid-phase microextraction (SPME) technique [32]. In this technique a reagent is initially adsorbed in the SPME fiber, followed by exposure to the analytes. The derivatized analytes are further desorbed in the injection port of the GC and analyzed using a detector such as MS. For example, formaldehyde from air can be analyzed using a polydimethylsiloxane (PDMS) fiber containing
Connected and Automated Vehicles (CAVs) are nowadays the area of extensive research and there are premises to suspect that the introduction CAVs may revolutionize the whole transportation area [1]. There is no lack of predictions stating that CAVs will solve many of the current problems experienced on roads today, such as congestion, traffic accidents and lost time [2].
Traffic state prediction and traffic control are two key modules in transportation systems with CAVs [3]. Traffic states such as flow, speed, congestion, etc., plays vital roles in traffic management, public service and traffic control [4]. By predicting the evolution of traffic state timely and accurately, decision-maker and traffic controller can make effective policy and control input to avoid traffic congestion ahead of time and thus ITS (Intelligent Transportation Systems), advanced traffic management systems and traveler information systems rely on real-time traffic state prediction. Traffic control can be divided into a decision-making module and a vehicle control module. The former is used to optimize the mobility, safety and energy consumption by using the vehicle trajectory prediction results to calculate vehicle platoon sizes, speed, flow, density, traffic merging, diverging flow and traffic signals, while the latter is used for vehicle path control, vehicle fleet control and steering wheel, throttle, brake, and other actuator control by using onboard units based on the control commands [3]. How to timely and accurately predict the future traffic state and deliver an effective traffic control strategy are fundamental issues in ITS.
Traffic state prediction approaches can be broadly divided into two parts: parametric and non-parametric approaches [5]. Parametric approaches utilize parametric models that capture all the information about its predictions within a finite set of parameters. The popular techniques in parametric approaches include ARIMA (Autoregressive Integrated Moving Average) [6, 7, 8, 9], linear regression [10] and Kalman Filter (KF) based method [11], which are linear models and able to have high accuracy with linear characteristics of traffic data. ARIMA model is based on the assumption that the future data will resemble the past and widely used in time series analysis, which can be made to be stationary by differencing. It can be specified three values that represent the order of autoregressive (
Non-parametric models such as DL (Deep Learning) outperform parametric models because of stochastic, indeterministic, non-linear and multidimensional characteristics of traffic data [5]. DL is a subset of machine learning (ML) which is based on the concept of deep neural network (DNN) and it has been widely used for data classification, natural language processing (NLP) and object recognition [5]. The most popular DL models used for traffic state prediction includes Convolution Neural Network (CNN) [12, 13, 14], Deep Belief Network (DBN) [15, 16], Recurrent Neural Network (RNN) [17, 18, 19] and Autoencoder (AE) [20] etc. CNN is useful for traffic prediction because of the two-dimensional characteristics of traffic data and its ability to extract the spatial feature. CNN is only connected to a smaller subset of input and thus decreases the computational complexity of the training process. DBN is a stacking of multiple RBMs (Restricted Boltzmann Machines), which can be used to estimate the probability distribution of the input traffic data. LSTM is the special type of RNN, which can capture the temporal feature of traffic data, and LSTM can overcome the gradient vanishing problem caused by the standard RNN.
Traffic control strategies can be generally divided into classical methods and learning-based methods. Classical methods develop traffic controller based on control theory or optimization-based techniques, which include dynamic traffic assignment based nonlinear controller [21], standard proportional-integral (PI) controller [22, 23], robust PI controller [24], model-based predictive control (MPC) [25, 26], linear quadratic controller [27], mixed-integer non-linear programming (MINLP) [28], multi-objective optimization based decision-making model [29]. Learning-based methods refer to the utilization of artificial intelligence technologies to achieve decision-making and control for CAVs, which can be further divided into three categories: statistic learning-based method, deep learning-based (DL) method and reinforcement learning-based (RL) method. The RL-based method is currently one of the most commonly used learning-based techniques for traffic control and decision-making because RL can solve complex control problems by using the Markov decision process (MDP) to describe the interaction states of agent and environment [4]. The most popular RL-based methods include Q-learning for adaptive traffic signal control [30, 31], multi-agent RL approaches [32, 33, 34, 35], Nash Q-learning strategy [36]. Many other RL-based approaches are also available in the literature. Q-learning based traffic signal control aims to minimize the average accumulated travel time by greedily selecting action at each iteration. Multi-agent RL approaches are more popularly used in network signal optimization and can be generally divided into centralized RL and decentralized RL, while the former considers the whole system as a single agent and the latter distributes the global control to each agent. Nash Q-learning strategy is a decentralized multi-agent RL strategy, which performs iterated updates based on assuming Nash equilibrium behavior over the current Q-values. It can be shown that traffic signal control using the Nash Q-learning strategy can converge to at least one Nash equilibrium for stationary control policies. However, Nash Q-learning is unable to achieve the Pareto Optimality without consideration of cooperation among different agents.
This chapter provides a comprehensive survey about state-of-the-art traffic state prediction and traffic control techniques. It is organized as follows: In Section 2, we firstly introduce the fundamental structure and main characteristics of two important DL models: CNN and LSTM (Long Short-Term Memory), as well as their advantages in traffic state prediction, then we introduce how to realize hybrid traffic state prediction by combining two models to achieve better accuracy. In Section 3, we detail RL fundamentals and introduce how it can be applied in traffic control and decision-making. We focus on multi-agent RL approaches. Pros and cons are discussed. Section 4 gives the summary of this chapter.
In this section, we first briefly overview the machine learning and deep learning concept. Then, we focus on introducing the architectures of two DL models: CNN and LSTM, which show good performance in processing high-dimensional and temporal correlated data. Finally, a hybrid model of CNN and LSTM is described and the research potential is about how to improve the prediction accuracy by incorporating spatio-temporal correlation.
ML approaches are broadly classified into two categories, i.e., Supervised Learning and Unsupervised Learning [5]. Supervised Learning requires input data to be clearly labeled. It involves a function
DL is a branch of ML which aims to construct a computational model with multiple processing layers to support high-level data abstraction. It can automatically extract the feature from data, without any human interference to explore hidden data relationships among different attributes of the dataset [37]. Concepts of DL are inspired by the thinking process of the human brain. Hence, the majority of DL architectures are using the framework of Artificial Neural Network (ANN), which consists of input, hidden and output layers with nonlinear computational elements (neurons and processing units). The network depth (the number of layers) can be adjusted according to the feature dimensions and complexity of the data. The number of neurons at the input layer is equal to the number of independent variables, while the number of neurons at the output layer is equal to the number of dependent variables, which can be single or multiple. Neurons of two successive layers are connected by weights which are updated while training the model. The neurons at each layer receive the output from the previous layer, which is generated by a weighted summation over inputs and then passed to an activation function (Figure 1).
(left) ANN with one input layer, two hidden layers and one output layer,
Let us take the four-layer ANN in Figure 2 for example. During the training process, the value of
CNN structure.
where
In this section, we examine two popular DL architectures: CNN and LSTM, which are used popularly for multidimensional and time sequential dataset. CNNs have been extensively applied in various fields, including traffic flow prediction [14, 40, 41], computer vision [42], Face Recognition [43], etc., while LSTMs are special kinds of RNNs, which are mainly applied in the area of temporal data processing, such as traffic state prediction [34, 44], speech processing [45] and NLP (Natural Language Processing) contexts [46], etc.
