珍珠层粉氨基酸指纹图谱的构建及氨基酸含量测定的研究

Construction of Amino Acid Fingerprint and Determination of Amino Acid Content of Pearl Layer Powder

目的 建立中药珍珠层粉HPLC氨基酸指纹图谱,进行指纹图谱的模式识别研究,并测定珍珠层粉的氨基酸含量,为珍珠层粉质量评价提供依据;方法 采用Venusi1-AA氨基酸分析柱(4.6 mm×250 mm,5μm,柱号:AA952505-0),柱温:28℃,进样量。10μL,波长:254 nm,流速:0.9 mL·min-1,以pH为6.5的醋酸钠水溶液为流动相A,以80%乙腈水溶液为流动相B。运用SAS和SPSS统计软件进行聚类分析和主成分分析,对珍珠层粉指纹图谱进行模式识别研究。结果 建立了珍珠层粉氨基酸指纹图谱,以及氨基酸指纹图谱的共有模式,检测出23个共有峰,并指认出其中15个共有峰相似度分析的结果与聚类分析和主成分分析保持一致,不同产地的珍珠层粉氨基酸成分差异不大,但不同基源的珍珠层粉氨基酸成分差异明显。本研究测定17个批次珍珠层粉中17种氨基酸(Asp天门冬氨酸、Glu谷氨酸、Ser丝氨酸、Gly甘氨酸、His组氨酸、Arg精氨酸、Thr苏氨酸、Ala丙氨酸、Pro脯氨酸、Tyr酪氨酸、Val缬氨酸、Met蛋氨酸、Cys胱氨酸、Ile异亮氨酸、Leu亮氨酸、Phe苯丙氨酸、Lys赖氨酸)的含量。结论 本研究建立的珍珠层粉氨基酸指纹图谱分析方法专属性强,及氨基酸含量测定方法简便易行、准确可靠,可对珍珠层粉的质量评价提供依据。

Objective To establish the fingerprint of amino acid of Chinese medicine pearl layer powder by HPLC,to study the pattern recognition of fingerprint,and to determine the content of amino acid in pearl layer powder to provide basis for the quality evaluation of pearl layer powder.Methods Venusi 1-AA amino acid analysis column(4.6 mm×250 mm,5μm,column number:AA952505-0),column temperature:28℃,sample volume:10μL,wavelength:254 nm,flow rate:0.9 mL·min-1,sodium acetate solution with pH 6.5 as mobile phase A and 80%acetonitrile solution as mobile phase B were used.Cluster analysis and principal component analysis were carried out by SAS and SPSS statistical software to study the pattern recognition of pearl layer powder fingerprint.Results The amino acid fingerprint of pearl layer powder and the common pattern of amino acid fingerprint were established.23 common peaks were obtained and 15 of them were identified.The results of similarity analysis are consistent with those of cluster analysis and principal component analysis.There was no significant difference in amino acid composition of pearl layer powder from different habitats,but amino acid composition of pearl layer powder with different origins was obviously different.The contents of 17 kinds of amino acids(Asp,Glu,Ser,Gly,His,Arg,Thr,Ala,Pro,Tyr,Val,Met,Cys,Ile,Leu,Phe,Lys)in 17 batches of pearl layer powder are obtained.Conclusion The established fingerprint analysis method of amino acids in pearl layer powder has strong specificity.Content determination method of the amino acid is simple,accurate and reliable.It can provide a basis for the quality evaluation of pearl layer powder.