Dilution mixing and incubation time.
\r\n\t
\r\n\tSince its discovery in the mid-50’s, a wide range of applications from low to high voltage appeared, putting polyimide as a key material to design more performing and reliable electrical devices and systems. On another hand, polyimide appears also essential for the development of new electronic devices where further considerations such as high power density, integration, higher temperature, thermal conduction management, energy storage, reliability or flexibility are required in order to sustain the growing electrical energy consumption needs of the global society.
\r\n\tConsequently, polyimide materials have and will have to face new exciting fundamental, technological and environmental challenges among which:
\r\n\t• a better understanding of its intrinsic electrical properties to identify current limitations and propose new advanced device designs,
\r\n\t• the development of innovative composites and nanocomposites structuration to tailor its physical properties by involving classical and original nanoparticles such as graphene layer, carbon nanotubes, metal, silicates, nitrides, etc.,
\r\n\t• the development of polyimide composites for energy storage, thermal management, reinforced nanodielectrics and corona-resistant nanocomposites,
\r\n\t• the development of new low and ultra-low dielectric constant polyimide for microelectronics (fluorinated polyimides, nanoporous, mesoporous),
\r\n\t• the development of new higher temperature reliable polyimide (high glass transition, high degradation temperature),
\r\n\t• the emergence of solvent-free processes to fit with environmental purposes
\r\n\tMoreover, many challenges regarding the aging mechanisms understanding under single or multiple constraints and the realistic lifetime prediction using robust physical modelling is a ubiquitous questioning in most of the electronic industries.
\r\n\tThis book will target to review both the state-of-the-art and new researches on Polyimide for Electronic and Electrical Engineering Applications. It will present interdisciplinary chapters on the state of knowledge of each topic under consideration through a combination of overviews and original unpublished research. Chapter proposals related to one of the following topics and their keywords (but not only restricted to them) are very welcome to be submitted for this book publication project:
\r\n\t• General Considerations and Technological Processes of Polyimide for Electronics and Electrical Systems
\r\n\tProcessability, Photosensitive and non-photosensitive polyimide, Curing temperature,
\r\n\tSpin-coating, Dip-coating, Extruded enameled wires, Other casting methods
\r\n\t• Polyimide in Microelectronic Applications
\r\n\tDielectric properties, Intermetal layer, Ultra Large-Scale Integration (ULSI), Low-k dielectrics, Fluorinated polyimide, Nanoporous polyimide, Flexible substrates, Thin film transistors, LCD devices, sensors and actuators (gas, humidity, pressure, tactile…)
\r\n\t• Polyimide in Medium and High Voltage Applications
\r\n\tElectrical insulation properties (conduction, breakdown), Digital isolators, Power electronics and devices, Power modules, Power integration, Passivation, Packaging, High voltage power systems, Enameled wires for fed-inverter rotating machine
Endotoxins, a type of pyrogen, are natural compounds found in the outer cell membrane of Gram-negative bacteria and can impact over 30 biological activities. Endotoxin can lead to cell death by initiating complement activation. The Limulus amebocyte lysate (LAL) test was commercially introduced in the 1970s. LAL is derived from the blood cells, or amebocytes, of the horseshoe crab,
Limulus amebocyte lysate test is an aqueous extract of blood cells (amoebocytes) which obtain from the horseshoe crab (
Activation of inflammation in body [
Horseshoe crab [
Among the most well-known and important applications of the LAL test are the ones related to the pharmaceutical industry. It can be said that the most common pyrogens in pharmaceutical products are endotoxins, which is why the pyrogen tests on rabbits have been replaced by the LAL test according to the recommendations of the international pharmacopeia. One of the reasons that has made the LAL test prevail in the pharmaceutical industry is the careful avoidance by the LAL manufacturers of bringing harm to live animals during both production and testing. It is important to clarify that the crabs, from which part of the hemolymph used for the LAL test was extracted, are returned to alive to their natural habitat with no lasting problems after the extraction.
Add volume of lysate to a volume of product dilution. Incubating the reaction mixture at 37.5°C. Endotoxin in the reaction would activate the LAL reagent. Cleave small chromogenic peptides and liberates pNA. pNA, color is yellow and absorbs light at 405 nm. For samples that absorb at 405–410 nm, Diazo-coupling agent modification may be used. In this method, pNA reacted with nitrite in hydrochloric acid, ammonium sulfamate and N-(1-naphthyl)-ethylenediamine (NEDA). Absorbs at a range between 540 and 550 nm. A standard curve is used to establish concentrations in product specimens.
