4. Hossam M. A., Ismail Z. (2013), Isolation and characterization of triterpenes from the leaves of Orthosiphon siamineiis, Arabian Journal of Chemistry, 6, 295-298. 5. Srinivasan G. V., Ranjith C , Vijayan K. K. (2008), Identification of chemical compounds from the leaves of Leea indica. Acta Pharm, 58, 207-214. 6. Nizami A. N., Rahman M. A., Ahmed N. U. et ai (2012), Whole Leea macropl^lla cthanolic extract normalizes kidney deposits and recovers renal impairments in an ethylene glycol-induced urolithiasis model of rats, Asian Pacific Journal of Tropical Medicine, 5, 533-538. 7, Gohari A. R.. Saeidnia S., Hadjiakhoondi A. el al. (2009), Isolation and quantificative analysis of oleanolic acid from Satureja mutica Fisch. & C. A.
Mey., Journal of Medicinal Plants, 8, 65-69. 8. Pal M., Singh V., Agrawal J. et at (2013), Chemical constituents of Viburnum betulifolium. Chemistry of Natural Compounds, 49, 390-391. 9. Ragasa C. Y., Lim K. (2005), Secondary metabolites from Schefflera odorata Blanco, Philippine Journal of Science, 134, 63-67.
Journal of Medicinal Materials, 2014, VoL 19, No. 5 (pp.3I0-3I3)
PRELIMINARY STUDY ON THE CHEMICAL COMPOSITION 0¥ ARDISIA INSULARIS BELONGING TO THE FAMILY MYRSINACEAE IN VIETNAM
Trinh Anh Vien^, Nguyen Thi Hong Van''*, Pham Quoc Long", Luu Tuan Anh' 'institute of Natural Products Chemistry, Vietnam Academy of Science and Technology;
^Industrial University of Ho Chi Minh city at Quang Tam, Thanh Hoa, Vietnam 'Corresponding author: [email protected]
(Received September 4*, 2014) Summary
Preliminary Study on the Chemical Composition otArdisia insularis Belonging to the Family of Myrsinaceae in Vietnam
Three compounds, gallic acid (1), methyl gallate (2) together one megastigmane glucoside, (35, 5^, 6R, IE, 95)- megastigman-7-ene-3,S,6,9-tetrot 3-0-^-D-giucopyranoside (3) have been isolated from the aqueous fraction of the leaves of Ardisia insularis Mez. belonging to the family of Myrsinaceae. The structure elucidation of these compounds was performed based on spectroscopic data (MS, ID-NMR, 2D-NMR). This is the first time (35, 5R, 6R, IE, 95)-megasligman-7-ene- 3,5,6,9-tetrol 3-0-/!-D-glucopyranoside was isolated from the genus Ardisia.
Keywords: Ardisia insularis Mez. Kfyrsinaceae, (3S.5R.6R, 7E9S)-megastigman-7-ene-3.5.6.9-tetrol 3-0-^I>-glucif)yranoside.
1. Introduction identified by Dr. Nguyen Quoc Binh, Vietnam The genus Ardisia is the largest in the family National Museum of Nature, Vietnam Acdemy of Myrsinaceae, and approximately 100 species are Science and Technology. Voucher specimen widely distiibuted in Vietiiam, especially in the (BMN-B0001532) was deposited at the Institute midland plains [1]. The plants belonging to of Natural Products Chemistry, Vietiiam Academy Ardisia have been used in the folk medicine to of Science and Technology,
treat various diseases such as arthritis, rheumatism, General experimental procedures: Thin sore throat, diarrhea, gonorrhea, malaria, enteritis, layer chromatography was carried out on pre- gastric ulcer and liver diseases [2, 3]. Phytochemical coated silica gel DC-Alufolien 60 F254 (Merck) studies of Ardisia species led to the isolation of or RPl 8 F254s (Merck) plates. Column chromatography saponins, coumarins, quinones [4]. However, the was performed on silica gel (Kieselgel 60, 70 - chemical composition of Ardisia insularis Mez. 230 mesh and 2 3 0 - 4 0 0 mesh, Merck) and YMC has not been studied yet. In this paper, we RP-18 resins.'H-NMR (400 MHz) and'^C-NMR describe the isolation and stiiictural elucidation (100 MHz) specti^ were recorded on an Agilent of three compounds from the leaves of A. 400-MR NMR spectrometer and TMS was used insularis Mez. growing in Vietnam. as an intemal standard. Data processing was carried 2. Material and methods out with the MesflleNova 6.0.2 program. HRESIMS Plant material: The leaves of A. insularis Mez. spectra were olrtained using an Agilent 6550 iFunnel were collected at Quang Khe, Dak Glong, Dak Q-TOF LC/MS system. GC was recorded on a Nong, Tay Nguyen in March, 2012, and Shimadzu GC-20I0 plus. Electrospray ionization 310 Journal of Medicinal Materials, 2014, Vol 19, No 5
mass spectra (ESI-MS) were performed on an AGILENT 1100 LC-MSD Trap spectiometer.
