中药龟甲HPLC指纹图谱研究

HPLC Fingerprint of Carapax et Plastrum Testudinis

目的:建立中药龟甲的HPLC指纹图谱,以达到鉴别龟甲与其混伪品的目的。方法:收集不同产地的龟甲及其混伪品21批,按上下甲进行分类,共42个样品。通过盐酸水解、脱磷、脱钙等前处理,以6-氨基喹啉基-N-羟基琥珀酰亚胺基-氨基甲酸酯(AQC)进行衍生化反应,采用C18色谱柱(3. 9 mm×150 mm,3μm),柱温37℃,以乙腈,水,缓冲盐为流动相进行梯度洗脱,流速1. 00 m L/min,检测波长248 nm。采用"中药指纹图谱相似度评价系统"(《中华人民共和国药典》2012版)对正品龟甲上甲与下甲、正品与常见混伪品进行相似度分析。结果:所建立的指纹图谱具有较好的精密度、重现性和稳定性。正品龟甲的上下甲相似度为0. 932～0. 995。正品与混伪品的相似度在0. 90以下者占90. 91%。结论:正品上、下甲的HPLC图谱差异较小,正品与混伪品差异较明显。所建立的HPLC指纹图谱可用于龟甲的质量评价。

Objective:To establish HPLC(High Performance Liquid Chromatography)fingerprint of Carapax et Plastrum Testudinis by identifying tortoiseshells and their adulterants.Methods:A total of 21 batches of tortoiseshells and their adulterants from different habitats was collected,and 42 samples were classified by the carapace and plastron.The pre-treatment of acid hydrolysis,dephosphorization and decalcification pretreatment with 6-aminoquinolyl-N-hydroxysuccinimidyl-carbamate(AQC)derivatization reaction was carried out using the C 18 column(3.9×150 mm,3μm),the column temperature of 37℃.Acetonitrile,water,salt buffer were used as mobile phase for gradient elution at a flow rate of 1.00 mL/min and a the detection wavelength of 248 nm.The system of similarity evaluation(Pharmacopoeia 2012)was used to analyze tortoiseshells by the carapace and plastron,tortoisshells and their common adulterants.Results:The fingerprint had good precision,reproducibility and stability.The similarity of tortoiseshells was between 0.932-0.995.The percentage was less than 90.91%under the similarity of 0.90 between tortoiseshells of their adulterants.Conclusion:The difference of HPLC is relatively small between tortoiseshells′carapaces and plastrons,and the difference between tortoise shells and their adulterants is obvious.The established HPLC fingerprint can be used for the quality evaluation of tortoiseshells.