Phytochemical Screening and GC-MS Analysis of Leaf Extract of Pergularia daemia (Forssk) Chiov

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Dss A and Kumari Pushpa Rani TP Department f Micrbilgy, Kamaraj Cllege, Tuticrin, Tamil adu, India ABSTRACT This study was implement t actuate the chemical cmpnents f Pergularia daemia leaves using

1 Available nline at Asian Jurnal f Plant Science and Research, 2017, 7(1):9-15 ISS : CDE (USA): AJPSKY Phytchemical Screening and GC-MS Analysis f Leaf Extract f Pergularia daemia (Frssk) Chiv Rukshana MS, Dss A and Kumari Pushpa Rani TP Department f Micrbilgy, Kamaraj Cllege, Tuticrin, Tamil adu, India ABSTRACT This study was implement t actuate the chemical cmpnents f Pergularia daemia leaves using Perkin- Elmer Gas Chrmatgraphy Mass Spectrmetry, ur results f GCMS cmpunds in the extract was relevant t the atinal Institute f Standards and Technlgy (IST) library. GC/MS analysis f ethanlic extract f Pergularia daemia leaves cnfess the presence exadecanic acid, methyl ester (33.42), Pentadecanic acid, 14-methyl-methyl ester (36.23, Ethyl 9,12,15-ctadecatrienate (33.12 and 4-(4-Chlrbenzyl)-1-cyclhexyl-5-tsylamin-1-1,2,3-triazle (31.24). Qualitative phytchemical screening f the ethanlic extracts f the leaves revealed the presence f many cmpunds such as flavnids, tannins, alkalids, terpenids, sterids and phenls. This study result will make a way fr the prductin f herbal medicines fr varius ailments by using Pergularia daemia leaves.. Keywrds: Biactive cmpunds, Phyt drugs, erbal medicine ITRDUCTI In lden days itself the imprtance f medicinal plants have been discvered. At that time there was n synthetic medicines, they have been using nly the herbal medicines t treat all diseases. Frm this we can understand that plants are rich in medicinal prperties and they are very useful in human health and wellbeing. Bilgical studies are essential t find mre medicinal prperties f the plants [1]. But still the many medicinal plants and their medicinal prperties are unexplred. Treating f diseases thrugh natural medicine is the mst ancient treatment knwn t mankind [2,3]. In India the medical systems using medicinal plants are Ayurveda, ddha, mepathy, etc., t treat varius ailments [4]. Traditinal plant based medicines fr primary healthcare need is fllwed in underdevelped cuntries f abut 80% f wrld s ppulatin (W) [5]. Recently many f the research were being carried ut in medicinal plants. The main reasn was that the synthetic drugs which was nw taking up by the human have many side effects that ften lead t serius cmplicatins. The develpment f herbal medicine was dne by the primary screening f the cmpunds in the plant extracts. Cmparing t mdern medicine the herbal medicine was the lifesaving drug. Amng 4,00,000 plant species nly 6% f the plants are studied fr their bilgical activity and nly few have been phytchemically investigated. This shws that the investigatin is needed fr many medicinal plants fr its activity and pharmaclgical prperties. Pergularia daemia (Figure 1) is a plant that was named as Veliparuthi in Tamil and Utranajutuka in indi. Pergularia daemia dried leaf was used as an effective agent in treating brnchitis, asthma, rheumatic fever, amenrrhea, dysmenrrheal and wunds [6]. It als has many anti-micrbial, anti-inflammatry, anti-rheumatic, anti-arthritic, anti-fungal prperties [7,8]. They als serve as an anti-prliferative and a best anti-xidant [9,10]. All these medicinal prperties f Pergularia daemia is due t the presence f its phytchemical cnstituents which is nt yet explred thrughly [11]. In this study the Gas Chrmatgram Mass Spectrmetric methd (GCMS) was carried ut in the ethanlic extract f dried leaves fr phytchemical analysis fllwed by qualitative and quantitative determinatin f the cmpunds. The nn-nutritive plant chemicals are called as phytchemicals which have the prperties t prtect r prevent diseases. 9

Plant prduces these chemicals t prtect themselves but the research have shwn that they have the capacity t treat human diseases in an effective way [12].

