HPLC波长切换法同时测定宝宝乐中四种成分的含量

Simultaneous determination of the content of four constituents in Baobaole by HPLC wavelength switching method

目的建立HPLC波长切换法同时测定宝宝乐中芍药内酯苷、芍药苷、毛蕊异黄酮葡萄糖苷和芒柄花素的含量。方法采用Venusil MP C_(18)(250 mm×4.6 mm,5.0μm),以乙腈-0.1%磷酸溶液为流动相,进行梯度洗脱,检测波长分别为230 nm(芍药内酯苷、芍药苷)和254 nm(毛蕊异黄酮葡萄糖苷、芒柄花素),流速0.9mL/min,柱温30℃,进样量为10μL。结果芍药内酯苷、芍药苷、毛蕊异黄酮葡萄糖苷和芒柄花素4个成分的质量浓度分别在4.18~83.60μg/mL(r=0.999 8)、5.14~102.80μg/mL(r=0.999 5)、5.60~112.00μg/mL(r=0.999 9)、4.72~94.40μg/mL(r=0.999 3)范围内与峰面积呈良好的线性关系;平均加样回收率及相应的RSD分别为99.66%(0.96%)、98.17%(1.39%)、97.00%(1.48%)、99.92%(0.58%);精密度良好,RSD≤1.06%;重复性良好,RSD≤1.69%。供试品溶液在室温条件下12 h内稳定,RSD≤1.12%。结论所建立的方法灵敏度高、快速、专属性好、准确度高,为宝宝乐的质量控制提供了依据。

Objective To develop an HPLC wavelength switching method for simultaneous determination of the content of alibiflorin, paeoniflorin, calycosinT-O-β-D-glucopyranoside and formononetin in Baobaole. Methods The Venusil MP C18 （250 mm× 4. 6 mm ,5.0 μm）chromatographic column was adopted;the mobile phase was acetoni- trile-0. 1% phosphoric acid solution with gradient elution, at a flow rate of 0. 9 mL/min. The detection wavelengths were 230 nm（ determination of alibiflorin and paeoniflorin） , and 254 nm（ determination of calycosinT-O-β-D-glucopyr- anoside and formononetin）. The column temperature was set at 30 ℃, and the sample quantity was 10 μL. Results The concentration of alibiflorin, paeoniflorin, calycosinT-O-β-D-glucopyranoside and formononetin was respectively 4. 18 -83.60 μg/mL（r =0. 999 8） ,S. 14 - 102. 80 μg/mL（r =0. 999 5） ,5.60 - 112. 00 μg/mL（r =0. 999 9） and 4. 72 - 94. 40 μg/mL （ r = 0. 999 3 ）. The average recoveries and the corresponding RSD were 99. 66% （ 0. 96% ） , 98. 17% （ 1.39% ） ,97. 00% （ 1.48% ） and 99. 92% （0. 58% ） ,respectively. The precision was good,RSD≤ 1.06% ; the repeatability was good RSD ≤1.69%. Test solution was stable at room temperature within 12 h, RSD ≤ 1. 12%. Conclusion The established method is rapid, with high sensitivity, good specificity and high accuracy, which can be applied to the quality control of Baobaole.