目的 建立豪猪刺特征图谱及18种氨基酸成分含量测定方法,为豪猪刺化学成分研究以及质量控制提供参考。方法 采用异硫氰酸苯酯(PITC)进行柱前衍生化,对16批不同产地的豪猪刺进行高效液相色谱法(HPLC)分析,以0.1 mol·L-1乙酸钠(体积分数36%乙酸调至pH 6.5)-乙腈(93∶7)为流动相A,乙腈-水(4∶1)为流动相B,进行梯度洗脱;运用《中药色谱指纹图谱相似度评价系统》(2012版)进行指纹图谱相似度评价,通过SPSS 26.0,SIMCA14.1软件进行聚类分析、主成分分析,同时测定了18种氨基酸成分的含量。结果 建立了豪猪刺氨基酸柱前衍生化-HPLC指纹图谱,其相似度≥0.950,共标定了24个共有峰,指认了天冬氨酸、谷氨酸、丝氨酸等18个氨基酸类成分;聚类分析和主成分分析将16批样品分为4类。16批豪猪刺总氨基酸含量在34.00%~92.30%之间。结论 本研究建立的豪猪刺氨基酸柱前衍生-HPLC指纹图谱以及18种氨基酸含量测定的方法简便,稳定可靠,可为豪猪刺药材的品质鉴定及质量控制提供一定的参考。
Abstract
:OBJECTIVE To establish a characteristic map of porcupine spines and a method for the determination of 18 amino acid components, and provide reference for the study of chemical components and quality control of porcupine spines. METHODS Sixteen batches of porcupine quines from different producing areas were analyzed by HPLC using pre-column derivatization with phenyl isothiocyanate(PITC). Gradient elution was carried out with 0.1 mol·L-1 sodium acetate(36% acetic acid adjusted to pH 6.5)-acetonitrile (93∶7) as mobile phase A and acetonitrile-water(4∶1) as mobile phase B. The similarity of fingerprint was evaluated by Similarity Evaluation System of Chromatographic Fingerprint of Traditional Chinese medicine(2012 edition). Cluster analysis and principal component analysis were performed by SPSS 26.0 and SIMCA 14.1 software, and the contents of 18 kinds of amino acids were determined. RESULTS The similarity of HPLC fingerprint was greater than or equal to 0.950, 24 common peaks were identified, and 18 amino acids such as aspartic acid, glutamic acid and serine were identified. Cluster analysis and principal component analysis divided 16 batches of samples into four categories. The total amino acid content of 16 batches of porcupine spines ranged from 34.00% to 92.30%. CONCLUSION The HPLC fingerprint and assay for 18 kinds of amino acids are established in this study, which are simple, stable and reliable, and can provide reference for the quality identification and quality control of porcupine spines.
关键词
豪猪刺 /
氨基酸 /
指纹图谱 /
化学计量学 /
质量评价
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Key words
porcupine spines /
amino acid /
fingerprint /
stoichiometry /
quality evaluation
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中图分类号:
R284
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脚注
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基金
贵州省教育厅青年科技人才成长项目资助(黔教合KY字193号);贵州省基础研究计划(科学技术基金)项目资助(黔科合基础1Y387)
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