Marker Profiling: An Approach for Quality Evaluation ...

natural health products

Pulok K. Mukherjee, PhD, FRSC School of Natural Product Studies, Department of Pharmaceutical Technology, Jadavpur University, Kolkata, India S. Ponnusankar, MPharm School of Natural Product Studies, Department of Pharmaceutical Technology, Jadavpur University, Kolkata, India P. Venkatesh, MPharm School of Natural Product Studies, Department of Pharmaceutical Technology, Jadavpur University, Kolkata, India Arunava Gantait, MPharm School of Natural Product Studies, Department of Pharmaceutical Technology, Jadavpur University, Kolkata, India Bikas C. Pal, PhD School of Natural Product Studies, Department of Pharmaceutical Technology, Jadavpur University, Kolkata, India Key Words Quality control; Indian medicinal plants; Marker profiling Correspondence Address Pulok K. Mukherjee, Director, School of Natural Product Studies, Department of Pharmaceutical Technology, Jadavpur University, Kolkata 700032, India (email: naturalproductm @gmail.com).

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Marker Profiling: An Approach for Quality Evaluation of Indian Medicinal Plants A key point in choosing the appropriate method to guarantee the quality and safety of herbal products is to develop suitable methodology for their standardization with a sufficient degree of specificity in ensuring the desired endpoint. The unique differences in the constituents of herbal drugs create distinct challenges based on their identity, quality, and consistency of efficacy. The differentiation of conventional medicines and herbal drugs can be minimally considered from regulatory, economic, and technical perspectives. Health risks associated with herbal products are considered in three categories: extrinsic (accidental, deliberate), intrinsic (bioavailability, pharmacokinetic, pharmacodynamic), and consumer-dependent causative factors

I NTRODUCT I ON The use of plant-derived products in disease management was an important breakthrough in the history of humankind. Many populations in developing countries, including India, use plants or plant-derived products as medicines for their primary health care. The Indian system of medicine (ISM) consists of several major components such as Ayurveda, Siddha, Unani, and homeopathy, which provide major health care for a large part of the population in India. The materia medica of India provides lots of information on the folklore and traditional practices of therapeutically important natural products. The evidence for the therapeutic actions of herbal drugs is documented in Indian, Chinese, European, and African systems of medicine (1). The reappearance of interest in green products throughout the world has created an expanding market for plant-based products, which should satisfy all the specifications related to their quality, safety, and efficacy (2). Chemical profiling of herbal products can help in distinguishing several constituents

(therapeutic failure, adverse drug reaction, hypersensitivity). A change in these categories through regulation using designated approaches can minimize the health risks. Chemical fingerprinting of natural products and their ability to interact with physiological substrates of the human body are the mainstay of their therapeutic efficacy. This requires a multidisciplinary approach, involving analytical techniques and methodologies common to ethnomedicine, botany, pharmacology, pharmacotherapy, toxicology, and pharmacoepidemiology. An attempt has been made through this article to highlight the use of marker profiling of natural products with special reference to Indian herbal medicine.

present therein. This can help in development of evidence based on their traditional uses as described in ISM. The evaluation of the quantity of these constituents can help to quantify the biologically active molecules so as to standardize the raw materials and finished products (3). Marker profiling in essence of chemoprofiling can help in establishing the quality of the raw material and finished products and can also help to note the batch-to-batch consistency of formulations made from those raw materials.

M AR K ERS I N NATURAL HEALTH PRODUCTS Natural products derived from botanicals are mostly available from wild sources and present the greatest challenges for ensuring consistent product quality. Environmental factors like soil conditions, availability of light and water, temperature variations, nutrients, and geographical location affect the accumulation or percentage of phytochemicals or phytoconstituents present in plants. Further, cultivation and harvesting techniques, postharvest processing, and storage methods also influence the physical ap-

Drug Information Journal, Vol. 45, pp. 1–14, 2011  •  0092-8615/2011 Printed in the USA. All rights reserved. Copyright © 2011 Drug Information Association, Inc.

Submitted for Publication: April 22, 2010 Accepted for Publication: October 20, 2010

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Figure 1 Application of marker analysis.