10 × 75 mm fully depyrogenated borosilicate glass culture tubes (Associates of Cape Cod, Inc. catalog numbers TB050).
Optical reader is capable of reading at 405 nm, or at 540–550 nm for the diazo method. Incubator is able to maintaining 37 ± 1°C. A water bath can be used for the endpoint test tube method. Both devices should have a uniform heat distribution. Test tube racks to hold the tubes and/or incubate dilution and reaction tubes. Micropipettes or disposable pipette tips free of interfering endotoxins and glucans are recommended. Vortex-type mixer, Para film (American National Can™) and hot-air oven with the capacity to heat to at least 250°C for depyrogenation of glassware.
Limulus amebocyte lysate (LAL), LAL reconstitution buffer, control standard endotoxins (CSE), solution 1 (nitrite), solution 1A (0.1 N hydrochloric acid), solution 2 (ammonium sulfamate), solution 3 (N-(1-naphthyl)-ethylenediamine (NEDA)), LRW.
The endotoxins limit for USP/BP sterile WFI is only 0.25 EU/ml; therefore, sterile WFI may contain detectable endotoxins and be unsuitable for use. Use certified LRW to make dilutions of standards, and to prepare positive controls.
Collect aseptically containers that are free of detectable endotoxins in depyrogenated glassware apparatus.
The pH must be 6–8. Adjust the pH of the product specimen with dilute HCl, NaOH, or buffer (free of endotoxins). Dilute concentrated HCl or NaOH with LRW. Use a volume that will not lead to significant dilution of the test specimen. Dilution (LRW) alone can overcome the issue sometimes.
Gently tap the vial of lysate. Loose material fall to the bottom. Break the vacuum by lifting the gray stopper. Do not contaminate the mouth of the vial. Remove and discard the stopper. Start the reconstituted lysate with 3.2 ml buffer. Avoid vigorous mixing that may cause excessive foaming and a loss of sensitivity. Wrap the vials with parafilm and store in a cold place (2–8°C) when not in use and use within 8 h of reconstitution.
This is relatively well stable and, if stored properly, will retain full activity through the expiration date on the label. Store the product at 2–8°C. Excess temperature over 37°C cause rapid deterioration, loss of sensitivity and distinct yellowing.
Each vial of control standard endotoxins (CSE) contains 10 ng of endotoxins. Reconstitute CSE with the volume mentioned on the Certificate of Analysis (CA, which gives the potency of the CSE). Gently knocks the vial of control standard endotoxins (CSE) to cause loose material to fall to the bottom. Break the vacuum by lifting the gray stopper. Do not contaminate the mouth of the vial. Remove the stopper and place it in a cold place aseptically for reuse.
Reconstitute CSE with the volume specified on the Certificate of Analysis (CA, which gives the potency of the CSE) and as directed in the package insert. Place the stopper. Vortex the vial for 40–60 s to form a homogenous mixture. Discard solution if not used immediately, vortex the vial for 30 s prior to use.
Read the tubes UV/visible spectrophotometers (Table 1).
CSE + lysate | Incubation time (min) |
---|---|
50 μl of 0.50 EU/ml + 50 μl | 30 |
50 μl of 0.250 EU/ml + 50 μl | 30 |
50 μl of 0.125 EU/ml + 50 μl | 30 |
50 μl of 0.0625 EU/ml + 50 μl | 30 |
Dilution mixing and incubation time.
Sample + lysate | Incubation (min) |
---|---|
50 μl of sample + 50 μl | 30 |
Stop the reaction by adding 50% acetic acid. Add 0.025 ml (25 μl) read the optical density (OD) at 405 nm read the test.
Reconstitute vial 1 with entire contents of vial, reconstitute vial 2 with 4 ml of water, reconstitute vial 3 with 4 ml of water. Add 0.05 ml (50 μl) of solution 1 (sodium nitrite reconstituted with dilute HCL). Add 0.05 ml (50 μl) of solution 2 (ammonium sulfamate). Add 0.05 ml (50 μl) of solution 3 (NEDA) use new pipette tip agitate the plate to mix. Full color (magenta) should develop immediately. Read the test at 540–550 nm.
