Solation, Purification and Identification of Antioxidant Compositions in Melilotus officinalis L.
MA Cui-xia, LIU Yu-ting, MIAO Lei, ZHAO Yu, SHAO Shuai*, YAN Ming-ming*
Traditional Chinese Medicine Health Food Science and Technology Innovation Center of Jilin Province, Changchun University of Chinese Medicine, Changchun 130117, China
Abstract：OBJECTIVE To study the chemical constituents from Melilotus officinalis(L.) and their anti-oxidant activities. METHODS DPPH Assay and β-carotene bleaching assay were used for screening the fraction of anti-oxidant activity in the 70% ethanol extract of Melilotus officinalis(L.). The isolation and purification were performed by silica,Sephadex LH-20 and preparative columns. The structures of the obtained compounds were identified by physicochemical properties and spectral data. Then their anti-oxidant activities were evaluated. RESULTS =The ethyl acetate fraction from Melilotus officinalis(L.) showed the most significant anti-oxidant activity. Thirteen compounds were isolated and identified as kaempferol(1), luteolin(2), quercetin(3), hesperidin(4), rutin(5), apigenin(6), salicylic acid(7), coumarin(8), scoparone(9), 7-methoxycoumarin(10), fumaric acid(11), caffeic acid(12), and p-hydroxybenzoate-4-O-α-D-mannopyranosyl-(1→3)-α-L-rhamnoside(13).Compounds 2, 4, 7 and 12 exhibited strong scavenging activities against DPPH free radical with the IC50 values of 25.89, 14.09, 28.03, and 23.04 μg·mL-1, respectively; compounds 4, 7, 12 and 13 exhibited strong scavenging activities against ABTS free radical with the IC50 values of 10.48, 23.77, 22.79, and 25.20 μg·mL-1, respectively. CONCLUSION Compounds 3, 7, 9, 10, 11, 12, and 13 are isolated from this plant for the first time. Compound 13 is a new compound not reported in the literature; compounds 2, 4, 7, and 12 have strong ability to scavenge DPPH free radicals; compounds 4, 7, 12, and 13 have strong ability to scavenge ABTS free radicals.
MACIAS F A, SIMONET A M, GALINDO J C G, et al. Bioactive polar triterpenoids from Melilotus messanensis. Phytochemistry, 1998, 49(3):709-717.
[3]
TANG C N. Extracing and identificating of total flavonoids from Melilotus. Anhui Chem Ind(安徽化工), 2014, 40(1):27-28.
[4]
YANG J, WANG L L, ZHANG T J, et al. Research progress on chemical constituents in plants of Melilotus Linn. and their pharmacological activities. Chin Tradit Herb Drugs(中草药), 2014, 45(3):447-454.
[5]
PLESCA-MANEA L, PARVU A E, PARUU M, et al. Effects of Melilotus officinalis on acute inflammation. Phytotherapy Res, 2002, 16(4):316-319.
[6]
GU B Q. Melilotus officinalis L. infusing tablets and Chinese medicine fumigation to prevent postoperative complications of mixed hemorrhoids. Zhejiang J Integr Tradit Chinese West Med (浙江中西医结合杂志), 2006, 16(10):607-608.
[7]
YAN M M, WU C Y, WEI Z X, et al. Anti-cancer effect of external saponins from Melilotus officinalis L. . Jilin J Tradit Chin Med (吉林中医药), 2015, 35(2):191-192.
[8]
CHENG C J, WANG Q, LI X. Research progress on antioxidant and bacteriostatic activities of flavonoids. J China Pharm(中国药房), 2011, 22(35):3346-3348.
[9]
XU Y D, ZHANG K F, ZHONG M L, et al. Research progress on chemical constituents and pharmacological effects of Dicliptera chinensis(L.) Juss. J China Pharm(中国药房), 2015, 26(34):4863-4864.
[10]
CONFORTI F, MARRELLI M, STATTI G, et al. Antioxidant and cytotoxic activities of methanolic extract and fractions from Senecio gibbosus subsp. gibbosus (GUSS) DC. Nat Prod Res, 2006, 20(9):805-812.
[11]
LI L, SUN J, SUN J Y, et al. Study on the chemical constituents of pinus massonianalamb. Chin Tradit Herb Drugs(中草药),2010,41(4):530-532.