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Authenticate the botanical source

Identify the botanical source Achieve more consistent quality

Detect adulteration and substitution

Quantify the active principle

Evaluate characteristic fingerprints

Ensure product efficacy Evaluate material grade

Confirm uncommon chemical entities Validation of therapeutic doses

Application of marker analysis

Confirm the potency

pearance of the plant and its chemical qualities (2,4). This means quality parameters should be set not only for the plant material but also for plant extracts and final dosage forms. Further, botanicals do not have a consistent, standard chemical composition in respect to their different parts like roots, leaves, stems, flowers, and fruits. Each part needs individual chemoprofiling based on different phytoconstituents. This variability with a range of extraction techniques and processing steps results in commercial products with wide quality differences (5–7). Evidence of safety and efficacy then becomes extract specific. The challenge for herbal manufacturers is to characterize and monitor the phytoconstituents in a way that enables consistent production and efficacious finished products. The use of markers, standardization, chemical and DNA fingerprinting, bioassays, and the emerging field of phytomics provide mechanisms for assuring consistent quality. According to the Natural Health Product Directorate of Canada, “marker compounds are a constituent that occurs naturally in the material and that is selected for special attention (ex: for identification or standardization) by a researcher or manufacturer” (eg, markers used in commercial products include hypericins [1] and stabilized hyperforins [2] from St. John’s

Verify the labeled claim Stability studies

wort, silymarin [3] in milk thistle, ginsenosides [4] from ginseng, etc). Marker selection may be based upon a variety of different factors including stability, ease of analysis, time and cost of analysis, relevance to therapeutic effect, indicator of product quality or stability, or previous use by other manufacturers or researchers (8). To identify or authenticate the source of the material, markers play an important role. They can be used in several ways (Figure 1) to evaluate quality, so as to ensure the efficacy and safety of the natural health products (NHPs) (5). There are several contexts of defining the concept of markers. Marker compounds are not necessarily pharmacologically active all the time but their presence is well established in products with characteristic chemical features (9). Marker components may be classified as active principles, active markers, analytical markers, and negative markers (10), while biomarkers may be defined as those markers with known pharmacological activity. However, there are several challenges to isolating and identifying a marker for an NHP. The points that need to be analyzed in this context are described in Figure 2. The herbal manufacturers and researchers need to address these critical questions to aid in the harmonization of specifications and analytical methodologies for natural products.

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Quality Evaluation of Indian Medicinal Plants

Combination of chemical constituents used? What is the appropriate marker for that botanical? At what level?

Does the marker have reference standards?

Chemical markers

Is there any validated method to detect and quantify the marker?

Figure 2 What criteria for use should be considered?

Can an agreement be derived about the use of a specific marker?

Stability of the marker during extraction and processing? How does the marker relate to potency and efficacy?

Adhatoda vasica (Acanthaceae) Adhatoda vasica is a well-known plant in Ayurveda, commonly known as malabar nut, and belongs to the family Acanthaceae. It has been most frequently used for the treatment of respiratory diseases like cough, asthma, and cold. Methanol extracts of the aerial part of the plant have been reported to possess antiallergic and antiasthmatic activities in the guinea pig after inhalation or intragastric administration at doses of 2.5 g/kg, respectively (11). The frequent use of A. vasica has resulted in its inclusion in the WHO manual for the use of traditional medicine in primary health care, which is intended for health workers in Southeast Asia to keep them informed of the therapeutic utility of their surrounding flora (12). This plant is the source of vasicine [5], a pyrralazoquinazoline alkaloid, which is the active principle obtained from this plant used against asthma. In addition, it contains other alkaloids like vasicol, adhatonine, vasicinone, and so on. Ephedra sinensis (Ephedraceae) Ephedra sinensis, also known as ma huang, is the common source of ephedra products. It is a popular dietary supplement that includes ephedrine alkaloids [6], which are active prinDrug Information Journal

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ciples (6–8%) found in this plant (13). Ephedra is found in at least 200 over-the-counter products, which are consumed in the treatment of weight loss, weight management, or energy enhancement. The dose-dependent pharmacological activity of ephedrine has been extensively investigated in human and animal studies, including central nervous system stimulants, adrenergic receptors, bronchodilators, cardiovascular and psychophysiological effects, and so on (14–18).

