HPLC法同时测定山药中的植物甾醇
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摘要
建立了一种高效液相色谱法(HPLC)同时测定山药中的3种植物甾醇(菜油甾醇、豆甾醇和β-谷甾醇)含量的方法。样品使用甲醇提取,经皂化、萃取后用HPLC进行分析。色谱条件:SunFire C8色谱柱(4.6 mm×250 mm,5μm),流动相乙腈-水(92.5:7.5),检测波长205 nm。结果表明:山药中3种植物甾醇在一定浓度范围内呈现良好的线性关系,相关系数均不小于0.9997,样品检出限为0.50～0.52μg/mL,样品平均加标回收率在93.5%～97.2%,相对标准偏差均小于3.6%。经测定,山药中的植物甾醇含量为菜油甾醇67.1～79.8μg/g,豆甾醇28.8～36.7μg/g,β-谷甾醇129.6～154.5μg/g。该方法操作简便、结果准确度高且重复性好,可作为山药中植物甾醇含量同时测定的方法。
A high performance liquid chromatographic(HPLC) method was established for the determination of three phytosterols(campesterol, stigmasterol and β-sitosterol) in Dioscorea opposita Thunb. The samples were extracted with methanol, then saponification and extraction followed by HPLC analysis with a SunFire C8 Column(4.6 mm×250 mm,5 μm) column. The mobile phase consisted of acetonitrile and water(92.5:7.5). The detection UV wavelength was 205 nm. The results showed that the concentration and peak area of the three phytosterols showed a good linear relationship with correlation coefficients not less than 0.9997. The limits of detection were in the ranges of 0.50～0.52 μg/mL. The mean recoveries for the samples at three spiked levels were in range of 93.5%～97.2%. By the test, the phytosterol contents in Dioscorea opposita Thunb were 67.1～79.8 μg/g of campesterol, 28.8～36.7 μg/g of stigmasterol, 129.6～154.5 μg/g of β-sitosterol. The method is simple, accurate and reproducible, and can be used for the determination of phytosterol contents in Dioscorea opposita Thunb.
A high performance liquid chromatographic(HPLC) method was established for the determination of three phytosterols(campesterol, stigmasterol and β-sitosterol) in Dioscorea opposita Thunb. The samples were extracted with methanol, then saponification and extraction followed by HPLC analysis with a SunFire C8 Column(4.6 mm×250 mm,5 μm) column. The mobile phase consisted of acetonitrile and water(92.5:7.5). The detection UV wavelength was 205 nm. The results showed that the concentration and peak area of the three phytosterols showed a good linear relationship with correlation coefficients not less than 0.9997. The limits of detection were in the ranges of 0.50～0.52 μg/mL. The mean recoveries for the samples at three spiked levels were in range of 93.5%～97.2%. By the test, the phytosterol contents in Dioscorea opposita Thunb were 67.1～79.8 μg/g of campesterol, 28.8～36.7 μg/g of stigmasterol, 129.6～154.5 μg/g of β-sitosterol. The method is simple, accurate and reproducible, and can be used for the determination of phytosterol contents in Dioscorea opposita Thunb.
引文
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[3]FAN Yijun,HE Qinyi,LUO Aoshuang,et al.Characterization and antihyperglycemic activity of a polysaccharide from Dioscorea opposita Thunb roots[J].International Journal of Molecular Sciences.2015,16(3):6391-6401.
[4]ZHAO Guohua,KAN Jianquan,LI Zhixiao,et al.Structural features and immunological activity of a polysaccharide from Dioscorea opposita Thunb roots[J].Carbohydrate Polymers.2005,61:125-131.
[5]白冰,李明静,王勇,等.怀山药化学成分研究[J].中国中药杂志,2008,33(11):1272-1274.
[6]白冰,刘绣华,王勇,等.怀山药化学成分研究(Ⅱ)[J].化学研究,2008,19(3):67-69.
[7]ZHANG Lin,BAI Bing,LIU Xiuhua,et al.α-Glucosidase inhibitors from Chinese yam(Dioscorea opposita Thunb.)[J].Food Chemistry,2011,126(1):203-206.
[8]刘绣华,白冰.怀山药与同属植物甾体成分差异性比较研究[J].河南大学学报(自然科学版),2012,(5):548-553.
[9]王玉,刘滨城,任运宏,等.植物甾醇酯对高脂血症小鼠的降血脂作用研究[J].食品科学,2011,32(17):326-329.
[10]张蕾,陈庆森,阎亚丽,等.植物甾醇对心血管疾病作用的研究现状[J].食品科学,2013,34(23):344-350.
[11]HU Qiaoling,ZHUO Zhao,FANG Shenglin,et al.Phytosterols improve immunity and exert antiinflammatory activity in weaned piglets[J].J Sci Food Agric,2017,97(12):4103-4109.
[12]MIRAS-Moreno B,SABATER-Jara A B,Pedre?o M A,et al.Bioactivity of phytosterols and their production in plant in vitro cultures[J].J Agric Food Chem,2016,64(38):7049-7058.
[13]PIIRONEN V,MIETTINEN T A,TOIVO J,et al.Plant sterols:biosynthesis,biological function and their importance to human nutrition[J].J Sci Food Agric,2000,7(7):939-966.
[14]SZALIELA A,GRABARCZYKA M,PACZKOWSKIC,et al.Comparison of the profiles of non-glycosylated triterpenoids from leaves of plants of selected species of genus Dioscorea[J].Phytochemistry Letters,2017,20:350-355.
[15]LU T J,YU C W,LIN Y C,et al.Simultaneous analysis of free/bound phytosterols and Diosgenin in Taiwanese yams[J].Food Anal Methods,2017,(10):1172-1180.
[16]LAGARDA M J,GARCIA-Llatas G,FARRE R.Analysis of phytosterols in foods[J].J Pharm Biomed Anal,2006,41(5):1486-1496.
[17]GACHUMI G,El-ANEED A.Mass spectrometric approaches for the analysis of phytosterols in biological samples[J].J Agric Food Chem,2017,65(47):10141-10156.