HPLC-CAD法检测小建中合剂中6个活性成分及稳定性研究

Determination of six active ingredients in Xiaojianzhong Mixture by HPLC-CAD and stability study

目的利用高效液相色谱联合电喷雾检测器(HPLC-CAD),建立同时测定小建中合剂中芍药内酯苷、芍药苷、甘草苷、甘草酸、桂皮醛及6-姜辣素含量的检测方法,并对小建中合剂稳定性进行研究。方法采用Thermo Hypersil GOLD C18色谱柱(250 mm×4.6 mm,5μm),以80%乙腈(A)-10 mmol·L^(-1)甲酸铵(甲酸调pH值至4.0)(B)为流动相,梯度洗脱,体积流量1.0 mL·min-1,柱温35℃,CAD雾化器温度为35℃,过滤常数为5.0,采集频率为10 Hz,进样量20μL。同时测定小建中合剂在加速及长期试验下各成分含量的变化。结果各成分分离度均良好,阴性无干扰;芍药内酯苷、芍药苷、甘草苷、甘草酸、桂皮醛及6-姜辣素的线性范围分别为2.01~40.16、4.21~84.20、1.18~23.64、0.60~12.04、0.70~14.06及1.30~25.90μg·mL^(-1)(r≥0.9991);平均加样回收率(n=6)为97.55%~98.46%,RSD≤3.0%。稳定性研究结果表明,芍药内酯苷、芍药苷、甘草苷、桂皮醛及6-姜辣素在加速条件下各成分质量浓度有明显下降,提示本品种不宜在高温下长时间放置。结论经方法学验证,建立的HPLC-CAD法具有良好的灵敏度及准确度,为小建中合剂的全面质量控制提供科学依据,本品应置阴凉处贮藏。

Objective To develop an HPLC-CAD method for simultaneous determination of six active ingredients of albiflorin,paeoniflorin,liquiritin,glycyrrhizic acid,cinnamaldehyde and 6-gingerol in Xiaojianzhong Mixture.Methods The chromatographic separation was performed on a Thermo Hypersil GOLD C18 column(250 mm×4.6 mm,5μm)with 80%acetonitrile(A)-10 mmol·L^(-1) ammonium acetate(formic acid tone pH 4.0)(B)as the mobile phase for gradient elution,the flow rate was 1.0 mL·min-1,the column temperature was maintained at 35℃,the nebulization temperature of detector was 35℃,the filtration constant was 5.0,the acquisition frequency was 10 Hz,and injection volume was 20μL.At the same time,the content changes of each component of Xiaojianzhong Mixture under accelerated and long-term test were determined to provide a basis for the stability and storage conditions of the variety.Results The separation of each component was good,negative and no interference.The linear ranges of albiflorin,paeoniflorin,liquiritin,glycyrrhizic acid,cinnamaldehyde and 6-gingerol were 2.01-40.16,4.21-84.20,1.18-23.64,0.60-12.04,0.70-14.06 and 1.30-25.90μg·mL^(-1)(r≥0.9991),respectively.The average recoveries were 97.55%-98.46%,and the RSD≤3.0%.The results of stability investigation showed that the contents of albiflorin,paeoniflorin,liquiritin,glycyrrhizic acid,cinnamaldehyde and 6-gingerol decreased rapidly under accelerated condition,implied that the storage time of Xiaojianzhong Mixture under high temperature condition should not be too long.Conclusion The method is sensitive and accurate by method validation,which can be used for the overall assessment of the quality of Xiaojianzhong Mixture,this variety should be stored in a cool place.