Diversity of Natural Bioactive Compound in Plant Origin

Abstract

Recent studies have claimed that people are now greatly relying on synthetic drug without considering any side effect; however, all the synthetic drugs have been formulated commercially by following the invention of an authentic source of crude drug; hence, people are still directly or indirectly dependent on natural source of medicine. Recently, I have completed a research work on black pepper (Piper nigrum), and piperine (in a crystal form) was isolated as a mother bioactive compound from black pepper through a plenty of in vitro investigations. After that, I have experimented some in vitro analysis to evaluate the antioxidant power of that pure compound, and it was found that the crystal compound has strong antioxidant power. After doing some theoretical analysis, it has been identified that piperine may exist in other medicinal plants also, and many plants belonging to the same species can able to show multiple types of biological activities, which actually reflected the diversity of bioactive compounds in nature. People can benefit from different types of bioactive compounds, such as piperine, if we biosysthesize and use them commercially.

1. Introduction

Medicinal plants or herbal medicine has been utilized to prevent and cure diseases since the ancient period of time, and it has played a significant role in drug discovery [1, 2]. The earlier evidences had been declared that the existing medicinal plants were consumed from 60,000 year ago. Recently, a 5000-year-old Sumerian clay slab was invented by utilization of medicinal plants for the manufacture of drug [3]; moreover, the current study is showing that more than 50% of marketed drugs have been derived from medicinal plants [4, 5].

The safety, efficacy, and quality of this active constituents greatly rely on the source, cross-contamination, and simultaneously the formulation procedure of finished products. The popularity of medicinal plants had been raised from fifteenth to seventeenth century, and the descriptions of herbal medicine had started to be available in various languages. In the eighteenth century, a scheme for classifying plant species had been initiated by Linnaeus [6].

Primarily, the medicinal plants were used in casual pharmaceutical preparations such as macerations, infusions, and decoctions; however, in between sixteenth and eighteenth centuries, the compounded drug was on demand. The early nineteenth century was a crucial point in the development of knowledge about the consumption of medicinal plants and the drug discovery, substantiation, and screening of alkaloids from the poppy as well as quinine, ipecacuanha, pomegranate also under trial. Scientific pharmacy was initiated followed by the isolation of glycosides from other medicinal plants, and the upgradation of chemical methodologies to isolate tannins, saponins, hormones, and vitamins was started [7, 8].

In the late nineteenth and early twentieth centuries, a steady fall in the therapeutic use of herbal medicines has been noticed. Many authors have claimed that drugs obtained from the medicinal plants may be possessed shortcomings due to degradative action of different types of enzymes. In the early twentieth century, stabilization method for the preparation and utilization of fresh medicinal plants had been proposed. After that, several steps has been taken to cultivate medicinal plants [9] as they have offered more authentic natural sources of active pharmacological ingredients [10].

1.1 Blessings of medicinal plants

Recently, due to the prominent side effects of updated synthetic drugs and escalating contraindications about their consumptions, a great attempt has been made to enhance the utilization of diverse medicinal plants [11]. Medicinal plants have played a vital and integral role in healthcare system from the earliest period of time, and some diverse types of instances of this blessing are described below [12].

Analgesic:

Opioids (morphine) can bind with cerebral opioid receptor and can modify the pain sensation by exerting receptor-mediated function and eventually will show analgesic effect [13, 14] and exert their analgesic effect. All the cannabis and cannabinoids had been consumed to relieve pain [15].

Anticancer:

Medicinal-plant-derived constituents such as vinblastine, vincristine from alkaloids Vinca rosea demonstrate anticancer activity [16].

Antihypertensive:

A plenty of medicinal plants were suggested by the ancient communities for the management of hypertension that may introduce a new area of research on the antihypertensive activity, [17, 18] such as reserpine from Rauwolfia serpentine.

Antidiabetic:

Medicinal plants such as Acacia arabica, Eucalyptus globulus may improve insulin secretion from pancreatic beta cell, thereby capable of exhibiting antidiabetic property [19].