Positive control must be included to verify that it is appropriate to use the parameters of a previous (archived) standard curve to calculate endotoxin concentrations.
LRW negative controls should be included with each test
1: Equation of straight line (results)
y = mx + c
m = slop
x = endotoxin concentration,
c = y-intercept and
y = mean absorbance
X = y-c/m
Example calculation
Prepare sample solutions by dissolving or diluting drugs (pH 6.0–8.0). The pH may be adjusted by the use of acid, base, or suitable buffers as recommended. Do not exceed the MVD or MCV while making dilutions and adjusting the pH.
MVD = (endotoxin limit × concentration of sample solution)/(λ)
Endotoxin limit given in USP, concentration of a sample of the label, λ: the labeled lysate sensitivity in the gel-clot technique (IU/ml) or the lowest concentration used in the standard curve for the turbidimetric or chromogenic techniques.
MVC = λ/endotoxin limit
λ: the labeled lysate sensitivity in the gel-clot technique (IU/ml) or the lowest concentration used in the standard curve for the turbidimetric or chromogenic techniques.
Sample 1
Endotoxin limit: 0.5 EU/ml
Concentration of sample: 100 mg/ml
λ: 0.06 EU/ml
MVD = 0.5 EU/ml × 100 mg/ml/0.06 EU/ml
MVD = 833
Add 1 ml of sample 1 in to 832 ml of LRW. Prepare sample 2 in using the same method.
Using 10-fold and 2-fold dilution methods prepare the following dilutions of control standard endotoxins (CSE)
0.5 EU/ml
0.25 EU/ml
0.125 EU/ml
0.0625 EU/ml
Reconstitute the lysate with 3.2 ml of buffer provided with it. Follow the standard procedure for reconstitution.
Stop reaction.
For sample 1 and sample 2:
Stop the reaction by adding 50% acetic acid. Add 0.025 ml (25 μl) (Tables 2 and 3).
CSE + lysate | Incubation (min) |
---|---|
50 μl of 0.50 EU/ml + 50 μl | 30 |
50 μl of 0.250 EU/ml + 50 μl | 30 |
50 μl of 0.125 EU/ml + 50 μl | 30 |
50 μl of 0.0625 EU/ml + 50 μl | 30 |
50 μl of sample 1 + 50 μl | 30 |
50 μl of sample 2 + 50 μl | 30 |
Different dilution of CSE and lysate.
Make two replicates of each CSE and sample preparation to reduce any errors.
Use Microsoft word for further calculations and results. Make standard curve and endotoxin concentration (Figure 3).
Validation of standard curve.
R2 = coefficient of determination
R = correlation coefficient
R ≥ 0.98
R2 = 0.99
R = √R2 = 0.99
Equation of straight line
y = mx + c
m = slop
x = endotoxin concentration
c = y intercept
y = mean absorbance
Equation of straight line
Y = 1.019X − 0.026
X = Y + 0.026/1.019
m = slop = 1.019,
C = y intercept = 0.026,
Y = mean absorbance,
X = endotoxin concentration
X = Y + 0.026/1.019
Y = 0.300, X = 0.300 + 0.026/1.019, X = 0.319EU/ml
X = Y + 0.026/1.019
Y = 0.335, X = 0.335 + 0.026/1.019, X = 0.354 EU/ml
Gel Clot LAL provides a simple positive/negative result and is most often mentioned in pharmacopeial monographs as the official referee test.
This is very easy to perform.
This is not time consuming.
Accuracy is 100 percent.
The LAL Gel-Clot assay, gives a more quantitative measurement of endotoxin over a range of concentrations.
Gel Clot lysate for 20 test, Gel Clot standard 0.5 EU/Vial, LAL reagent water (LRW 50 ml).
Lysate: add 2 mL LRW and mix it slowly. Do not shake and avoid foaming. Transfer 0.1 ml in 20 test tubes. Store it at –degree (in freezer).
Standard: Add 2 mL of LRW in the vial and mix it well for 15 min. Store the vial at 2–8°C. Storage life is 15 days.
Take three test tubes and mark them as test, positive control and negative control [1].