The significant difference between fully connected ANN and CNN is that CNN neurons are only connected to a smaller subset of input which decreases the total parameters in the network [47]. CNNs have the ability to extract important and distinctive features from multidimensional by making use of filtering operations. A commonly used type of CNNs, which is similar to multi-layer perception (MLP), consists of numerous convolution layers preceding pooling layers and fully connected layers. CNN structure is illustrated in Figure 2, where it consists of the input layer, convolution layer, pooling layer and fully connected layer. Convolution layer outputs higher abstraction of the feature. Each convolution layer uses several filters, which are designed to have a distinct set of weights. Filters used by the convolution layer have the smaller dimensions compared to the data size. In the training phase, filter weights are automatically determined according to an assigned task. The filters of each convolution layer are applied through the input layer by computing the sum of the product of input and filter, leading to a feature map of each filter. Each feature map detects a distinct high-level feature which is then processed by a pooling layer and a fully connected layer. ReLU activation function is applied to remove all negative values in the feature map.
The benefits of CNNs over other statistical learning methods and DL methods are listed followings [48]:
CNNs have the weight sharing feature, which reduces the number of trainable network parameters and in turn helps the network speed up the training process and avoid overfitting.
Concurrently learning the feature extraction layers and the classification layer causes the model output to be both highly organized and highly reliant on the extracted features.
Large-scale network implementation is much easier with CNN than with other neural networks.
CNN and other kinds of ANNs such has MLP are not designed for sequences and time series data because they do not have memory element. In such cases, RNN can deliver more accurate results. RNNs are widely used in traffic state prediction because traffic data has spatiotemporal characteristics, which cannot be captured by CNN or other kinds of ANNs. RNN structure is illustrated in Figure 3, where RNNs involve an internal memory element that memorizes the previous output. The current output
RNN structure.
where
LSTM is firstly proposed in [49] to overcome the gradient vanishing problems generated by other RNNs. A typical LSTM network consists of an input layer, a recursive hidden layer and an output layer. In the recursive hidden layer, each neuron is made up of four structures: a forget gate, an input gate, an output gate and a memory block. The state of the memory cell reflects the features of the input, while the three gates can read, update and delete features stored in the cell. The LSTM structure is illustrated in Figure 4.
LSTM structure.
The past information carried by the cell state
where
Generally, LSTM can address the vanishing gradient problem that makes network training difficult for a long-sequence temporal data. The long-term dependencies in the data can be learned to improve the prediction accuracy.
Although, CNN and LSTM have advantages in dealing with traffic data with spatiotemporal dependencies, due to the complex and non-linear models of traffic data, it is hard to predict accurate results by using a single model [5]. Some literature proposed that prediction accuracy can be improved by hybrid modeling such as combining CNN and LSTM [50, 51, 52, 53].
The spatial and temporal features can be fully extracted by hybrid models, where CNN in this model is used to capture spatial features of traffic data whereas LSTM is used to extract temporal features. Suppose that we have traffic state data of
Note that
There are mainly two hybridization manners: the first one is to extract spatio-temporal features by concatenating CNN and LSTM, that is, each column of
model to capture the temporal features; the second one is to parallelize CNN and LSTM modeling process by considering the extracted spatial and temporal features are of the same importance, that is, the same traffic state data is input into two models, the final prediction is obtained by passing the output of two models through a FC (Fully Connected) layer. The structure of the two hybridizations is illustrated as follows (Figure 5).
(left) concatenated hybrid model; (right) parallelized hybrid model.
For concatenated hybrid models, the real-time measured data matrix
The high-level spatial feature map output by the one-dimensional CNN can be expressed by
where
where
To extract the temporal features, the high-level spatial feature vector for single or multiple time instants will be selected for the input of each LSTM, which is denoted as
where
where
where
Concatenated hybrid models utilize a one-dimensional CNN to obtain a smaller range of spatial features, in addition, they do not contain a fully connected layer at the output of LSTM models, and thus concatenated hybrid models are with low learning complexity. However, the temporal features delivered by LSTM have a strong correlation with the spatial features output by CNN, which needs some special assumptions about the raw data.
For parallelized hybrid models, the historical data matrix
where
A LSTM is utilized to obtain the high-level temporal feature map. The output of the
By posing a fully connected layer to the output of the
where
In parallelized hybrid models, the spatial and temporal feature maps are considered to be of the same importance, and thus are extracted independently. The fully connected layer merges the output of CNN and LSTM without any special assumptions about the high-level spatial and temporal features.
Traffic state has strong periodic features because people get used to repeating some similar or same behaviors on the same time period of different days or the same day of different weeks, e.g., most people routinely go to work in the morning and go home in the evening during the peak hour [53]; most people routinely go for shopping on weekends rather than weekdays, etc. The periodic features can be used as supplementary information to predict the future traffic state. For the short-term traffic state prediction, the real-time data only contains the data before the prediction time instant, but the historical data on previous days or weeks contain the full data of that period, that means, traffic state information after the inspected time instant on previous days or weeks can be utilized to get the prediction about that on the inspected time instant. Suppose we use parallelized hybrid models, the complete prediction structure should contain CNN and LSTM for the real-time data, CNN and bidirectional LSTM for the historical data, which are connected by using a fully connected layer.
The bidirectional LSTM is composed of two independent forward and backward LSTMs, whose inputs are the time series before and after the inspected time instant. The final prediction of bidirectional LSTM is obtained by concatenating the forward and backward LSTMs. The structure of bidirectional LSTM is depicted in Figure 6.
Bidirectional LSTM structure.
Suppose that additionally, we have historical traffic state data
where
Using Eq. (12), the output of the
Then, the
An accurate and efficient traffic state prediction can provide continuous and precise traffic status and vehicle states based on past information. How to utilize the current and predicted traffic states to make a real-time optimum decision is the main task of the traffic signal control module in ITS. The objectives of traffic signal control include minimizing the average waiting time at multiple intersections, reducing traffic congestion and maximizing network capacity. There exist real-time linear feedback control approaches and MPC (Model-based Predictive Control) that are specifically designed for traffic signal control systems to achieve the targets. The drawback of linear feedback control techniques that have been tried is that the system should always remain in the linear region at all times for the controller. Although, MPC has some advantages such as imposing constraints, the main shortcoming is it needs an accurate dynamic model, which is difficult to be obtained for traffic control systems. Data-driven approaches such as DRL (Deep Reinforcement Learning) based traffic control techniques are widely presented for ITS in recent years because RL can solve complex control problems and deep learning helps to approximate highly nonlinear functions from the complex datasets. In this section, we firstly briefly review the fundamental principles of RL. Then, we focus on multi-agent DRL based traffic signal control techniques such as decentralized multi-agent advantage actor-critic, which can converge to the local optimum and overcome the scalability issue by considering the non-stationarity of MDP transition caused by policy update of the neighborhood; and Nash Q-learning strategy, which can converge to Nash equilibrium by only considering the competition among agents.
