phytochemical investigation of methanolic rhizome ...

WORLD JOURNAL OF PHARMACY AND PHARMACEUTICAL SCIENCES Jadhao et al.

World Journal of Pharmacy and Pharmaceutical Sciences

SJIF Impact Factor 6.647

Volume 6, Issue 9, XXX-XXX

Research Article

ISSN 2278 – 4357

PHYTOCHEMICAL INVESTIGATION OF METHANOLIC RHIZOME EXTRACT OF CURCUMA ANGUSTIFOLIA ROXB. BY HRLC-MS METHOD Anand S. Jadhao* and Anil S. Bhuktar Vivekanand Arts, Sardar Dalipsingh Commerce and Science College, Samarth Nagar, Aurangabad, Maharashtra - 431001. Article Received on 24 July 2017, Revised on 14 August 2017, Accepted on 03 Sept. 2017, DOI: 10.20959/wjpps20179-10168

ABSTRACT The aim of the present study was to characterize Curcuma angustifolia Roxb. for the presence of biologically active phytochemical constituents using methanol extracts of the rhizome by High Resolution Liquid Chromatography - Mass Spectroscopy (HRLC-MS),

*Corresponding Author

while mass spectra of the compounds found in the extract was matched

Anand S. Jadhao

with the Metlin database library, results confirmed the presence of

Vivekanand Arts, Sardar Dalipsingh Commerce and

therapeutically potent compounds in C. angustifolia rhizome extract.

Science College, Samarth

Metabolites analysis by ESI-Q-TOF-MS revealed the presence of 10

Nagar, Aurangabad,

major abundant compounds viz. 1alpha-hydroxy-23- [3-(1-hydroxy-1-

Maharashtra – 431001.

methylethyl) phenyl]-22,22,23,23-tetradehydro-24,25,26,27-tetranorv,

[email protected]

5,6-Dihydrouridine, 1-methyl-N-(9-methyl-9-Azabicyclo [3.3.1] non-

[email protected],

3-yl)-7-(sulfooxy)-,endo glucuronide, 2-Methoxyestrone 3glucuronide, Anandamide (18:3, n-6), 27-nor-5b-cholestane-3a, 7a, 12a, 24, 25-pentol, Madecassic acid, Vecuronium, Chaulmosulfone, Descladinosyl-6-Omethylerythromycin. This report is the first of its kind to analyze the chemical constituents of C. angustifolia Roxb., using HRLC-MS. KEYWORDS: Curcuma angustifolia Roxb., HRLC- MS, phytochemical. INTRODUCTION C. angustifolia Roxb, belongs to Zingiberaceae, it is commonly known as ‘narrow-leaved turmeric (English), yaipan (Manipuri), tikhur (Hindi) and Koova (Malayalam). Different ethnic communities of Koraput district, Orissa uses leaf and rhizomes of C. angustifolia in

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Jadhao et al.


the treatment of constipation. [1] Mache people of Jhapa district, Eastern Nepal use rhizome of C. angustifolia along with Allium sativum bulb crushed and mixed with oil of Brassica campestris has found potent on wound hilling and act as an antiseptic.[2] Rhizome juice was recorded potent on Jaundice[3] Methanolic rhizome extract of C. angustifolia shows potential antioxidant, antimicrobial and anti-cancerous activity towards human cervical cancer HeLa cells.[4] Herbs, root fiber numerous, slender; root tubers small, ellipsoid, pale yellow, white inside, 13 × 0.5-1-3 cm. Rhizomes ovoid or conical, 1.2-5.5 × 0.5-1.7 cm, white inside, having pungent smell. Leafy shoot 50-120 cm tall; leaves 3-4, elliptic, oblong, acuminate, narrowed at both ends, 30-65 × 9-15 cm; petiole with sheath, 10-45 cm long. Flower in terminal or lateral spikes 9-15 × 3-5 cm long; 3-3.5 cm, yellow, 1-2 in each bracts; peduncle 15-30 cm long; floral bracts ovate- oblong, 3-4 cm long, obtuse or rounded at tips with dark purple spot; coma bracts oblong, purple, base white, minutely pubescent, 4-5.5 cm long; calyx white, 3 lobed, hairy outside, 1 cm long, lobes oblong, obtuse; corolla tubes funnel shape, white, 2.5- 3 cm, lip yellow obovate, acuminate as long as corolla; lateral staminodes obovate - cuneate; stamens 5-7 mm, anther thecae parallel 4-5 mm long, spurred. Ovary trilocular, hairy, ovules 2 in each locules; capsule ovoid, 7-10 mm long, seed globose, reddish brown. MATERIAL AND METHODS The plant material of Curcuma angustifolia Roxb was collected from Sitakhandi, district Nanded of Maharashtra and identified by using floristic literature