Add your sample in test tube marked as sample. Add standard in test tube marked as Positive control. Add LRW in test tube marked as negative control. Incubate the test tubes at 37 + 2°C for 60 min. After an incubation, check for the gel by inverting the test tube. If the material remains firm in the bottom of the test tube, it means gel has formed. This positive if the material gets the flow down, it means gel has not formed. This means negative.
Take similarly three test tubes as above and add water for injection (WFI) in test tube marked as sample. And proceed as above. The results should be as follows (Table 4):
Sample | Positive control | Negative control | Result |
---|---|---|---|
−ve (gel not formed) | +ve | −ve | Pass |
Sample | Positive control | Negative control | Result |
---|---|---|---|
+ve (gel formed) | +ve | −ve | Fail |
Results shown sample pass or not.
We have to make dilution.
Example: If the product endotoxin limit is 1 EU/ml, then we have to make the dilution as follows:
Since we are using 0.25 EU/ml, this is called lambda. Divide the endotoxin limit of product with lambda
1/0.25 = 1:4
As per USP, we have to test 3 test as follows:
One test tube | 1:3 | The result should be positive |
Second | 1:4 | The result may be positive or negative |
Third | 1:5 | The result should be negative |
This means the product is passed.
Chromogenic lysate [2],
Respective endotoxin standard,
Diazo coupling reagent (set of four bottles).
Note: All reagents must be stored in refrigerator at 2–8°C.
Dissolve 45.6 ml of acetic acid in 1 liter of distilled water. The final concentration of acetic acid is 0.8 M. This solution can be stored for 3 months.
Remove the plastic cover. Wipe off with 70% alcohol around the rubber cap and top portion of every vial. Remove the aluminum cap with sterile and pyrogen free forceps and then cover with depyrogenated aluminum foil to avoid any Endotoxin contamination. (2.8 ml LAL water vial is provided with Endotoxin vial, concentration is mentioned on the label). Pour whole quantity of LAL water into the ET vial and cover with foil. Mix vigorously for at least 10 s by vortexer. During stirring solution must not touch the foil.
Note: Stir every time vigorously before use.
Toxicolor lysate
(Buffer vial 0.35 ml and LAL water are provided with Lysate. Sensitivity is mentioned on the certificate). After taking from the refrigerator, pour whole quantity of buffer and 0.35 ml LAL water into the lysate vial as soon as possible, covers with foil. Then quickly stir to dissolve. Avoid air bubbling during stirring. Place the vial in ice water bath for 2–3 min before use.
Note: Be sure that the reagent is completely dissolved. This reagent must be reconstituted just before use. The reagent is extremely sensitive and must be consumed at one time. Storage should be avoided, but can be stored at −20°C in 0.1 ml dispensed quantities in small test tubes. Use stored lysate if the color is not changed. Reconstituted lysate may only be deep frozen once.
Four bottles are provided with one set, marked as 7, 8, 9 and 10s respectively. Transfer whole quantity of bottle no. 7 s into bottle no. 8 s. Then add 12 ml distilled water into each of bottle no. 9 s and 10s. Ultimately, we will have three bottles 8, 9, and 10 s, which are used stepwise to block the reaction.
The pH of the sample is adjusted by pyrogen free 0.1 N NaOH or 0.1 N HCl. The pH of the sample should be between 6.0 and 8.0.
Arrange test tubes in two stands as under; stand 1—test tubes for sample and standard dilutions; stand 2—test tubes for reaction.
Take 0.05 ml well-mixed sample into small test tubes. If required, make 1/10 dilution of the sample with Pyrogen free water as Below, Take 4.5 ml of pyrogen free water in the test tube. Then add 0.5 ml of well-mixed sample. Vortex mixing for a few seconds.
Take 0.1 ml into a small test tube for further process.
Make a dilution of the endotoxin (concentration 0.470 EU/ml) according to the product limit. For making 0.235 EU/ml (if the product limit is 0.25) proceed as follows;
Take 0.05 ml of the reconstituted endotoxin in the test tube after stirring. Add 0.05 ml of pyrogen free water and vortex to mix. Now the final dilution is 0.235 EU/ml.
Take 0.05 ml of step 2 into a small test tube for further process.
Pour 0.05 ml of pyrogen free water (being used in the test) in small test tube as a blank for further process.
