5. Fingerprint Profile of an important therapeutic plant of …abap.co.in/sites/default/files/Paper - 5 colour.pdf · fingerprints of herbal drugs. Fingerprint analysis approach using

257Current Trends in Biotechnology and PharmacyVol. 12 (3) 257-264 July 2018, ISSN 0973-8916 (Print), 2230-7303 (Online)

AbstractNature is crucial source of drugs. Plants

have played important role in identifying importantdrugs and are basis of various modernpharmaceuticals. Therefore ethnomedicinal plantsmust be exploited to identify lead compounds. C.acuminatum is an important plant of Astavarga(combination of 8 drugs, most commonly used inAyurveda) C. acuminatum is used in breathingdisorders, burning sensation, cough, decrease inbone tissue, bleeding disorders, blood disorders,tuberculosis, insect bites and rheumatism. It isrefrigerant, febrifuge and aphrodisiac. It is utilizedas tonic and in general debility. HPTLC is avaluable analytical tool for the investigation ofherbal products and drugs. Hence, HPTLCFingerprint profile has been developed for this plant.

Objective: The present study was aimedto develop the fingerprint profile by HPTLC ofCrepidium acuminatum (D. Don) Szlach.

Materials and Methods: HPTLC System(CAMAG, Switzerland), equipped with linomat 5applicator, development chamber, scanner,derivatizer, vision Cats software, was used. Theplate was scanned at 580 nm using Tungstenlamp and images were captured at visible light,UV 254 nm and UV 366 nm.

Results: HPTLC method for separation ofphytoconstituents using different solvent systemhas been developed for C. acuminatum. The studyrevealed the presence of 2 saponins, 3 bitter

principles, 2 steroids, 2 Sterols, 2 essential oils,1 anthraquinones, 2 coumarin and 5 flavonoids inmethanol extract of pseudobulbs of this plant.

Conclusion: The HPTLC fingerprint profiledeveloped for methanol extract of C. acuminatumcan be used for routine quality control of herbalformulations comprising of this plant and serveas a base for qualitative, quantitative analysis andstandardization of the drug ‘Jeevak’. It will alsohelp in identification and quantification of active/marker compounds. By isolating and identifyingmarker compounds, new drugs can be formulatedto treat various diseases.

Keywords: HPTLC, Fingerprint profile, C.acuminatum (Jeevak), Malaxis acuminataAstavarga, Phytochemical analysis

IntroductionTraditional plant medicine is becoming an

area of ever-increasing importance in the healthcare systems. Since times immemorial, plantsform the basis of various traditional therapeuticsystems like, Ayurveda, Unanai, Sidha. Uses ofplant based remedies in healthcare preparationshave been reported in Vedas and the Bible. Plantsproduce a diverse group of bioactive molecules,making them a rich source of different types ofmedicines (1). These days medicinal andneutraceutical herbs are receiving immensescientific attention for their holistic effects (2).Thus, natural products with pharmacological or

5. Fingerprint Profile of an important therapeutic plant ofAstavarga Crepidium acuminatum (D. Don) Szlach by

HPTLCMamta Arora*1, Anupama Mahajan2, Jaspreet K. Sembi3

1 Research Scholar of IKG, PTU Jallandhar, Faculty of Biotechnology, ASBASJSM College,Bela Rupnagar (Punjab)

2. Assistant Professor in Biotechnology, SUSCET, Tangori, Mohali, (Punjab)3. Assistant Professor in Botany, Panjab University, Chandigarh

*[email protected]

Fingerprint Profile of an important therapeutic plant


biological activities are playing a very importantrole in medicine (3). World Health Organization(WHO) has confirmed that herbal medicines areserving the health needs of about 80 percent ofthe world’s population especially in rural areas ofdeveloping countries. Attention has also been paiddue to the side effects of most modern drugs. Ithas been estimated that in the mid-1990s over200 companies and research organizationsworldwide were screening plant and animalcompounds for medicinal properties (4, 5).Important drugs like vinblastine, vincristine,topotecan, taxol, teniposide, etopside, irinotecanetc. have come from plant sources. Curiosity isescalating in the overall fitness & wellness of mandue to nutraceutical plants.