Besides this overall pharmacological coverage, medicinal plants may have thousands of phytoconstituents, for instances, compound atropine from Atropa belladonna exhibits antispasmodic activity, ephidrine from Ephedra vulgaris might have bronchodialating effect.

Our present study is to represent the blessings of medicinal plants, which are readily available in our nature. I have already discussed a little bit about diversity of bioactive compounds in the above section, and it actually clarifies that nature may conserve every type of remedy related to the pathological state of human body. Actually, most of marketed drugs were being explored with the help of the concept of biodiversity of medicinal plants. As disease state does not consider any class of people such as rich, poor, middle-class, allergic people (to synthetic drug), suburb, or any city belonging people, so we should have developed our dependency on bioactive constituents containing medicinal plants, which will be economically affordable rather than synthetic drug. My research work was related to identifying the medicinal plant having great biological activity, which will be very cheap in source and obviously included to our daily food supplement. I have chosen black pepper (BP) (Piper nigrum) shown in Figure 1 as my research topic where my prime concern was to identify bioactive compound (antioxidant activity containing) and correlate the biodiversity of that compound in nature.

Figure 1.

Black pepper (Piper nigrum) [20].

1.2 Black pepper

1.2.1 Scientific name: Piper nigrum Linn

1.2.2 Etymology of black pepper

The etymological background of black pepper was so complex, some believe that the word pepper has come from old English pipor, Latin pipor, and Sanskrit pippali for “long pepper.” Besides this, people were using the word pepper to indicate the unrelated new word chili pepper (genous capsicum) during sixteenth century [21].

1.2.3 History of black pepper

Black peppercorns were explored stuffed in the nostril of Ramesses II, placed there as a part of the mummification rituals shortly after his death in 1213 BOE [22]. Someone has perceived that the black pepper was used in ancient Egypt, and it arrived in the Nile from south Asia. Black pepper is mainly popular in south Asia, southeast India, and is native to Kerala, a southwestern coast of India [23, 24].

1.2.4 Production and trade of black pepper (BP)

Ethiopia was the world’s largest manufacturer and exporter of BP in 2019 producing 374,413 tonnes or 34 % of the world’s total according to Figure 2. The rest of the major producers were Vietnam, Brazil, Indonesia, India, China, and Malaysia. Internationally pepper production has varied annually, which is dependent on the crop management, disease, and environmental impact [26, 27].

Figure 2.

Graphical representation of black pepper production, 2019 [25].

1.2.5 Perspective regarding antioxidant

I have tried my level best to isolate pure compound from black pepper and investigated a lot to identify antioxidant activity of that pure compound through a variety of in vitro evaluation processes.

The main target of any type of antioxidant or reductant is unpaired electron containing free radicals. Free radicals are a group of unstable chemicals, which can readily bind with biological components to be stable by gaining electrons resulting in destructive damage of cellular materials [28].

Typically, free radicals are formed inside the human body as a result of casual metabolic process, which can be both beneficial and harmful for human physiology; however, uncontrolled level of free radical production may initiate different types of diseases [29].

Antioxidant serves a prominent role in guarding our body from disease by neutralizing the series of oxidative stress produced by ROS [30]. During my research, my thought was to explore the free radical scavenging activity of sample and measure the extend of antioxidant power of that plant material as recent investigations have revealed that the plant-derived antioxidants with free-radical scavenging properties may have great therapeutic significance in diseases mediated by the free radicals such as neurodegenerative disease, autoimmune disorder, inflammatory disorder, cancer, and so on. To get relieved from these diseases, a variety of synthetic drugs are available in the market; however, these drugs may have some unavoidable side effects also. If people can put more concern on plant-based antioxidants, they will definitely get expected pharmacological effect with less or no side effects [31].