2 Plant prduces these chemicals t prtect themselves but the research have shwn that they have the capacity t treat human diseases in an effective way [12]. There are thusands f phytchemicals, each have their pharmaclgical prperties f their wn [13]. The plant pssesses varius medicinal prperties; the aim f this study was t identify the phytcmpunds in the ethanlic leaf extract f Pergularia daemia by qualitative screening f phytchemicals and t identify each specific cmpund with their cncentratins by Gas Chrmatgraphy Mass Spectrum (GC- MS) analysis [14]. MATERIALS AD METDS Cllectin f the plant The Pergularia daemia (Figure 1) plant leaves were cllected frm the villages f Tirunelvelli district, Tamilnadu n August 17 at AM. The cllected plant was verified by Dr. V. andagapalan, Assciate Prfessr, PG and Research Department f Btany, atinal Cllege, Trichy, Tamil adu. Preparatin f extract The leaf was shadw dried and it was made as a pwder using electrical blender. This pwder was treated with 95% ethanl and kept fr 48 h. Then it was filtered by using Whatman filter.1. This extract was used fr the phytchemical analysis and GC-MS analysis t find the biactive cmpnents. Phytchemical analysis Phytchemical screening f crude extracts f Pergularia daemia was carried ut accrding t the methds described by Trease and Evans [15,16]. Phytchemical analysis f crude pwder f the samples fr the evaluatin f phytchemicals such as a tannins, alkalid, sterid, phenls, glycsides [17,18] and terpenid, flavnid, etc. [18,19]. GCMS analysis GCMS analysis is a cmmn cnfirmatin test. It is best used t make an effective chemical analysis. This analysis will prvide a representative spectral utput f all the cmpunds that get separated frm the sample. The first step f GCMS was started by injecting the sample t the injected prt f the Gas chrmatgraphy (GC) device. The GC instrument vaprizes the sample and then separates and analyzes f the varius cmpnents. Each cmpnent was ideally prduces a specific spectral peak that may be recrded n a paper chart electrnically. The time elapsed between elutin and injectin is called the "retentin time". Differentiate between sme cmpunds was identified using the Retentin time. The peak is measured frm the base t the tip f the peak. GCMS analysis was dne by The Suth Indian Textile Research Assciatin, Cimbatre. Identificatin f biactive cnstituents by GCMS GCMS analysis f the ethanl extract f leaf f the plant was perfrmed using a TERM Gas Chrmatgraphy- TRACE ULTRA VER: 5.0 [20]. The ven temperature is maintained at 220ºC at a rate f 6ºC/min; the carrier gas with a flw rate f 1 ml/min. The split sampling technique was used t inject the sample in the rati f 1:10. Retentin indices (RI) f the cmpunds were determined by cmparing the retentin times f a series and identificatin f each cmpnent was cnfirmed by cmparisn f its retentin index with data in the literature. Interpretatin f Figure 1: Pergularia daemia 10

3 Mass-Spectrum was carried ut by using the database f atinal institute Standard and Technlgy (IST) having mre than 62,000 patterns. The spectrum f the unknwn cmpnents was cmpared with the spectrum f knwn cmpnents which was stred in the IST library. The mlecular weight, name, chemical structure and mlecular frmula f the cmpnents f the test materials were ascertained. The peak in GCMS f ethanl extract f leaf f Pergularia daemia shwed the presence f the secndary phytchemical cmpunds like phenlic and fatty acids and its esters (Figure 2). RESULTS The phytchemical cmpnents have been analyzed qualitatively in ur lab by the cmmn methds using chemicals. Flavnids, glycsides, tannins, terpenids, sterids, alkalids and carbhydrates were fund in the leaf extract f this plant (Table 1). GC-MS is the best techniques t identify the cnstituents f vlatile matter, lng chain, branched chain hydrcarbns, alchls acids, esters, etc. Peak area, retentin time and mlecular frmula were used fr the cnfirmatin f phytchemical cmpunds. The active principles with their Retentin time (RT), Mlecular frmula, Mlecular weight (MW) and peak area in percentage are presented. GC/MS analysis f ethanlic extract f Pergularia daemia leaves revealed the existence f exadecanic acid, methyl ester (33.42), Pentadecanic acid, 14-methyl-methyl ester (36.23, Ethyl 9,12,15-ctadecatrienate (33.12 and4-(4-chlrbenzyl)-1-cyclhexyl-5- tsylamin-1-1,2,3-triazle (31.24) (Table 2). Frm the GC-MS analysis f Pergularia daemia leaves the presence f seventeen cmpunds (phytchemical cnstituents) were revealed the medicinal quality f the plant (Table 3). DISCUSSI Metablite prfiling in plant species was dne by gas chrmatgraphy mass-spectrmetry methd fr last few years. Figure 2: GCMS spectral chrmatgram Table 1: Phytchemical cmpnents in the ethanl extract f P. daemia S.. Phytchemical Cmpnents Ethanlic extract f P. daemia 1 Flavnid + 2 Tannins + 3 Alkalids + 4 Glycsides + 5 Terpenids + 6 Sterids + 7 Phenls + 11