Silybum marianum (Asteraceae) The extracts of the flowers and leaves of Silybum marianum (milk thistle, family Asteraceae) have been used for centuries to treat liver, spleen, and gallbladder disorders (19). The biologically active principles of silymarin [3], a flavonolignan mixture, were studied extensively. Recently, oxidized derivatives of silybin (the major component forming 70–80% of silymarin) and their antiradical and antioxidant activity were studied (20). Further antioxidant, anti-inflammatory, and anticarcinogenic properties were also demonstrated (21). Recent studies have focused on mechanisms regarding the possible molecular targets of silymarin for cancer prevention (22).

Challenges associated with the use of chemical markers.

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Allium sativum (Liliaceae) Allium sativum (garlic) is a common food for flavor and spice and it is one of the herbs most commonly used in modern folkloric medicine. Garlic is proven to possess many therapeutic benefits due to its effective compounds, which exhibit anticoagulant (antithrombotic), antioxidant, antibiotic, hypocholesterolemic, hypoglycemic, and hypotensive activity (23,24). Garlic’s strong odor is largely due to sulfurcontaining compounds (eg, S-allylcysteine sulfoxide), which are believed to account for most of its medicinal properties (25). Allicin has been shown to be important in many health effects of garlic. However, the anticancer effect of garlic might be shared between allicin and other unidentified compounds (26). Garlic contains about 1% alliin [7], which is converted enzymatically by allicinase to allicin and other sulfur-containing compounds (25,27). Ocimum sanctum (Lamiaceae) Ocimum sanctum, commonly known as Tulsi, has been well documented for its therapeutic potential in Ayurveda including its effect as “Dashemani Shwasaharni” (antiasthmatic). Eugenol [8] is an essential oil extracted from different parts of this plant (28). O. sanctum has been reported to protect against histamine, pollen-induced bronchospasm in guinea pigs, and to inhibit antigen-induced histamine release from sensitized mast cells (29). Other plants of genus Ocimum are known for their therapeutic potentials, such as Ocimum gratissimum L. (Ram Tulsi), Ocimum canum L. (Dulal Tulsi), and Ocimum basilicum L. (Ban Tulsi). It is reported to contain several other constituents like carvacrol, caryophyllene, nerol and camphene, eugenol, eugenol methyl ether, methyl chavicol, cineole, and linalool (30). Hypericum perforatum (Clusiaceae) The hypericin [1] and hyperforin [2] content in St. John’s wort is another example of active markers. It has gained popularity as an alternative treatment for mild to moderate depression. Hypericum contains at least 10 classes of biolog-

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ically active detectable compounds, including hypericin (0.09%) (naphthodianthrones) and hyperforin (2–4.5%) (phloroglucinols) (31). Several types of pharmacological activity of this plant have been demonstrated and this activity has been attributed to hyperforin, hypericin, and flavonic compounds. Further, antimicrobial, in vitro cytotoxicity, and antitumor properties of Hypericum mysorense and Hypericum patulum were also elucidated (32–34). Some of the most important Indian medicinal plants, along with their known analytical markers, are shown in Table 1 (35–44).

Echinacea angustifolia (Asteraceae) Different alkylamides [19] present in the roots of Echinacea angustifolia and E. purpurea may be used as suitable markers. Echinacea (purple coneflower) is one of the most popular herbal supplements used to alleviate cold, sore throat, cough, and other respiratory infections (45). The most common constituents present in echinacea include alkylamides, caffeic acid derivatives, and so on. Several studies have described the effect of E. purpurea on the immune system, upper respiratory tract infections, and so on (46,47). Acorus calamus (Acoraceae) Acorus calamus is the source of β-asarone [9], a yellow aromatic volatile oil obtained from the dry rhizomes of this plant. This plant has potential use against central nervous system disorders in Ayurveda (48). Its other constituents include α-asarone, calamenol, calamine, and eugenol. β-asarone has toxic effects even though it is a major constituent, which is the negative marker in this well-reported plant. Ginkgo biloba (Ginkgoaceae) Ginkgo is referred to as a living fossil, since it can live for as long as 1,000 years. Ginkgo contains many bioactive compounds including flavonols and flavone glycosides, terpenoids, and ginkgolides. Many studies report its activity including antioxidant, vasoacting properties, inhibition of platelet-activating factor, and neu-

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Some Medicinally Important Indian Plants and Their Known Phytomarkers Sl. No.