2. Experimental methodology with explaining physicochemical nature of that crystal compound

2.1 Extraction process

The extraction process has been completed based on the method described by Alam et al. [32]. About 500 g of sun and shade dried black pepper has been converted into powdered material by using a blender. Powdered black pepper was then transferred into an opaque bottle, and it has been filled with 1.5 L of methanol as a mother solvent for this experiment. The plant materials were kept in the bottle for 7 days with occasional shaking for better extraction (it can be called crude black pepper extract). The extracts of black pepper have been filtered through a fresh cotton plug and finally with Whatman No.1 filter papers. The filtrates found from the extraction process had been sent into the rotary evaporator (Bibby Sterlin Ltd, UK) to get a concentrated mass of sample under reduced pressure at 50 °C.

2.2 Fractionation (in between liquid-liquid solvent) process

The obtained concentrated mass of black pepper had been partitioned in between liquid solvent as described in Figure 3, where ultimately, a crystal seed fraction has been found.

Figure 3.

Overall fractionation process of crude extract of black pepper in between liquid-liquid solvent.

2.3 Recrystallization process

Recrystallization is a process through which we can easily eradicate our desired compound from a mixture of undesired one by forming crystal of that compound in a favorable solvent, and simply it could be designated as an efficient method of purification [33]. As we have found a crystal seed fraction from black pepper, so we have utilized this recrystallization technique mentioned in Figure 4 to purify crystal.

Figure 4.

Recrystallization technique for crystal seed produced from chloroform fraction.

The flow diagram we have mentioned above had been followed to purify our crystal seed. The crystal seed was washed with KOH to remove resin-like compound or isomer of the crystal seed, and it was then filtered through filter paper for preliminary purification. Now it has been treated with petroleum ether for recrystallization and warmed up the combination to dissolve it at 50 °C. After that, the mixture was cooled to form crystal. The crystal is filtered for final purification.

2.4 Thin-layer chromatography (TLC)

It is the most convenient way to eradicate individual constituents from a mixture of compounds by using stationary phase and mobile phase, and the formation of a single spot actually indicates the presence of pure compound [34].

The spotted TLC paper was placed into the TLC chamber in such a way that the spot can stay just above the surface of the solvent system. After running of the solvent through a specified limit, then the operation was discontinued, and the TLC paper is dried in air. By treating with specified vanillin-sulfuric acid spray reagent, the paper was observed under UV light of different wavelengths to locate spots, so that can easily explore different types of medicinal constituents [35]. The paper was inserted into an iodine chamber to confirm about the spot is formed, and at last, I identified only single spot for the obtained crystal, which was examined under UV light and iodine chamber (Figure 5).

Figure 5.

TLC of crystal compound (CC) after spraying vanillin-sulfuric acid spray reagent.

2.5 Calculating the R_f value of crystal

In TLC study, the retention factor has been utilized to screen and analyze the phytocompounds. The R_f value is a ratio of the distance moved by the compound and the distance moved by the solvent. The obtained R_f value for crystal compound was found as 0.43.

2.6 Visual identification crystal

The crystal was light brown in color, and visually it was a needle-like structure, which has been shown in Figure 6.

Figure 6.

Isolated crystal compound found in black pepper.

2.7 Solubility of crystal compound

After doing a plethora of experiments, it has been found, as seen Table 1 that the crystal compound was readily soluble in chloroform and likely insoluble in water.

Solvent name	Solubility
Water	42 mg/L (likely insoluble)
Alcohol (ethanol)	1.5 gm/14.4 ml
Di-ethyl-ether	1 gm/35 ml
Chloroform	1 gm/1.5 ml

Table 1.

Solubility of crystal compound found through experimental method.

2.8 Melting point

The melting point of that crystal was near 127$°C$.

2.9 Boiling point

The crystal was so sensitive to increased level of heat, and it had been totally destructed during checking boiling point. So, the crystal material does not have any fixed boiling point.

2.10 Phytochemical contents

Alkaloid test for crystal (found from chloroform fraction of crude black pepper extract) had shown strong positive result according to Table 2, which clarifies that the crystal may have alkaloid compounds [36, 37, 38].

Phytochemical test	Crystal compound (found from chloroform extract of black pepper)
Alkaloids	+++
Glycosides	$-$
Tannins	++
Steroids	$-$
Saponins	$-$
Carbohydrates	+
Terpenoids	+

Table 2.