Crepidium acuminatum (D.Don) Szlach(Syn. Malaxis acuminata) is having immenseethnomedicinal potential. The dried pseudobulbsknown as ‘jeevak’ are important ingredients ofvarious formulations and a polyherbal immune-booster nutraceutical ‘Chyavanprash’. This drughas been stated in various ayurvedic formulationslike Astavarga churna, chyavanprash Rasayan,Ghrita, Taila, Gutika, Agada etc. (6). C.acuminatum is used in breathing disorders,burning sensation, Cough, decrease in bonetissue, bleeding disorders, blood disorders,tuberculosis, insect bites, rheumatism. It isreported to be refrigerant therefore used to reducefever (Febrifuge). It has been described asaphrodisiac and used in emaciation, seminalweakness. It is utilized as tonic and in generaldebility (7, 8, 9) Although its antimicrobial (10),antioxidant (11) essential oil analysis (12), andanti-inflammatory activities (13) have been reportedbut work has not been described on fingerprintprofile of this plant by HPTLC.

High Performance Thin LayerChromatography (HPTLC) is a sophisticatedanalytical technique pedestal on the full potentialof thin layer chromatography. Automation,scanning, full optimization, selective detectionprinciple, minimum sample preparation,hyphenation enable it to be a powerful tool in

modern research for qualitative and quantitativeanalysis of complex mixtures of bio molecules(14). It has gained popularity to become a leadingtype of analysis in fingerprinting of herbal drugs.Although thin layer chromatography (TLC) iscommonly used for the analysis of herbal drugssince long time back. Various pharmacopoeiassuch as American Herbal Pharmacopoeia (AHP),Chinese Drug Monographs and Analysis,Ayurvedic pharmacopeia of India (API)Pharmacopoeia of People’s Republic of China,etc. still use TLC to provide first characteristicfingerprints of herbal drugs. Fingerprint analysisapproach using chromatography is becoming themost powerful tool for identification, authenticationand quality control of herbal products. For qualitycontrol, the concept of phyto-equivalence isutilized. Chromatographic fingerprint candemonstrate ‘sameness’ and ‘differences’between various formulations and theauthentication and identification of herbalmedicines can be accurately carried out even ifthe number and concentration of chemicallycharacteristic components are not very similar indifferent samples of herbal formulations (16). Asper World Health Organization (WHO), the quality,quantity, safety and efficacy data on traditionalmedicine are not sufficient and there is still a lackof adequate/accepted research methodology forevaluating traditional medicine till date. In thissituation, HPTLC is playing pivot role for thefingerprinting of medicinal plants. It is a realisticalternative to meet the need for effectual andpowerful assessment of herbal products. HPTLCis mainly used for expressing various patternswhich are preserved as ‘databases’ known as‘fingerprints’ for future studies. In such a situation,there is dire need that chromatographic fingerprint,for a herbal product should be constructed. Hence,this technique was adopted for deriving thefingerprint patterns of the crude drug of Crepidiumacuminatum.

Materials and MethodsPseudobulbs of the plant were screened

for profiling. Pseudobulbs were shade dried in airat room temperature, powdered and stored in air

Mamta Arora et al


tight container. It was authenticated and identifiedas Crepidium acuminatum (D. Don) Szlach fromNational Institute of Science Communication andInformation Resources (NISCAIR), New Delhi anddeposited in herbarium of Panjab University,Chandigarh with PAN 21262.

For analysis 500mg of powder wasdissolved in 10 ml of methanol with occasionalshaking for duration of 2 hrs., then filtration wasdone by using membrane filter. Sample of 1, 2,4, 6 μl aliquot were loaded as 8 mm band lengthon a 5 X10 silica gel 60 F254 TLC plate usingLINOMAT 5 auto sampler instrument (CAMAG,Muttenz, Switzerland). The samples-loaded plates

were kept in TLC twin trough developing chamberafter saturation, (saturation time 5 min) withsolvent vapour using respective mobile phases(as shown in table 1) and the plates weredeveloped in the respective mobile phases up to70 mm. The developed plates were dried by hotair to evaporate solvents from the plate. The plateswere kept in a photo-documentation chamber(CAMAG) and the images were captured.Software Visioncats-serv, version 2.4.17207.2 wasused for data analysis.

