HPLC法同时测定植物样品中3种甾醇含量
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摘要
建立高效液相色谱法(HPLC)同时测定植物样品中麦角甾醇、胆甾醇和豆甾醇的方法。样品经皂化、萃取后,用紫外检测器进行测定。优化后的色谱条件为:Shim-pack CLC ODS色谱柱(250 mm×4.6 mm, 5.0μm);流动相采用纯甲醇;检测波长205 nm和282 nm,流速1.0 mL/min;柱温35℃;进样量10μL。检测方法显示, 3种甾醇在1.00～200 mg/L的线性范围内相关系数R~2>0.999,方法精密度RSD<0.5%(N=6),在加标浓度为20.0～200 mg/kg条件下,加标回收率93.6%～103.4%(N=6),检出限(S/N=3) 3.0～6.0 mg/kg,定量限(S/N=10) 10～20 mg/kg。
A HPLC method was established to determine the contents of ergosterol, cholesterol and stigmasterol in plant sample. The plant sample was detected by ultraviolet detector after the process of saponification and extraction. The optimized HPLC method was that a shim-pack CLC ODS column(250 mm×4.6 mm, 5.0 μm) used as chromatographic column, methanol used as mobile phase, 205 nm and 282 nm used as detection wavelength, flow rate 1.0 mL/min, column temperature 35 ℃ and injection volume 10 μL. The detection method revealed that the correlation coefficients(R~2) of the three sterols were greater than 0.999 in their linear range of 1.00-200 mg/L; The precision of the method(RSD) was less than 0.5%(N=6); The recovery of the three sterols was between 93.6% and 103.4% in the added concentration range of 20.0-200 mg/kg(N=6); The limit of detection(S/N=3) was between 3.0 mg/kg and 6.0 mg/kg; The limit of quantitation(S/N=10) was between 10 mg/kg to 20 mg/kg.
A HPLC method was established to determine the contents of ergosterol, cholesterol and stigmasterol in plant sample. The plant sample was detected by ultraviolet detector after the process of saponification and extraction. The optimized HPLC method was that a shim-pack CLC ODS column(250 mm×4.6 mm, 5.0 μm) used as chromatographic column, methanol used as mobile phase, 205 nm and 282 nm used as detection wavelength, flow rate 1.0 mL/min, column temperature 35 ℃ and injection volume 10 μL. The detection method revealed that the correlation coefficients(R~2) of the three sterols were greater than 0.999 in their linear range of 1.00-200 mg/L; The precision of the method(RSD) was less than 0.5%(N=6); The recovery of the three sterols was between 93.6% and 103.4% in the added concentration range of 20.0-200 mg/kg(N=6); The limit of detection(S/N=3) was between 3.0 mg/kg and 6.0 mg/kg; The limit of quantitation(S/N=10) was between 10 mg/kg to 20 mg/kg.
引文
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[7]高虹,谷文英.三波长光度法测定姬松茸中麦角甾醇含量[J].分析化学, 2007, 35(4):586-588.
[8]刘海霞,仇农学,王峰,等.苹果籽油中植物甾醇含量的薄层色谱-分光光度法测定[J].中国油脂, 2008, 33(11):76-79.
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[11]陈月晓,何涛,唐凌轩,等.气相色谱法同时测定食品中的胆固醇和植物甾醇[J].食品科学, 2016, 37(14):180-183.
[12] FENG S, LIU S, LUO Z, et al. Direct saponification preparation and analysis of free and conjugated phytosterols in sugarcane(Saccharum officinarum L)by reversed-phase high-performance liquid chromatography[J]. Food Chemistry,2015(181):9-14.
[13] YUAN C X, JU Y J, JIN R S, et al. Simultaneous HPLCDAD analysis of tocopherols, phytosterols, and squalene in vegetable oil deodorizer distillates[J]. Chromatographia, 2015,78(3/4):273-278.
[14]王梦丽,冯亚男,谢红英,等. RP-HPLC法同时测定12种植物油中的3种甾醇含量[J].中国测试, 2016, 42(4):60-64.