Isolation. Dried leaves of ^. insularis Mez.
(2.0 kg) were extracted with MeOH three times at room temperature to yield 150 g of a dark solid extract, which was then suspended in water and successively partitioned with «-hexane and ethyl acetate (EtOAc) to obtain n-hexane (All, 40.0 g), EtOAc (AI2, 45.0 g), and water (AI3, 65.0 g) extracts after removal solvent in vacuo. The water soluble fraction (AD, 65.0 g) was chromatographed on a Diaion HP-20 column eluting with water containing increasing concentiations of MeOH (0, 25, 50, 75, and 100 %) to obtain five fractions ADA (30.0 g), AI3B (9.0 g), ADC (13.0 g), ADD (7.0 g), and AOE (6.0 g). Fraction ADB (9.0 g) was chromatographed on a silica gel column eluting with CHCI3 - MeOH - water (5 : I : 0.1, v/v/v) to give four fractions, ADBl (3.0 g), ADB2 (2.4 g), ADB3 (2.1 g), and ADB4 (1.0 g). Fraction AI3B2 was chromatographed on an YMC RP-18 column eluting with MeOH - water (1 ; 1, v/v) to yield 1 (8.0 mg) and 2 (9.0 mg).
Fraction ADD was separated on silica gel CC eluting with CHCl3:MeOH:H20 (4:1:0.1, v/v/v) solvent to give 5 fractions, AI3D1-ADD5.
Compound 3 was purified from fraction AI3D3 by column chromatography silica gel CC YMC RP-18 eluting the with MeOH:H20 (1:3, v/v).
Compound 1 (gallic acid): White needles crystals; Mp.: 237 - 238 "C; ESI-MS: m/z 171.1 [M+H]*, CHeOs. 'H-NMR (CD3OD, 400 MHz) 5 (ppm): 7.00 (2H, s, H-2, H-6). '^C-NMR (CD3OD, 100 MHz) 5 (ppm): 126.39 (C-l>, 110.07 (C-2, C-6), 146.01 (C-3, C-5), 138.0 (C-4), 173.6 (C-7).
Compound 2 (methyl gallate): White powder.
ESI-MS: m/z 185 [M+H]*, CgHgOs ' H - N M R (CD3OD, 400 MHZ) 5(ppm): 7.10 (2H, s, H-2, H- 6), 3.32 (3H, s, H-8). '^C-NMR (CD3OD, 100 MHz) 5 (ppm): 121.54 (C-l), 110.32 (C-2, C-6), 145.92 (C-3, C-5), 139.35 (C-4), 170.59 (C-7), 49.83 (C-8).
Compound 3 ((353*,6fl,7£^5>megastigman- 7-ene-3,5,6,9-tetrol 3-0-^-D-glucopyranoside):
White powder. EI-MS: m/z 406 [M]*, C19H34O9.