Lysate addition
Place the tube stand for small test tubes (containing the tubes of blank, standard and diluted samples) in ice water bath or suitable ice water container. Add 0.05 ml of lysate to all of the tubes as soon as possible. Stir the contents of every tube soon after the addition of lysate for a few seconds. Avoid foaming.
Soon after the addition of lysate, place the test tube rack in the incubator set at 32.5 + 2.5°C for 30 min. The tube rack can be placed in the water container placed in the incubator.
After completion of the incubation period, place tube rack in ice water bath, then blocks the reaction immediately from one of the two methods mentioned below:
By acetic acid
Add 0.4 ml of 0.8 M acetic acid into each tube and stir to mix.
By diazo coupling reagent
Three bottles of the reagent are used as under;
Add 0.5 ml from bottle no 8 s to each tube and stir to mix.
Add 0.5 ml from bottle no 9 s to each tube and stir to mix.
Add 0.5 ml from bottle no 10s to each tube and stir to mix.
Absorbance reading (using spectrophotometer) measurement at 405 nm
If 0.8 M acetic acid is used to block the reaction, then absorbance reading is taken at 405 nm.
Note: The readings. Glass photocell is used for reading at 405 nm. Because the volume of the tube content is not sufficient, the distilled water is added to each tube and is stirred to mix.
When Diazo coupling reagent is used for blockage of the reaction then the reading is taken at 545 nm. Note all the readings.
Note: Distilled water is used for reference in both cases.
All the absorbance readings are fed in the “Software reader for window version 1.51” to collect the results.
Following Formula is used to calculate the results
Calculations of MVC, MVD and ELC.
Where the lowest sensitivity of lysate, M is the maximum dose/kg body weight and K is constant having value equal to 5.
The author(s) confirm that this chapter content has no conflict of interest.
"I work with IntechOpen for a number of reasons: their professionalism, their mission in support of Open Access publishing, and the quality of their peer-reviewed publications, but also because they believe in equality. Throughout the world, we are seeing progress in attracting, retaining, and promoting women in STEMM. IntechOpen are certainly supporting this work globally by empowering all scientists and ensuring that women are encouraged and enabled to publish and take leading roles within the scientific community." Dr. Catrin Rutland, University of Nottingham, UK
",metaTitle:"Advantages of Publishing with IntechOpen",metaDescription:"We have more than a decade of experience in Open Access publishing. \n\n ",metaKeywords:null,canonicalURL:null,contentRaw:'[{"type":"htmlEditorComponent","content":"We have more than a decade of experience in Open Access publishing. The advantages of publishing with IntechOpen include:
\\n\\nOur platform – IntechOpen is the world’s leading publisher of OA books, built by scientists, for scientists.
\\n\\nOur reputation – Everything we publish goes through a two-stage peer review process. We’re proud to count Nobel laureates among our esteemed authors. We meet European Commission standards for funding, and the research we’ve published has been funded by the Bill and Melinda Gates Foundation and the Wellcome Trust, among others. IntechOpen is a member of all relevant trade associations (including the STM Association and the Association of Learned and Professional Society Publishers) and has a selection of books indexed in Web of Science's Book Citation Index.
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\n\nOur platform – IntechOpen is the world’s leading publisher of OA books, built by scientists, for scientists.
\n\nOur reputation – Everything we publish goes through a two-stage peer review process. We’re proud to count Nobel laureates among our esteemed authors. We meet European Commission standards for funding, and the research we’ve published has been funded by the Bill and Melinda Gates Foundation and the Wellcome Trust, among others. IntechOpen is a member of all relevant trade associations (including the STM Association and the Association of Learned and Professional Society Publishers) and has a selection of books indexed in Web of Science's Book Citation Index.
\n\nOur expertise – We’ve published more than 4,500 books by more than 118,000 authors and editors.
\n\nOur reach – Our books have more than 130 million downloads and more than 146,150 Web of Science citations. We increase citations via indexing in all the major databases, including the Book Citation Index at Web of Science and Google Scholar.
\n\nOur services – The support we offer our authors and editors is second to none. Each book in our program receives the following:
\n\nOur end-to-end publishing service frees our authors and editors to focus on what matters: research. We empower them to shape their fields and connect with the global scientific community.
