HPLC波长切换法同时测定理气散结颗粒中4个成分的含量

目的建立同时测定理气散结颗粒中氧化芍药苷、芍药内酯苷、芍药苷和α-香附酮的HPLC波长切换联合梯度洗脱法。方法采用Venusil MP C18色谱柱(4.6 mm×250 mm,5μm),以乙腈(A)-0.1%磷酸溶液(B)为流动相,梯度洗脱(0~15 min,12.0%A;15~31 min,12.0%A→26.0%A;31~39 min,26.0%A→40.0%A;39~45 min,40.0%A→12.0%A),流速0.9 mL/min,波长切换(0~31 min,在230 nm波长下检测氧化芍药苷、芍药内酯苷和芍药苷;31~45 min,在242 nm波长下检测α-香附酮),柱温30℃,进样量为20μL。结果氧化芍药苷、芍药内酯苷、芍药苷和α-香附酮4个成分的质量浓度分别在5.15~103.00μg/mL(r=0.999 6)、8.99~179.80μg/mL(r=0.999 5)、19.83~396.60μg/mL(r=0.999 9)、6.35~127.00μg/mL(r=0.999 3)呈良好线性关系;平均加样回收率及相应的RSD(n=6)分别为96.83%(0.93%)、97.85%(1.31%)、99.71%(0.80%)、98.77%(1.59%)。结论所建立的方法灵敏度高、快速、专属性好、准确度高,为理气散结颗粒的质量控制提供了依据。

Simultaneous determination of 4 components in Liqi Sanjie Keli by HPLC wave- length switching method

Objective To develop a joint gradient elution HPLC wavelength switching method for determina-tion of the content of oxypaeoniflorin,alibiflorin,paeoniflorin andα-cyperone in Liqi Sanjie Keli simultaneously. Meth-ods The Venusil MP C18 (4. 6 mm × 250 mm,5 μm) chromatographic column was adopted;the mobile phase was acetonitrile (A)-0. 1% phosphoric acid solution (B) with gradient elution(0~15 min,12. 0% A;15~31 min,12. 0%A→26. 0% A;31~39 min,26. 0% A→40. 0% A;39 ~45 min,40. 0% A→12. 0% A),at a flow rate of 0. 9 mL/min;the detection wavelengths were set at 230 nm in 0 ~31 min to determine oxypaeoniflorin,alibiflorin and paeoni-florin,and 242 nm in 31~45 min to determine α-cyperone;the column temperature was set at 30 ℃;sample quantity was 20 μL. Results The quality concentration of oxypaeoniflorin,alibiflorin,paeoniflorin andα-cyperone was in good linear relationship within 5. 15~103. 00 μg/mL (r=0. 9996),8. 99~179. 80 μg/mL (r=0. 9995),19. 83~396. 60μg/mL (r=0. 9999) and 6. 35 ~127. 00 μg/mL(r=0. 9993) respectively. The average recoveries and the corre-sponding RSD were 96. 83%(0. 93%),97. 85%(1. 31%),99. 71%(0. 80%) and 98. 77%(1. 59%),respectively. Conclusion The established method is highly sensitive and rapid with good specificity and high accuracy. It can be ap-plied to the quality control of Liqi Sanjie Keli.