ResultsThe present study, which was performed to

develop fingerprint of the drug by using HPTLCtechnique showed valuable results. 2 saponins, 3


1.


bitter principles, 2 steroids, 2 sterols, 2 essentialoils, 1 anthraquinones, 2 coumarin and 5 flavonoidswere observed in methanol extract.

Methanol extract was subjected to differentcomposition of the mobile phase (Table 1) toseparate different secondary metabolites. Afterderivatization with appropriate reagents, the colourdevelopment was noted. Based on colourdevelopment and fluorescence (15), the secondarymetabolite were differentiated and Rf values werecalculated and presented in figure Best solventsystem for phytcohemical analysis are presentedin table 1.

HPTLC fingerprinting studies of methanolextract showed distinct band pattern before andafter spraying with derivatizing reagent. Rf valuesunder different wavelengths before and afterderivatization are taken and presented in figure 1-8 and Table1. HPTLC, now a days is applied notonly to obtain “Fingerprint” patterns of herbalformulations, quantification of active ingredientsbut also for the detection of adulterant.

DiscussionThe preliminary phytochemical screening of

crude drug indicated the presence of saponins,essential oils, anthraquinones, Sterols,coumarins, flavonoids, steroids, tannins andglycosides by HPTLC.

Saponins are a large family ofphytochemicals which are structurally relatedcompounds. ’Sapo’ is a Latin word for soap. Theypossess soap like properties and form lather inaqueous solution. They possess surfactantproperties and are used as natural detergent.Chemically they are glycosides of steroids (C27)and triterpenes (C30). Saponin = glycone +aglycone. Glycone is sugar moiety, which is polarin nature and consists of hexoses/pentoses/uronicacid, whereas aglycone part, is known assapogenin, steroidal/triterpene. They have receivedindustrial, commercial, pharmaceutical attention.They are used as food additives, as ingredients inphotographic emulsions, in fire extinguishers etc.(17) Biological effects of saponins are in the

membrane-permeabilising, immunostimulant,hypocholesterolaemic and anticarcinogenicproperties. They have also been found to affectsignificantly reproduction in animals. Thesestructurally diverse compounds have also beenobserved to kill protozoans and molluscs, to havean effect on cold blooded animals, also to havethe analgesic, antinociceptive, antioxidant activity,to impair the digestion of protein, to causehypoglycemia and to act as antifungal and antiviralagents (18). Investigations are going on towardsthe development of new natural medicines andprove the efficacy of traditional herbal medicine.The plant under investigation showed two typesof saponins at Rf 0.80 and 0.50 which can beisolated and characterized for further studies.

The term ‘essential oil’ derives its name fromthe drug Quinta essentia, named by Paracelsusvon Hohenheim of Switzerland in sixteen century.Essential oils are ‘essences’ which areresponsible for different scents that plants emit.They are odours, flammable, volatile productswhich have tendency to evaporate on exposure toair even at ambient conditions and therefore alsoreferred to as volatile oils or ethereal oils. Essentialoils are indispensable in food, cosmetic andhuman health field. They are extensively used inperfumery and aromatherapy (therapeutictechnique including massage, inhalations, or bathsusing these volatile oils). They serve as chemicalsignals allowing the plant to control or regulateits environment (ecological role), attraction ofpollinating insects, repellent to predators, inhibitionof seed germination, and communication betweenplants. They have antibacterial, antioxidant, anti-inflammatory, cancer chemoprotective activity,antifungal or insecticide and deterrent activitiesetc. (19, 20).

