乌鸡白凤软胶囊中氨基酸类含量测定与特征性寡肽类成分分析  

Content determination of amino acids and analysis of discriminal oligopeptides in Wuji Baifeng soft capsule

作　　者：李莎 王洋[2] 洪艳 曾万溪 关艳贞 梁柳艳 LI Sha;WANG Yang;HONG Yan;ZENG Wanxi;GUAN Yanzhen;LIANG Liuyan(Kinhoo Pharmaceutical Co.,Ltd.,Zhuhai 519041,China;School of Pharmacy,Shenyang Pharmaceutical University,Shenyang 110016,China)

机构地区：[1]金鸿药业股份有限公司,广东珠海519041 [2]沈阳药科大学药学院,辽宁沈阳110016

出　　处：《沈阳药科大学学报》2025年第3期232-242,共11页Journal of Shenyang Pharmaceutical University

摘　　要：目的采用超高效液相色谱-质谱联用技术对乌鸡白凤软胶囊中氨基酸类成分进行含量测定,对特征性寡肽类成分进行成分分析,完善乌鸡白凤软胶囊质量控制指标。方法氨基酸含量测定和寡肽类成分分析均采用Acquity BEH Hilic色谱柱(100 mm×2.1 mm,1.7μm),以乙腈和体积分数0.1%甲酸水溶液(含10 mmol·L-1甲酸铵)为流动相进行梯度洗脱。寡肽类成分的分析中采用偏最小二乘判别分析法(PLS-DA)对木瓜蛋白酶水解的乌鸡提取物和未经木瓜蛋白酶水解的乌鸡提取物样品中寡肽类成分进行分析,找出其主要特征性寡肽类成分。结果所建立的氨基酸含量测定方法在考察范围内线性关系良好(r≥0.9970),具有较好的重复性和稳定性,加样回收率在86.0%~91.5%内,其中含量较高氨基酸为天冬氨酸、谷氨酸、亮氨酸和异亮氨酸(两氨基酸含量之和)、酪氨酸;寡肽类成分的分析中生成的得分图将3组样品很好的区分,其中根据VIP结果筛选出木瓜蛋白酶水解的乌鸡提取物特征性寡肽类成分为Asp-His-Ile-Val、Arg-Ile-Thr-Tyr、Arg-Ala、Gly-Glu-Pro-Arg、Arg-Gly-Pro-Tyr、His-Ala-Val-Val、Cyclo(-Ser-Tyr)、His-Phe-Ala、Asp-Gly-Ser-Trp、Gly-Lys、Gly-Pro-Ala、Gly-Gly-Phe、Ala-Leu-Gly、Lys-Leu、Ala-Asn-His-Thr、Pro-Glu-Leu-Phe、Pro-Glu-Leu-Phe、Lys-Lys-Lys-Val。结论所建立的方法和实验结果为乌鸡白凤软胶囊药效物质基础的确定和后续研究提供了数据支持。Objective To improve quality control indicators,the content of amino acids in Wuji Baifeng soft capsule was determined by ultra high-performance liquid chromatography-tandem quadrupole mass spectrometry and the oligopeptides were analyzed by ultra high-performance liquid chromatography-orbital trap high resolution mass spectrometry.Methods The content determinations of amino acids and analysis of oligopeptides were both exerted using Acquity BEH HILIC chromatographic column(100 mm×2.1 mm,1.7μm),the mobile phase was acetonitrile and 0.1%formic acid aqueous solution(including 10 mmol·L-1 ammonium formate)with gradient elution.The MS was based on turbo ion spray ionization in positive ion mode using MRM modes for quantification of amino acids and scanning modes for analysis of oligopeptides.Partial least squares discriminant analysis(PLS-DA)was used to screen out distinguishing small peptide components among black chicken extract hydrolyzed by papain,black chicken extract without hydrolysis group.Results The established method for the determination of amino acid content exhibited good linearity(r≥0.9970),good repeatability and stability,and the recovery rates were within 86.0%-91.5%,and the contents of aspartic acid,glutamic acid,the sum of leucine and isoleucine and tyrosine were higher.In the analysis of small peptide components,the discriminal small peptide components were selected according to the VIP results including Asp-His-Ile-Val,Arg-Ile-Thr-Tyr,Arg-Ala,Gly-Glu-Pro-Arg,Arg-Gly-Pro-Tyr,His-Ala-Val-Val,Cyclo(-Ser-Tyr),His-Phe-Ala,Asp-Gly-Ser-Trp,Gly-Lys,Gly-Pro-Ala,Gly-Gly-Phe,Ala-Leu-Gly,Lys-Leu,Ala-Asn-His-Thr,Pro-Glu-Leu-Phe,Pro-Glu-Leu-Phe and Lys-Lys-Lys-Val.Conclusion The established method and obtained results provide data support for confirmation of bioactive components and the follow-up study of Wuji Baifeng soft capsule.

关 键 词：乌鸡白凤软胶囊 寡肽 氨基酸 高分辨质谱 串联四级杆质谱 

分 类 号：R917[医药卫生—药物分析学]