[5, 6, 7]

. The voucher

herbarium specimen (ASJ 7615) deposited in VH Herbarium, department of Botany, Vivekanand Arts, Sardar Dalipsingh Commerce and Science College, Samarth Nagar, Aurangabad. Preparation of crude extracts The dried rhizome powder was extracted through Soxhlet using methanol as solvent and heated at 650C for 18-24 hours, extract was kept for evaporation and the sample was stored in amber coloured bottle and further, the phytochemical analysis was carried out using HRLCMS technique.

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HRLC- MS analysis Instruments and chromatographic conditions Equipment and conditions for identification of metabolites from an active sub-fraction of methanol extract was carried out at SAIF, IIT, Bombay. Samples were analyzed on a LCESI-Q-TOF-MS (Agilent Technologies 6550 i-Funnel) system equipped with a G4220B pump, G4226A auto sampler and G1316C, and a diode array detector (DAD). The elution solvent consisted of a gradient system of 0.1% formic acid in water (A) and acetonitrile (B) at a flow rate of 0.3 ml/min. The gradient system started with 95% A: 5% B reaching 5% A: 95% B in 50 min., then back to initial composition 95% A: 5% B in 10 min which was held at same composition for 5 min. The MS analysis was carried out by ESI positive ionization mode. MS source conditions were as follows: capillary voltage 3500 V, Gas temperature 250 C, drying gas flow 13 L/min, sheath Gas temp 300, sheath Gas Flow 11, nebulizing gas pressure 35 (psig), fragmentor 175 V, Skimmer 65 V, Octopole RF Peak 750 V, and mass range m/z 50–1000. The resolution was 40,000 FWHM. Metlin database was used to structure confirmation. RESULTS AND DISCUSSION The results pertaining to HRLC-MS analysis led to the identification of number of compounds from the HR fractions of the methanol extract of Curcuma angustifolia Roxb. These compounds were identified through mass spectrometry attached with HRLC; the results of the present study were tabulated in (Table 1). Metabolites analysis by ESI-Q-TOFMS revealed the presence of major abundant metabolites identified in methanol rhizome extract fraction of C. angustifolia Roxb., were 1alpha-hydroxy-23- [3-(1-hydroxy-1methylethyl) phenyl]-22,22,23,23-tetradehydro-24,25,26,27-tetranorv, 5,6-Dihydrouridine, 1methyl-N-(9-methyl-9-Azabicyclo [3.3.1] non-3-yl)-7-(sulfooxy)-,endo glucuronide, 2Methoxyestrone

3-glucuronide,

Anandamide

(18:3,

n-6),

27-nor-5b-cholestane-

3a,7a,12a,24,25-pentol, Madecassic acid, Vecuronium, Chaulmosulfone, Descladinosyl-6Omethylerythromycin (spectra 1-10). The retention time, m/z value, mass, molecular formula and the DB difference (ppm) of the major 10 abundant metabolites are shown in (Table 1), the spectra showed counts versus mass to charge (m/z) ratio (fig. 1).

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Table 1: Phytocomponents identified in methanol rhizome extracts of C. angustifolia Roxb. Sr. Name of compounds no. 1alpha-hydroxy-23-[3-(1- hydroxy-11 methylethyl)phenyl]- 22,22,23,23tetradehydro- 24,25,26,27-tetranorv 2 5,6-Dihydrouridine 3 Glu Ala 1-methyl-N-(9-methyl-94 azabicyclo[3.3.1]non-3-yl)-7(sulfooxy)-, endo- glucuronide 6 2,3-Didehydrogibberellin A9 7 CANDESARTAN CILEXTIL 8 13,14-dihydro-PGF2alpha 9 2-Methoxyestrone 3- glucuronide 10 Anandamide (18:3, n-6) 11 Leu Phe Val 12 3alpha-androstanediol glucuronide 13 DAIDZEIN 14 GPEtn(12:0/0:0) 15 Cys Gly Ala 16 SECURININE 27-nor-5b-cholestane- 3a,7a,12a,24,2517 pentol 18 MADECASSIC ACID 20 Megalomicin C1 21 OH-Spheroidenone 22 Vecuronium 1,2 di-(9Z,12Z,15Z-octadecatrienoyl)-323 O-Beta-D- galactosyl-sn-glycerol 24 CHAULMOSULFONE 25 Descladinosyl-6-O-methylerythromycin

