HPLC-DAD法同时测定栀芩清热合剂中多指标成分含量

目的：建立高效液相色谱法同时测定栀芩清热合剂中京尼平苷酸、京尼平龙胆双糖苷、京尼平苷、黄芩苷、汉黄芩苷、甘草酸、黄芩素、绿原酸8个成分的含量。方法：采用Platisil ODS色谱柱（250 mm×4.6 mm，5 μm），以乙腈（A）-0.1%磷酸水溶液（B）为流动相，梯度洗脱（0~20 min，10% A→16% A；20~30 min，16% A→40% A；30~50 min，40% A→80% A），流速1 mL·min^-1，柱温30℃，变换波长检测（240 nm处检测京尼平苷酸、京尼平龙胆双糖苷、京尼平苷、黄芩苷、汉黄芩苷、甘草酸、黄芩素；327 nm处检测绿原酸）。结果：京尼平苷酸、京尼平龙胆双糖苷、京尼平苷、黄芩苷、汉黄芩苷、甘草酸、黄芩素、绿原酸的线性范围分别为0.081 9~4.914 μg（r=0.999 4）、0.005~0.3 μg（r=0.999 0）、0.016 2~0.972 μg（r=0.999 8）、0.057~3.42 μg（r=0.999 5）、0.002 1~0.126 μg（r=0.999 9）、0.005 5~0.33 μg（r=0.999 7）、5.45~87.17 μg（r=0.999 7）、0.004 1~0.246 0 μg（r=0.999 4），平均回收率（n=9）分别为98.7%（RSD=1.4%）、97.6%（RSD=1.35%）、102.0%（RSD=1.5%）、97.5%（RSD=1.3%）、96.6%（RSD=1.4%）、98.5%（RSD=1.6%）、98.10%（RSD=1.5%）、101.5%（RSD=1.9%）。3批样品中京尼平苷酸、京尼平龙胆双糖苷、京尼平苷、黄芩苷、汉黄芩苷、甘草酸、黄芩素、绿原酸的含量分别为1.745 5~1.804 3、1.123 6~1.185 2、10.128 3~10.448 5、10.743 8~11.421 9、2.676 1~2.810 2、0.998 9~1.025 1、0.969 7~0.980 8、0.178 3~0.186 9 mg·g^-1。结论：经方法学验证，本法可作为栀芩清热合剂质量控制的一个有效的方法。

Objective: To establish an HPLC method for simultaneous determination of geniposidic acid, genipin 1-gentiobioside, geniposide, baicalin, wogonoside, glycyrrhizic acid, baicalein and chlorogenic acid in Zhiqin Qingre mixture. Methods: The separation was performed on a Platisil ODS column (250 mm×4.6 mm, 5 μm)with gradient elution (0-20 min, 10%A→16%A;20-30 min, 16%A→40%A;30-50 min, 40%A→80%A)using acetonitrile and 0.1% phosphoric acid aqueous solution as mobile phase. The flow rate was 1 mL·min^-1 and the detection wavelengths were 240 nm for geniposidic acid, genipin 1-gentiobioside, geniposide, baicalin, wogonoside, glycyrrhizic acid and baicalein;and 327 nm for chlorogenic acid, respectively. Results: The linear ranges of geniposidic acid, genipin 1-gentiobioside, geniposide, baicalin, wogonoside, glycyrrhizic acid, baicalein and chlorogenic acid were 0.081 9-4.914 μg (r=0.999 4), 0.005-0.3 μg (r=0.999 0), 0.016 2~0.972 μg (r=0.999 8), 0.057-3.42 μg (r=0.999 5), 0.002 1-0.126 μg (r=0.999 9), 0.005 5-0.33 μg (r=0.999 7), 5.45-87.17 μg (r=0.999 7)and 0.004 1-0.246 0 μg (r=0.999 4), respectively. Their average recoveries were 98.7% (RSD=1.40%), 97.6% (RSD=1.4%), 102.0% (RSD=1.5%), 97.5% (RSD=1.3%), 96.6% (RSD=1.4%), 98.5% (RSD=1.6%), 98.1% (RSD=1.5%)and 101.5% (RSD=1.9%), respectively. The contents of geniposidic acid, genipin 1-gentiobioside, geniposide, baicalin, wogonoside, glycyrrhizic acid, baicalein and chlorogenic acid in three samples were 1.745 5-1.804 3 mg·g^-1, 1.123 6-1.185 2 mg·g^-1, 10.128 3-10.448 5 mg·g^-1, 10.743 8-11.421 9 mg·g^-1, 2.676 1-2.810 2 mg·g^-1, 0.998 9-1.025 1 mg·g^-1, 0.969 7-0.980 8 mg·g^-1 and 0.178 3-0.186 9 mg·g^-1, respectively. Conclusion: The HPLC method is proved by methodology validation that it can be used as quality control method of Zhiqin Qingre mixture.