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海藻及其混伪品中17种氨基酸的含量测定
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| 篇名： | 海藻及其混伪品中17种氨基酸的含量测定 |
| TITLE： | Content Determination of 17 Kinds of Amino Acids in Sargassum and Its Adulterants |
| 摘要： | 目的：建立海藻及其混伪品中17种氨基酸的含量测定方法并进行聚类分析，为海藻药材的质量控制提供依据。方法：取18批样品（S1～S6为真品、S7～S18为混伪品），经酸水解后，采用氨基酸全自动分析仪测定氨基酸含量。色谱柱为LCAK06/Na型磺酸基强酸性阳离子交换树脂分离柱，流动相为缓冲液-再生液系统（梯度洗脱），流速为洗脱泵0.45mL/min、衍生泵0.25mL/min，检测波长为440nm（脯氨酸）、570nm（其余氨基酸），进样量为50μL。采用PASWStatistics18.0软件，以“平方Euclidean距离”为度量标准使用组间联接的聚类方法对18批样品进行聚类分析。结果：17种氨基酸均分离良好，空白无干扰；其质量浓度与峰面积的线性关系良好（r均大于0.998），线性范围上下限分别为48.06μg/L（胱氨酸）、1.501μg/L（甘氨酸）；精密度、重复性、稳定性试验RSD均小于2％；平均加样回收率为90.60％～101.56％（RSD为0.88％～2.15％，n＝6）。海藻及其混伪品样品中均检出17种氨基酸，其中谷氨酸、天冬氨酸、亮氨酸、丙氨酸、甘氨酸、缬氨酸的含量相对较高。聚类分析结果显示，18批样品可以聚为4类，S1～S6为一类，S7～S9为一类，S10～S12、S16～S18为一类，S13～S15为一类，与海藻真品与其混伪品的鉴定结果相一致。结论：该方法操作简便、快速、准确度高、重复性好，可用于海藻及其混伪品的氨基酸定量分析及真伪区分。 |
| ABSTRACT： | OBJECTIVE：To establish the method for content determination of 17 kinds of amino acids in Sargassum and its adulterants，and to carry out cluster analysis ，so as to provide reference for quality control of Sargassum. METHODS ：Totally of 18 batches of sample （S1-S6 as certified product ，S7-S18 as adulterants ）were collected. After acid hydrolysis ，amino acids contents were detected by using automatic amino acid analyzer. The separation was performed on LCAK 06/Na sulfonic acid cation exchange resin column with mobile phase consisted of buffer-regeneration system （gradient elution ）at the flow rate of 0.45 mL/min （elution pump ）and 0.25 mL/min（derivative pump ）. The detection wavelengths were set at 440 nm（proline）and 570 nm（other amino acids ），and the sample size volume was 50 μL. PASW Statistics 18.0 software was used ，and cluster analysis was conducted by using group connection method of cluster analysis with “square Euclidean distance ”as the measurement standard. RESULTS ：17 kinds of amino acids were well separated without interference from blank sample. The linear relationship between mass concentration and peak area was good （all r were over 0.998），and the upper and lower limits of the linear range were 48.06 μg/L （cystine）and 1.501 μg/L（glycine），respectively；RSDs of precision ，reproducibility and stability tests were lower than 2％. The average recoveries were between 90.60％-101.56％（RSDs were 0.88％-2.15％，n＝6）. 17 kinds of amino acids were detected in Sargassum and its adulterants ，among which the contents of glutamic acid ，aspartic acid ，leucine，alanine，glycine and valine were relatively high . Results of cluster analysis showed that 18 batches of sample were clustered into 4 categories，i.e. S 1-S6 into one category；S7-S9 into one category ；S10-S12，S16-S18 into one category ；S13-S15 into one category ；which was consistent with the identification result of Sargassum and its adulterants . CONCLUSIONS ：The method is simple ， rapid， accurate and reproducible，and can be used for the quantitative analysis and identification of amino acids in Sargassum and adulterants. |
| 期刊： | 2020年第31卷第04期 |
| 作者： | 张晓萍，张文，戚鹏飞，朱仁愿，李洁，张彩霞，邱国玉 |
| AUTHORS： | ZHANG Xiaoping ，ZHANG Wen，QI Pengfei ，ZHU Renyuan ，LI Jie，ZHANG Caixia ，QIU Guoyu |
| 关键字： | 海藻；混伪品；阳离子交换色谱法；柱后衍生；氨基酸 |
| KEYWORDS： | Sargassum；Adulterants；Cation exchange chromatography ；Post-column derivatization ；Amino acids |
| 阅读数： | 324 次 |
| 本月下载数： | 7 次 |
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