4 Table 2: GC MS analysis f Pergularia daemia S.. Retentin Time Peak area % ame f the cmpund Mlecular frmula Mlecular weight Chemical structure Tetradecamethylcyclheptasilxane C C 3 C 3 C C 3 3C C3 C3 3 C C 3 C 3 C 3 C 3 3 C C ,9-Di-tert-butyl-1- xaspir(4,5)deca-6,9- diene-2,8 C dine Dcsenamide, (Z)- C Dcsenamide, (Z)- C C ,2-Benzenedicarbxylic acid, diethyl ester (CAS) C C exadecanic acid, ethyl ester (CAS) C Ethyl 2-dibrmmethyl-6-cyan-7- ethxy-5-phenyl-1,8-n C C aphthyridine-3-carbxylate SYAPTGEI B C Phytl C

5 Tetrakis (Dimethylsilylcarbdiimide) C Tris(trimethylsilyl), methyl ester derivative f 3,3,4,4-[24]-prstaglandin F2à C3056D (4-Chlrbenzyl)-1- cyclhexyl-5-tsylamin- 1-1,2,3-triazle C2223Cl43S 458 Cl S C Ethyl 9,12,15-ctadecatrienate C C Pentadecanic acid, 14-methyl-, methyl ester (CAS) C C C exadecanic acid, methyl ester (CAS) C C C 3 Table 3: Bilgical prperties f the phytcmpunds S.. Cmpunds Bilgical activity 1 Tetradecamethylcyclheptasilxa E Antixidant 2 7,9-Di-tert-butyl-1-xaspir(4,5)deca-6,9 diene-2,8-dine activity reprted 3 13-Dcsenamide, (Z)- Antimicrbial 4 13-Dcsenamide, (Z)- Antimicrbial 5 1,2-Benzenedicarbxylic acid, diethyl ester (CAS) Antiasthmatic 6 exadecanic acid, ethyl ester (CAS) Antimicrbial 7 Ethyl 2-dibrmmethyl-6-cyan-7-ethxy-5 phenyl-1,8-n aphthyridine-3-carbxylate Antidiabetic 8 SYAPTGEI B Antimicrbial 9 Phytl Antimicrbial, Antiasthmatic 10 Tetrakis( Dimethylsilylcarbdiimide) Antimicrbial 11 Tris(trimethylsilyl),methylester derivative f 3,3,4,4-[24]- prstaglandin F2à Antibacterial, Anti-infective 12 4-(4-Chlrbenzyl)-1-cyclhexyl-5-tsylamin-1-1,2,3-triazle activity reprted 13 Ethyl 9,12,15-ctadecatrienate Antimicrbial 14 Pentadecanic acid, 14-methyl-, methyl ester (CAS) Antibacterial, Antiallergic 15 exadecanic acid, methyl ester (CAS) Antibacterial, Antiallergic 13

