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目的:建立同时测定虎力散片剂中7个核苷(胞苷、尿苷、腺苷、2-脱氧尿苷、鸟苷、2-脱氧鸟苷、胸苷)和3个碱基(腺嘌呤、次黄嘌呤、黄嘌呤)类成分含量的HPLC方法。方法:采用Waters XselectTMHss C18(4.6 mm×250 mm,5μm)色谱柱,以甲醇(A)-0.1%磷酸水溶液(B)为流动相,梯度洗脱,流速0.6 m L·min-1,柱温30℃,检测波长260 nm。结果:腺嘌呤、胞苷、次黄嘌呤、黄嘌呤、尿苷、腺苷、2-脱氧尿苷、鸟苷、2-脱氧鸟苷、胸苷质量浓度分别在9.07~181.44、5.03~100.50、2.03~40.60、1.54~30.87、3.57~71.47、1.25~25.08、3.01~60.24、2.02~40.48、0.52~10.40、1.05~21.00μg·m L-1范围内与峰面积呈良好线性关系(r≥0.9994);平均回收率(n=6)为96.2%~103.5%,RSD≤2.6%。结论:本试验所建立的用于同时测定虎力散片剂中核苷及碱基类成分含量的分析方法简单、重复性好、准确高,可为虎力散片剂的质量控制提供依据。
Objective: To establish a method for the simultaneous determination of 7 nucleosides (cytidine, uridine, adenosine, 2-deoxyuridine, guanosine, 2-deoxyguanosine and thymidine) and 3 bases Purine, hypoxanthine, xanthine) class content of HPLC method. METHODS: The mobile phase was eluted with a gradient of methanol (A) - 0.1% phosphoric acid (B) with a flow rate of 0.6 m L · min-1 using a Waters XselectTM Hss C18 column (4.6 mm × 250 mm, 30 ℃, detection wavelength 260 nm. Results: The concentrations of adenine, cytidine, hypoxanthine, xanthine, uridine, adenosine, 2-deoxyuridine, guanosine, 2-deoxyguanosine and thymidine were 9.07-181.44 and 5.03-100.50, 2.03 ~ 40.60,1.54 ~ 30.87,3.57 ~ 71.47,1.25 ~ 25.08,3.01 ~ 60.24,2.02 ~ 40.48,0.52 ~ 10.40,1.05 ~ 21.00μg · m L-1 and the peak area showed a good linear relationship (r≥0.9994 ); Average recovery (n = 6) was 96.2% ~ 103.5%, RSD≤2.6%. Conclusion: The assay established in this study for the simultaneous determination of nucleosides and bases in Huli Powder tablets is simple, reproducible and accurate, which can provide the basis for the quality control of Hu Li San tablets.