Reinforcement Learning (RL) is a promising data-driven approach for decision-making and control in complex dynamic systems. RL methodology formally comes from a Markov Decision Process (MDP), which is a general mathematical framework sequential decision-making algorithms, and consists of five elements [54]:
A set of states
A set of actions
Transition probability
Reward function
The discount factor
RL aims to maximize a numerically defined reward by interacting with the environment to learn how to behave in an environment without any prior knowledge by learning. In traffic signal control systems, RL is used to find the best control policy
where
RL generally can be classified into model-based RL which knows or learns the transition model from state
where
The stochasticity in Eq. (21) comes from the control policy
where
The learning rate
Value-based RL does not work well for continuous control problems with infinite-dimensional action space or high-dimensional problems because it is difficult to explore all the states in a large and continuous space and store them in a table. In such a case, policy-based RL can provide better solutions than value-based RL. By treating the policy
The optimum policy parameters
Policy-based RL tries to select the optimum actions by using the gradient of the objective function with respect to
where
where
where
Actor-critic RL combines the characteristics of policy-based methods and value-based methods, in which an actor is used to control the agent’s behaviors based on policy, critic evaluates the taken action based on value function. From Eq. (27), the objective function can be rewritten as
The loss function for policy and value updating can be respectively defined as
where
Recall that
where
A real traffic network consists of multiple signalized intersections, each of which can be considered as an agent. The states for the
A set of states space of the
A set of action space of the
Transition probability
Control policy of the
The instantaneous reward function of the
The centralized multi-agent RL considers the multi-agent systems as a single-agent system with joint state space
Suppose we have a multi-intersection traffic network, which can be modeled as
where the global states and policies can be communicated from all other agents in the system as well as the neighborhood
We assume Eq. (33) has continuous state-action space and thus multi-agent A2C can be applied to search the optimum policy parameter. From Eq. (31), the Advantage value for the
where
If each agent follows Eqs. (35) and (36) in a decentralized manner, a local optimum policy
In practice, the information exchange among multiple intersections may not be synchronized and communication delay should be considered, which causes policy changing within the same episode and thus leads to non-stationarity. There is some research that try to stabilize convergence and relieve non-stationarity. Tesauro proposes a “Hyper-Q” learning, in which values of mixed strategies rather than base actions are learned and other agents’ strategies are estimated from observed actions via Bayesian inference [55]. Foerster et al. include low-dimensional fingerprints, such as
To relieve non-stationarity, the key is to keep policies from neighboring agents fixed within one episode. We can apply a DNN network to approximate the local policy
Then, the loss function for policy updating can be rewritten by
Even if the policies from the neighbors are fixed and are considered to be additional input, it is still difficult to approximate
where
Then, the cumulative discounted reward can be obtained by
and the local return and Advantage value
and Eq. (38) can be rewritten as
The loss function for value updating can be expressed as
The decreolized MA2C can overcome the scalability issue and achieve local optimum (Pareto Optimality). How to achieve the global optimum using a decentralized approach when the global reward function is non-convex in the future research direction.
Compared to decentralized MA2C, Nash Q-learning does not consider cooperation among agents and thus it has lower computational complexity but can only achieve the Nash equilibrium. Nash Q-learning aims to find the optimal global control policy
where
Eqs. (45) and (46) show that at each iteration
In traffic signal control application, the state space
By conducting a simulation on SUMO for a two-intersection case, we can observe in Figure 7 that the centralized DQN outperform the centralized Q-learning in terms of reward value (Average Waiting Time/s) and convergence rate (the Number of Iterations). When the number of agents is small (two, in this case), by using the centralized methods, the average waiting time can converge to the local optimum, which is more optimal than the Nash equilibrium delivered by Nash Q learning. However, the convergence rate of Nash Q learning is higher than that of centralized methods.
Comparison of different multi-agent RL methods for traffic signal control.
In this chapter, we introduced deep learning-based traffic state prediction technique, which can provide accurate future information for traffic control and decision making. The traffic state data depicts a strong correlation in the spatial and temporal domain, which can be utilized by applying CNN and LSTM techniques to improve the prediction accuracy. CNN technique is used to capture high-level spatial features while LSTM can provide excellent performance in dealing with time-sequential data by extracting high-level temporal features. We firstly reviewed the fundamentals of deep learning and presented the architecture of CNN and LSTM. Then, we introduced how to combine these two models to form concatenated hybrid models and parallelized hybrid models. Finally, we proposed bidirectional LSTM models to enhance prediction performance by learning additional high-level temporal features from the historical data in previous days.
Furthermore, we introduced the decentralized multi-agent advantage Actor-Critic technique and Nash Q learning for traffic signal control applications. We firstly briefly review the fundamental principles of RL. Then, we focus on multi-agent DRL-based traffic signal control techniques such as decentralized multi-agent advantage actor-critic, which can converge to the local optimum and overcome the scalability issue by considering the non-stationarity of MDP transition caused by policy update of the neighborhood.
The main contribution of this chapter can be summarized as followings:
We reviewed the state-of-the-art technique in traffic state prediction and traffic control strategies, and provide readers with a clear framework for understanding how to apply deep learning models to traffic state prediction and how to deal with multi-agent traffic control by using RL strategies.
We proposed the hybrid prediction models, which can utilize CNN and LSTM to capture the spatio-temporal feature of traffic data.
We proposed a multi-agent deep RL (MARL) strategy, which conducts in a decentralized manner and considers the cooperation among agents and thus can overcome the scalability issue and achieve local optimum.
We compared the centralized RL Q-learning, DQN to the Nash Q-learning strategy in terms of the reward value and convergence rate.