Phytochemical contents explored in crystal (found from chloroform fraction of crude black pepper extract).

After comparing the physicochemical properties of crystal compound with standard one [39], it could be predicted that the crystal may be piperine.

3. In-vitro free radical scavenging assay with results interpretation

3.1 DPPH radical scavenging activity of different fractions of plant materials

Free radical scavenging ability of different fractions of sample had been evaluated by DPPH radical scavenging assay according to Blois and Desmarchelier et al. [40, 41]. The hydrogen atom donating capability of the plant extractives was estimated by identifying the color change of samples with 2,2-diphenyl-1-picrylhydrazyl (DPPH). DPPH exhibits made violet/purple color in samples having antioxidant property, and it was then faded into yellowish color. 2.4 ml of DPPH solution was mixed with 1.6 ml of sample solution at different concentration. The reaction mixture has been vortexed and left it for 30 min in a dark place at room temperature. The absorbance was taken for individual fractions at different concentration via spectrophotometer at 517 nm. Butylated hydroxy toluene had been used as standard. The percentage of DPPH radical scavenging activity was calculated by the following equation:

$$\%\text{DPPH radical scavenging activity}=\left\{\left(\text{A0}-\mathrm{A}1\right)/\mathrm{A}0\right\}\times 100$$

where A₀ represents the absorbance of the control, and A₁ represents the absorbance of the sample materials. Then % of inhibition has been plotted against concentration, and the IC₅₀ was calculated graphically. The same experiment was repeated for three times with every fraction at each concentration.

3.1.1 Data interpretation of DPPH radical scavenging assay

According to Table 3, it could be easily assumed that the crystal compound may have a great antioxidant power as the data for that compound had shown a nearly similar value of standard catechin. The IC₅₀ value was so small, which was an indication for strong free radical scavenger.

Name of sample	Concentration ($\mathrm{\mu g}/\mathrm{ml}$)	% of scavenging (Mean$\pm$S.D)	IC50 ($\mathrm{\mu g}$/ml)
Catechin (Standard)	0.78	19.09 $\pm$0.01	3.2
	1.56	27.58$\pm$0.021
	3.125	45.46$\pm$0.015
	6.25	69.55$\pm$0.031
	12.5	87.86 $\pm$.02
	25	94.39 $\pm 0.0058$
	50	94.39$\pm 0.0058$
	100	94.39$\pm 0.0058$
Crystal compound (Pure)	1.56	27.67$\pm$2.006	4.1
	3.125	34.27 $\pm$0.657
	6.25	63.26 $\pm$2.75
	12.5	75.58$\pm$0.811
	25	85.30$\pm$1.109
	50	88.55 $\pm$1.017
	100	89.30$\pm$0.630

Table 3.

DPPH radical scavenging data analysis for standard catechin and crystal compound.

3.2 Hydroxyl radical scavenging activity of different fraction of plant materials

Hydroxyl radical scavenging activity of samples has been evaluated according to the method of Halliwell et al. [42]. Hydroxyl radical has been generated by the Fe³⁺-ascorbate-EDTA-H₂O₂ system, which is known as Fenton reaction. The overall reaction mixture was a combination of phosphate buffer solution, FeCl₃, EDTA, 2-deoxy-d-ribose, and sample extracts at different concentration. After that, it was placed into water bath at 36°C, and the reaction was initiated after addition of ascorbic acid and H₂O₂. The reaction had started, and it was sent for incubation at 36°C for maximum 1 hour, simultaneously thiobarbituric acid and HCl were added. The solution was heated for around 15 min at 100 °C followed by immediate cooling with water. The absorbance was taken at 532 nm, and it will reflect the free radical scavenging capability of samples by inhibiting the oxidation process in 2-deoxy-d-ribose in the presence of hydroxyl radical, and the ability was evaluated by using the following formula

$$\text{Percentage of hydroxyl radical scavenging activity}=[\mathrm{A}0-(\mathrm{A}1-\mathrm{A}2]\times 100/\text{A0}]$$

where A₀ represents the absorbance of control without any sample. A₁ represents the absorbance after incorporating the sample and 2-deoxy-D-ribose. A₂ represents the absorbance of the sample without adding 2-deoxy-d-ribose. IC₅₀ was investigated graphically after plotting percentage of inhibition against concentration into the graph, and the test had been repeated for three times for individual concentration of samples.