Anthraquinones, another class ofphytochemicals, are aromatic compounds havinganthracene ring with 2 keto groups (9, 10-dioxoanthracene). These are derivatives ofphenolic and glycosidic compounds but in livingplants they are generally found as glycosides.They are derived from anthracene and givingvariable groups based on the degree of oxidation

Mamta Arora et al


and position of double bonds in the polycyclicand side chain systems of sterols can be different.Generally, the sterols can be categorized intothree subclasses: (I) 4, 4 desmethylsterols (II)4a-methylsterols and (III) 4, 4-dimethylsterols.Phytosterols are important products for health andnutrition industries. They havehypocholesterolemic activities, and are used ascholesterol-lowering agents contributing towardscardiac health benefits. They are useful emulsifiersfor cosmetic manufacturers and used asprecursors for the production of hormones. Theyare known to inhibit oxidative deterioration of oils,therefore serving as potential antipolymerizationagents for frying oils and used as markers for theassessment of adulterated oils. Sterols are present

e.g. anthrones, anthranols, chrysophanol,poramide, luteolin, emodin etc.(21). They are mostcommomnly utilized as laxatives and possessantiviral and antifungal properties. Thesecompounds impart color to plants and have beenextensively employed as natural dyes (22). Oneanthraquinone has been observed in methanolextract of this plant in current study.

Sterols are a class of phyto-compounds,derived from hydroxylated polycyclicisopentenoids which is having a 1,2-cyclopentanophenthrene structure. Thesecompounds contain a total of 27-30 carbon atomsin which a side chain with carbon atoms isattached at the carbon 17 position. The number

Fig. 1: represents results for saponins captured afterderivatization 1 (a) under white light ; arrows at band Rf0.80 and Rf 0.50 observed under white light indicatespresence of saponins (b) same plate under UV 366 nm

Fig. 2: shows image representing results for bitterprinciples , visualized under 2 (a) white light and 2 (b)UV 366nm arrows at bands in 2 (a) corresponding to Rf0.37 , 0.60 and 0.90 of bluish violet color indicated presenceof bitter principles.

Fig. 3: Image obtained after derivatizing with vanillinand Sulphuric acid and observed under 3(a) whitelight and 3 (b) under UV 366 nm. Arrows pointing atbands corresponding to Rf 0.29 and 0.75 of violetcolor in 3 (a) indicated presence of differentessential oils (terpenoids).

Fig. 4. Image acquired after derivatizing with alcoholicKOH and taken under 4(a) white light and 49b) UV366 nm. Arrow at band Rf 0.65 in 4 (a) showingyellow florescence is pointing towardsanthraquinone


1 a 1 b

2 a 2 b 4 a 4 b

3 a 3 b


in the nonsaponifiable fraction of plant oils. Sincethese are of plant origin and not synthesized inhumans therefore are poorly absorbed and areexcreted faster than cholesterol. Therefore theyhave lipid lowering efficacy. Main phytosterolswhich are used in the human diet are sitosterol,stigmasterol, and campesterol. (23, 24). Two typesof sterols observed in the present study at Rf 0.10and 0.79 of reddish–violet color band confirmedthe presence of sterols.

‘Coumarins’ word is derived from‘Coumarou’, the vernacular name of the tonka bean(Dipteryx odorata), from which coumarin, was firsttime isolated in 1820. There are four classes ofcoumarin; simple coumarins; furanocoumarins;pyranocoumarins; and pyrone-substitutedcoumarins. The coumarins have bacteriostatic and

anti-tumor activity and these compounds are beingscreened as novel therapeutic agents (25). Twotypes of coumarins are first time reported inexisting study at Rf 0.05 & 0.71 showingfluorescence of pale yellow colour indicatedpresence of coumarins.

Flavonoids are polyphenolic compounds,ubiquitous in nature and are more than 4,000commonly occur in vegetables, fruits andbeverages like tea, coffee and fruit drinks. Theyoccur as aglycones, glucosides and methylatedderivatives. They are utilised as food andpharmaceutical supplements. The flavonoidsappear to have played a major role in successfulmedical treatments (26). Flavonoids are linked totheir potential cytotoxicity and their capacity tointeract with enzymes through protein

Fig. 5. Image taken after derivatizing the plateswith Anisaldehyde Sulphuric acid and observedunder 5 (a) white light and (b) UV 366 nm . Arrowsat Rf 0.10 and 0.79 in 5(a) of reddish–violet colorband confirmed the presence of sterols.

Fig. 6. Image taken after derivatization and observedunder (a) white light and (b) UV 366 nm . Arrowspointing towards bands at Rf 0.05 & 0.71 showingfluorescence of pale yellow colour indicatedpresence of coumarins.