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\n\nDr Alex Lazinica
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Builders",coverURL:"https://cdn.intechopen.com/books/images_new/6302.jpg",editedByType:"Edited by",editors:[{id:"182744",title:"Dr.",name:"Philip",middleName:null,surname:"Builders",slug:"philip-builders",fullName:"Philip Builders"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"5612",title:"Aromatic and Medicinal Plants",subtitle:"Back to Nature",isOpenForSubmission:!1,hash:"ccf7987200bfc541e2e56bb138de86f3",slug:"aromatic-and-medicinal-plants-back-to-nature",bookSignature:"Hany A. El-Shemy",coverURL:"https://cdn.intechopen.com/books/images_new/5612.jpg",editedByType:"Edited by",editors:[{id:"54719",title:"Prof.",name:"Hany",middleName:null,surname:"El-Shemy",slug:"hany-el-shemy",fullName:"Hany El-Shemy"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"5494",title:"Chinese Medical Therapies for Diabetes, Infertility, Silicosis and the Theoretical Basis",subtitle:null,isOpenForSubmission:!1,hash:"7b3b6a2700d7fd0511770bf77290a422",slug:"chinese-medical-therapies-for-diabetes-infertility-silicosis-and-the-theoretical-basis",bookSignature:"Xing-Tai Li",coverURL:"https://cdn.intechopen.com/books/images_new/5494.jpg",editedByType:"Edited by",editors:[{id:"73821",title:"Dr.",name:"Xing-Tai",middleName:null,surname:"Li",slug:"xing-tai-li",fullName:"Xing-Tai Li"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"5222",title:"Cannabinoids in Health and Disease",subtitle:null,isOpenForSubmission:!1,hash:"d684a703afd17dc97d18480a982e5316",slug:"cannabinoids-in-health-and-disease",bookSignature:"Rosaria Meccariello and Rosanna Chianese",coverURL:"https://cdn.intechopen.com/books/images_new/5222.jpg",editedByType:"Edited by",editors:[{id:"143980",title:"Prof.",name:"Rosaria",middleName:null,surname:"Meccariello",slug:"rosaria-meccariello",fullName:"Rosaria Meccariello"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"4625",title:"Complementary Therapies for the Body, Mind and Soul",subtitle:null,isOpenForSubmission:!1,hash:"48cd88cd7a6ffb4ade0088448e5ac56b",slug:"complementary-therapies-for-the-body-mind-and-soul",bookSignature:"Marcelo Saad",coverURL:"https://cdn.intechopen.com/books/images_new/4625.jpg",editedByType:"Edited by",editors:[{id:"51991",title:"Prof.",name:"Marcelo",middleName:null,surname:"Saad",slug:"marcelo-saad",fullName:"Marcelo Saad"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}}],booksByTopicTotal:16,seriesByTopicCollection:[],seriesByTopicTotal:0,mostCitedChapters:[{id:"61866",doi:"10.5772/intechopen.76139",title:"Plants Secondary Metabolites: The Key Drivers of the Pharmacological Actions of Medicinal Plants",slug:"plants-secondary-metabolites-the-key-drivers-of-the-pharmacological-actions-of-medicinal-plants",totalDownloads:8875,totalCrossrefCites:56,totalDimensionsCites:140,abstract:"The vast and versatile pharmacological effects of medicinal plants are basically dependent on their phytochemical constituents. Generally, the phytochemical constituents of plants fall into two categories based on their role in basic metabolic processes, namely primary and secondary metabolites. Primary plant metabolites are involved in basic life functions; therefore, they are more or less similar in all living cells. On the other hand, secondary plant metabolites are products of subsidiary pathways as the shikimic acid pathway. In the course of studying, the medicinal effect of herbals is oriented towards the secondary plant metabolites. Secondary plant metabolites played an important role in alleviating several aliments in the traditional medicine and folk uses. In modern medicine, they provided lead compounds for the production of medications for treating various diseases from migraine up to cancer. Secondary plant metabolites are classified according to their chemical structures into various classes. In this chapter, we will be presenting various classes of secondary plant metabolites, their distribution in different plant families and their important medicinal uses.",book:{id:"6302",slug:"herbal-medicine",title:"Herbal Medicine",fullTitle:"Herbal Medicine"},signatures:"Rehab A. Hussein and Amira A. El-Anssary",authors:[{id:"212117",title:"Dr.",name:"Rehab",middleName:null,surname:"Hussein",slug:"rehab-hussein",fullName:"Rehab Hussein"},{id:"221140",title:"Dr.",name:"Amira",middleName:null,surname:"El-Anssary",slug:"amira-el-anssary",fullName:"Amira El-Anssary"}]},{id:"64851",doi:"10.5772/intechopen.80348",title:"Herbal Medicines in African Traditional Medicine",slug:"herbal-medicines-in-african-traditional-medicine",totalDownloads:14206,totalCrossrefCites:30,totalDimensionsCites:52,abstract:"African traditional medicine is a form of holistic health care system organized into three levels of specialty, namely divination, spiritualism, and herbalism. The traditional healer provides health care services based on culture, religious background, knowledge, attitudes, and beliefs that are prevalent in his community. Illness is regarded as having both natural and supernatural causes and thus must be treated by both physical and spiritual means, using divination, incantations, animal sacrifice, exorcism, and herbs. Herbal medicine is the cornerstone of traditional medicine but may include minerals and animal parts. The adjustment is ok, but may be replaced with –‘ Herbal medicine was once termed primitive by western medicine but through scientific investigations there is a better understanding of its therapeutic activities such that many pharmaceuticals have been modeled on phytochemicals derived from it. Major obstacles to the use of African medicinal plants are their poor quality control and safety. Traditional medical practices are still shrouded with much secrecy, with few reports or documentations of adverse reactions. However, the future of African traditional medicine is bright if viewed in the context of service provision, increase of health care coverage, economic potential, and poverty reduction. Formal recognition and integration of traditional medicine into conventional medicine will hold much promise for the future.",book:{id:"6302",slug:"herbal-medicine",title:"Herbal Medicine",fullTitle:"Herbal Medicine"},signatures:"Ezekwesili-Ofili Josephine Ozioma and Okaka Antoinette Nwamaka\nChinwe",authors:[{id:"191264",title:"Prof.",name:"Josephine",middleName:"Ozioma",surname:"Ezekwesili-Ofili",slug:"josephine-ezekwesili-ofili",fullName:"Josephine Ezekwesili-Ofili"},{id:"211585",title:"Prof.",name:"Antoinette",middleName:null,surname:"Okaka",slug:"antoinette-okaka",fullName:"Antoinette Okaka"}]},{id:"54028",doi:"10.5772/67291",title:"Chemical Composition and Biological Activities of Mentha Species",slug:"chemical-composition-and-biological-activities-of-mentha-species",totalDownloads:7474,totalCrossrefCites:13,totalDimensionsCites:46,abstract:"The genus Mentha L. (Lamiaceae) is distributed all over the world and can be found in many environments. Mentha species, one of the world’s oldest and most popular herbs, are widely used in cooking, in cosmetics, and as alternative or complementary therapy, mainly for the treatment of gastrointestinal disorders like flatulence, indigestion, nausea, vomiting, anorexia, and ulcerative colitis. Furthermore, it is well documented that the essential oil and extracts of Mentha species possess antimicrobial, fungicidal, antiviral, insecticidal, and antioxidant properties. The economic importance of mints is also evident; mint oil and its constituents and derivatives are used as flavoring agents throughout the world in