UPLC法测定射干药材中10个异黄酮类成分的含量
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篇名: UPLC法测定射干药材中10个异黄酮类成分的含量
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摘要: 目的:建立同时测定射干药材中10个异黄酮类成分含量的测定方法,并用于评价不同产地射干药材中各成分的含量差异。方法:采用超高效液相色谱(UPLC)法。色谱柱为Waters ACQUITY UPLC BEH C18,流动相水相组成为含0.5%甲基-β-环糊精、0.1%磷酸的水溶液,有机相为乙腈,梯度洗脱,流速为0.2 mL/min,柱温为35 ℃,检测波长为265 nm,进样量为2 μL,分析时间为20 min。对来自于8个省份的26个样品中的10个异黄酮类成分射干苷、鸢尾甲苷A、鸢尾甲苷B、野鸢尾苷、鸢尾黄素、鸢尾甲黄素B、鸢尾甲黄素A、野鸢尾黄素、次野鸢尾黄素、白射干素进行含量测定。结果:射干苷、鸢尾甲苷A、鸢尾甲苷B、野鸢尾苷、鸢尾黄素、鸢尾甲黄素B、鸢尾甲黄素A、野鸢尾黄素、次野鸢尾黄素、白射干素检测质量浓度线性范围分别为8.569 5~342.78、0.643~25.72、1.119 8~44.79、2.187 8~87.51、0.770 3~30.81、0.421 3~16.85、0.288 5~11.54、1.795 3~71.81、0.560 8~22.43、0.086~3.44 μg/mL(r均≥0.999 6),定量限分别为0.015、0.102、0.096、0.013、0.036、0.088、0.102、0.019、0.067、0.092 μg/mL;精密度、稳定性(24 h)、重复性试验的RSD均<2.00%(n=6);加样回收率为95.30%~103.30%(RSD均≤2.33%,n=6)。在26个射干样品中,以射干苷含量最高(3.66%~57.79%),白射干素含量最低(0.09%~0.59%),次野鸢尾黄素含量为0.29~2.80 mg/g,且不同产地中各异黄酮类成分含量差异较大。结论:建立的含量测定方法灵敏、分析时间较短、重复性较好,可以用于同时测定射干药材中10个异黄酮类成分的含量及评价各成分含量差异。
ABSTRACT: OBJECTIVE: To establish a method for simultaneous determination of 10 isoflavones in Belamcanda chinensis, and to evaluate the differences of active ingredient content of B. chinensis from different areas. METHODS: UPLC method was adopted. The determination was performed on Waters ACQUITY UPLC BEH C18 column with mobile phase consisted of 0.5 % methyl-β-cyclodextrin and 0.1% phosphate as water phase, acetonitrile as organic phase (gradient elution) at the flow rate of 0.2 mL/min. The column temperature was set at 35 ℃, and the detection wavelength was set at 265 nm. The sample size was 2 μL, and analysis time was 20 min. The contents of 10 isoflavones in 26 samples from 8 provinces, including tectoridin, iristectorin A, iristectorin B, iridin, tectorigenin, iristectorigenin B, iristectorigenin A, irigenin, irisflorentin, dichotomitin, were determined. RESULTS: The linear ranges of tectoridin, iristectorin A, iristectorin B, iridin, tectorigenin, iristectorigenin B, iristectorigenin A, irigenin, irisflorentin, dichotomitin were 8.569 5-342.78, 0.643-25.72, 1.119 8-44.79, 2.187 8-87.51, 0.770 3-30.81, 0.421 3- 16.85, 0.288 5-11.54, 1.795 3-71.81, 0.560 8-22.43, 0.086-3.44 μg/mL(all r≥0.999 6). The limits of quantitation were 0.015, 0.102, 0.096, 0.013, 0.036, 0.088, 0.102, 0.019, 0.067, 0.092 μg/mL. RSDs of precision, stability(24 h)and reproducibility tests were lower than 2.00% (n=6). The recoveries ranged 95.30%-103.30% (all RSD≤2.33%, n=6). Among 26 samples of B. chinensis, the content of tectoridin was the highest (3.66%-57.79%), and the content of dichotomitin was the lowest (0.09%- 0.59%), the contents of irisflorentin were 0.29-2.80 mg/g. The contents of isoflavones in B. chinensis from different areas were different greatly.  CONCLUSIONS: The established method is sensitive, with short analysis time and good repeatability, and can be used to determine the content of 10 isoflavones and evaluate the content difference of each component.
期刊: 2019年第30卷第23期
作者: 姜鸿,王光函,辛旭阳,邹桂欣,李国信
AUTHORS: JIANG Hong,WANG Guanghan,XIN Xuyang,ZOU Guixin,LI Guoxin
关键字: 射干;不同产地;异黄酮类成分;含量测定;超高效液相色谱法
KEYWORDS: Belamcanda chinensis; Different producing areas; Isoflavone; Content determination; UPLC
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