3.2.1 Data interpretation of hydroxyl radical scavenging assay

The outcome of this assay was also close to standard one according to Table 4, where the IC₅₀ for standard catechin was 31.77 ($\mathit{\mu g}$/ml), and for crystal, it was 35.01 ($\mathit{\mu g}$/ml).

Name of sample	Concentration ($\mathit{\mu g}/\mathit{ml}$)	% of scavenging (Mean$\pm$S.D)	IC50 ($\mathit{\mu g}$/ml)
Catechin (Standard compound)	25	$49.10\pm$0.032	31.77
	50	$55.14\pm$0.14
	75	$58.38\pm$0.87
	100	68.44$\pm 1.21$
	125	73.51 $\pm$1.01
	150	80.69 $\pm$1.09
Crystal compound (Pure)	25	$47.98\pm 0.45$	35.01
	50	$54.90\pm 1.22$
	75	$57.11\pm 1.11$
	100	$66.80\pm$1.51
	125	$72.79\pm 0.53$
	150	$78.95\pm 0.29$

Table 4.

Hydroxyl radical scavenging data analysis for standard catechin and crystal compound.

3.3 Superoxide radical scavenging capability

This experiment had been completed based on the method mentioned in [43]. Superoxide radical scavenging assay actually estimated the scavenging efficiency of sample by reducing the nitro blue tetrazolium (NBT) by providing electron. The reaction mixture or solution was a combination of nonenzymatic phenazine methosulfate (PMS), nicotinamide adenine dinucleotide (NADH), phosphate buffer, and samples with various concentrations, and it was then incubated for 5 min at room temperature. The PMS-NADH system created free radical in the solution mixture. In the presence of sample solutions, the NBT was reduced into purple formazan. The overall activity has been evaluated by taking absorbance via spectrophotometer at 562 nm. All the tests had been operated for three times for better accuracy and blank has been determined to calculate the amount of formazan that has been produced, and quercetin has been taken as a standard for this experiment.

3.3.1 Data interpretation of superoxide radical scavenging assay

According to Table 5, the IC₅₀ value for catechin was 26.79 ($\mathit{\mu g}$/ml), and crystal compound was 29.04 ($\mathit{\mu g}$/ml), which was satisfactory result for this in vitro test.

Name of sample	Concentration ($\mathit{\mu g}/\mathit{ml}$)	% of scavenging (Mean$\pm$S.D)	IC50 ($\mathit{\mu g}$/ml)
Catechin (Standard compound)	1.25	$17.01\pm$0.91	26.79
	2.50	$19.25\pm$0.53
	5.00	$24.27\pm 1.91$
	10	28.06 $\pm 0.19$
	15	35.22 $\pm 0.66$
	20	41.44 $\pm 0.46$
Crystal compound (Pure)	1.25	$15.29\pm 0.29$	29.04
	2.50	$19.01\pm 0.53$
	5.00	$23.34\pm 0.26$
	10	$26.51\pm 1.03$
	15	$33.93\pm 1.13$
	20	$39.32\pm 0.85$

Table 5.

Superoxide radical scavenging data analysis for standard catechin and crystal compound.

3.4 Graphical presentation of cumulative data obtained from in vitro antioxidant assay

From the above graph, it can easily assumed that the crystal compound may have great antioxidant activity. We know that lower the value of IC₅₀ actually enhances the probability of having better antioxidant power of sample. When performing DPPH free radical scavenging activity, the sample had shown a very low value (4.1 $\mathit{\mu g}/\mathit{ml})$ of IC₅₀ close to standard catechin (3.2 $\mathit{\mu g}/\mathit{ml}$), which ultimately provided a confirmation about the antioxidant capability of plant material.

