绞股蓝中6种多酚化合物的HPLC检测及其醇提液的抗氧化活性
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摘要
采用高效液相色谱法分析野生(5叶、7叶和9叶3种生态类型)和人工种植(7叶)绞股蓝的叶及茎中6种多酚化合物的含量,并测定样品醇提液的总多酚含量及抗氧化活性。结果表明:色谱条件为色谱柱poroshell120 PFP(4.6 mm×100 mm,2.7μm),流动相1%甲酸(A)和乙腈(B),检测波长254 nm和350 nm,在此条件下绞股蓝样品中6种多酚单体可在35 min内得到较好分离,且重复性好(相对标准偏差(relative standard deviation,RSD)≤4.6%)、精密度高(RSD≤2.68%)、稳定性好(RSD≤2.45%)、加标回收结果准确可靠(平均回收率84%～99%,RSD≤4.02%)。6种多酚单体在不同的绞股蓝样品中含量差异较大,但均呈现出芦丁含量最高,对羟基苯甲酸含量最低。绞股蓝样品醇提液中总多酚及抗氧化活性在不同样品间差异较大,在野生绞股蓝(7叶)叶中最高(分别为75、39.65、70.48 mg/g),在其茎中最低(分别为2.81、2.1、3.7 mg/g)。总体而言,绞股蓝的叶中总多酚含量高于茎中,醇提液的抗氧化活性也强于茎。
A new high performance liquid chromatography(HPLC) method was used to analyze the contents of 6 selected phenolic compounds and total phenolics in the leaves and stems of wild(five-leaf, seven-leaf and nine-leaf) and cultivated(seven-leaf) Gynostemma pentaphyllum(Thumb.) Makino. The total phenolics and antioxidant activity of the methanol extracts of these plant samples were determined. The chromatographic column used was poroshell 120 PFP(4.6 mm × 100 mm, 2.7 μm) and the mobile phase was composed of 1% formic acid(A) and acetonitrile(B). The detection wavelengths were set at 254 and 350 nm. The results showed that the six phenolic compounds could be separated well within 35 min, with good repeatability(relative standard deviation(RSD) ≤ 4.6%), high precision(RSD ≤ 2.68%), good stability(RSD ≤2.45%) and reliable recovery(average recovery rate 84%–99%, RSD ≤ 4.02%). The contents of the phenolic compounds and total phenolics and antioxidant activities in G. pentaphyllum varied greatly among different varieties. For all the samples,rutin was the most dominant phenolic constituent whereas p-hydroxybenzoic acid was the least dominant one. Total phenols content and antioxidant activity were both highest in the leaves(respectively 75, 39.65, 70.48 mg/g) and lowest in the stems(respectively 2.81, 2.1, 3.7 mg/g) of wild G. pentaphyllum among the 8 samples. In general, the content of total phenols and antioxidant activity in the leaves were higher than in the stems of G. pentaphyllum.
A new high performance liquid chromatography(HPLC) method was used to analyze the contents of 6 selected phenolic compounds and total phenolics in the leaves and stems of wild(five-leaf, seven-leaf and nine-leaf) and cultivated(seven-leaf) Gynostemma pentaphyllum(Thumb.) Makino. The total phenolics and antioxidant activity of the methanol extracts of these plant samples were determined. The chromatographic column used was poroshell 120 PFP(4.6 mm × 100 mm, 2.7 μm) and the mobile phase was composed of 1% formic acid(A) and acetonitrile(B). The detection wavelengths were set at 254 and 350 nm. The results showed that the six phenolic compounds could be separated well within 35 min, with good repeatability(relative standard deviation(RSD) ≤ 4.6%), high precision(RSD ≤ 2.68%), good stability(RSD ≤2.45%) and reliable recovery(average recovery rate 84%–99%, RSD ≤ 4.02%). The contents of the phenolic compounds and total phenolics and antioxidant activities in G. pentaphyllum varied greatly among different varieties. For all the samples,rutin was the most dominant phenolic constituent whereas p-hydroxybenzoic acid was the least dominant one. Total phenols content and antioxidant activity were both highest in the leaves(respectively 75, 39.65, 70.48 mg/g) and lowest in the stems(respectively 2.81, 2.1, 3.7 mg/g) of wild G. pentaphyllum among the 8 samples. In general, the content of total phenols and antioxidant activity in the leaves were higher than in the stems of G. pentaphyllum.
