目的: 建立同时测定川射干中8个异黄酮成分(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)的一测多评含量测定方法(QAMS法)。方法: 采用高效液相色谱法,使用Agilent ZORBAX SB-C18色谱柱(250 mm×4.6 mm,5 μm),以乙腈为流动相A,以水(含0.55%甲基-β-环糊精和0.1%磷酸)为流动相B,以0.1%磷酸为流动相C,进行梯度洗脱,流速为1.0 mL·min-1,检测波长为266 nm,柱温为35 ℃。以鸢尾黄素为内参照物,采用多点校正法,分别建立射干苷、鸢尾甲苷A、鸢尾甲苷B、野鸢尾苷、鸢尾甲黄素B、鸢尾甲黄素A、野鸢尾黄素7个待测组分与鸢尾黄素的相对校正因子。采用校正因子计算8批川射干样品中7个待测物的含量,与外标法进行比较,验证QAMS法的准确性和可行性。结果: 8个待测组分的线性范围分别是25.00~800.00、2.50~80.00、3.06~100.00、1.53~50.00、6.12~200.00、1.53~50.00、1.53~50.00、1.53~50.00 μg·mL-1。以鸢尾黄素为内参照物,上述7个组分的相对校正因子分别是1.643、1.445、2.036、1.334、1.241、1.548、1.268,在不同的实验条件下重复性比较好。同时8个批次川射干中8个组分的含量范围分别为射干苷(48.53~109.94 mg·g-1)、鸢尾甲苷A(4.28~8.36 mg·g-1)、鸢尾甲苷B(9.47~14.78 mg·g-1)、野鸢尾苷(1.24~1.92 mg·g-1)、鸢尾黄素(11.74~34.29 mg·g-1)、鸢尾甲黄素B(3.00~7.07 mg·g-1)、鸢尾甲黄素A(1.94~5.36 mg·g-1)、野鸢尾黄素(1.02~2.28 mg·g-1)。结论: 以鸢尾黄素为内参物,建立了7个异黄酮类成分的一测多评定量方法,该方法简便、准确、可靠、可节省大量对照品使用,可用于川射干的质量评价及定量分析。
Objective: To establish a quantitative analysis of multi-components by single marker method for the determination of 8 isoflavones (tectoridin, iristectorin A, iristectorin B, tectorigenin, iridin, iristectorigenin B, iristectorigenin A and irigenin) in Iris tectorum Maxim. Methods: The Agilent ZORBAX SB-C18 column (250 mm×4.6 mm, 5 μm) was used with acetonitrile as mobile phase A, water (containing 0.55% methyl-β-cyclodextrin and 0.1% phosphoric acid) as mobile phase B, and 0.1% phosphoric acid as mobile phase C. Gradient elution was performed at a flow rate of 1.0 mL·min-1, a detection wavelength of 266 nm, and a column temperature of 35 ℃. Using tectorigenin as the reference substance, relative correction factors of other ingredients were calculated, and the contents were determined. At the same time, the standard curve method was used to determine the contents of 8 components, and the differences between the two methods were compared to verify the accuracy and feasibility. Results: The linear ranges of tectoridin, iristectorin A, iristectorin B, tectorigenin, iridin, iristectorigenin B, iristectorigenin A and irigenin were 25.00-800.00, 2.50-80.00, 3.06-100.00, 1.53-50.00, 6.12-200.00, 1.53-50.00, 1.53-50.00 and 1.53-50.00 μg·mL-1, respectively. The relative correction factors of tectoridin, iristectorin A, iristectorin B, iridin, iristectorigenin B, iristectorigenin A and irigenin were 1.643, 1.445, 2.036, 1.334, 1.241, 1.548 and 1.268, respectively. The repeatability was good under different experimental conditions. Conclusion: Using tectorigenin as an the internal reference, quantitative analysis of multi-components with single marker method for 7 isoflavones were established. The method is simple, accurate and reliable, and can be used for quality evaluation and quantitative analysis of I. tectorum.
[1] 中华人民共和国药典2020年版.一部[S].2020:43
ChP 2020. Vol Ⅰ [S].2020:43
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