Fig. 7. Image captured under UV 366 nm 7(a) beforederivatization and 7(b) after derivatization. Arrowspointing towards fluorescent bands at Rf 0.10,0.25,0.75,0.80 , 0.97 indicated presence offlavonoids.

Fig. 8. Image scanned under white light and UV 366nm after derivatization. Arrows at Rf 0.60 and 0.67bands depicting red-violet color proved the presenceof steroids.

Mamta Arora et al

8 a 8 b6 a 6 b

5 a 5 b 7 a 7 b


complexation (27). They have antioxidative activity,free-radical scavenging capacity, anticanceractivity and have role in coronary heart diseaseprevention and anti-human immunodeficiency virusfunctions They are reported to be hepatoprotective,anti-inflammatory, and antiviral also (28). Fivetypes of flavonoids are reported in presentinvestigation carried out.

Steroids have the fundamental structure offour carbon rings called the steroid nucleus. Theaddition of different chemical groups at differentpositions on backbone leads to the formation ofmany different types of steroidal compounds. Plantsteroids are synthesized by cyclisation of 2,3-epoxysqualene into cycloartenol, which are furthermetabolized to produce biologically activesteroids. Plant steroids classified in differentclasses based on their chemical structure,pharmacological activities and source from whichthey have been isolated. Plant steroids possessmany interesting medicinal, pharmaceutical andagrochemical activities like anti-tumor,immunosuppressive, hepatoprotective,antibacterial, plant growth hormone regulator, sexhormone, antihelminthic, cytotoxic and cardiotonicactivity (29).

Chemically, it is difficult to define tanninsas they include diverse number of oligomers andpolymers. Tannins are a heterogeneous group ofhigh molecular weight polyphenolic compoundswhich form reversible and irreversible complexeswith proteins, polysaccharides (cellulose,hemicellulose, pectin etc.), alkaloids, nucleicacids and minerals etc. Structurally, tannins canbe categorised into four groups: Gallotannins,ellagitannins, complex tannins, and condensedtannins. Gallotannins are all those tannins in whichgalloyl units or their derivatives are joined to polyol-, catechin-, or triterpenoid units. Ellagitannins arethose tannins in which at least two galloyl unitsare C–C coupled to each other, and do not containa glycosidically linked catechin unit. Complextannins are tannins in which a catechin unit isbound glycosidically to a gallotannin or anellagitannin unit. Condensed tannins are alloligomeric and polymeric proanthocyanidins.

Tannin-containing plant extracts are used asastringents, anti-inflammatory, as antiseptic, asdiuretics, antioxidant and haemostatic. They arealso used against diarrhoea and various types oftumours. Tannins are used in the dye stuff industryand also in the production of inks. In the foodindustry tannins are used to clarify wine, beer,and fruit juices. Other industrial uses of tanninsinclude textile dyes, as antioxidants in the fruitjuice, beer and wine industries, and as coagulantsin rubber production. Recently the tannins haveattracted scientific interest for treatment of AIDSand various cancers (25, 30)

Glycoside is a generic term used forphytochemicals that are bound to a sugar. Theyare the compounds that yield one or more sugarsupon hydrolysis, Hence the glycoside consistsof two parts: the sugar and the aglycone part.The aglycon may be a terpene, flavonoid,coumarine etc. Among the sugars found in naturalglycosides, D-glucose, L- rhamnose and L-fructose, L-arabinose are commonly found. Thesugar part can be disaccharide also. Theclassification of glycosides is a difficult matter.They are usually mixed acetals. The sugar moietyof a glycoside is joined to the aglycone, accordingto the chemical group of the aglycone involvedinto the acetal union, they are O-glycoside (OHgroup); S-glycoside (SH group), N-glycoside (NHgroup), C-glycoside (C group). The systematicnames are formed by replacing the ‘ose’ suffix ofthe parent sugar with “oside”. Classification canbe based on the sugar group for e.g., lucosidesand rhamnosides, classification can be based onaglycone group, Examples are lignan glycosides,alkaloidal glycosides etc. Glycosides which showsoap like properties are called saponin.Glycosides that release hydrocyanic acid onhydrolysis are known as cyanogenic glycosides.Based on functional group they can be Phenolicglycosides, Aldehyde glycosides, Anthraquinoneglycosides etc. (31)