' H - N M R (CD3OD, 400 MHz) 5 (ppm): 1.57 (IH, m, H-2a), 1.72 (IH, t, J = 12.0 Hz, H-2e), 4.18 (lH,m,H-3), l.74(lH,m,H-4a), 1.92(lH,t,y=
11.0 Hz, H-4b), 6.03 (d, J= 16.0 Hz, H-7), 5.76 (IH, dd, J = 5.6/16.0 Hz, H-8), 4.32 (IH, m, H- 9), 1.25 (3H, dd, J= 6.0/16.0 Hz, H-10), 0.86 (3H,s,H-ll), 1.20(3H, s, H-12), 1.09(3H,s,H- 13), 4.38 (IH, d, J= 8.0 Hz, H-l'), 3.12 (IH, m, H-2'), 3.31 (IH, m, H-3"), 3.28 (IH, m, H-4'), 3.25 (IH, m, H-5'), 3.65 (IH, dd, J= 5.0/12.0 Hz, Ha-6'), 3.82 (IH, d, J= 12.0 Hz, Hb-e"). '^C- NMR (CD3OD, 100 MHz) 5 (ppm): 40.76 (C-l), 44.48 (C-2), 73.18 (C-3), 42.28 (C-4), 77.05 (C-5), 79.16 (C-6), 130.91 (C-7), 136.11 (C-8), 69.56 (C-9), 24.10 (C-IO), 27.53 (C-11), 26.24 (C-12), 27.15 (C-13), 102.16 (C-l'), 75.66 (C-2'), 77.70 (C-3'), 71.57 (C-4'), 77.82 (C-5'), 62.67 (C-6').
3. Result and Discussion
Compound 1 was obtained as white needle crystals. The ' H - N M R spectium of 1 displayed only a singlet signal of arometic proton at 5H 7.00 (2H, s), suggesting for the presence of a symmetric four-subistitued benzene ring. The '^C-NMR spectrum showed six aromatic carbons including two methine groups and four quatemary carbons together one cacboxyl carbon atom at Sc 173.60 (C-7). The ESI-MS of 1 gave a pseudomolecular ion peak at m/z 171 [M+H]*, corresponding to the molecular formula of CjUeOs. According to the above spectral analyses, and comparison of these data with those in literature [5], compound 1 was idenfied as gallic acid. This compound was also found in A. balansana [6], A. elliptica [7], A.
compressa [8], A. colorata [9] and demonstiated many biological activities such as antibiotic, cytotoxic, and antioxidant activities, ect.
Compound 2 was obtained as a white powder.
The ' H - N M R and '^C-NMR spectral data of 2 were similar to those of gallic acid (1), except for the presence of a methoxy group that was established by signals at 5H 3.32 (3H, s, OMe) and 5c 49.83, indicating that 2 could be the methyl ester derivative of 1. The molecular formula of 2 was determined to be CgHgOs by ESI-MS ([M+H]* 185) that differed by a CH;
moiety from 1. Comparision of the spectral data of 2 with those in literature [5], this compound was identified as methyl gallate. Methyl gallate was isolated from a related plant, A. balanscma [6] and possesses potential antioxidant, cytotoxic, and anti-virus HIV activities [10,11].
Journal of Medicinal Materials, 2014, Vol 19, No 5 311
1:R=H 2. R=CH,
Fig. 1. Chemical structures of compounds 1,2 and 3 Compound 3 was isolated as a white powder.
The ' H - and '^C-NMR spectia of 3 showed signals attributable to a ^-D-glucopyranosyl unit, one secondary methyl, three tertiary methyls, two methylenes, two methines with hydroxyl groups, one trans double bond, two quatemary carbons with hydroxyl groups and one other quatemary carbon. These functionalities were the same as those of megastigmane glucoside [12]. In detail, the ' H - N M R spectmm of 3 showed signals of^- D-glucopyranoside moiety including an anomeric proton at 8H 4.38 (IH, d, y = 8.0 Hz, H-l'), two protons of methylene group at SH 3.65 (dd, J = 5.0, 12.0 Hz, H,-6'), 3.82 (d,J= 12.0 Hz, Hb-6'), four other protons of four oximethine groups at SH 3.12 - 3.31 ppm. The aglycole moiety was displayed one signal of the secondary methyl at 5H 1.25 (3H, d, y = 6.4 Hz, H-10); three signals of three tertiary methyls at SH 0.86 (3H, s, H-l I), 1.20 (3H, s, H-12), 1.08 (3H, >, H-13); two signals of two methylene groups at SH 1.57 (IH, m, H,-2), 1.73 (IH, m, Hb-2), 1.74 (IH, m, H,-4), 1.94 (IH, m, Hb-4), two signals of two methines with hydroxyl groups at SH 4.18 (IH, m, H-3) and 4.32 (IH, m, H-9) together two protons of one trans double bond at SH 6.03 (d, J = 16.0 Hz, H- 7), 5.76 (d, J = 5.6, 16.0 Hz, H-8). The '^C-NMR and DEPT spectia (Table 1) revealed 19 carbon signals for a megastigmane aglycone and a sugar moiety. Furtiiemore, the HMBC correlation between anomeric proton H-l' (5H 4.38) and C-3 (5c 73.4) suggested tiiat the^-D-glucopyranoside moiety were at C-3 position of aglycole.