Scientific and Local Name

Family

Parts Used

Marker Compound β-asarone [9]

  1.

Acorus calamus (35), sweet flag, Vacha Araceae Rhizome powder

  2.

Albizia lebbeck (36), Shirisha

Fabaceae Stem bark Catechin [10]

  3.

Clitoria ternatea (37), Aparajita, Gokarna

Fabaceae

  4.

Curcuma longa (38), Haridra

Zingiberaceae Rhizome Curcumin [12]

  5.

Glycyrrhiza glabra (39), Yasti madhu Leguminaceae Root Glycyrrhizin [13]

  6.

Murraya koeniggii (40), Karipatha Rutaceae Leaf

Mahanimbine [14]

  7.

Nelumbo nucifera (41), Indian lotus, padma Nymphaeceae Rhizome

Betulinic acid [15]

  8.

Terminalia chebula (42), Haritaki Combretaceae

  9.

Trigonellum foenum-graecum (43), Methi Leguminaceae Seed Trigonelline [17]

10.

Zingiber officinalis (44), Adraka

rotransmitter modulation (49). Due to its association with improvement in cognitive function, it is among the most consumed phytopharmaceuticals in the United States and Europe. Ginkgolic acids, a mixture of structurally related n-alkyl phenolic acid compounds in Ginkgo biloba L., are strong allergens that could cause severe allergic reactions (50). Besides allergic properties, they have been recognized to possess possible cytotoxic, mutagenic, carcinogenic, and genotoxic properties (51). Another study indicated that ginkgolic acids activated protein phosphatase 2C to induce neurotoxic effects in cultured chick embryonic neurons (52). Active principles are chemically characterized and defined chemicals with known clinical activity. Several examples of such active principles as major markers are explained in subsequent sections. Active markers are chemicals with known pharmacological activity contributing to efficacy, but that may or may not have proven clinical efficacy. Analytical markers are chemicals chosen for quantitative determination. However, they may or may not be biologically active. Further, they may aid in the positive identification of raw material and extracts or may be used to achieve the standardization profiles of the plant

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Flower Taraxerol [11]

Fruit Gallic acid [16]

Zingiberaceae Rhizome

6-gingerol [18]

either in the raw material or in finished formulations. Negative markers are chemically characterized compounds with allergenic or toxic principles or those that interfere with bioavailability.

CHE M I CAL F I NGERPR I NT I NG AND STANDARD I Z AT I ON O F B OTAN I CALS THROUGH M AR K ERS Botanicals are standardized based on the presence of a known active ingredient or specific markers when the active markers are not yet recognized. But this can help in establishing the product’s quality depending on the characteristic fingerprints. Plants contain several active substances in certain ratios and in standardized extracts. This ratio must be kept constant, within narrow limits, from one preparation to another (53). The unique processing methods followed for the manufacturing of ISM turn herbal ingredients into very complex mixtures, through which the separation, identification, and estimation of chemical components become more challenging in some cases. Moreover, herbals are known to contain several components and in many cases the absolute compound responsible for the pharmacological activity is unknown. Chemical analysis should be extended to several groups