The compounds which were reported earlierin C. acuminatum are alkaloid, carbohydrates,flavonoids, resin, saponin, steroids, tannins,whereas triterpenoids have been reported negative



qualitatively (32), whereas in same report proteinsare also documented as negative. Essential oilssuch as Limonene, Eugenol, citronellal,1-8-cineole,Piperitone and p-cymene were reportedby TLC (33),where as in another study beta –sitosterol, cetyl alcohol, glucose, rhamnose arereported. (34) Volatile oils are estimated as 0.54± 0.28(%v/w) in Tarikhet sample (32). In onerecent studies (35) dietary fatty acids, alpha –hydroxy acids, phenolic acids, sterols, aminoacids, sugars and glycoside are reported in thisplant by GC-MS. Another group of scientists (36)analysed quantitatively metal content and volatileconstituents of the plant by Atomic absorptionspectrophotometer and GC-MS. Alpha tocopheroland gamma tocopherol along with terpenoids arealso reported (36), whereas presence of Acidicpolysaccharides, Anthocyanins, Lignin, Phenolicsubstances, cutin, suberin, lignin and starch havebeen documented (37). Scientists (38) alsoconfirmed the presence of polyphenols, flavonoids,while synthesising gold nanoparticles of theextracts of pseudobulbs. Whereas comprehensivereport and fingerprint profilile has been presentedin this paper.These secondary metabolites areaccountable for the therapeutic activity of plants(39). Hence, great potential of Crepidiumacuminatum as a nutraceutical and herbal drugis confirmed in the present study.

Conclusion:C. acuminatum is an important plant of

Astavarga. It has a lot of therapeutic potential.The present study revealed the presence ofsaponins, bitter principles, steroids, Sterols,essential oils, anthraquinones, coumarin andflavonoids in methanol extract. The HPTLCfingerprint profile developed for methanol extractof C. acuminatum can be used for routine qualitycontrol of the drug and serve as a base forqualitative & quantitative analysis andstandardization of the drug. It will also help inidentification and quantification of active/markercompounds. By isolating and identifying markercompounds, new drugs can be formulated to treatvarious diseases.

Acknowledgement:Authors are thankful to all the members of

Anchrom, Mulund East, Mumbai (Maharashtra)for providing facilities. Authors duly acknowledgecooperation of Mr. Prashant Hande and Mr.Prateek Singal, Anchrom, Mumbai. Authors arealso thankful to IKG, PTU, jallandhar for support.

REFERENCES1. Durga RK, Karthikumar S, Jegatheesan K.

(2009) Isolation of potential Anti bacteria andantioxidant compounds from Acalypha indicaand Ocimum basilicum. Afr J PlantSci;4:163–16.

2. Cousins M.M., Adelberg J.W. (2009). Invitroplant and organ culture of medicinal andneutriceutical species in laboratory andindustrial scales. Acta PhysiologiaePlantarum; 31(5): 961-967

3. Cragg GM, Newmann DJ, Snader KM.(1997). Natural products in drug discoveryand development. J.Nat Prod. 60. 52-60.

4. Kong Jin-Ming, Goh Ngoh-Khang, Chia Lian-Sai1, Chiatet-Fatt. (2003). Recent advancesin traditional plant drugs and orchids ActaPharmacologica Sinica. 24(1): 7-21

5. Manako Yoshihiro, Wake Hitoshi, TanakaTakashi, Shimomura Koichiro, Ishimaru(2001) Phenanthropyran derivatives fromPhalaenopsis equestris. Phytochemistry.58:603-605

6. Anonymous. The Ayurvedic Pharmacopoeiaof India, Part 1, Vol. 5, 78-80 1st Ed., NewDelhi, Ministry of Health and Family Welfare,Department of AYUSH, Govt. of India, 2008.Available from http://www.ayurveda.hu/api/API-Vol-5.pdf

