高效液相色谱法测定浓缩乳腺丸有效成分

Determination of Active Components in Concentrated Breast Pills by HPLC

目的建立浓缩乳腺丸中有效成分吉马酮、芍药苷、柴胡皂苷(d)及阿魏酸的高效液相色谱(HPLC)测定方法。方法醇提并水提法制备浓缩乳腺丸。Diamonsil-C18(4.6 mm×200 mm,5μm)色谱柱,4种有效成分流动相分别为,吉马酮:乙腈∶磷酸水溶液(50∶50,pH 3.23);芍药苷:甲醇∶水(25∶75);柴胡皂苷(d):甲醇∶乙腈∶水(20∶35∶45);阿魏酸:甲醇∶水∶冰醋酸(32∶68∶0.5)。流速均为1.0 mL.min-1,检测波长(λ)分别为210,230,201,323 nm。进样体积均为20μL,柱温分别为35,25,25,25℃。结果 4种有效成分在其对应线性范围内与峰面积呈良好的线性关系,r依次为0.999 9,0.999 8,0.999 5,0.999 5;平均提取回收率分别为75.91%,82.38%,75.04%,67.67%。结论该实验所建立的HPLC法可用于浓缩乳腺丸中相关有效成分的含量测定,方法灵敏,操作简便,测定结果准确可靠,可用于浓缩乳腺丸的质量控制。

Objective To establish a method about high-performance liquid chromatography （HPLC） to determine concentration of germacrone, peoniflorin, saikoside （d）, and ferulie acid in concentrated breast pills. Methods Concentrated breast pills were prepared by ethanol and water extraction. Diamonsil-Cls column （4.6 min×200mm, 5μm） was used. Mobile phases of the four active components were as follows ： germacrone, methyl cyanides ： phosphoric acid solution （ 50 ： 50, pH 3.23） ; peoniflorin, methanol： water （25 ： 75） ; saikoside （d）, methanol： methyl cyanides： water （20 ： 35 ： 45） ; ferulic acid, methanol： water： glacial acetic acid （32 ： 68 ： 0.5）. Flow rate was 1.0 mL ·min-1. Wavelength was 210, 230, 201 and 323 nm, respectively. Injection volume was 20 μL. Column temperature was 35, 25, 25 and 25℃, respectively. Results The peak area of each of the active component showed a good linear relationship with the concentration in the corresponding linear range, r values were ： 0. 999 9, 0. 999 85, 0. 999 5, 0. 999 5. The average extraction recoveries were ： 75. 91%, 82.38% , 75.04% , 67.67%. Conclusion This method is simple and reliable; it can be used for the determination of active components in the concentrated breast pills.