Like DPPH test, the hydroxyl free radical scavenging assay and superoxide radical scavenging assay also convey messages regarding the free radical scavenging power and oxidant neutralizing potency of plant materials or samples. The IC₅₀ value for hydroxyl radical scavenging assay of crystal compound was found as 35.01 $(\mathit{\mu g}/\mathit{ml}$), which was near the value of standard catechin of 31.77 $\left(\mathit{\mu g}/\mathit{ml}\right)$; similarly the IC₅₀ value was found as 29.04 and 26.79 for crystal compound and standard catechin, respectively, during super oxide radical scavenging assay (Figure 7).

Figure 7.

Graphical representation for in vitro data analysis (DPPH radical scavenging test, hydroxyl radical scavenging test, superoxide radical scavenging test) regarding crystal compound found in black pepper and standard catechin.

3.5 Reducing power capacity assessment of crystal compound isolated from black pepper

The reducing power of plant material and standard material had been evaluated through the method of Oyaizu (1986) [44]. During this assay, the color of sample has altered to various shades of green and blue color based on the reducing capacity of the samples. The antioxidant compound actually served as reductant, and it induces reduction of the ferriccyanide to the ferrous form by releasing an electron. The overall reaction has been examined by monitoring the appearance of Perl^’s Prussian blue at 700 nm.

$${\mathrm{Fe}}^{3+}-\text{ferricyanide}+\mathrm{e}\to {\mathrm{Fe}}^{2+}-\text{ferrouscyanide}$$

Reducing power was investigated using the method developed by Oyaizu (1986). 2.5 ml of phosphate buffer (200Mm, p^H 6.6) and 2.5 ml 1% potassium ferricyanide had been incorporated into 2.5 ml of sample (crystal compound). The mixture was transferred into a water bath for maximum 20 min at 50$°\mathrm{C}$ followed by rapid cooling after warm-up. After that, 2.5 ml of 10% trichloro acetic acid was added, and it has centrifuged at 3000 rpm for 10 min. From the supernatant, 5 ml had been collected and being dispersed it into 5 ml of distilled water and 1 ml of ferric chloride, and absorbance has been taken, which reflects that higher the absorbance value, stronger the reducing capability. According to Table 6, the absorbance of crystal has increased with increasing concentration, which was close to similar with standard ascorbic acid.

Name of sample	Concentration ($\mathit{\mu g}/\mathit{ml}$)	Absorbance Mean$\pm$S.D
Ascorbic acid (Standard compound)	5	1.326 $\pm$0.116
	10	2.119$\pm$0.116
	20	2.455$\pm$0.116
	40	2.5973 $\pm$0.116
	80	2.756$\pm$0.1160
Crystal compound (Pure)	5	0.526 $\pm$0.045
	10	0.842$\pm$0.127
	20	1.326$\pm$0.116
	40	2.119$\pm$0.1160
	80	2.59 $\pm$0.106

Table 6.

Reducing power capacity assessment data analysis for standard ascorbic acid and crystal compound.

3.5.1 Graphically presenting the data obtained from the reducing power assessment

It has been identified that the reducing power assay of crystal compound was slightly discriminable with standard ascorbic acid at low concentration; however, the difference between plant material and standard has diminished with escalating concentration with giving high absorbance value as shown in Figure 8.

Figure 8.

Graphical representation of in vitro data analysis (reducing power capacity assessment) regarding standard ascorbic acid and crystal compound isolated from black pepper.

4. Discussions

4.1 Discussions regarding in vitro test

So, the overall in vitro antioxidant assay could be concluded that the obtained crystal compound (may be piperine) may have strong antioxidant power. By comparing with standard compound (catechin, ascorbic acid), the prediction could be extended that the black pepper is a great source of antioxidant compound with specifying the crystal constituent (may be piperine) as a prominent free radical scavenger.