Additional, tiie EI-MS of 3 gave a ion peak at m/z 406 [M]', corresponding to the molecular formula
of C|9H3<09. Consequently, the above spectral data of compound 3 were in good agreement with those reported in the literature [12], so compound 3 was determined to be (3S, 5R, 6R, IE, 95>
megastigman - 7 - ene - 3,5,6,9 - tetrol 3 - 0-fi-D- glucopyranoside. This is the first time 3 was isolated from a plant of the genus Ardisia.
Table I: "C-NMR data for compound 3 in DMSO Position Sc|12| Sr-^ 5H*^(mult.,yinHz) Aglycone
1 2 3 4 5 6 7 8 9 10 II 12 13
40.9 44.6 73.4 42.5 77.8 79.2 131.0 136.2 69.2 24.0 27.6 26.3 27.2
40.76 44.48 73.18 42.28 78.05 79.16 130.93 136.11 69.56 24.10 27.53 26.24 27.15
1.57 1.73 4.18 1.74 1.92
- -
6.03 <d, 16.0) 5.76(dd, 5.6.16.0) 4.32 1.25 (d, 6.4) 0.86 (s) 1.20 (s) 1.08 (s) 0-/J-D-glucopyranosyl
2' r
3' 4' 5' 6'
102.3 75.2 78.2 71.8 77.9 62.8
102.16 75.66 77.70 71.57 77.82 62.67
4.38 ( 4 8.0) 3.12 3.31 3.28 3.25 3.65 (dd, 5.0, 12.0) 3.82 (d, 12.0)
"recorded in DMSO, ''100 MHz. '=400MHz 4. Conclusion
From the aqueous fraction of the leaves A.
insularis, three compounds including gallic acid (1), methyl gallate (2) and (35, 5R, 6R, 7E, 95)- megastigman - 7 - ene - 3,5,6,9 - tetrol 3-O-fi-D-
312 Journal of Medicinal Materials, 2014, Vol 19, No 5
g l u c o p y r a n o s i d e ( 3 ) w e r e isolated a n d identified. Viemam National Foundation for Science and T h i s is t h e first time ( 3 5 , 5R, 6R, IE, 9 5 ) - Technology Development (NAFOSTED) under grant m e g a s t i g m a n - 7 - e n e - 3,5,6,9 - teti-ol 3 - O - ^ - D - number 104.01-2011.20. The authors are grateful to glucopyranoside w a s found in the g e n u s Ardisia. Dr. Nguyen Quoc Binh for collection and identification
Acknowledgments: This research was funded by of the plant.
References
1. Ph?m HoJng HQ (1999), CSy co Vi?t Nam, Nh4 xuil ban uc. Quy&i 1. pp. 674-710. 2. D 5 Tat Uri (2001), NhOng cay thu6c vi vj Uiu6c Vt§t Nam, nxb Y hpc, pp. 129,167,265.3. V5 Van Chi (1997), Tir diln cay thu6c Vi?t Nam, nxb Y hpc, p.
244, 315, 623, 1271. 4. Kobayashi H., de Mejia E. (2005), The genus Ardisia: A novel source of health-promoting compounds and phytophamiaceuticals. Journal of Ethnopharmacology. (96), 347-354. 5. Yoshida T., Scno K., Takama Y..