food, pharmaceutical, herbal, perfumery, and flavoring industry. To provide a scientific basis for their traditional uses, several studies have been conducted to determine the chemical composition of mints and assess their biological activities. This chapter describes the therapeutic effects and uses of Mentha species and their constituents, particularly essential oils and phenolic compounds; some additional biological activities will also be considered.",book:{id:"5612",slug:"aromatic-and-medicinal-plants-back-to-nature",title:"Aromatic and Medicinal Plants",fullTitle:"Aromatic and Medicinal Plants - Back to Nature"},signatures:"Fatiha Brahmi, Madani Khodir, Chibane Mohamed and Duez Pierre",authors:[{id:"193281",title:"Dr.",name:"Fatiha",middleName:null,surname:"Brahmi",slug:"fatiha-brahmi",fullName:"Fatiha Brahmi"},{id:"199693",title:"Prof.",name:"Khodir",middleName:null,surname:"Madani",slug:"khodir-madani",fullName:"Khodir Madani"},{id:"199694",title:"Prof.",name:"Pierre",middleName:null,surname:"Duez",slug:"pierre-duez",fullName:"Pierre Duez"},{id:"203738",title:"Prof.",name:"Mohamed",middleName:null,surname:"Chibane",slug:"mohamed-chibane",fullName:"Mohamed Chibane"}]},{id:"58270",doi:"10.5772/intechopen.72437",title:"Toxicity and Safety Implications of Herbal Medicines Used in Africa",slug:"toxicity-and-safety-implications-of-herbal-medicines-used-in-africa",totalDownloads:3376,totalCrossrefCites:16,totalDimensionsCites:39,abstract:"The use of herbal medicines has seen a great upsurge globally. In developing countries, many patronize them largely due to cultural acceptability, availability and cost. In developed countries, they are used because they are natural and therefore assumed to be safer than allopathic medicines. In recent times, however, there has been a growing concern about their safety. This has created a situation of ambivalence in discussions regarding their use. Some medicinal plants are intrinsically toxic by virtue of their constituents and can cause adverse reactions if inappropriately used. Other factors such as herb-drug interactions, lack of adherence to good manufacturing practice (GMP), poor regulatory measures and adulteration may also lead to adverse events in their use. Many in vivo tests on aqueous extracts largely support the safety of herbal medicines, whereas most in vitro tests on isolated single cells mostly with extracts other than aqueous ones show contrary results and thus continue the debate on herbal medicine safety. It is expected that toxicity studies concerning herbal medicine should reflect their traditional use to allow for rational discussions regarding their safety for their beneficial use. While various attempts continue to establish the safety of various herbal medicines in man, their cautious and responsible use is required.",book:{id:"6302",slug:"herbal-medicine",title:"Herbal Medicine",fullTitle:"Herbal Medicine"},signatures:"Merlin L.K. Mensah, Gustav Komlaga, Arnold D. Forkuo, Caleb\nFirempong, Alexander K. Anning and Rita A. Dickson",authors:[{id:"190435",title:"Dr.",name:"Caleb",middleName:null,surname:"Firempong",slug:"caleb-firempong",fullName:"Caleb Firempong"},{id:"212111",title:"Dr.",name:"Gustav",middleName:null,surname:"Komlaga",slug:"gustav-komlaga",fullName:"Gustav Komlaga"},{id:"217045",title:"Dr.",name:"Arnold Forkuo",middleName:null,surname:"Donkor",slug:"arnold-forkuo-donkor",fullName:"Arnold Forkuo Donkor"},{id:"217049",title:"Prof.",name:"Merlin Lincoln Kwao",middleName:null,surname:"Mensah",slug:"merlin-lincoln-kwao-mensah",fullName:"Merlin Lincoln Kwao Mensah"},{id:"217488",title:"Dr.",name:"Alexander K.",middleName:null,surname:"Anning",slug:"alexander-k.-anning",fullName:"Alexander K. Anning"},{id:"223959",title:"Prof.",name:"Akosua Rita",middleName:null,surname:"Dickson",slug:"akosua-rita-dickson",fullName:"Akosua Rita Dickson"}]},{id:"26489",doi:"10.5772/28224",title:"Alternative and Traditional Medicines Systems in Pakistan: History, Regulation, Trends, Usefulness, Challenges, Prospects and Limitations",slug:"alternative-and-traditional-medicines-systems-in-pakistan-history-regulation-trends-usefulness-chall",totalDownloads:9199,totalCrossrefCites:9,totalDimensionsCites:21,abstract:null,book:{id:"542",slug:"a-compendium-of-essays-on-alternative-therapy",title:"A Compendium of Essays on Alternative Therapy",fullTitle:"A Compendium of Essays on Alternative Therapy"},signatures:"Shahzad Hussain, Farnaz Malik, Nadeem Khalid, Muhammad Abdul Qayyum and Humayun Riaz",authors:[{id:"73162",title:"Dr.",name:"Shahzad",middleName:null,surname:"Hussain",slug:"shahzad-hussain",fullName:"Shahzad Hussain"},{id:"82266",title:"Dr.",name:"Farnaz",middleName:null,surname:"Malik",slug:"farnaz-malik",fullName:"Farnaz Malik"},{id:"124185",title:"Dr.",name:"Humayun",middleName:null,surname:"Riaz",slug:"humayun-riaz",fullName:"Humayun Riaz"},{id:"124186",title:"Mr.",name:"Muhammad Abdul",middleName:null,surname:"Qayyum",slug:"muhammad-abdul-qayyum",fullName:"Muhammad Abdul Qayyum"},{id:"125340",title:"Mr.",name:"Nadeem",middleName:null,surname:"Khalid",slug:"nadeem-khalid",fullName:"Nadeem Khalid"}]}],mostDownloadedChaptersLast30Days:[{id:"64851",title:"Herbal Medicines in African Traditional Medicine",slug:"herbal-medicines-in-african-traditional-medicine",totalDownloads:14207,totalCrossrefCites:30,totalDimensionsCites:52,abstract:"African traditional medicine is a form of holistic health care system organized into three levels of specialty, namely divination, spiritualism, and herbalism. The traditional healer provides health care services based on culture, religious background, knowledge, attitudes, and beliefs that are prevalent in his community. Illness is regarded as having both natural and supernatural causes and thus must be treated by both physical and spiritual means, using divination, incantations, animal sacrifice, exorcism, and herbs. Herbal medicine is the cornerstone of traditional medicine but may include minerals and animal parts. The adjustment is ok, but may be replaced with –‘ Herbal medicine was once termed primitive by western medicine but through scientific investigations there is a better understanding of its therapeutic activities such that many pharmaceuticals have been modeled on phytochemicals derived from it. Major obstacles to the use of African medicinal plants are their poor quality control and safety. Traditional medical practices are still shrouded with much secrecy, with few reports or documentations of adverse reactions. However, the future of African traditional medicine is bright if viewed in the context of service provision, increase of health care coverage, economic potential, and poverty reduction. Formal recognition and integration of traditional medicine into conventional medicine will hold much promise for the future.",book:{id:"6302",slug:"herbal-medicine",title:"Herbal Medicine",fullTitle:"Herbal Medicine"},signatures:"Ezekwesili-Ofili Josephine Ozioma and Okaka Antoinette Nwamaka\nChinwe",authors:[{id:"191264",title:"Prof.",name:"Josephine",middleName:"Ozioma",surname:"Ezekwesili-Ofili",slug:"josephine-ezekwesili-ofili",fullName:"Josephine Ezekwesili-Ofili"},{id:"211585",title:"Prof.",name:"Antoinette",middleName:null,surname:"Okaka",slug:"antoinette-okaka",fullName:"Antoinette Okaka"}]},{id:"61866",title:"Plants Secondary Metabolites: The Key Drivers of the Pharmacological Actions of Medicinal Plants",slug:"plants-secondary-metabolites-the-key-drivers-of-the-pharmacological-actions-of-medicinal-plants",totalDownloads:8875,totalCrossrefCites:56,totalDimensionsCites:140,abstract:"The vast and