4.2 Discussions regarding biodiversity of black pepper

The diversity of plant-derived medicinal constituents assists in a plenty of ways to enrich our research for instances in drug design and development. My current research was focused on merely to explore the free radical scavenger; however, my theoretical research through existing research article has claimed that piperine (crystal material) may exist in a lot of medicinal plants such as Piper longum, Piper officinarum rather than Piper nigrum (black pepper) also [45, 46].

Besides the antioxidant capability of piperine, the existing research has mentioned that it has a cluster of pharmacological implications also, which actually introduce the biodiversity of the piperine found in black pepper.

Li S, Wang C et al., 2007 show that piperine could be effective against corticosterone-mediated depression.

Taqvi SI, Shah AJ et al., 2008 have identified antihypertensive effect of piperine.

Bang JS, oh da H et al., 2009 have observed that piperine found in black pepper may exhibit both antiarthritic and anti-inflammatory activity.

Manoharan S, Balakrishnan S et al., 2009 have found that probably the piperine of piper nigrum has anticancer activity.

Parganiha R, Verma S et al., 2011 invented that Piper nigrum may have in vitro antiasthmatic activity.

Hussain A, Naz S et al., 2011 found that piperine may enhance saliva and pancreatic enzyme secretion, thereby accelerating the digestion process by reducing gastric emptying time.

Kumar KP, Vanaja M et al., 2014 have explored that the leaf and stem of piper nigrum might possess antibacterial coverage against plant microorganisms.

4.3 Discussions regarding biodiversity of Piperaceae family or piper species

I have already mentioned that black pepper (Piper nigrum) is included in piperaceae family; simultaneously a lot of medicinal plants are also included in same family possessing diverse type of biological activity. The biodiversity of medicinal compound is a blessing for us due to the presence of a variety of bioactive constituents, which makes individual plants able to show different type of implications against several types of diseases or pathological conditions, and some of these are explained below.

Piper longum L. utilized as an antidote to snake bite, scorpion stings. Besides this, it can be applied in chronic bronchitis, cough, and cold [47].

Piper betle L. has a lot of implications such as in cuts, boils, scabies, mouth odor, cough remedy, bronchitis, and nosebleed [48].

Piper aduncum L. may have utilization in stomach aches, vaginitis, influenza, rheumatism, cough, fever, and general infections [49].

Piper aborescens Roxb. could be used in rheumatism, cytotoxic activity, and antiplatelet aggregation [50].

Piper capense L.F. might have application in renal disorder, gonorrhoea, syphilis, abdominal pain, enteritis, and asthma [51].

Piper ovatum Vahl possesses anti-inflammatory and analgesic activity [52].

Piper retrofractum Vahl can be consumed as digestive aid, stimulant, carminative, and in intestinal disorders, postpartum treatment (women) [53].

Piper tuberculatum Jacq. had been used as antidiuretic, analgesic, sedative, and antidote for snake bites [54].

5. Conclusions

Nowadays, we are suffering from several types of physiological abnormalities due to environmental impact or changes in lifestyle, which ultimately develop a lot of diseases that lead to acute or chronic effect on health. Now we are really blessed as nature has provided a plethora of medicinal plants in our surroundings, and we just need to identify them with their perfect implication. My overall research work was based on one type of plant with identifying bioactive compound with its pharmacological application; moreover, I have tried my best to explain how diversity of bioactive compound varies from one medicinal plant to another.

From the above analysis, it is clear that same type of plant may be applicable in different types of diseases, which is called biodiversity of medicinal plant. The invention of every type of synthetic drugs directly or indirectly relieson natural sources as the basis of production of synthetic drug comes from medicinal plants. With increasing diseases, the demand of synthetic drug has also proliferated, which may be life-threatening for humans as formulated drugs can cause damage of vital organs with frequent consumption. To diminish the side effects or dependency on synthetic drugs, we should raise awareness as early as possible.

Our future plan could be implemented by cultivating medicinal plants commercially and procuring the identified bioactive compound with maintaining all the precautions and after that it will be possible to mitigate the demand on synthetic drug. This is the ultimate way to get beneficial effect from medicinal plants and the adverse effect of marketed drug might be avoided.