Okuda T. (1982), Bergenin derivatives from Mallotus japonicas, Phytochemistry. 21(S),1180-I182. 6. Luu Tuin Anh, Nguyen Thi H6ng Van, VO Dinh Hoang, Trinh Anh Vien, Pham Qu6c Long (2013), Preliminary study on the chemical constituents of Ardisia balansana belonging to the fomily Myrsinaceae in Vietnam, Tap chi Hoa hgc (51) (6ABC), 99-102. 7.
Methin Phadungkit, Omboon Luanratana (2006), Anti-salmonella activity of constituents of Ardisia elliptica Thunb., Natural Product Research, 20 (7), 693-696.8. Sumino M., Sekine T., Ruangningsi N., Igarashi K., Ikegami F. (2002), Ardisiphenols and other antioxidant principles from the fruits of Ardisia colorata. Chemical and Pharmaceutical Bulletin , 50 (11), 1484- 1487. 9. Hsu F. L., Huang W. J., Wu T. H., Lee M. H., Chen L. C , Lu H. J., Hou W. C , Lm M. H. (2012). Evaluation of antioxidant and free radical scavenging capacities of polyphenolics from pods of Caesalpinia pulcherrima, international Journal of Molecular Sciences, 13(5), 6073-6088. ID. Khurana S., Hollingsworth A., Pichc M., Venkataraman K., Kumar A , Ross GM.. Tai TC. (2014), Antiapoplotic actions of methyl gallate on neonatal rat cardiac myocytes exposed to HjOj, Oxidative Medicine and Cellular Longevity., Epub 2014 Jan 12. doi. 10.1155/2014/657512. I I . Wang C. R., Zhou R., Ng T.
B., Wong J. H., Qiao W. T., Liu F. (2014), First report on isolation of methyl gallate with antioxidant, anti-HIV-1 and HIV-1 enzyme inhibitoiy activities from a mushroom (Phohota adiposa). Environmental Toxicology and Pharmacology, 37(2), 626- 637.12. Otsuka H.. Hirata E., Shinzato T„ Takeda Y. (2003), Stereochemistry of megastigmane glucosides from Glochidion zeylanicum and Alangium premnifolium, Phytochemistry (62). 763-768.
Journal of Medicinal Materials, 2014, VoL 19, No. 5 (pp.313-3I6)
EFFECTS OF ETHANOLIC LEAF EXTRACT OF MANGIFERA INDICA L.
ON BLOOD GLUCOSE LEVEL IN IWSG-INDUCED TYPE 2 DIABETIC MICE
Hoang Le Son *, To Ngoc Kim Thanh, Tran Van Minh International University, Vietnam National University Ho Chi Minh City
(Corresponding author: [email protected] (Received August I5^ 2014)
Summary
Effect of Ethanolic Leaf Extract of Afofl^fyera/mffcuL. on Blood Glucose Level in MSG-Induced type 2 Diabetic Mice The aim of this study was to investigate the anti-hyperglycemic property of ethanolic leaf extract of Mangifera indica in monosodium glutamate (MSG)-induced type 2 diabetic mouse model. The results showed that the ethanolic leaf extract of Mangifera indica at a dose of 250 mg/kg body weight significantly reduced blood glucose level up to 25.4% in MSG-induced type 2 diabetic mice after 4 hours of administration, higher than that of negative control (8.83%). Meanwhile, the hypoglycemic effect of the positive control (Metformin) was recorded up to 26.26% after 4 hours of treatmenL
Keywords: Mangifera indica, Anti-hyperglycemic, MSG-induced Mouse modeL
1. Introduction associated with older age, obesity and physical Diabetes mellitus has broken out rapidly in inactivity [I]. Remedy for type 2 diabetes has developed countries as well as developing much progress in anti-diabetic medications such countries in the recent two decades. It is as Sulphonylureas, Metformin, Glinides, and characterized by metabolic disorder associated Thiazolidinedione. However, it takes lots of with blood glucose level elevation. The type 2 effort and expense to get along with long-term diabetes accounts for 90% of cases and is usually treatment. Moreover, patients inevitably suffer
Journal of Medicinal Materials, 2014, Vol 19, No 5 313
