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High Resolution GC/MS Analysis of the Holoptelea Integrifoli's Leaves And Biocatalysis and Agricultural Biotechnology 22 (2019) 101405 Contents lists available at ScienceDirect Biocatalysis and Agricultural Biotechnology High resolution GC/ MS analysis of the Holoptelea integrifoli's leaves and their medicinal qualities Vijay Kumar a , * , 1, Simranjeet Singh b, c, d, 1, Sapna Avinash Kondalkar e, 1, Bhavana Srivastava a, Brijesh Singh Sisodia f, Bandana Barthi a, Ravindara Singh g, Om Prakash h a Department of Chemistry, Regional Ayurveda Research Institute for Drug Development, Aamkho, Gwalior, M. P, 474009, India b Department of Biotechnology, Lovely Professional University, Jalandhar, Punjab, 144111, India c Regional Advanced Water Testing Laboratory, Mohali, Punjab, 160059, India d Punjab Biotechnology Incubators, Mohali, Punjab, 160059, India e Department of Pharmacognosy, Regional Ayurveda Research Institute for Drug Development, Aamkho, Gwalior, M. P, 474009, India f Department of Biochemistry, Regional Ayurveda Research Institute for Drug Development, Aamkho, Gwalior, M. P, 474009, India g Department of Chemistry, Central Council for Research in Ayurvedic Sciences (CCRAS), New Delhi, 110058, India h Department of Ayurveda, Regional Ayurveda Research Institute for Drug Development, Aamkho, Gwalior, M. P, 474009, India ARTICLE INFO ABSTRACT Keywords : Plants and their sec ondary mol e cules are con sid ered as unique gift of God to humans. Secondary mol e cules of Holoptelea integrifolia plants are con sid ered as major ther apeu tic agents to cure di verse classes of dis ease. The aim of the present Antioxidant activity study was to iden tify the sec ondary mol e cules of the leaf ex tract of Holopte lea in tegri fo lia (HI). Iden ti fica tion of Anti - diabetic activity bio mol e cules us ing the fourier trans forma tion in frared (FTIR) and gas chro matog ra phy cou pled with high res - Anti - inflammatory activity Antimicrobial activity olu tion mass spec trome ter (GC - HRMS) have con firmed the presence of seven prominent mol e cules in clud ing four new mol e cules. The four new mol e cules iden ti fied though GC - HRMS were (Z) - octadeca - 9,17 - dienal (13.17%), (9Z,12Z) - octadeca - 9,12 - dienoyl chlo ride (3.22%), 1,3 - dihydroxypropan - 2 - yl palmi tate (6.01%), and 17 - acetyl - 14 - hydroxy - 10,13 - dimethyl - decahydro - 2H- cyclopenta [α]phenanthrene - 3,12,15 (4H,14H,16H) - tri - one. The aque ous ex tract of HI leaves ex hibited the an tiox i dant po ten tial (>59%), anti - diabetic (78.45 ± 5.55%), anti - inflammatory (value of bind ing con stants (k) was 1.83 ± 0.06 × 10 −4 μM −1) and an - timicro bial (MIC val ues 20– 30 mg/ ml) qual ities. More over, the observed val ues of var i ous physic ochem i cal pa rame ters in clud ing pH, loss on dry ing, to tal ash, acid in sol u ble ash, wa ter ex trac tive val ues, and al cohol ex - trac tive stud ies were 7.07 ± 0.37, 8.55 ± 0.75%, 18.12 ± 1.21%, 10.07 ± 0.87%, 37.97 ± 2.31%, and 29.36 ± 1.27% respec tively. Also, pharma cog nos tic analy ses were performed. The re sults of present study could con tribute to the iso la tion of new com pounds those may have med i c i nal qual i ties. 1 . Introduction genus Holoptela and family ( Ulmaceae ) of al mond plants. It is a tree with height 15– 25 m having yellow ish and light gray col ored bark Holoptelea in tegri fo lia (HI) is a well doc u mented plant in Ayurvedic ( Benjamin and Christopher, 2009 ; Kumar et al., 2012a & 2012b; Sys tem of Med icine ( Padmaa and Durga, 2011 ; Reddy et al., 2008 ; Mahmud et al., 2010 ). Of fensive odour and irri ta tive taste are the ma- Sahoo et al., 2010 ; Srinivas et al., 2008 ; Vinod et al., 2010 ). It is con - jor traits of HI. Asia and Africa are the major cul tiva tors and dis trib- sumed as folk medi cine to treat arthritis, in flamma tion, anaemia, skin uter of HI. In In dia, it is abun dantly found throughout the whole Hi- dis eases, liver dis or ders etc ( Ahmad et al., 2012 ; Ganie and Yadav, malayan belt ( Kumar et al., 2012a & 2012b; Mahmud et al., 2010 ; 2014 , 2015 ; Joshi et al., 2013 ; Kavitha and Narasu, 2014 ). In India, it Padmaja et al., 2009 ; Prajapati and Patel, 2010 ; Sharma et al., 2010a, is well known plant for its pollen al lergenic proper ties ( Acharya, b). 2008 ; Sharma et al., 2005 ; Singh and Kumar, 2003 ). It is from the * Corresponding author. E- mail address: vijaychem99@ gmail. com, vijay. kr1984@ gov. in (V. Kumar). 