摘要
目的:建立美洲大蠊抗肝纤维化活性部位(ML-HB)氨基酸类指纹图谱,评价不同批次ML-HB的质量,并测定氨基酸的含量。方法:采用盐酸水解法制备水解氨基酸样品液,领苯二甲醛柱前衍生化后进行HPLC分析。色谱柱为AgilentZORBAXEclipseXDB-C18(4.6mm×250mm,5μm);柱温35℃;检测波长338nm;流动相为甲醇-17.6mmol/L的磷酸二氢钠溶液(pH7.80),进行梯度洗脱。依据相似度评价、聚类分析和主成分分析结果进行不同批次ML-HB质量评价。结果:建立了10批ML-HB指纹图谱,标定共有峰15个,指认出13个峰,10批样品相似度均大于0.980;测定了10批样品中13种氨基酸的含量。结论:所建立的ML-HB氨基酸类指纹图谱及含量测定方法稳定性、重复性好,可为ML-HB的质量控制和评价提供参考。
Objective:To establish an amino acid fingerprint of the active site of anti-hepatic fibrosis of Periplaneta americana(MLHB), evaluate the quality of different batches of ML-HB, and determine the amino acid content. Methods:Hydrolyzed amino acid sample solution was prepared by hydrochloric acid hydrolysis method, and HPLC analysis was performed after precolumn derivatization of o-phthalaldehyde. The column was Agilent ZORBAX Eclipse XDB-C18(4.6 mm × 250 mm, 5 μm);column temperature was 35 °C;detection wavelength was 338 nm;mobile phase was methanol-17.6 mmol/L NaH2PO4 solution(pH 7.80), and gradient elution was applied. According to the similarity evaluation, cluster analysis and principal component analysis results, the quality of different batches of ML-HB was evaluated. Results:Ten batches of ML-HB fingerprints were established, 15 common peaks were identified, 13 peaks were identified, and the similarity of 10 batches was greater than 0.980. The content of 13 amino acids in 10 batches was determined. Conclusion:The established ML-HB amino acid fingerprints and content determination methods have good stability and repeatability, which can provide reference for quality control and evaluation of ML-HB.
作者
付文鹏
谢静静
赵艳雯
杨永寿
肖培云
Fu Wenpeng;Xie Jingjing;Zhao Yanwen;Yang Yongshou;Xiao Peiyun(College of Pharmacy and Chemistry, Dali University, Dali, Yunnan 671000, China;Yunnan Provincial Key Laboratory of Entomological Biopharmaceutical R&D, Dali, Yunnan 671000, China)
出处
《大理大学学报》
CAS
2019年第8期27-32,共6页
Journal of Dali University
基金
国家自然科学基金资助项目(81360634
81560634)
关键词
美洲大蠊
抗肝纤维化活性部位
氨基酸
指纹图谱
衍生化
Periplaneta americana
the active site of anti-hepatic fibrosis
amino acid
fingerprint
derivatization
