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目的建立用HPLC双波长法同时测定清热凉血丸中黄芩苷、毛蕊花糖苷2种成分含量的方法。方法采用安捷伦TC-C_(18)(4.6×250mm,5μm)为色谱柱,以乙腈为流动相A,0.1%磷酸溶液为流动相B,梯度洗脱(0~30 min,16%A;30~50 min,16%A→47%A;50~60min,47%A),流速0.8 m L·min~(-1);黄芩苷检测波长为280nm、毛蕊花糖苷检测波长为334nm。结果黄芩苷、毛蕊花糖苷线性范围分别为0.3896~3.896μg(r=0.9999)、0.03626~0.3626μg(r=0.9998);平均加样回收率(n=6)均在95.6%~98.8%范围内;RSD均小于2.0%。结论本文建立的方法符合方法学验证要求,适用于同时测定清热凉血丸中毛蕊花糖苷、黄芩苷的含量。
OBJECTIVE To establish a method for simultaneous determination of two components baicalin and verbascoside in Qingre Liangxue Wan using HPLC dual wavelength method. Methods Agilent TC-C 18 column (4.6 × 250 mm, 5 μm) was used as mobile phase A with acetonitrile as mobile phase A and 0.1% phosphoric acid as mobile phase B with gradient elution (0-30 min, 16% A; 30 ~ 50 min, 16% A → 47% A; 50 ~ 60min, 47% A) at a flow rate of 0.8 m L · min -1. The detection wavelength of baicalin was 280 nm and the detection wavelength of verbascoside was 334 nm. Results The linear ranges of baicalin and verbascoside were 0.3896 ~ 3.896μg (r = 0.9999) and 0.03626 ~ 0.3626μg (r = 0.9998), respectively. The average recoveries were 95.6% ~ 98.8% RSD are less than 2.0%. Conclusion The method established in this paper meets the validation requirements of methodology and is suitable for the simultaneous determination of verbascoside and baicalin in Qingre Liangxue Wan.