versatile pharmacological effects of medicinal plants are basically dependent on their phytochemical constituents. Generally, the phytochemical constituents of plants fall into two categories based on their role in basic metabolic processes, namely primary and secondary metabolites. Primary plant metabolites are involved in basic life functions; therefore, they are more or less similar in all living cells. On the other hand, secondary plant metabolites are products of subsidiary pathways as the shikimic acid pathway. In the course of studying, the medicinal effect of herbals is oriented towards the secondary plant metabolites. Secondary plant metabolites played an important role in alleviating several aliments in the traditional medicine and folk uses. In modern medicine, they provided lead compounds for the production of medications for treating various diseases from migraine up to cancer. Secondary plant metabolites are classified according to their chemical structures into various classes. In this chapter, we will be presenting various classes of secondary plant metabolites, their distribution in different plant families and their important medicinal uses.",book:{id:"6302",slug:"herbal-medicine",title:"Herbal Medicine",fullTitle:"Herbal Medicine"},signatures:"Rehab A. Hussein and Amira A. El-Anssary",authors:[{id:"212117",title:"Dr.",name:"Rehab",middleName:null,surname:"Hussein",slug:"rehab-hussein",fullName:"Rehab Hussein"},{id:"221140",title:"Dr.",name:"Amira",middleName:null,surname:"El-Anssary",slug:"amira-el-anssary",fullName:"Amira El-Anssary"}]},{id:"77433",title:"Extraction of Bioactive Compounds from Medicinal Plants and Herbs",slug:"extraction-of-bioactive-compounds-from-medicinal-plants-and-herbs",totalDownloads:1266,totalCrossrefCites:2,totalDimensionsCites:5,abstract:"Human beings have relied on herbs and medicinal plants as sources of food and remedy from time immemorial. Bioactive compounds from plants are currently the subject of much research interest, but their extraction as part of phytochemical and/or biological investigations present specific challenges. Herbalists or scientists have developed many protocols of extraction of bioactive ingredients to ensure the effectiveness and the efficacy of crude drugs that were used to get relief from sickness. With the advent of new leads from plants such as morphine, quinine, taxol, artemisinin, and alkaloids from Voacanga species, a lot of attention is paid to the mode of extraction of active phytochemicals to limit the cost linked to the synthesis and isolation. Thus, the extraction of active compounds from plants needs appropriate extraction methods and techniques that provide bioactive ingredients-rich extracts and fractions. The extraction procedures, therefore, play a critical role in the yield, the nature of phytochemical content, etc. This chapter aims to present, describe, and compare extraction procedures of bioactive compounds from herbs and medicinal plants.",book:{id:"10356",slug:"natural-medicinal-plants",title:"Natural Medicinal Plants",fullTitle:"Natural Medicinal Plants"},signatures:"Fongang Fotsing Yannick Stéphane, Bankeu Kezetas Jean Jules, Gaber El-Saber Batiha, Iftikhar Ali and Lenta Ndjakou Bruno",authors:[{id:"224515",title:"Dr.",name:"Fongang Fotsing",middleName:null,surname:"Yannick Stéphane",slug:"fongang-fotsing-yannick-stephane",fullName:"Fongang Fotsing Yannick Stéphane"},{id:"227816",title:"Dr.",name:"Bankeu Kezetas",middleName:null,surname:"Jean Jules",slug:"bankeu-kezetas-jean-jules",fullName:"Bankeu Kezetas Jean Jules"},{id:"227817",title:"Prof.",name:"Lenta Ndjakou",middleName:null,surname:"Bruno",slug:"lenta-ndjakou-bruno",fullName:"Lenta Ndjakou Bruno"},{id:"349790",title:"Prof.",name:"Gaber",middleName:null,surname:"El-Saber Batiha",slug:"gaber-el-saber-batiha",fullName:"Gaber El-Saber Batiha"},{id:"357350",title:"Dr.",name:"Iftikhar",middleName:null,surname:"Ali",slug:"iftikhar-ali",fullName:"Iftikhar Ali"}]},{id:"26491",title:"Homeopathy: Treatment of Cancer with the Banerji Protocols",slug:"homeopathy-treatment-of-cancer-with-the-banerji-protocols",totalDownloads:54047,totalCrossrefCites:1,totalDimensionsCites:2,abstract:null,book:{id:"542",slug:"a-compendium-of-essays-on-alternative-therapy",title:"A Compendium of Essays on Alternative Therapy",fullTitle:"A Compendium of Essays on Alternative Therapy"},signatures:"Prasanta Banerji and Pratip Banerji",authors:[{id:"79939",title:"Dr",name:"Prasanta",middleName:null,surname:"Banerji",slug:"prasanta-banerji",fullName:"Prasanta Banerji"},{id:"79943",title:"Dr.",name:"Pratip",middleName:null,surname:"Banerji",slug:"pratip-banerji",fullName:"Pratip Banerji"}]},{id:"54028",title:"Chemical Composition and Biological Activities of Mentha Species",slug:"chemical-composition-and-biological-activities-of-mentha-species",totalDownloads:7474,totalCrossrefCites:13,totalDimensionsCites:46,abstract:"The genus Mentha L. (Lamiaceae) is distributed all over the world and can be found in many environments. Mentha species, one of the world’s oldest and most popular herbs, are widely used in cooking, in cosmetics, and as alternative or complementary therapy, mainly for the treatment of gastrointestinal disorders like flatulence, indigestion, nausea, vomiting, anorexia, and ulcerative colitis. Furthermore, it is well documented that the essential oil and extracts of Mentha species possess antimicrobial, fungicidal, antiviral, insecticidal, and antioxidant properties. The economic importance of mints is also evident; mint oil and its constituents and derivatives are used as flavoring agents throughout the world in food, pharmaceutical, herbal, perfumery, and flavoring industry. To provide a scientific basis for their traditional uses, several studies have been conducted to determine the chemical composition of mints and assess their biological activities. This chapter describes the therapeutic effects and uses of Mentha species and their constituents, particularly essential oils and phenolic compounds; some additional biological activities will also be considered.",book:{id:"5612",slug:"aromatic-and-medicinal-plants-back-to-nature",title:"Aromatic and Medicinal Plants",fullTitle:"Aromatic and Medicinal Plants - Back to Nature"},signatures:"Fatiha Brahmi, Madani Khodir, Chibane Mohamed and Duez Pierre",authors:[{id:"193281",title:"Dr.",name:"Fatiha",middleName:null,surname:"Brahmi",slug:"fatiha-brahmi",fullName:"Fatiha Brahmi"},{id:"199693",title:"Prof.",name:"Khodir",middleName:null,surname:"Madani",slug:"khodir-madani",fullName:"Khodir Madani"},{id:"199694",title:"Prof.",name:"Pierre",middleName:null,surname:"Duez",slug:"pierre-duez",fullName:"Pierre Duez"},{id:"203738",title:"Prof.",name:"Mohamed",middleName:null,surname:"Chibane",slug:"mohamed-chibane",fullName:"Mohamed Chibane"}]}],onlineFirstChaptersFilter:{topicId:"172",limit:6,offset:0},onlineFirstChaptersCollection:[{id:"82022",title:"Vibration Therapy for Health Promotion",slug:"vibration-therapy-for-health-promotion",totalDownloads:9,totalDimensionsCites:0,doi:"10.5772/intechopen.105024",abstract:"Vibration therapy has been used as a clinical intervention, in which mechanical vibration is transmitted to a part or to the whole body of the individual. It is very important to point out that mechanical vibration is a natural stimulus that is part of the daily life of all living beings and is periodically added to the organism due to a movement of the body. When, for several reasons, the person cannot add mechanical vibration to the body, the mechanical vibration generated by a device can be transmitted to the person