7. Nambiar VPK. Indian Medicinal Plants: ACompendium of 500 species 1993 ;3.Available from https://books.google.co.in/books?id=mBB_aRjQIo0C&source=gbs_book_other_versions

8. Teoh ES. Medicinal Orchids Of Asia. SpringerInternational Publishing 2016;(1):212-./3 doi/

Mamta Arora et al


10.1007/978-3-319-24274-3

9. Arora M, Singh S, Mahajan A, Sembi JK.Propagation and Phytochemical Analysis ofCrepidium acuminatum (D.Don) SzlachIOSR Journal of Pharmacy and BiologicalSciences2017. 12 (3);14-20

10. Arora M, kaur G, KahlonPS, Mahajan A,SembiJK. Pharmacognostic Evaluation &Antimicrobial Activity of EndangeredEthnomedicinal Plant Crepidiumacuminatum (D. Don) SzlachPharmacognosy Journal, Vol 9, Issue 6(Suppl), Nov-Dec, 2017

11. Arora M, Kaur G, Singh S, Mahajan A SembiJK. TPC, TFC & Antioxidant activities ofCrepidium acuminatum (D. Don) SzlachJournal of Pharmacognosy andPhytochemistry 2017; 6(4): 811-

12. Arora M, Mahajan A, 2,. Sembi JK Essentialoils analysis of pseudobulbs of Crepidiumacuminatum (D.DON) SZlach by GC-MSAsian Pac. J. Health Sci., 2017; 4(3):198-204

13. Bose B, Choudhury H, Tandon P, KumariaS. Studies on secondary metabolite profiling,anti-inflammatory potential, in vitrophotoprotective and skin-aging relatedenzyme inhibitory activities of Malaxisacuminata, a threatened orchid ofnutraceutical importance. J PhotochemPhotobiol B. 2017 Aug;173:686-695. doi:10.1016/j.jphotobiol.2017.07.010. Epub 2017Jul 12.

14. Srivastava MM (ed.), High-Performance Thin-Layer Chromatography (HPTLC), DOI10.1007/978-3-642-14025-9_7, # Springer-Verlag Berlin Heidelberg 2011

15. Wagner H, Baldt S, Zgainski EM. Plant druganalysis-A thin layer chromatography AtlasNew York, Berlin, Springer, 1996.

16. Lalit Giri, Harish C. Andola, Vijay KantPurohit, M.S.M. Rawat, R.S. Rawal andI.D. Bhatt, 2010. Chromatographic and

Spectral Fingerprinting Standardization ofTraditional Medicines: An Overview asModern Tools. Research Journal ofPhytochemistry, 4: 234-241.

17. Makkar H.P.S., Siddhuraju P., Becker K.(2007) Saponins. In: Plant SecondaryMetabol i tes. Methods in MolecularBiology™, vol 393. Humana Press https://doi.org/10.1007/978-1-59745-425-4_16

18. Desai SD, Desai DG, Kaur H. Saponins andtheir biological activities Saponins and theirBiological Activities. 2017;(January). PharmaTimes - Vol 41 - No. 3 - March 2009

19. Essential Oils’ Chemical Characterizationand Investigation of Some BiologicalActivities: A Critical Review Medicines 2016,3, 25; doi:10.3390/medicines3040025www.mdpi.com/journal/medicines

20. Doughari JH. Phytochemicals: Extractionmethods, basic structures and mode ofaction as potential chemotherapeuticagents. Phytochem – A Glob Perspect theirrole Nutr Heal [Internet]. 2012;1–32.Available from: http://cdn.intechopen.com/pdfs/32936/InTech-Phytochemicals_extraction_methods_basic_s tructures_and_ mode_ of_action_ as_ potential_chemotherapeutic_agents.pdf

21. D. Simpson, S. Amos, Other PlantMetabolites in Pharmacognosy, 2017Pharmacognosy Fundamentals,Applications and Strategies Editor SimoneBadal McCreath and Rupika Delgoda ISBN:978-0-12-802104-0 Pages 267–280

22. Raju MP, Babu DGA, Rakesh B, RajashekarCH. The Role of Phytosterols EnrichedFoods-A Review. 2013;7(6):40–7.

23. Abidi, SL Chromatographic analysis ofplant sterols In foods and vegetable oilsJournal of Chromatography A, 935 (2001)173-201

24. Jain PK, Joshi H. Coumarin/ : Chemical andPharmacological Profile. Journal of Applied



Pharmaceutical Science 02 (06); 2012: 236-240.

