HPLC法测定五种产地地龙中次黄嘌呤和肌苷的含量  

Determination of Hypoxanthine and Inosine in Five Origins of Geosaurus by HPLC Method

作　　者：王晖[1] 王脉 梅艳飞 史玉洁 WANG Hui;WANG Mai;MEI Yan-fei;SHI Yu-jie(Hebei North University,Zhangjiakou,075000,China;Zhangjiakou First Hospital,Zhangjiakou,075000,China)

机构地区：[1]河北北方学院,张家口075000 [2]张家口市第一医院,张家口075000

出　　处：《神经药理学报》2024年第4期18-22,28,共6页Acta Neuropharmacologica

基　　金：张家口市基础研究和人才培养计划项目(No.2221012A);张家口市基础研究和人才培养计划项目(No.2221019A)。

摘　　要：目的:建立地龙中次黄嘌呤与肌苷含量的测定方法,测定五种产地地龙成分,为地龙成分的提取工艺研究、质量分析探索新型高效的研究方法。方法:建立HPLC高效液相色谱法测定分析方法。色谱柱Agilent extend C18色谱柱,4.6 mm×250 mm,粒径:5μm。流动相为甲醇-0.2%磷酸,波长选择254 nm,进样量选择20μL,柱温为30℃,使用梯度洗脱法,建立次黄嘌呤和肌苷的标准曲线,进行精密度试验,稳定性试验,重复性试验,加样回收试验等方法学考察。结果:次黄嘌呤标准曲线:y=78.819x+245.92,R2=0.9999,在10.08~161.28μg·mL^(-1)范围内呈良好线性关系;肌苷标准曲线:y=56.566x+68.333,R2=0.9995,在20.08~321.28μg·mL^(-1)范围内呈良好线性关系。实验精密度、稳定性、重复性良好,计算得到次黄嘌呤的平均回收率为95.43%,RSD值为1.52%(n=6);肌苷的平均回收率为99.86%,RSD值为2.36%(n=6),广东、河南、海南、上海、广西五种产地地龙的次黄嘌呤含量分别为:0.5335、0.3468、0.6874、0.5694、0.7945 mg·g^(-1);肌苷含量为:1.5625、0.5674、1.1267、0.8436、1.2849 mg·g^(-1)。结论:建立的HPLC高效液相色谱法,梯度洗脱效果较好,实验准确度良好,精密度良好,稳定性良好,重复性高,是一种高效的分析测定方法。Objective:To establish a method for the determination of hypoxanthine and inosine content in Geosaurus,and to determine the constituents of Geosaurus of five origins,so as to explore a new and efficient analytical research method for the study of extraction process and quality analysis of Geosaurus's constituents.Methods:A high performance liquid chromatographic(HPLC)method was established for the determination of hypoxanthine and inosine.The separation was carried out on an Agilent extend C18 column,4.6 mm×250 mm,particle size:5µm,with the mobile phase of methanol-0.2%phosphoric acid,injection volume of 20μL,wavelength of 254 nm,column temperature of 30℃,and gradient elution,to establish the standard curves of hypoxanthine and inosine and carry out the methodology of precision,stability,reproducibility,and sample recovery.The methodology of precision test,stability test,repeatability test and sample recovery test was investigated.Results:The standard curve of hypoxanthine:y=78.819x+245.92,R2=0.9999,with good linearity in the range of 10.08~161.28μg·mL^(-1);the standard curve of inosine:y=56.566x+68.333,R2=0.9995,with good linearity in the range of 20.08~321.28μg·mL^(-1);the standard curve of inosine:y=56.566x+68.333,R2=0.9995,with good linearity in the range of 20.08~321.28μg·mL^(-1).The linear relationship was good in the range of 20.08~321.28μg·mL^(-1).The average recovery of hypoxanthine was 95.43%with the RSD value of 1.52%(n=6);the average recovery of inosine was 99.86%with the RSD value of 2.36%(n=6);and the contents of hypoxanthine in the five types of Geosaurus of Guangdong,Henan,Hainan,Shanghai and Guangxi were 0.5335,0.3468,0.6874,0.6874,0.566x+68.333,R2=0.9995,with good linear relationship in the range of 20.08~321.28μg·mL^(-1).The contents of hypoxanthine in Geosaurus from Guangdong,Henan,Hainan,Shanghai and Guangxi were 0.5335,0.3468,0.6874,0.5694,0.7945(mg·g^(-1)),and the contents of inosine were 1.5625,0.5674,1.1267,0.8436,1.2849(mg·g^(-1)).Conclusion:The established HPLC method o

关 键 词：HPLC 地龙 蚯蚓 次黄嘌呤 肌苷 

分 类 号：R917[医药卫生—药物分析学]