[15]贾蔓箐,唐辉,李翠华,等.棉油皂脚中β-谷甾醇与菜油甾醇的HPLC分析测定[J].食品科技, 2013, 38(6):300-302.
[16]鄢庆祥,高明焱,陆冬艳,等.高效液相色谱法同时测定西瓜藤中豆甾醇和β-谷甾醇含量[J].亚太传统医药, 2017,13(13):19-21.
[17]赵英永,赵晔,林瑞超,等. RP-HPLC法测定猪苓中麦角甾醇的含量[J].药物分析杂志, 2009, 29(6):898-900.
[18]李丽萍,赵榕,赵海燕,等.超高效液相色谱法测定食品中胆固醇的方法改进[J].中国卫生检验杂志, 2015, 25(17):2893-2895.
[2]RUDZINSKAM,HASSANEINMMM,ABDELRAZEK A G, et al. Blends of rapeseed oil with black cumin and rice bran oils for increasing the oxidative stability[J]. Journal of Food Science&Technology, 2016, 53(2):1055-1062.
[3] FU Y Q, ZHANG Y, HU H Y, et al. Design and straightforward synthesis of novel galloyl phytosterols with excellent antioxidant activity[J]. Food Chemistry, 2014, 163(20):171-177.
[4] SRIRAMAN S, RAMANUJAM G M, RAMASAMY M K, et al. Identification of beta-sitosterol and stigmasterol in Bambusa bambos(L)voss leaf extract using HPLC and its estrogenic effect in vitro[J]. Journal of Pharmaceutical&Biomedical Analysis, 2015(115):55-61.
[5]葛志伟,罗自生.食品中植物甾醇的色谱分析方法研究进展[J].食品安全质量检测学报, 2017, 8(3):773-778.
[6]石馨,魏雪冬,刘波,等.植物甾醇的应用及检测方法研究进展[J].中国乳业, 2014, 150(6):59-61.
[7]高虹,谷文英.三波长光度法测定姬松茸中麦角甾醇含量[J].分析化学, 2007, 35(4):586-588.
[8]刘海霞,仇农学,王峰,等.苹果籽油中植物甾醇含量的薄层色谱-分光光度法测定[J].中国油脂, 2008, 33(11):76-79.
[9] DU M, AHN D U. Simultaneous analysis of tocopherols,cholesterol, and phytosterols using gas chromatography[J].Journal of Food Science, 2010, 67(5):1696-1700.
[10]王多娇,周玮,颜春荣,等.油脂中胆固醇及植物甾醇组成及含量分析[J].中国油脂, 2016, 41(2):106-109.
[11]陈月晓,何涛,唐凌轩,等.气相色谱法同时测定食品中的胆固醇和植物甾醇[J].食品科学, 2016, 37(14):180-183.
[12] FENG S, LIU S, LUO Z, et al. Direct saponification preparation and analysis of free and conjugated phytosterols in sugarcane(Saccharum officinarum L)by reversed-phase high-performance liquid chromatography[J]. Food Chemistry,2015(181):9-14.
[13] YUAN C X, JU Y J, JIN R S, et al. Simultaneous HPLCDAD analysis of tocopherols, phytosterols, and squalene in vegetable oil deodorizer distillates[J]. Chromatographia, 2015,78(3/4):273-278.
[14]王梦丽,冯亚男,谢红英,等. RP-HPLC法同时测定12种植物油中的3种甾醇含量[J].中国测试, 2016, 42(4):60-64.
[15]贾蔓箐,唐辉,李翠华,等.棉油皂脚中β-谷甾醇与菜油甾醇的HPLC分析测定[J].食品科技, 2013, 38(6):300-302.
[16]鄢庆祥,高明焱,陆冬艳,等.高效液相色谱法同时测定西瓜藤中豆甾醇和β-谷甾醇含量[J].亚太传统医药, 2017,13(13):19-21.
[17]赵英永,赵晔,林瑞超,等. RP-HPLC法测定猪苓中麦角甾醇的含量[J].药物分析杂志, 2009, 29(6):898-900.
[18]李丽萍,赵榕,赵海燕,等.超高效液相色谱法测定食品中胆固醇的方法改进[J].中国卫生检验杂志, 2015, 25(17):2893-2895.