UFLC法同时测定黄芪桂枝五物汤中4个活性成分的含量

目的：建立超快速液相色谱法同时测定黄芪桂枝五物汤（黄芪、桂枝、芍药、生姜、大枣）中毛蕊异黄酮苷、芍药苷、芍药内酯苷和桂皮酸的含量，为黄芪桂枝五物汤的质量控制提供依据。方法：采用Shim-Pack XR-ODS色谱柱（75 mm×3.0 mm，2.2 μm）；流动相为0.1%磷酸水溶液（A）-乙腈（B），梯度洗脱（0~5 min，10% B→20% B；5~10 min，20% B→25% B；10~15 min，25% B→35% B；15~18 min，35% B→ 43% B），平衡时间为5 min；流速为0.4 mL·min^-1；柱温为35℃；检测波长分别为232 nm（毛蕊异黄酮苷、芍药苷、芍药内酯苷）和290 nm（桂皮酸）；进样量为5 μL。结果：毛蕊异黄酮苷、芍药苷、芍药内酯苷和桂皮酸质量浓度分别在1~50 μg·mL^-1（r=0.999 7）、10~500 μg·mL^-1（r=0.999 8）、2.5~125 μg·mL^-1（r=0.999 8）和5~250 μg·mL^-1（r=0.999 7）范围内与峰面积呈良好的线性关系；平均回收率分别为96.9%、99.4%、98.5%和98.9%。6批样品中上述4个成分的含量范围分别为0.185~0.221、18.80~23.49、4.00~4.92和0.644~0.681 mg·mL^-1。结论：本法可用于黄芪桂枝五物汤的质量控制。

Objective:To develop an ultra-fast liquid chromatography (UFLC) method for simultaneous determination of four active constituents(calycosin-7-O-β-D-glucoside, paeoniflorin, albiflorin and cinnamic acid) in Huangqi Guizhi Wuwu decoction. Methods:Chromatographic separation was achieved on a Shim-Pack XR-ODS column (75 mm×3.0 mm, 2.2 μm). The mobile phase consisted of water (containing 0.1% phosphoric acid) (A) and acetonitrile(B) with gradient elution(0-5 min, 10%B→20%B;5-10 min, 20%B→25%B; 10-15 min, 25%B→35%B;15-18 min, 35%B→43%B) at a flow rate of 0.4 mL·min^-1 and the equilibration time was 5 min. Column temperature was maintained at 35℃ and the injection volume was 5 μL. Calycosin-7-O-β-D-glucoside, paeoniflorin and albiflorin were determined at 232 nm, and cinnamic acid was determined at 290 nm. Results:The linear ranges were 1.0-50 μg·mL^-1(r=0.999 7) for calycosin-7-O-β-D-glucoside, 10-500 μg·mL^-1(r=0.999 9) for paeoniflorin, 2.5-125 μg·mL^-1(r=0.999 8) for albiflorin and 5.0-250 μg·mL^-1(r=0.999 7) for cinnamic acid. The average recoveries (n=9) of calycosin-7-O-β-D-glucoside, paeoniflorin, albiflorin and cinnamic acid were 96.9%, 99.4%, 98.5% and 98.9%, respectively. The content ranges of four components in six batches of samples were 0.185-0.221 mg·mL^-1 for calycosin-7-O-β-D-glucoside, 18.80-23.49 mg·mL^-1 for paeoniflorin, 4.00-4.92 mg·mL^-1 for albiflorin and 0.644-0.681 mg·mL^-1 for cinnamic acid. Conclusion:The developed method is suitable for the quality control of Huangqi Guizhi Wuwu decoction.