in contact with it. When the intervention aims to treat a complaint referring to a specific anatomical segment, it is called local or segmental vibration therapy. However, when mechanical vibration is transmitted to the whole person’s body, aiming for an improvement in the performance, or as a clinical intervention, the procedure is called Systemic vibration therapy. The biological effects would be due to the mechano-transduction mechanism by which cells convert mechanical stimulus into biological activity, releasing various hormones and other substances. This form of mechano-transduction is important to physiological processes in the body, including proprioception, effects on bone mineral density, muscle, balance, and functionality, promoting the modulation of biological effects through specific signaling pathways. In this chapter, the use of mechanical vibration as an intervention aiming to improve and optimize daily life is discussed, either as a local or systemic application, targeting a specific part of the body or the whole body, respectively.",book:{id:"11042",title:"Complementary Therapies",coverURL:"https://cdn.intechopen.com/books/images_new/11042.jpg"},signatures:"Danúbia da Cunha de Sá-Caputo, Adérito Seixas, Redha Taiar and Mario Bernardo-Filho"},{id:"81876",title:"An Introduction to Chiropractic BioPhysics® (CBP®) Technique: A Full Spine Rehabilitation Approach to Reducing Spine Deformities",slug:"an-introduction-to-chiropractic-biophysics-cbp-technique-a-full-spine-rehabilitation-approach-to-red",totalDownloads:76,totalDimensionsCites:0,doi:"10.5772/intechopen.102686",abstract:"Chiropractic Biophysics® (CBP®) technique is a full-spine and posture correcting method that incorporates mathematical principles into a unique approach to treat spinal disorders. It considers that the identification of postural rotations and translations of human postures are first evaluated and compared to the radiographic assessment of the spine alignment. Mirror image® postural positions and movements are utilized including spinal extension positions to improve the spine and posture towards a normal/ideal alignment. Specifically, corrective exercises, corrective traction and chiropractic adjustments are performed encompassing a multimodal rehabilitation program with the goal of improving the posture and spine alignment. CBP Rehabilitation programs are typically performed in-office with supportive at-home measures. Repeat assessment including radiographs are used to quantify and monitor structural improvements. CBP technique is an evidence-based approach to treat spine deformities and is supported by all forms of clinical evidence including systematic literature reviews, randomized controlled trials, non-randomized controlled trials, case reports/series as well as is supported by biomechanical posture-spine coupling validity, radiographic and posture analysis reliability/repeatability and use of a validated biomechanical spinal model as the outcome goal of care. CBP technique is a proven method to improve pain, disability and quality of life in those with structural deformities.",book:{id:"11042",title:"Complementary Therapies",coverURL:"https://cdn.intechopen.com/books/images_new/11042.jpg"},signatures:"Deed E. Harrison and Paul A. Oakley"},{id:"80903",title:"Methods and Tools for Assessing Muscle Asymmetry in the Analysis of Electromyographic Signals",slug:"methods-and-tools-for-assessing-muscle-asymmetry-in-the-analysis-of-electromyographic-signals",totalDownloads:33,totalDimensionsCites:0,doi:"10.5772/intechopen.103061",abstract:"The generalized information about the possibilities of assessing asymmetry and the prospects of research tools is presented. The important role of the choice of different methods for processing electromyographic signals, the results of which can be considered as an objective criterion for assessing the asymmetry of the muscles of the extremities, is noted, such as the asymmetry coefficient, a widely used parameter in statistical analysis, which characterizes the asymmetry of the statistical distribution. Also applied is the segmental method of studying the body to obtain estimates of the composition and differences between individual body segments. The isokinetic test method, which makes it possible to assess asymmetry in measuring muscle strength, relies on the randomness of the dynamic processes of the biological system. Use of nonlinear dynamics, the theory of dynamic chaos, and fractal analysis allows for determining the fractal properties of biosignals, and from the classical methods used correlation analysis.",book:{id:"11042",title:"Complementary Therapies",coverURL:"https://cdn.intechopen.com/books/images_new/11042.jpg"},signatures:"Kamala Pashayeva and Namiq Abdullayev"},{id:"80124",title:"Non-pharmacological Therapies in Integrative Rehabilitation and Physiotherapy",slug:"non-pharmacological-therapies-in-integrative-rehabilitation-and-physiotherapy",totalDownloads:49,totalDimensionsCites:0,doi:"10.5772/intechopen.101920",abstract:"Although pharmacological therapy is a resource to be taken into account in rehabilitation medicine and physiotherapy (RMP), a series of therapies, techniques, and empirical practices have been available since time immemorial, some with scientific publications, which play an important role in healthcare current health worldwide. Some millennials like Traditional Chinese Medicine (TCM) and medicinal plants (MP), and other centenarians like Homeopathic Medicine (HM) and Osteopathic Medicine (OM). In the twentieth century, numerous medical techniques were developed, framed under the term of Biological Medicine in Europe such as Antihomotoxic Medicine (AM), Neural Therapy (NT), Catalytic Oligotherapy (CO). Throughout the twentieth and twenty-first centuries, the wealth of experiences and knowledge on the application of Growth Factors in therapeutics, Oxygen-Ozone Therapy, and other related techniques that have enriched medical practice has increased. The objective of this chapter is to highlight the convenience of incorporating unconventional therapies (UT) techniques with scientific evidence into RMP that enjoy efficacy, efficiency, and economic and environmental sustainability. It would be desirable to allocate more financial resources to continue researching these therapies, implement them in Health Sciences studies and continue working to comply with the maxim of medicine that is “primum non nocere” (first do no harm).",book:{id:"11042",title:"Complementary Therapies",coverURL:"https://cdn.intechopen.com/books/images_new/11042.jpg"},signatures:"Andrés J. Ursa Herguedas"},{id:"79981",title:"Traditional Islamic Herbal Medicine and Complementary Therapies",slug:"traditional-islamic-herbal-medicine-and-complementary-therapies",totalDownloads:116,totalDimensionsCites:0,doi:"10.5772/intechopen.101927",abstract:"Herbal products found abundantly in several plants are the rich source of phytochemicals with a wide range of pharmacological activities and few adverse effects. Medicinal plants contain active ingredients that assist the body in reestablishing its natural balance and healing itself. Various herbs, which are commonly used in traditional Islamic medicine, can have an impact on human body systems. Natural products are primary sources of effective drugs with novel structures and distinct mechanisms of action for the treatment of various types of complications as well as the drug discovery process. The various pharmacological properties such as antimicrobial, anticancer, antioxidant, antihypertensive, immunomodulatory, anti-inflammatory