1 Equal con tribu tion. https://doi.org/10.1016/j.bcab.2019.101405 Received 16 November 2018; Received in revised form 1 March 2019; Accepted 26 October 2019 Available online 31 October 2019 1878-8181/© 2019 Elsevier Ltd. All rights reserved. V. Kumar et al. Biocatalysis and Agricultural Biotechnology 22 (2019) 101405 HI is renowned for its medi c inal proper ties like an timicro bial (SA), napthyl diamine dichlo ride (NED), bovine serum al bumin (BSA), ( Ahmad et al., 2012 ; Ganie and Yadav, 2014 & 2015; Joshi et al., sodium car bonate, alu minium chlo ride salt, phos phomolyb de num 2013 ; Kavitha and Narasu, 2014 ; Padmaa and Durga, 2011 ; Reddy et reagent, ferrozine, ferrous ammo nium sul phate salt, α- amylase, and al., 2008 ; Sahoo et al., 2010 ; Srinivas et al., 2008 ; Vinod et al., 2010 ), dini tro - salicylic acid were of AR grade and used without any purifi ca - antivi ral, an tiox i dant ( Ganie and Yadav, 2014 & 2015; Kavitha and tion. Narasu, 2015 ; Reddy et al., 2008 ; Saraswathy et al., 2008 ; Srinivas et al., 2008 ; Srivastava et al., 2013 ), anti -obesity ( Bambhole and 2.2 . Pharmacognostical analysis Jiddewar, 1985 ; Ganie and Yadav, 2014 ; Kumar et al, 2012b ), wound - healing ( Reddy et al., 2008 ; Srinivas et al., 2008 ), hy polipi daemic Organolep tic (color, odour, taste etc.) and micro scopic (cu ticle, ( Subash and Augustine, 2013 ), anal gesic ( Rizwani et al., 2012 ), anti - epider mis, tri chomes, stom ata etc. i.e. his to log i cal study) pa rame ters diabetic ( Sharma et al., 2010a, b & 2012 ), anti - inflammatory ( Kalpana were performed as per reported methods ( Kumar et al., 2012a & and Upadhyay, 2010 ; Sharma et al., 2009 ), anti -tumour ( Lakshmi et 2012b). al., 2010 ; Soujanya et al., 2011 ), anti- tubercular ( Joshi et al., 2013 ), lar vi ci dal ( Singha et al., 2012 ), anthelmintic ( Durga and Paarakh, 2.3 . Physicochemical analysis 2010 ; Kaur et al., 2010 ), anti- neoplastic ( Guo et al., 2013 ), and adap - to genic ac tiv ities ( Puri et al., 2011 ). Physico chemi cal proper ties like pH, loss on dry ing, to tal ash, acid It is well es tab lished fact that, the medi c i nal ac tivi ties of medi c i nal in sol u ble ash, water extrac tive val ues, and al co hol ex tractive val ues plants de pend on the matrix of sec ondary metabolites present in were tested ac cording to reported methods ( Kumar et al., 2018c ). plants ( Padmaa and Durga, 2011 ; Reddy et al., 2008 ; Sahoo et al., Stud ies were performed in tripli cates. 2010 ; Srinivas et al., 2008 ; Vinod et al., 2010 ). Qual ita tive phyto - chemi cal analy sis revealed that, HI con tained sev eral classes of chemi - 2.4 . Extraction cal con stituents like phenol, ter penoids, gly cosides, flavanoids, sterols and saponins ( Benjamin and Christopher, 2009 ; Padmaja et al., 2009 ; Exper i ment was performed in two phase, in first phase, 4 gm of Prajapati and Patel, 2010 ). The iso lated and doc u mented com pounds powder was put into 100 ml dis tilled water, shaken mechan ically and of HI are 1, 4 - naphthalenedione, hexacosanol, stig masterol, β- amyrin, the su pernatant was filtered af ter 24 h by us ing the Whatman filter betulin, betulinic acid, epifriedlin, octa cosanol, friedlin, holoptelin - A paper No. 1. A green color so lu tion was obtained and su pernatant of and holoptelin -B ( Ganie and Yadav, 2014 ; Kumar et al., 2012a & first ex peri ment was stored at 4– 10 °C for biochem i cal analy sis. 2012b ; Sutar et al, 2014a, b; Vinod et al., 2010 ). In sec ond phase of ex peri ment, initially above steps were repeated To iden tify the sec ondary metabo lites of HI, var ious researchers and vol ume of pure aqueous ex tract of HI was reduced to semi solid have performed HPTLC analy sis ( Reddy et al., 2008 ), where they have mass by evapo rating at 60 – 65 °C. Further, semi- solid mass was recon - quan tified the phytos terols ( Sutar et al, 2014a, b), flavonoids ( Sutar et sti tuted with ethanol for GC -HRMS quan tifica tion and an timicro bial al, 2014a, b), al kaloids ( Sutar et al., 2016 ), and 1,4 - naphthalene dione as say. To perform the FTIR study, a disc was prepared by mixing ( Vinod et al., 2010 ). Sub ash and Augus tine have applied liquid chro- semisolid mass with KBr. matographic mass spectro scopic (LC -MS) technique for the identi fi ca - tion and iso la tion of chemi cal compound, 3 - (7 -ethoxy -4 - methyl- 2 - oxo - 2.5 . Phytochemical analysis 2H- chromen- 3 -yl) propanoate having signif i cant hy polip idaemic effect ( Subash and Augustine, 2013 ). Methanol, ethyl ac etate, ethanol and Qual ita tive analy sis of phyto chemi cals in clud ing car bohy drates, hexane ex tracts of HI have been stud ied by appling the gas chromato - gly cosides, proteins, amino acids, al kaloids, terpenoids, flavanoids, graphic mass spectrom e try (GC - MS) tech nique, where n- hexadecanoic saponins, tan nins, pheno lics and steroids, present in the aque ous ex - acid, phytols and some other fatty acids were observed and reported tract of HI were performed as per the reported methods ( Kumar et al., ( Kavitha and Narasu, 2014 ).
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