TA B LE 1

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or classes of constituents and a chemical fingerprint should better be established to evaluate the phytoconstituents present therein. Chemical fingerprints can be used to authenticate plant material, identify the quantification of active compounds, and relate the chemical composition to biological activity for product standardization. A cautionary principle also applies to standardization; that is, manufacturers do not use the same markers or methods to obtain standardization. Thus, similar products may not be standardized to the same marker compound; or if to the same marker, to different levels. In addition, marker content may not necessarily vary in direct proportion to the content of other important constituents or to its therapeutic effectiveness. Standardization requires control of both raw material quality and manufacturing processes to final product. Different chromatographic techniques are most frequently used for the identification and quality control (QC) of herbal medicines or products. While there are multitudes of chromatographic techniques to achieve separation, the common thread is the separation of compounds through the use of variations in mobile and stationary phases. These include thin layer chromatography (TLC), high-performance thin layer chromatography (HPTLC), high-performance liquid chromatography (HPLC), gas chromatography (GC), and several hyphenated techniques like HPLC-MS, GC-MS, and so on. All these techniques can help to quantify the phytochemicals present in complex mixtures of herbal products (54,55). Method development and analytical evaluation of these constituents through chemical fingerprinting is a challenging task, but due to its popularity and several advantages it is gaining momentum for rapid characterizations and evaluation of compounds extracted from herbs (56). Chemical fingerprinting does have some disadvantages, such as using only a small quantity of chemicals in the plant as costs, time, and limits of current technologies preclude all chemicals from being examined. Some molecules like phenolics and sterols may overlap, making it difficult to separate individual components.

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PHAR M ACOPOE I AL M ETHODS F OR STANDARD I Z AT I ON O F HER B S AND HER B AL PRODUCTS Most of the regulatory guidelines and pharmacopoeias in India recommend macroscopic and microscopic evaluation and chemical profiling of the herbs for QC and standardization. Chemical profiling is much more characteristic where a chemical pattern for a crude drug, its fractions, or extracts is identified. TLC and HPTLC are very important tools for qualitative as well as quantitative determination of the presence or absence of a particular compound in the herb. Many other analytical techniques such as volumetric analysis, gravimetric determinations, GC, column chromatography, HPLC, and spectrophotometric methods are regularly used for QC and standardization. But for quantitative studies, use of specific markers and chemical profiling to distinguish between different herbs remains a preferred option. These markers may or may not be therapeutically active but they should ideally be neutral to environmental effects (57). Marker analysis can be used in identification of chemical entities, detection of adulteration, QC of mixtures, shelf life of herbal formulations, and control of pharmacological activity for the evaluation of clinical trial data. By use of markers in the clinical trial data, the material used in the trial can be characterized as thoroughly as possible, so that the result of the trial can be extrapolated to other batches of the same products and to products of similar botanical identity. The Indian Pharmacopoeia 2007 (IP) includes pharmacopoeial specifications with monographs for 58 individual herbs being used in therapy. The specifications include the name of the drug (along with its common name), its biological source (Latin name), the part of the plant under consideration, its description, macroscopic and microscopic study, identification, several QC parameters, and assay in respect to the phytochemical reference standards or botanical reference standards (58). The Ayurvedic Pharmacopoeia of India (API) is another official compendium published by

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Quality Evaluation of Indian Medicinal Plants

Limit test for arsenic

Systematic study of crude drugs

Limit test for chlorides Limit test for iron

Microscopic method of examining crude vegetable drugs

Limit test for lead Limit test for sulfates Limit test for heavy metals

Determination of stomatal index Determination of palisade ratio Determination of vein islet number

Sulfated ash

Morphological study of drugs

Limit tests Several aspects of quality control of herbal medicine

Determination of quantitative data of vegetable drugs

Sampling of drugs Determination of foreign matter Determination of total ash Determination of acid insoluble ash Determination of moisture content Determination of water soluble ash Determination of fixed oil content Determination of volatile oil Special processes used in alkaloidal assays

the Ministry of Health and Family Welfare, Government of India (59). This describes different methods for QC and standardization of herbs and herbal products. Several specifications for quality evaluation of natural products as prescribed in the Ayurvedic Pharmacopoeia include morphological study, determination of quantitative data (eg, extractive values, foreign matter, etc), limit tests, and different physical tests (eg, boiling range, refractive index, pH, etc). A schematic presentation of these parameters is shown in Figure 3.

