HPLC法分别测定大苞荆芥中2种黄酮及其糖苷的含量

目的:建立HPLC法分别定量测定大苞荆芥中木犀草素、芹菜素及其糖苷木犀草素-7-O-β-D-葡萄糖醛酸苷（LGCRP）、芹菜素-7-O-葡萄糖醛酸苷（AGCRP）的方法。方法:采用 C₁₈ 柱（250 mm×4.6 mm, 5 μm），以乙腈-0.1%磷酸水溶液（0.1:99.9，v/v）为流动相进行梯度洗脱，检测波长347 nm，流速1 mL/min，柱温30 ℃；优化了提取溶剂、溶剂加入量、提取时间等药材前处理方法；通过系统适用性试验，线性关系、检出限及定量限考察，精密度、重复性、稳定性及加标回收率试验，验证方法的适用性。结果:木犀草素、芹菜素和LGCRP、AGCRP分别在3.080~12.321、4.753~19.010、5.67~22.68、9.97~39.86 μg/mL范围内与各自峰面积呈良好的线性关系，r值均大于0.9993；检出限分别为0.11、0.08、0.03、0.03 μg/mL，定量限分别为0.35、0.23、0.09、0.10 μg/mL，精密度、稳定性和重复性均良好，加标回收率范围分别为98.8%~102.8%、92.9%~98.1%、93.9%~97.9%、93.7%~97.3%。结论:两套方法操作简便、灵敏度高、稳定性好、准确度高、适用性强，可用于大苞荆芥中2个黄酮及其糖苷类成分的定量测定，从而为大苞荆芥药材质量标准的制定提供依据。

Individual Determination of Two Kinds of Flavonoids and Their Glycosides in Nepeta bracteata by HPLC

Objective: To establish two HPLC methods for determination content of luteolin, apigenin, and their glucosides such as luteolin-7-O-β-D-glucuronide (LGCRP), apigenin-7-O-glucuronide (AGCRP) in Nepeta bracteata. Methods: The determination was performed on C₁₈ column（250 mm×4.6 mm, 5 μm）with mobile phase consisted of acetonitrile-0.1% phosphoric acid solution (0.1:99.9, v/v) at the flow rate of 1.0 mL/min. The detection wavelength was 347 nm, and the column temperature was 30 ℃. The methods of pretreatment such as extraction solvent, amount of solvent, extraction time were optimized. Then, applicability of methods was investigated by system suitability test, linear relationship, limit of detection and limit of quantification inspection, precision, repeatability, stability, recovery test. Results: The linear ranges of luteolin, apigenin, LGCRP, AGCRP were 3.080~12.321, 4.753~19.010, 5.67~22.68, 9.97~39.86 μg/mL, r≥0.9993, the limit of detection were 0.11, 0.08, 0.03, 0.03 μg/mL, and the limit of quantification were 0.35, 0.23, 0.09, 0.10 μg/mL. The precision, repeatability and stability were good, the recovery were in the range of 98.8%~102.8%, 92.9%~98.1%, 93.9%~97.9%, 93.7%~97.3%. Conclusion: Both methods can be used for the quantitative determination of two flavonoids and their glycosides in Nepeta bracteata because there are simple operation, high sensitivity, good stability, high accuracy and strong applicability. Thus, it provide a basis for the formulation of the quality standard of Nepeta bracteata.