瓜蒌皮注射液及其中间体中17种氨基酸含量测定及其变化规律研究  

Content determination of seventeen amino acids in Gualoupi Injection and its intermediates and research on their change laws

作　　者：陶香 张静娴 胡青 孙健 董莹 丁金国 于泓 沈盈盈 毛秀红 季申[1,2] TAO Xiang;ZHANG Jing-xian;HU Qing;SUN Jian;DONG Ying;DING Jin-guo;YU Hong;SHEN Ying-ying;MAO Xiu-hong;JI Shen(Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China;National Medical Products Administration Key Laboratory for Quality Control of Traditional Chinese Medicine,Shanghai Municipal Institute for Food and Drug Control,Shanghai 201203,China;SPH No.1 Biochemical&Pharmaceutical Co.,Ltd.,Shanghai 200240,China)

机构地区：[1]上海中医药大学,上海201203 [2]上海市食品药品检验研究院,国家药品监督管理局中药质量控制重点实验室,上海201203 [3]上海第一生化药业有限公司,上海200240

出　　处：《中成药》2024年第3期709-717,共9页Chinese Traditional Patent Medicine

基　　金：上海市中药和保健食品品质与安全检测专业技术服务平台(21DZ2290200);上海市人才发展资金(2021085)。

摘　　要：目的测定瓜蒌皮注射液及其中间体中天冬氨酸、谷氨酸、丝氨酸、甘氨酸、苏氨酸、瓜氨酸、精氨酸、丙氨酸、γ-氨基丁酸、酪氨酸、缬氨酸、苯丙氨酸、异亮氨酸、鸟氨酸、亮氨酸、赖氨酸、脯氨酸的含量,并分析其变化规律。方法OPA-FMOC在线衍生化分析采用Waters XBridge C_(18)色谱柱(4.6 mm×100 mm,3.5μm);流动相磷酸盐缓冲液-[甲醇-乙腈-水(45∶45∶10)],梯度洗脱;体积流量1 mL/min;柱温45℃;检测波长262、338 nm。主成分分析、热图分析对10道工序相应中间体的6批样品进行化学模式识别。结果17种氨基酸在各自范围内线性关系良好(R^(2)>0.9980),平均加样回收率83.4%~119.5%,RSD 0.91%~7.94%。相同工序不同批次样品较聚集,不同工序相应中间体聚为3组。醇沉和阳离子交换柱对氨基酸组成影响最大。结论本实验可为瓜蒌皮注射液质量控制的关键因素提供重要参考,以保证终产品的稳定性和均一性。AIM To determine the contents of aspartic acid,glutamic acid,serine,glycine,threonine,citrulline,arginine,alanine,γ-amino-butyric acid,tyrosine,valine,phenlalanine,isoleucine,ornithine,leucine,lysine and proline in Gualoupi Injection and its intermediates,and to analyze their change laws.METHODS The OPA-FMOC online derivatization analysis was performed on a 45℃thermostatic Waters XBridge C_(18) column(4.6 mm×100 mm,3.5μm),with the mobile phase comprising of phosphate buffer solution-[methanol-acetonitrile-water(45∶45∶10)]flowing at 1 mL/min in a gradient elution manner,and the detection wavelengths were set at 262,338 nm.Principal component analysis and heatmap analysis were adopted in chemical pattern recognition for the corresponding intermediates in ten processes of six batches of samples.RESULTS Seventeen amino acids showed good linear relationships within their own ranges(R^(2)>0.9980),whose average recoveries were 83.4%-119.5%with the RSDs of 0.91%-7.94%.Different batches of samples in the same process were clustered,and the corresponding intermediates in different processed were clustered into three groups.Alcohol precipitation and cation exchange column demonstrated the biggest influences on amino acid composition.CONCLUSION This experiment can provide important references for the critical factors on quality control of Gualoupi Injection,thus ensure the stability and uniformity of final product.

关 键 词：瓜蒌皮注射液 中间体 氨基酸 含量测定 变化规律 OPA-FMOC在线衍生化 主成分分析 热图分析 

分 类 号：R927.2[医药卫生—药学]