[12]
FENG W S, CHEN W J, ZHENG X K, et al. Flavonoids from the flowers of Chrysanthemum morifolium Ramat. . Chin Pharm J(中国药学杂志), 2017, 52(17):1497-1502.
[13]
CAO J Q, WANG Y N, ZHOU Y Z, et al. Isolation and identification of the chemical constituents from Blumea riparia DC. (Ⅱ). Chin J Med Chem (中国药物化学杂志),2008, 18(6):449-451,
[14]
WU B. Studies on chemical constituents and physical and chemieal identification of Smilx falbra. Shenyang: Shenyang Pharmaeeutieal University, 2009.
[15]
ZHANG J, HE X W, CAO J C, et al. Study on chemical constituents from Alpinia tonkinensis. Chin Pharm J (中国药学杂志), 2003, 38(7):23-24.
[16]
SHI L, JI Z Q, YU Q Y, et al. Chemical constituents in methanol parts of Toddalia asiatica (Linn) Lam. China Pharm(中国药师), 2014, 17(4):534-537.
[17]
WU K, LAI Y H. Evision of the thin layer identification method of hesperidin in Xiangweiweian tablets. J Chin Med Mater (中药材), 2007, 30(2):153-154.
[18]
QIAN Y C, LIU J D, HUANG F, et al. The identification of rutin in the flower of Sophora japonica L. f. pendula Loud. Arb. Brit. by TLC and UV. Lishizhen Med Mater Med Res(时珍国医国药), 2003, 14(2):86-87.
[19]
GUO S J, YANG Y L, MA R J. Studies on hydrophilic chemical constituents of aster souliei. Chin Pharm J (中国药学杂志), 2006,41(18):1376-1378.
[20]
XIANG Z B, CHEN H S, JIN Y S, et al. Phenolic constituents of Canarium album. Chemi Nat Compd, 2010, 46(1):119-120
[21]
LI J. Study on chemical constituents of coumarins Flavonoids from Melilotus officinalis L. Hubei University, 2007.
[22]
ZOU Z J, CUI H H, YU Z C, et al. Chemical constituents of Dicliptera chinensis (L. ) Nees. Chin Tradit Pat Med (中成药),2012, 34(7):1309-1311.
[23]
WANG G D, WANG G K, LIN S S, et al. Studies on the chemical constituents from Gueldenstaedtia stenophylla Bunge. China J Chin Mater Med (中国中药杂志),2012, 37(10):1412-1416.
[24]
SONG Z Z, JIA Z J. Studies on the chemical constituents of Bupleurum sibiricum Ves. J Lanzhou Univ(兰州大学学报),1992,28 (8):99-103.
[25]
FAN Q, ZHANG Y, ZHOU X L, et al. Chemical constituents from the aerial parts of Codonopsis nervosa. Chin Pharm J (中国药学杂志), 2011, 46(4):256-259.
[26]
KISIEL W, ZIELINSKA K. Sesquiterpenoids and phenolics from Lactuca perennis. Fitoterapia,2000,71(1):86-87.
[27]
ZHOU L Y, ZHANG X H, CHEN C X. Chemical study on Rhodiola from Lijiang. Nat Prod Res Dev(天然产物研究与开发), 2004,16(5):410-414.
[28]
YU D Q, YANG J S. Hand Book of Analtical Chemistry(分析化学手册). Vol 5, Beijing:Chemical Industry Press,1999:902.
[29]
MILIAUSKAS G, VENSKUTONIS P, VAN BEEK T A, et al. Screening of radical scavenging activity of some medicinal and aromatic plant extracts . Food Chem, 2003, 85(2):231-237.
[30]
XU T T, LI Y F, JI R M, et al. Preliminary research of Radix Phytolaccae Decoction on the renal injury in rats. Chin Pharm J (中国药学杂志), 2015, 50(5):403-407.
[31]
CHEN H L, KUANG W, SHI F. Study on protective and repairing effects of Sargassum Fusiforme Polysaccharides on oxidative injured vascular endothelial cell in vitro. Chin Pharm J (中国药学杂志), 2007,42(11):829-831.
[32]
LU X Y. Research progress on natural antioxidant pycnogenol. Chin Pharm J(中国药学杂志), 2005, 39(6):403-405.
2019, Vol. 54 