引文
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[26]徐国前,张振文,郭安鹊,等.植物多酚抗逆生态作用研究进展[J].西北植物学报,2011,31(2):423-430.
[27]王永涛.白皮松植物多酚的地理变异及其与环境因子的耦合关系[D].杨凌:西北农林科技大学,2016.
[28]DUDT J F,SHURE D J.The influence of light and nutrients on foliar phenolics and insect herbivory[J].Ecology,1994,75(1):86-98.DOI:10.2307/1939385.
[29]陈嘉景,彭昭欣,石梅艳,等.柑橘中类黄酮的组成与代谢研究进展[J].园艺学报,2016,43(2):384-400.DOI:10.16420/j.issn.0513-353x.2015-0689.
[30]LOPIERO A R.The state of the art in biosynthesis of anthocyanins and its regulation in pigmented sweet oranges[(Citrus sinensis)L.Osbeck][J].Journal of Agricultural and Food Chemistry,2015,63(16):4031-4041.DOI:10.1021/acs.jafc.5b01123.
[31]伍国怡.药用植物绞股蓝的研究进展[J].中国民族民间医药,2016,25(12):60-63.
[32]彭亮,李诒光,陈杰,等.不同产地、不同品种绞股蓝总黄酮含量比较研究[J].亚太传统医药,2015,11(21):33-35.DOI:10.11954/YTCTYY.201521014.
[2]刘世彪.绞股蓝结构和发育与其总皂甙积累相关性的研究[D].西安:西北大学,2005:6.
[3]中药大辞典编委会.中药大辞典[M].2版.上海:上海科学技术出版社,2015.
[4]张景凯.绞股蓝皂苷对HepG2细胞的凋亡、转移侵袭的影响[D].天津:天津科技大学,2015.
[5]YEO J Y,KANG Y J,JEONUN S M,et al.Potential hypoglycemic effect of an ethanol extract of Gynostemma pentaphyllum in C57BL/KsJ-db/db mice[J].Journal of Medicinal Food,2008,11(4):709-716.DOI:10.1089/jmf.2007.0148.
[6]MEGALLI S,AKTAN F,DAVIES N M,et al.Phytopreventative antihyperlipidemic effects of Gynostemma pentaphyllum in rats[J].Journal of Pharmaceutical Sciences,2005,8(3):507-515.
[7]CHEN T,LI B,LI Y Y,et al.Catalytic synthesis and antitumor activities of sulfated polysaccharide from Gynostemma pentaphyllum Makino[J].Carbohydrate Polymers,2011,83(2):554-560.DOI:10.1016/j.carbpol.2010.08.024.
[8]SUNTARARUKS S,YOOPAN N,RANGKADILOK N R,et al.Immunomodulatory effects of cadmium and Gynostemma a pentaphyllum herbal tea on rat splenocyte proliferation[J].Journal of Agricultural and Food Chemistry,2008,56(19):9305-9311.DOI:10.1021/jf801062z.
[9]CIRCOSA C,DE PASQUALE R,OCCHIUTO F.Cardiovascular effects of the aqueous extract of Gynostemma pentaphyllum Makino[J].Phytomedicine,2005,12(9):638-643.DOI:10.1016/j.phymed.2004.06.023.
[10]肖小河,陈士林,尹国萍,等.四川绞股蓝属的生态分布及资源利用[J].中药材,1991,14(3):16-19.DOI:10.13863/j.issn1001-4454.1991.03.007.
[11]金莹,孙爱东.多酚的食物来源及生物有效性[J].食品与发酵工业,2006,32(9):101-106.DOI:10.13995/j.cnki.11-1802/ts.2006.09.026.
[12]WAIN R L,TAYLOR H F.Phenols as plant growth regulators[J].Nature,1965,207:167-169.DOI:10.1038/207167a0.
[13]WASTON R R.Polyphenols in plants:isolation,purification and extract preparation[M].2nd ed.Salt Lake City:Academic Press,2019:45-58.DOI:10.1016/B978-0-12-813768-0.00003-7.
[14]ARTS I C W,HOLLMAN P C H.Polyphenols and disease risk in epidemiologic studies[J].The American Journal of Clinical Nutrition,2005,81(1):317S-325S.DOI:10.1093/ajcn/81.1.317S.
