高效离子色谱法测定骨瓜提取物注射液中核苷及核苷酸有关物质的含量

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ThermoFisher色谱柱(Dionex IonPac AS16,250 mm×4 mm),保护柱(Dionex IonPac 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 mL·min^-1,柱温为35℃,紫外检测器(PDA),检测波长260 nm,进样体积为50 μL。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1.0~50.0 μg·mL^-1范围内线性关系良好,回归方程的线性相关系数均大于0.9997;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 develop a method for determining contents of related substances of three nucleosides and four nucleotides in osseous and cucumis extract injection by using high performance ion chromatography. Methods:The ion chromatographic column (Dionex IonPac AS16,250 mm×4 mm) and the guard column (Dionex IonPac AG16,50 mm×4 mm) were adopted, the mobile phase was water-250 mmol·L-1 sodium hydroxide solution with gradient elution at a flow rate of 1.0 mL·min^-1, the column temperature was 35℃, the photo-diode array detector (PDA) was used with the detection wavelength of 260 nm, and the injection volume was 50 μL. Results:In 35 minutes, this method simultaneously determined nucleosides and nucleotides, including uridine, adenosine, inosine, cytidine monophosphate, adenosine 5'-monophosphate, uridylic acid, and inosine 5'-monophosphate. The linear relation of each component was excellent within the range of 1.0-50.0 μg·mL^-1 (r≥0.999 7), the recoveries (n=9) were 105.7% for uridine, 96.0% for adenosine, 99.4% for inosine, 100.1% for adenosine 5'-monophosphate, 97.9% for uridylic acid, 98.0% for inosine 5'-monophosphate with RSD of 1.5%, 0.8%, 5.1%, 1.3%, 2.2%, 2.7% respectively. Conclusion: This method was simple,rapid and accurate in determining nucleosides and nucleotides in osseous and cucumis extract injection.