\n\n"In developing countries until now, advancement in science has been very limited, because insufficient economic resources are dedicated to science and education. These limitations are more marked when the scientists are women. In order to develop science in the poorest countries and decrease the gender gap that exists in scientific fields, Open Access networks like IntechOpen are essential. Free access to scientific research could contribute to ameliorating difficult life conditions and breaking down barriers." Marquidia Pacheco, National Institute for Nuclear Research (ININ), Mexico
\n\nInterested? Contact Ana Pantar (book.idea@intechopen.com) for more information.
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I am also a member of the team in charge for the supervision of Ph.D. students in the fields of development of silicon based planar waveguide sensor devices, study of inelastic electron tunnelling in planar tunnelling nanostructures for sensing applications and development of organotellurium(IV) compounds for semiconductor applications. I am a specialist in data analysis techniques and nanosurface structure. I have served as the editor for many books, been a member of the editorial board in science journals, have published many papers and hold many patents.",institutionString:null,institution:{name:"Sheffield Hallam University",country:{name:"United Kingdom"}}},{id:"54525",title:"Prof.",name:"Abdul Latif",middleName:null,surname:"Ahmad",slug:"abdul-latif-ahmad",fullName:"Abdul Latif Ahmad",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:null},{id:"20567",title:"Prof.",name:"Ado",middleName:null,surname:"Jorio",slug:"ado-jorio",fullName:"Ado Jorio",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"Universidade Federal de Minas Gerais",country:{name:"Brazil"}}},{id:"47940",title:"Dr.",name:"Alberto",middleName:null,surname:"Mantovani",slug:"alberto-mantovani",fullName:"Alberto Mantovani",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:null},{id:"12392",title:"Mr.",name:"Alex",middleName:null,surname:"Lazinica",slug:"alex-lazinica",fullName:"Alex Lazinica",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/12392/images/7282_n.png",biography:"Alex Lazinica is the founder and CEO of IntechOpen. After obtaining a Master's degree in Mechanical Engineering, he continued his PhD studies in Robotics at the Vienna University of Technology. Here he worked as a robotic researcher with the university's Intelligent Manufacturing Systems Group as well as a guest researcher at various European universities, including the Swiss Federal Institute of Technology Lausanne (EPFL). During this time he published more than 20 scientific papers, gave presentations, served as a reviewer for major robotic journals and conferences and most importantly he co-founded and built the International Journal of Advanced Robotic Systems- world's first Open Access journal in the field of robotics. Starting this journal was a pivotal point in his career, since it was a pathway to founding IntechOpen - Open Access publisher focused on addressing academic researchers needs. Alex is a personification of IntechOpen key values being trusted, open and entrepreneurial. Today his focus is on defining the growth and development strategy for the company.",institutionString:null,institution:{name:"TU Wien",country:{name:"Austria"}}},{id:"19816",title:"Prof.",name:"Alexander",middleName:null,surname:"Kokorin",slug:"alexander-kokorin",fullName:"Alexander Kokorin",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/19816/images/1607_n.jpg",biography:"Alexander I. Kokorin: born: 1947, Moscow; DSc., PhD; Principal Research Fellow (Research Professor) of Department of Kinetics and Catalysis, N. Semenov Institute of Chemical Physics, Russian Academy of Sciences, Moscow.\r\nArea of research interests: physical chemistry of complex-organized molecular and nanosized systems, including polymer-metal complexes; the surface of doped oxide semiconductors. He is an expert in structural, absorptive, catalytic and photocatalytic properties, in structural organization and dynamic features of ionic liquids, in magnetic interactions between paramagnetic centers. The author or co-author of 3 books, over 200 articles and reviews in scientific journals and books. He is an actual member of the International EPR/ESR Society, European Society on Quantum Solar Energy Conversion, Moscow House of Scientists, of the Board of Moscow Physical Society.",institutionString:null,institution:{name:"Semenov Institute of Chemical Physics",country:{name:"Russia"}}},{id:"62389",title:"PhD.",name:"Ali Demir",middleName:null,surname:"Sezer",slug:"ali-demir-sezer",fullName:"Ali Demir Sezer",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/62389/images/3413_n.jpg",biography:"Dr. Ali Demir Sezer has a Ph.D. from Pharmaceutical Biotechnology at the Faculty of Pharmacy, University of Marmara (Turkey). 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