25. Saxena Mamta, Jyoti Saxena, RajeevNema, Dharmendra Singh and AbhishekGupta Phytochemistry of medicinal plants.Med Plants Cent Asia Uzb Kyrg.2013;1(6):13–4.

26. Falcone Ferreyra ML, Rius SP and CasatiP (2012) Flavonoids: biosynthesis, biologicalfunctions, and biotechnological applications.Front. Plant Sci. 3:222. doi: 10.3389/fpls.2012.00222

27. Yao LH1, Jiang YM, Shi J, Tomás-BarberánFA, Datta N, Singanusong R, Chen SSFlavonoids in food and their health benefitsPlant Foods Hum Nutr. 2004Summer;59(3):113-22.

28. Kumar S, Pandey AK. Chemistry andBiological Activities of Flavonoids/ : AnOverview. 2013;2013. Hindawi PublishingCorporation The ScientificWorld JournalVolume 2013, Article ID 162750, 16 pageshttp://dx.doi.org/10.1155/2013/162750

29. Patel SS, Savjani JK. Systematic review ofplant steroids as potential anti- inflammatoryagents/ : Current status and futureperspectives. The Journal ofPhytopharmacology 2015; 4(2): 121-125

30. Khanbabace Karamali, Ree VT. Tannins/ :Classification and Definition Tannins/ : Classifi cation and De fi nition. 2001 18, 641-649Natural products report DOI 10.1039/B101061L

31. M. Bartnik, P.C. Facey2Glycosides,Pharmacognosy Fundamentals,Applications and Strategies 2017, Pages101–161Academic press

32. Sharma, A,Rao,CV,Tiwari RK, Tyagi LK, KoriML, Shankar Comparative Study onPhysicochemical Variation of Microstyliswallichii/ : A Drug Used in Ayurveda. Acad JPlant Sci. 2009;2(1):4–8.

33. Gupta, R., M. Agarwal and R.K. Baslas,1978. Chromatographic separation andidentification of various constituents ofessential oil from the bulb of M. accuminata.Indian Perfume, 22(4): 287-288.

34. Bhatnagar, J.K., S.S. Handa and S.C.Duggal, 1970. Chemical investigation onMicrostylis wallichii. Planta Medica, 20:157-161

35. Bose B, Choudhury H, Tandon P, KumariaS. Studies on secondary metabolite profiling,anti-inflammatory potential, in vitrophotoprotective and skin-aging relatedenzyme inhibitory activities of Malaxisacuminata, a threatened orchid ofnutraceutical importance. J PhotochemPhotobiol B. 2017 Aug;173:686-695. doi:10.1016/j.jphotobiol.2017.07.010. Epub 2017Jul 12

36. Lohani N, Tewari LM, Joshi GC, et al.Chemical composition of Microstyliswallichii Lindl. from Western Himalaya. JMed Plants Res. 2013;7(31):2289-92.doi:10.5897/JMPR11.1754.

37. ADAMS, Sebastian John et al. Distribution,morphology, anatomy and histochemistry ofCrepidium acuminatum. ModernPhytomorphology, [S.l.], v. 12, p. 15-32, mar.2018. ISSN 2227-9555. Available at: <https://ojs.phytomorphology.org/index.php/MP/article/view/513>. Date accessed: 11 may2018. doi: https://doi.org/10.5281/zenodo.1195691.

38. Bag Braja Gopal, Dash Shib Shankar andPatra Sumit Kumar. Study of AntioxidantProperty of the Pseudobulb Extract ofCrepidium acuminatum (Jeevak) and its usein the Green Synthesis of Goldnanoparticles. Int. J. Res. Chem. Environ.Vol. 4 Issue 3 (133-138) July 2014

39. Rabe T and J Vanstoden 2000 Isolation ofan antimicrobial sesquiterpenoid fromWarbugie salutaris; J. Ethnopharcol. 93 171-174.

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