美洲大蠊抗肿瘤活性部位CII-3指纹图谱的建立及多成分含量测定

Establishment of fingerprint and multi-content determination in CII-3 of Periplaneta americana antitumor active site

建立美洲大蠊抗肿瘤活性部位CII-3的HPLC指纹图谱及其核苷类成分的含量测定方法。采用Phenomenex Gemini C_(18) 110A(4.6 mm×250 mm,5μm)色谱柱,以乙腈(A)-0.01%甲酸水(B)为流动相,梯度洗脱,测定16批供试品。采用“中药指纹图谱相似度评价系统(2012版)”进行相似度评价,并对结果进行聚类分析(HCA)、主成分分析(PCA)、正交偏最小二乘判别分析(OPLS-DA)。16批CII-3的HPLC指纹图谱中共有峰为16个,相似度介于0.870~0.982之间,HCA将CII-3分为3类,结合PCA、OPLS-DA分析筛选出3个导致批次间质量差异的标志物。指认出8种核苷类成分并测定了其含量,分别为胞嘧啶(0.50~1.17 mg·g^(-1))、腺嘌呤(0.54~2.10 mg·g^(-1))、鸟嘌呤(0.08~1.07 mg·g^(-1))、黄嘌呤(0.74~4.67 mg·g^(-1))、胸腺嘧啶(0.32~2.46 mg·g^(-1))、腺苷(1.02~7.67 mg·g^(-1))、肌苷(4.02~37.35 mg·g^(-1))、胸腺嘧啶核苷(0.56~1.86 mg·g^(-1)),8种核苷类成分在16批供试品中含量差异较大。该方法简单、准确,可为美洲大蠊抗肿瘤活性部位CII-3的质量控制提供理论依据。

To establish the HPLC fingerprint and determine the content of nucleoside components in CⅡ-3 of anti-tumor active site form Periplaneta americana.A Phenomenex Gemini C18 110A(4.6 mm × 250 mm,5 μm) column with acetonitrile(A)-0.01%formic acid(B) as mobile phase,16 batches of samples were determined by gradient elution.The similarity was evaluated by " Similarity Evaluation System for Chromatographic Fingerprint of Traditional Chinese Medicine(2012 Edition)",and the results were analyzed by HCA,PCA and OPLS-DA.The results showed that there were 16 peaks in the HPLC fingerprints of 16 batches of CⅡ-3,and the similarity were between 0.870~0.982.HCA could classify CⅡ-3 into three categories,PCA and OPLS-DA analysis screened out three markers that caused quality differences among batches.Eight nucleoside were identified and determined contents,namely cytosine(0.50~1.17 mg·g^(-1)),adenine(0.54~2.10 mg·g^(-1)),guanine(0.08~1.07 mg·g^(-1)),xanthine(0.74~4.67 mg·g^(-1)),thymine(0.32~2.46 mg·g^(-1)),adenosine(1.02~7.67 mg·g^(-1)),inosine(4.02~37.35 mg·g^(-1)) and thymidine(0.56~1.86mg·g^(-1)),there were significant differences among 8 nucleosides in 16 batches.The method was simple and accurate,and could provide a theoretical basis for the quality control of anti-tumor active site CⅡ-3 of Periplaneta americana.