[15]MANACH C,SCALBERT A,MORAND C,et al.Polyphenols:food sources and bioavailability[J].The American Journal of Clinical Nutrition,2004,79(5):727-747.DOI:10.1093/ajcn/79.5.727.
[16]WENG C J,YEN G C.Chemopreventive effects of dietary phytochemicals against cancer invasion and metastasis:phenolic acids,monophenol,polyphenol,and their derivatives[J].Cancer Treatment Reviews,2012,38(1):76-87.DOI:10.1016/j.ctrv.2011.03.001.
[17]NARA K.Spores of ectomycorrhizal fungi:ecological strategies for germination and dormancy[J].New Phytologist,2009,181(2):245-248.DOI:10.1111/j.1469-8137.2008.02691.x.
[18]REMSBERG C M,Yá?EZ J A,OHGAMI Y,et al.Pharmacometrics of pterostilbene:preclinical pharmacokinetics and metabolism,anticancer,antiinflammatory,antioxidant and analgesic activity[J].Phytotherapy Research,2008,22(2):169-179.DOI:10.1002/ptr.2277.
[19]SABIR A,RAFI M,DARUSMAN L K,Discrimination of red and white rice bran from Indonesia using HPLC fingerprint analysis combined with chemometrics[J].Food Chemistry,2017,221:1717-1722.DOI:10.1016/j.foodchem.2016.10.114.
[20]何志勇,夏文水.Folin-Ciocalteu比色法测定橄榄中多酚含量的研究[J].林产化学与工业,2006,26(4):15-18.
[21]YU J,WANG L M,WAZLEM R L,et al.Antioxidant activity of citrus limonoids,flavonoids,and coumarins[J].Journal of Agricultural and Food Chemistry,2005,53(6):2009-2014.DOI:10.1021/jf0484632.
[22]CHENG H R,FENG S L,JIA X Y,et al.Structural characterization and antioxidant activities of polysaccharides extracted from Epimedium acuminatum[J].Carbohydrate Polymers,2013,92(1):63-68.DOI:10.1016/j.carbpol.2012.09.051.
[23]CAI Y Z,LUO Q,SUN M,et al.Antioxidant activity and phenolic compounds of 112 traditional Chinese medicinal plants associated with anticancer[J].Life Sciences,2004,74(17):2157-2184.DOI:10.1016/j.lfs.2003.09.047.
[24]WRIGHT D M,JORDAN G J,LEE W G,et al.Do leaves of plants on phosphorus-impoverished soils contain high concentrations of phenolic defence compounds?[J].Functional Ecology,2010,24(1):52-61.DOI:10.1111/j.1365-2435.2009.01597.x.
[25]?AMEC D,VALEK-?ULJ L,MARTINEZ S,et al.Phenolic acids significantly contribute to antioxidant potency of Gynostemma pentaphyllum,aqueous and methanol extracts[J].Industrial Crops and Products,2016,84:104-107.DOI:10.1016/j.indcrop.2016.01.035.
[26]徐国前,张振文,郭安鹊,等.植物多酚抗逆生态作用研究进展[J].西北植物学报,2011,31(2):423-430.
[27]王永涛.白皮松植物多酚的地理变异及其与环境因子的耦合关系[D].杨凌:西北农林科技大学,2016.
[28]DUDT J F,SHURE D J.The influence of light and nutrients on foliar phenolics and insect herbivory[J].Ecology,1994,75(1):86-98.DOI:10.2307/1939385.
[29]陈嘉景,彭昭欣,石梅艳,等.柑橘中类黄酮的组成与代谢研究进展[J].园艺学报,2016,43(2):384-400.DOI:10.16420/j.issn.0513-353x.2015-0689.
[30]LOPIERO A R.The state of the art in biosynthesis of anthocyanins and its regulation in pigmented sweet oranges[(Citrus sinensis)L.Osbeck][J].Journal of Agricultural and Food Chemistry,2015,63(16):4031-4041.DOI:10.1021/acs.jafc.5b01123.
[31]伍国怡.药用植物绞股蓝的研究进展[J].中国民族民间医药,2016,25(12):60-63.
[32]彭亮,李诒光,陈杰,等.不同产地、不同品种绞股蓝总黄酮含量比较研究[J].亚太传统医药,2015,11(21):33-35.DOI:10.11954/YTCTYY.201521014.