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 C18色谱柱(4.6 mm×100 mm,3.5 μm);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(R2＞0.998 0),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.
Abstract
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 C18 column(4.6 mm×100 mm,3.5 μm),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(R2>0.998 0),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.
基金项目
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项目(21DZ2290200)
上海市人才发展基金(2021085)
维普
万方数据