高效液相色谱梯度洗脱法同时测定关节痛丸中β-蜕皮甾酮、豨莶苷、奇壬醇和豨莶精醇的含量

Simultaneous determination of β-ecdysterone, darutoside, kirenol and darutigenol in Guanjietong pills by HPLC gradient elution

目的建立HPLC梯度洗脱法同时测定关节痛丸中β-蜕皮甾酮、豨莶苷、奇壬醇和豨莶精醇的含量。方法采用Diamonsil C18(4.6 mm×200 mm,5μm),以乙腈-0.1%甲酸溶液为流动相,进行梯度洗脱,流速1.0 m L·min^(-1),柱温30℃,波长切换法检测(0~17 min,250 nm,β-蜕皮甾酮;17~60min,215 nm,豨莶苷、奇壬醇和豨莶精醇),进样量为10μL。结果β-蜕皮甾酮、豨莶苷、奇壬醇和豨莶精醇4个成分的线性范围分别为4.35~87.00μg·m L^(-1)(r=0.9993)、3.78~75.60μg·m L^(-1)(r=0.9999)、8.51~170.20μg·m L^(-1)(r=0.9994)、5.35~107.00μg·m L^(-1)(r=0.9997);平均加样回收率及相应的RSD分别为98.8%(1.4%)、99.3%(1.3%)、97.4%(1.0%)、99.0%(1.2%)。结论本文建立的方法,方法专属性强,测定结果准确,可有效保障关节痛丸临床用药安全。

Objective To develop an HPLC gradient elution method to determinate the content of 4 components （β-ecdysterone, darutoside, kirenol and darutigenol） in Guanjietong pills simultaneously. Methods The Diamonsil C18 （4.6 mm × 200 mm, 5 μm） chromatographic column was adopted; the mobile phase was acetonitrile- 0.1% formic acid solution with gradient elution at 1.0 mL . min^-1, the column temperature was 30 ℃ , and the detection wavelengths were 250 nm for β-ecdysterone, 215 nm for darutoside, kirenol and darutigenol. The sample volumn was 10 μL. Results β-ecdysterone, darutoside, kirenol and darutigenol had good linearities at 4.35 - 87.00 μg . mL-1 （r = 0.9993）, 3.78 - 75.60 μg . mL-1 （r = 0.9999）, 8.51 - 170.20 μg .mL-1 （r = 0.9994）, and 5.35 - 107.00 μg . mL-1 （r = 0.9997）, respectively. The average recoveries and the RSDs were 98.8% （1.4%）, 99.3% （1.3%）, 97.4% （1.0%）, and 99.0% （1.2%）. Conclusion The method is specific and the measurements are accurate. It is helpful to ensure clinical drug safety of Guanjietong pills.