HPLC-MS法测定双黄连口服液中木犀草苷和绿原酸的含量并评价二者对成品质量的影响

Content Determination of Galuteolin and Chlorogenic Acid in Shuanghuanglian Oral Solution by HPLC-MS and Evaluation of the Effects of Them on the Quality of Products

目的:建立测定双黄连口服液中主要成分木犀草苷含量的方法,并探讨木犀草苷和绿原酸对成品质量控制的影响。方法:木犀草苷含量测定采用高效液相色谱-质谱联用法。色谱柱为Agilent HC-C18柱,流动相为0.2%磷酸(A)-乙腈(B)(梯度洗脱),流速为1mL·min-1,检测波长为350nm,进样量为10μL,柱温为25℃;绿原酸的含量测定采用2010年版《中国药典》(一部)双黄连口服液项下方法。通过测定9批双黄连口服液样品和4个模拟样品中木犀草苷和绿原酸的含量分析二者对成品质量的影响。结果:木犀草苷检测浓度在3.9～39.0μg·mL-1范围内与峰面积积分值呈良好线性关系(r=0.9999),平均加样回收率为99.5%,RSD=3.6%。9批双黄连口服液中木犀草苷的含量为5.56～30.20μg·mL-1,有较大差异;绿原酸的含量为0.71～0.94μg·mL-1,差异不大。4个模拟样品中木犀草苷的含量为0～33.28μg·mL-1,有较大差异;绿原酸的含量为1.07～1.23μg·mL-1,差异不大。结论:仅测定绿原酸来表征双黄莲口服液中金银花的含量有失准确、专属和有效,建议在双黄连口服液质量标准中增加木犀草苷的含量测定。

OBJECTIVE： To establish the method for the content determination of galuteolin in Shuanghuanglian oral solution, and to investigate the effects of galuteolin and chlorogenic acid on the quality control of products. METHODS：The content of galuteolin was determined by HPLC-MS. The Agilent HC-C18 column was adopted with mobile phase consisted of 0.2% phosphoric acid（A）-acetonitrile（B） with gradient elution at the flow rate of 1.0 mL·min-1. The column temperature was 25 ℃ and the injection volume is 10 μL. The detection wavelength was set at 350 nm. The method for the content determination of chlorogenic acid was same to the method of content determination of Shuanghuanglian oral solution stated in the appendix Ⅰof Chinese Pharmacopeia （2010 edition）. The effects of galuteolin and chlorogenic acid on the quality control of products were analyzed by determining the contents of galuteolin and chlorogenic acid in 9 batches of Shuanghuanglian oral solution and 4 batches of simulated sample. RESULTS： The linear range of galuteolin was 3.9～39.0 μg·mL^-1 （r=0.999 9） with an average recovery of 99.5% （RSD=3.6%）. The contents of galuteolin in 9 batches of Shuanghuanglian oral solution were 5.56～30.20 μg·mL^-1, there was significant difference. The contents of chlorogenic acid were 0.71～0.94 μg·mL^-1, there was no significant difference. The contents of galuteolin in 4 batches of simulated sample were 0～33.28μg·mL^-1, there was significant difference. The contents of chlorogenic acid were 1.07～1.23 μg·mL^-1, there was no significant difference. CONCLUSION： It’s not accurate, specific and effective enough just to inflect the content of Shuanghuanglian oral solution with chlorogenic acid. It is suggested that the content determination of galuteolin is included in the quality control of product.