6 But nly a limited number f plant research labratries have gas chrmatgraphy mass-spectrmetry. The identified cmpunds ccupy many bilgical prperties. GC-MS analysis f phytcnstituents in plants gives a clear picture f the pharmaceutical value f that plant. Thus, this type f GC-MS analysis is the first step twards understanding the nature f medicinal prperties in this medicinal plant and this type f study will be helpful fr further detailed study. Further investigatin is needed t identify the pharmaclgical imprtance and phytchemistry f Pergularia daemia. Phytl is ne amng fifteen cmpunds f the present study. Phytl is a key acyclic diterpene alchl that is a precursr fr vitamins E and K1. milarly presence f phytl was bserved in the leaves f Lantana camara [17] and in Mimsa pudica leaves [21]. milar result was als bserved in the leaves f Lantana camara [22]. Phytl was bserved t have antibacterial activities against Staphylcccus aureus by causing damage t cell membranes [23]. Phytl, Phenl, 2, 4-bis (1-phenylethyl) - which are all have medicinal prperties. Ptential antixidant and anticancer activities were fund in these cmpunds. Alchl extract f the leaves f Kigelia pinnata has earlier been reprted fr the presence f exadenic acid cmpund [24] and in Melissa fficinalis [22,25]. 17 cmpunds with n-exadecanic acid and ctadecanic acid was identified as the majr cmpunds in the leaves f Cleistanthus cllinus [26]. GC-MS analysis f ethyl acetate extract f Gnithalamus umbrsus revealed the presence f n-exadecanic acid [27], exadecanic acid, 9, 12 - ctadecadienic acid, n-hexadecanic acid, 9, 12, 15-ctadecatrienic acid, Squalene and Phytl were identified in the ethanl leaf extract f Ale vera [28]. These reprts are in accrdance with the result f this study. The surce f many plants (herbs and spices) can be ften identified frm the peak pattern f the chrmatgrams btained directly frm headspace analysis. milarly, particular qualitative and quantitative patterns frm a GC analysis will shw all the cmpunds in the leaf extract. Pergularia daemia have many bilgical prperties which can be used in varius purpsed t treat many diseases. The cmpunds identified by the initial qualitative analysis and GCMS analysis have many uses in medical field. Each cmpunds identified have their unique character t treat varius diseases. Further studies needed t reveal its imprtance in specific field t treat the diseases prperly. CCLUSI GCMS analysis f the ethanlic extract f leaves f Pergularia daemia reveals the presence f medicinally valued biactive cmpnents like sapnins, tannins, alkalids, terpenid, sterids and flavnids. As the medicinal value f similar cmpnents in ther plant extracts are already prved, n wnder if these cmpnents in Pergularia daemia may als have equally effective. The wrk is in prgress t ascertain its bilgical activity and brighten the pharmaclgical prfile f it in the arena f traditinal medicine. REFERECES [1] Rastgi RP, Mehrtra B. Cmpendium f Indian medicinal plants. (Vl 2), ew Delhi, India: Central Drug Research, Lucknw and ISCAIR, 1979, 521. [2] Cragg GM, Snadder KM. atural prducts as surces f new drugs ver the ewman perid, J at Prd, 2003, 66: [3] Gill LS, Akinwumi C. igerian flk medicine: Practices and beliefs f the nd peple. J Ethnpharmacl, 1986, 18: [4] Pushpangadan P, Atal CK. Ethn-medic-btanical investigatins in Kerala I. Sme primitive tribal f Western Ghats and their herbal medicine. J Ethnpharmacl, 1984, 11: [5] W, IUC, WWF. Guidelines n the cnservatin f medicinal plants. Switzerland: IUC Gland, [6] Bhaskar V, Balakrishnan. Veliparuthi (Pergularia daemia (Frsk.) Chiv.) As a phytmedicine: A review. Int J PharmTech Res, 2009, 1: [7] Kakrani K, Saluja AK. Traditinal treatment thrugh herbs in Kutch district, Gujarat state, India. Part II. Analgesic, antiinflammatry, antirheumatic, antiarthritic plants. Fitterapia, 1994, 65: [8] Kaushik JC, Sanjay A, Tripathi, Arya S. Antifungal prperties f sme plant extracts against the damping ff fungi f frest nurseries. Indian Jurnal f Frestry, 2002, 25:

7 [9] Lamprnti I, Saab AM, Gambari R. Antiprliferative activity f essential ils derived frm plants belnging t the magnliphyta divisin. Int J ncl, 2006, 29: [10] Lee SJ, Uman K, Shibamt T, Lee KG. Identificatin f vlatile cmpnents in basil (cimum basilicum L.) and thyme leaves (Thymus vulgaris L) and their antixidant prperties. Fd Chem, 2005, 91: [11] Elang V, Ambujavalli L, Amala Basker E, Sulchana. Pharmaclgical and micrbilgical studies n Pergularia extensa. Fitterapia, 1985, 56: [12] Dutta A, Ghsh S. Chemical examinatin f Daemia extensa. J Am Pharm Assc, 1947a, 36: [13] Anjaneyulu AS, Raju DVS, Srinivasa Ra S. Chemical evaluatin f Pergularia daemia. Indian J Chem, 1998, 37: [14] Dhar ML, Dhar MM, Dhawan B, Mehrta B, Ray C. Screening f Indian plants fr bilgical activity. Indian J Exp Bil, 1968, 11: [15] Evans WC, Trease E. Pharmacgnsy W.B. Saunders Cmpany Ltd., Lndn, (14th Edn), 2000, [16] Maria Jancy Rani P, Kannan PSM, Kumaravel S. GC-MS analysis f Lantana camara L. leaves. JPRD, 2011, 2: [17] Mittal P, Tammz C, Reichstein T. Glycsides and aglycns. CCXXVII. The glycsides f Pergularia extensa. elv Chim Acta, 1962, 45: 907. [18] Velmurugan P, Kamaraj M, Prema D. Phytchemical cnstituents f Cadaba trifliata Rxb. rt extract. Int J Phytmedicine, 2010, 2: [19] Victr, Chidi. Phytchemical cnstituents f sme selected medicinal plants, Afr J Pure Appl Chem, 2009, 3: [20] aznagy-radnal E, Czigle S, Mathe I. TLC and GC analysis f the essential ils f Stachys species. J Planar Chrmatgr, 2007; 20: [21] Sridharan S, Meenaa V, Kavitha V, ayagam AAJ. GC-MS study and phytchemical prfiling f Mimsa pudica Linn. J Pharm Res, 2011, 4: [22] Kumar SM, Manimegalai S. Evaluatin f larvicidal effect f Lantana camara Linn against msquit species Aedes aegypti and Culex quinquefasciatus. Adv Bil Res, 2008, 2: [23] Inue Y, ada TA, Shiraishi K, irre, amashima, et al. Biphasic effects f Geranylgeranil, Terpenne and Phytl n the grwth f Staphylcccus aureus. Antimicrb Agents Chemther, 2005, 49: [24] Grace M, Light ME, Lindsey KL, Mhlland DA, Staden JV, et al. Antibacterial activity and islatin f antibacterial cmpunds frm fruit f the traditinal African medicinal plant, Kigelia africana. S Afr J Bt, 2002, 68: [25] Sharafzadeh S, Mrteza Khsh-Khui, Javidnia K. Arma prfile f leaf and stem f lemn balm (Melissa fficinalis L.) grwn under greenhuse cnditins. Adv Envirn Bil, 2011, 5: [26] Parasuraman S, Raveendran R, Madhavra C. GC-MS analysis f leaf extracts f Cleistanthus cllinus Rxb. (Euphrbiaceae). Int J Ph Sci, 2009, 1: [27] Ibraham SA, Bustamam AA, Mhammed ME, Syam MI, Mhamed Yusif M, et al. GC-MS determinatin f biactive cmpnents and antibacterial prperties f Gnithalamus umbrsus extracts. Afr J Bitechnl, 2009, 8: [28] Arunkumar S, Muthuselvam M. Analysis f phytchemical cnstituents and antimicrbial activities f Ale vera L. against clinical pathgens. Wrld J Agric Sci, 2009, 5:

ANTIMICROBIAL ACTIVITY OF AQUEOUS & METHANOLIC EXTRACT OF YOUNG & MATURE LEAVES OF PSIDIUM GUAJAVA L. (GUAVA)

J. Indian bt. Sc. Vl. 92 (1&2) 2013 : 4751 ANTIMICROBIAL ACTIVITY OF AQUEOUS & METHANOLIC EXTRACT OF YOUNG & MATURE LEAVES OF PSIDIUM GUAJAVA L. (GUAVA) MEENAKSHI VAIDYA Department f Btany, Mithibai Cllege,