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目的建立鹿茸血酒中嘌呤类物质的高效液相色谱分析方法。方法用高氯酸水解过滤制备测试样,以0.02mol/LKH2PO4-H3PO4的缓冲溶液(pH=4.00)为流动相,试样流经EclipseXDB-C18反相色谱柱,在流速为1.00ml/min、检测紫外波长为254.4nm、柱温为25℃时的条件下,用高效液相色谱法(HPLC)分析测定鹿茸血酒中的嘌呤类物质(鸟嘌呤、腺嘌呤、黄嘌呤、次黄嘌呤、5’-鸟苷酸和5’-腺苷酸)。结果嘌呤类物质(鸟嘌呤、腺嘌呤、黄嘌呤、次黄嘌呤、5’-鸟苷酸和5’-腺苷酸)在0.1～50mg/L浓度范围内,线性关系良好(r≥0.995)。方法的最低检出限(LOQ)和定量限(LOD):鸟嘌呤为2.42mg/L和7.26mg/L、腺嘌呤为4.01mg/L和12.03mg/L、黄嘌呤为1.04mg/L和3.12mg/L、次黄嘌呤为1.99mg/L和5.97mg/L、5’-鸟苷酸为1.54mg/L和4.62mg/L、5’-腺苷酸为1.10mg/L和3.30mg/L,各组份精密度(n=10)为3.9%～4.9%,加标回收率:5’-腺苷酸(5’-AMP)为95.0%～101.0%、5’-鸟苷酸(5’-GMP)为93.5%～99.3%、腺嘌呤为93.8%～101.8%,鸟嘌呤为94.9%～101.0%,黄嘌呤为92.0%～99.5%,次黄嘌呤为92.6%～99.6%。结论该方法具有操作高效,快速,准确的特点,适用于鹿茸血酒中嘌呤类物质的定量测定。
Objective To establish a method for the determination of purines in antler blood wine by high performance liquid chromatography. Methods The samples were prepared by perchloric acid hydrolysis and filtration. The mobile phase was 0.02mol / L KH2PO4-H3PO4 buffer solution (pH = 4.00). The sample was passed through EclipseXDB-C18 reversed-phase column and the flow rate was 1.00ml / The detection of purine (guanine, adenine, xanthine, hypoxanthine, and xanthine) in antler blood wine by high performance liquid chromatography (HPLC) under the condition of UV wavelength 254.4nm and column temperature 25 ℃. 5’-guanylate and 5’-adenylate). Results The purine compounds (guanine, adenine, xanthine, hypoxanthine, 5’-guanylic acid and 5’-adenylate) showed good linearity (r≥0.995) in the range of 0.1-50 mg / . The limits of detection (LOQ) and the limit of quantification (LOD) of the method were 2.42 mg / L and 7.26 mg / L for guanine, 4.01 and 12.03 mg / L for adenine and 1.04 mg / L for xanthine and 3.12 mg / L, hypoxanthine 1.99 mg / L and 5.97 mg / L, 5’-guanylic acid 1.54 mg / L and 4.62 mg / L, 5’-adenosine acid 1.10 mg / L and 3.30 mg / L, and the precision of each component (n = 10) was 3.9% ~ 4.9%. The spiked recoveries were 95.0% ~ 101.0% for 5’-AMP and 5’-guanylic acid (5’-GMP) of 93.5% -99.3%, adenine of 93.8% -101.8%, guanine of 94.9% -101.0%, xanthine of 92.0% -99.5% and hypoxanthine of 92.6% -99.6%. Conclusion The method has the characteristics of high efficiency, rapidity and accuracy. It is suitable for the quantitative determination of purines in antler blood wine.