M AR K ER PRO F I L I NG O F SO M E I ND I AN M ED I C I NAL PLANTS The detection of markers or bioactive metabolites is the starting point for a strategic approach in the search for potentially useful compounds. Many new natural product–oriented Drug Information Journal

Physical tests and determinations

Determination of boiling range Determination of pH value Determination of melting range Specific optical rotation Power fineness Refractive fineness Weight per mL and specific gravity

bioactive compounds effective in treating several diseases have been isolated and evaluated at our laboratory. Development of lead compounds from these Indian medicinal plants and their evaluation may help to promote natural products based on their quality, efficacy, and safety. Marker analysis of several herbal drugs including polyherbal formulations from ISM has been performed and reported by our research group. The fingerprint profiles of catechin [10] from Albizia lebbeck (36), betulinic acid [15] from Nelumbo nucifera (immunomodulatory agent) (42), β-asarone [9] from Acorus calamus L. (35) (AChE inhibitor), mahanimbine [14] from Murrya koenigii (AChE inhibitor) (41), and tilianin from Semecarpus anacardium (hepatoprotective) (60) and their pharmacological activities have been reported.

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Figure 3 Several aspects of quality control of herbal medicines.

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HPTLC Chromatogram of Andrographis paniculata Extract With Standard Andrographolide

Figure 4 Marker profile of andrographolide.

Andrographis paniculata

Andrographolide Standard

3D Overlay Solvent SystemChloroform: Acetone: Benzene (20:20:10)

Marker analysis of glycyrrhizin [13] from Glycyrrhiza glabra has been reported through HPTLC densitometry (39). This is a validated method per ICH guidelines. The method was validated in terms of specificity, linearity, precision, detection limit, and quantification limit (39). A method for the estimation of the content of taraxerol in extracts of Clitoria ternatea was developed and validated. Taraxerol [11] was isolated from C. ternatea and using this compound as the marker, standardization of the plant was performed using hexane and ethyl acetate (80:20 v/v) as the mobile phase. Scanning was done at 420 nm after spraying the plate with anisaldehyde reagent. Rf of taraxerol was found to be 0.53. Linearity range was found to be 100–1,200 ng and recovery studies showed an average recovery of 99.65–99.74%. Limits of detection and quantification were determined as 31 and 105 ng/spot (37). An HPTLC method was also developed for standardization of a polyherbal formulation comprising Emblica officinalis, Terminalia chebula, and Terminalia belerica. This formulation is very popular in India and is known as Triphala in Ayurvedic medicine as a laxative and immunomodulatory agent. Gallic acid was used as a

Andrographis paniculata Raw Material

Andrographolide

S1−Andrographolide (Standard) T1, T2−Test Samples

phytochemical marker for this formulation, as this compound was found to be present in all three fruits used in this formulation. A method was developed with toluene: ethyl acetate: glacial acetic acid: formic acid (20:45:20:05) as mobile phase and scanned at 254 nm (42). A rapid, simple, and accurate analytical method has been developed to standardize Triphala and its individual components using gallic acid as a marker. Zingiber officinale is a very well known medicinal plant. The 6-gingerol [18] content in ginger was determined through a sensitive and accurate HPTLC method (44). Methanol extracts of the rhizomes of ginger were analyzed using a mobile phase comprising n-hexane: diethyl ether (40:60). This method also showed good reproducibility and accuracy. The developed method permits reliable quantification of 6-gingerol and good resolution and separation of 6-gingerol from other constituents of ginger (44). The marker profiling of some hepatoprotective Indian medicinal plants like Andrographis paniculata, Berberis aristata, and Phyllanthus amarus has been reported (61). Andrographis paniculata is a well-known hepatoprotective plant in

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Quality Evaluation of Indian Medicinal Plants

ISM and has been analytically standardized using andrographolide [21] as a marker compound, which has been developed using a solvent system consisting of chloroform: methanol (70:10) (Figure 4). A list of several phytomarkers from therapeutically potent plant species is shown in Figure 5. With the global increase in the demand for plant-derived medicines, there is a call for ensuring the quality of herbal drugs using modern analytical techniques. Technical drawbacks associated with natural product research have been narrowed, and there are better opportunities to investigate the biological activity of previously inaccessible sources of natural products. Various methods of standardization and testing are required immediately. With the increasing acceptance that the chemical diversity of natural products is well suited to provide the

OH

O

core scaffolds for future drugs, there will be additional developments in the use of novel natural products and chemical libraries based on natural products in drug discovery campaigns.