and anti-diabetic properties of several natural products are well documented in the Ayurveda and Unani system of medicine. Some of the natural products’ active ingredients have been documented, but the majority are still being researched as complementary medicine. As a result, more research is required to investigate their complementary medicine system. The present chapter provides a comprehensive update on selected traditional Islamic medicinal plants and their bioactive products mentioned in Islamic scriptures as complementary therapies to various diseases. The chapter also provides an in-depth update of pharmacological and clinical studies of natural products with special emphasis on cancer and diabetes.",book:{id:"11042",title:"Complementary Therapies",coverURL:"https://cdn.intechopen.com/books/images_new/11042.jpg"},signatures:"Sahabjada Siddiqui, Afsana Khatoon, Khursheed Ahmad, Shivbrat Upadhyay, Aditi Srivastava, Anchal Trivedi, Ishrat Husain, Rumana Ahmad, Mohsin Ali Khan and Md Arshad"},{id:"79924",title:"Cognitive Hypnotherapy and EMDR: Two Effective Psychodynamic Therapies for the Rapid Reduction of Cognitive Anxiety",slug:"cognitive-hypnotherapy-and-emdr-two-effective-psychodynamic-therapies-for-the-rapid-reduction-of-cog",totalDownloads:115,totalDimensionsCites:0,doi:"10.5772/intechopen.101770",abstract:"In the main research into cognitive anxiety has focused on the conscious mind. The aim of this chapter is to review two psychodynamic psychotherapies, cognitive hypnotherapy (CH) and eye movement desensitisation and reprocessing (EMDR). Both therapies focus on implicit or unconscious processes for the rapid relief of cognitive anxiety. The objective is to give credence to CH and EMDR both in the scientific and medical domains. The philosophy is concerned with changing negative cognitions and dysfunctional feelings through a process of desensitisation and reprocessing, utilising positive imagery. CH and EMDR were investigated in an intervention study with advanced pianists (n = 46). Participants were of mixed gender aged 18–26 and were randomly assigned to a therapy or control group. The therapy groups received two therapies of either CH or EMDR during a two week period between two concerts. Quantitative data were collected through the Spielberger State-Trait Anxiety Inventory. Results showed that both therapy groups (but not the control) experienced a significant reduction in state anxiety post-therapy and trait anxiety decreased significantly below baseline levels in the EMDR group. This chapter further reviews research into CH and EMDR documented through a case study allowing for qualitative assessment of the therapies where two sessions only were required to effect positive change.",book:{id:"11042",title:"Complementary Therapies",coverURL:"https://cdn.intechopen.com/books/images_new/11042.jpg"},signatures:"Elizabeth Brooker"}],onlineFirstChaptersTotal:12},preDownload:{success:null,errors:{}},subscriptionForm:{success:null,errors:{}},aboutIntechopen:{},privacyPolicy:{},peerReviewing:{},howOpenAccessPublishingWithIntechopenWorks:{},sponsorshipBooks:{sponsorshipBooks:[],offset:0,limit:8,total:null},allSeries:{pteSeriesList:[{id:"14",title:"Artificial Intelligence",numberOfPublishedBooks:9,numberOfPublishedChapters:89,numberOfOpenTopics:6,numberOfUpcomingTopics:0,issn:"2633-1403",doi:"10.5772/intechopen.79920",isOpenForSubmission:!0},{id:"7",title:"Biomedical Engineering",numberOfPublishedBooks:12,numberOfPublishedChapters:104,numberOfOpenTopics:3,numberOfUpcomingTopics:0,issn:"2631-5343",doi:"10.5772/intechopen.71985",isOpenForSubmission:!0}],lsSeriesList:[{id:"11",title:"Biochemistry",numberOfPublishedBooks:32,numberOfPublishedChapters:318,numberOfOpenTopics:4,numberOfUpcomingTopics:0,issn:"2632-0983",doi:"10.5772/intechopen.72877",isOpenForSubmission:!0},{id:"25",title:"Environmental Sciences",numberOfPublishedBooks:1,numberOfPublishedChapters:12,numberOfOpenTopics:4,numberOfUpcomingTopics:0,issn:"2754-6713",doi:"10.5772/intechopen.100362",isOpenForSubmission:!0},{id:"10",title:"Physiology",numberOfPublishedBooks:11,numberOfPublishedChapters:141,numberOfOpenTopics:4,numberOfUpcomingTopics:0,issn:"2631-8261",doi:"10.5772/intechopen.72796",isOpenForSubmission:!0}],hsSeriesList:[{id:"3",title:"Dentistry",numberOfPublishedBooks:8,numberOfPublishedChapters:129,numberOfOpenTopics:2,numberOfUpcomingTopics:0,issn:"2631-6218",doi:"10.5772/intechopen.71199",isOpenForSubmission:!0},{id:"6",title:"Infectious Diseases",numberOfPublishedBooks:13,numberOfPublishedChapters:113,numberOfOpenTopics:3,numberOfUpcomingTopics:1,issn:"2631-6188",doi:"10.5772/intechopen.71852",isOpenForSubmission:!0},{id:"13",title:"Veterinary Medicine and Science",numberOfPublishedBooks:11,numberOfPublishedChapters:106,numberOfOpenTopics:3,numberOfUpcomingTopics:0,issn:"2632-0517",doi:"10.5772/intechopen.73681",isOpenForSubmission:!0}],sshSeriesList:[{id:"22",title:"Business, Management and Economics",numberOfPublishedBooks:1,numberOfPublishedChapters:19,numberOfOpenTopics:3,numberOfUpcomingTopics:0,issn:"2753-894X",doi:"10.5772/intechopen.100359",isOpenForSubmission:!0},{id:"23",title:"Education and Human Development",numberOfPublishedBooks:0,numberOfPublishedChapters:5,numberOfOpenTopics:1,numberOfUpcomingTopics:1,issn:null,doi:"10.5772/intechopen.100360",isOpenForSubmission:!0},{id:"24",title:"Sustainable Development",numberOfPublishedBooks:0,numberOfPublishedChapters:15,numberOfOpenTopics:5,numberOfUpcomingTopics:0,issn:null,doi:"10.5772/intechopen.100361",isOpenForSubmission:!0}],testimonialsList:[{id:"13",text:"The collaboration with and support of the technical staff of IntechOpen is fantastic. The whole process of submitting an article and editing of the submitted article goes extremely smooth and fast, the number of reads and downloads of chapters is high, and the contributions are also frequently cited.",author:{id:"55578",name:"Antonio",surname:"Jurado-Navas",institutionString:null,profilePictureURL:"https://s3.us-east-1.amazonaws.com/intech-files/0030O00002bRisIQAS/Profile_Picture_1626166543950",slug:"antonio-jurado-navas",institution:{id:"720",name:"University of Malaga",country:{id:null,name:"Spain"}}}},{id:"6",text:"It is great to work with the IntechOpen to produce a worthwhile collection of research that also becomes a great educational resource and guide for future research endeavors.",author:{id:"259298",name:"Edward",surname:"Narayan",institutionString:null,profilePictureURL:"https://mts.intechopen.com/storage/users/259298/images/system/259298.jpeg",slug:"edward-narayan",institution:{id:"3",name:"University of Queensland",country:{id:null,name:"Australia"}}}}]},series:{item:{id:"25",title:"Environmental Sciences",doi:"10.5772/intechopen.100362",issn:"2754-6713",scope:"\r\n\tScientists have long researched to understand the environment and man’s place in it. The search for this knowledge grows in importance as rapid increases in population and economic development intensify humans’ stresses on ecosystems. Fortunately, rapid increases in multiple scientific areas are advancing our understanding of environmental sciences. Breakthroughs in computing, molecular biology, ecology, and sustainability science are enhancing our ability to utilize environmental sciences to address real-world problems.
\r\n\tThe four topics of this book series - Pollution; Environmental Resilience and Management; Ecosystems and Biodiversity; and Water Science - will address important areas of advancement in the environmental sciences. They will represent an excellent initial grouping of published works on these critical topics.