RT

Mass

Formula

5.901

474.3174

C32 H42 O3

6.295 6.299

246.0807 218.0852

C9 H14 N2 O6 C8 H14 N2 O5

7.261

518.2333

C25 H34 N4 O8

7.964 8.089 8.916 10.317 10.373 10.653 12.388 12.742 17.629 17.636 17.636

314.1529 610.2614 356.256 476.2042 321.2691 377.2354 468.2779 254.0608 397.2246 249.0797 217.1129

C19 H22 O4 C33 H34 N6 O6 C20 H36 O5 C25 H32 O9 C20 H35 N O2 C20 H31 N3 O4 C25 H40 O8 C15 H10 O4 C17 H36 N O7 P C8 H15 N3 O4 S C13 H15 N O2

18.075

438.3335

C26 H46 O5

18.198 21.956 22.184 22.229

504.3414 960.5774 600.4555 557.4379

C30 H48 O6 C48 H84 N2 O17 C41 H60 O3 C34 H57 N2 O4

22.683

774.5272

C45 H74 O10

24.331 25.876

776.543 605.3839

C48 H76 N2 O4 S C30 H55 N O11

Fig. 1: A chromatogram of methanolic extract of rhizome (C. angustifolia Roxb.).

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Curcuma Angustifolia Roxb In the present study chemical profile of methanol rhizome extract of C. angustifolia Roxb., using HRLC-MS spectra, the chromatogram showed the relative concentrations of various compounds getting eluted as a function of retention time. The heights of the peak indicate the relative concentrations of the components present in the plant. The mass spectrometer analyzes the compounds eluted at different times to identify the nature and structure of the compounds. The large compound fragments into small compounds giving rise to appearance of peaks at different m/z ratios. These mass spectra are fingerprint of that compound which can be identified from the data library. In addition to this, the results of HRMS profile can be used as pharmacognostical tool for the identification of the plant. CONCLUSION The presence of various bioactive compounds i.e. 1alpha-hydroxy-23- [3-(1-hydroxy-1methylethyl) phenyl]-22,22,23,23-tetradehydro-24,25,26,27-tetranorv, 5,6-Dihydrouridine, 1methyl-N-(9-methyl-9-Azabicyclo [3.3.1] non-3-yl)-7-(sulfooxy)-,endo glucuronide, 2Methoxyestrone

3-glucuronide,

Anandamide

(18:3,

n-6),

27-nor-5b-cholestane-

3a,7a,12a,24,25-pentol, Madecassic acid, Vecuronium, Chaulmosulfone, Descladinosyl-6Omethylerythromycin was inferred by using HRLC-MS technique and showed with different chemical structures. However, isolation of individual phytochemical constituents may proceed to find a novel drug. ACKNOWLEDGEMENT Authors are thankful to principal of Vivekanand Arts, Sardar Dalipsingh Commerce and Science College, Samarth Nagar, Aurangabad for providing laboratory facilities for the present research work.

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REFERENCES 1. Misra MK, and Dash S S. Ethnomedicinal Plants of Koraput district Orisa and their conservation. Perspective of Plant Biodiversity, Bishen Singh Mahendra Pal Singh Pub., Dehra Dun, 2002; 621-634. 2. Rai SK,. Medicinal plants used by Meche people of Jhapa district, Eastern Nepal. Our Nature, 2004; (2): 27-32. 3. Abhyankar RK, and Upadhyay R,. Ethno-medicinal studies of tubers of Hoshangabad M. P. Bulletin of Environment, Pharmacology & Life Sciences, 2011; 1(1): 57- 59. 4. Nayak S, Jena A K, and Sucharita S, In vitro bioactivity studies of wild Curcuma angustifolia rhizome extract against (Hela) Human Cervical Carcinoma cells. World Journal of Pharmacy and Pharmaceutical Sciences, 2013; 2(6): 4972 - 498. 5. Naik VN, Flora of Marathawada., Vol. II, Amrut Prakashan, 1998; 830 - 832. 6. Sharma BD, Karthikeyan S, Singh NP, and Lakshminarsimhan P., Flora of Maharashtra state Monocotyledons. B.S.I., 1996; 72-77. 7. Sabu M, Zingiberaceae and Costaceae of South India, Indian Association for Angiosperm Taxonomy, Calicut University, India, 2006.

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