CONCLUS I ON ISM is a highly evolved ancient science-based medicine system, which functions on its own unique fundamental principles. The principles of Ayurveda have been developed on a philosophical background with scientific reasoning. However, ISM needs a new methodology for its development in the realm of existing science and technology. Existing concepts of ISM are to be revalued through modern concept strategies that are more logical and acceptable to the world community. With the emerging worldwide interest in adopting and studying Indian traditional medicines and in exploiting their poten-

OH

Figure 5

HO HO

OH

O

Me Me

HO O

OH

O

OH

Hypericin [1] from Hypericum perforatum

Hyperforin [2] from Hypericum perforatum

HO O H

O

O

H

OH

H

CH2OH OCH3

H

OH

Silymarin [3] from Silybum marianum

OH

O H

H

O

O

H

O

OH

OH

OH

H

OH

HO

HO

H HO HO

O

O O O

OH

9

OH

OH

OH

Ginsenoside [4] from Ginseng

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OH

CH3

N N Vasicine [5] from Adhatoda vasica

NH

CH3

(−) Ephedrine [6] from Ephedra sinensis

O

NH2

S Alliin [7] from Allium sativum

COOH

Important phytomarkers of Indian medicinal plants. (Figure continues on next 2 pages)

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Figure 5 OH OH

(Continued)

O

O

OH CH3

C H3

HO O O

Eugenol [8] from Ocimum sanctum

β-Asarone [9] from Acorus calamus

OH OH Catechin [10] from Albizia lebbeck

HO O

Curcumin [12] from Curcuma longa

O

OH

O

HO

OH

HO

Taraxerol [11] from Clitoria ternatea

OH

OH O OH O

HO O

O

OH

O

O

2+

OH

O

C H2 CH 2

O

O

2+

O

H N O

O

O

Glycyrrhizin [13] from Glycyrrhiza glabra

Mahanimbine [14] from Murrya koenigii

O OH

HO

O

C-N HH3 C-N 3

++

HO

HO

CH C H2 2

OH

--

O O

OH Betulinic acid [15] from Nelumbo nucifera

Gallic acid [16] from Terminalia chebula

Trigonelline [17] from Trigonella foenum-graecum

O O

OH OH 33C C

O

OH OH CH

O

C H3 3

N H

HO

HO

6-gingerol [18] from Zingiber officinale

N H

Isomer pair dodeca-2E, 4E, 8Z, 10E-tetraenoic acid isobutylamide and dodeca-2E, 4E, 8Z, 10Z-tetraenoic acid isobutylamide Alkylamide [19] from Echinacea angustifolia

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Quality Evaluation of Indian Medicinal Plants

Figure 5

HO

OH (Continued)

COOH &2 2 + &+ & & + & + & + ) CH ) +CHCH(CH (CH 2 7 2 5 3

+2 HO

OH

O O

Andrographolide [21] from Andrographis paniculata

Ginkgolic acid [20] from Ginkgo biloba

tial based on different health care systems, the Government of India has initiated several approaches through its department of AYUSH (Ayurveda, Yoga, Unani, Siddha, Homeopathy), which functions through individual councils for all these systems of medicine. The National Medicinal Plant Board has taken special interest in the development of medicinal plants in general and the protection of biodiversity of medicinal plants in particular through its various projects. Beside these, the Council for Scientific and Industrial Research (CSIR) has taken several initiatives including Traditional Knowledge Digital Library, CSIR NMITILI projects, and others for promotion of Indian natural products. These approaches by different councils for evaluation of the various facets of ISM have enabled validation of their therapeutic potentials as well as generation of data to put these systems of medicine in national health care programs. Though the development of new technologies helps in rapid isolation, identification, and characterization of molecules, the use of different plants for treating ailments in ISM is based on the fact that the additive or synergistic effects of the phytoconstituents present in those plants can enhance the therapeutic viability of the plant products. Combining the strengths of the knowledge base of ISM with the combinatorial chemistry and high throughput screening will help in generation of leads for different diseases. However, a concept of understanding the complex principles of ISM must be developed through marker profiling and related approach-

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es so as to develop evidence-based practice of ISM.

Acknowledgments—The authors are thankful to the National Medicinal Plants Board (NMPB), New Delhi: F. No.: Z. 18017/187/CSS/R&D/WB-01/2009-10MPB for providing support to the School of Natural Product Studies, Jadavpur University, Kolkata.

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