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目的:建立高效离子色谱法测定骨瓜提取物注射液中3种核苷及4种核苷酸有关物质的含量。方法:使用Thermo Fisher色谱柱(Dionex Ion Pac AS16,250 mm×4 mm),保护柱(Dionex Ion Pac AG16,50 mm×4 mm);流动相为水-250 mmol·L-1氢氧化钠溶液,梯度洗脱,流速1.0 m L·min-1,柱温为35℃,紫外检测器(PDA),检测波长260 nm,进样体积为50μL。结果:该方法可以在35 min内,同时定量测定尿嘧啶核苷、腺嘌呤核苷、次黄嘌呤核苷、胞嘧啶核苷酸、腺嘌呤核苷酸、尿嘧啶核苷酸、次黄嘌呤核苷酸;各待测物质量浓度在1.0~50.0μg·m L-1范围内线性关系良好,回归方程的线性相关系数均大于0.999 7;平均回收率(n=9)分别为尿嘧啶核苷105.7%、腺嘌呤核苷96.0%、次黄嘌呤核苷99.4%、腺嘌呤核苷酸100.1%、尿嘧啶核苷酸97.9%、次黄嘌呤核苷酸98.0%,回收率RSD分别为1.5%、0.8%、5.1%、1.3%、2.2%、2.7%。结论:该方法简单、快速、准确,可用于骨瓜提取物注射液中核苷及核苷酸的测定。
Objective: To establish an HPLC method for the determination of three kinds of nucleosides and four kinds of nucleotides-related substances in the extract of bone pumpkin extract. METHODS: Thermo Fisher columns (Dionex Ion Pac AS16, 250 mm × 4 mm) and guard column (Dionex Ion Pac AG16, 50 mm × 4 mm) were used. The mobile phase was water-250 mmol·L -1 sodium hydroxide solution , Gradient elution at a flow rate of 1.0 m L · min-1 with a column temperature of 35 ° C and a UV detector (PDA) at a detection wavelength of 260 nm with a sample volume of 50 μL. Results: This method can simultaneously measure uridine, adenosine, inosine, cytosine, adenine, uracil, hypoxanthine The linear correlation coefficients of regression equation were all above 0.999 7. The average recoveries (n = 9) were uracil nucleus Glycosides 105.7%, adenosine 96.0%, inosine 99.4%, adenine nucleotide 100.1%, uracil nucleotide 97.9%, and inosine 98.0%. The recoveries RSD were 1.5 %, 0.8%, 5.1%, 1.3%, 2.2%, 2.7%. Conclusion: The method is simple, rapid and accurate and can be used for the determination of nucleosides and nucleotides in the extract of Bonegua extract.