HPLC同时测定百令胶囊中5种核苷类成分的含量

Simultaneous Determination of Five Nucleosides in Bailing Capsule by HPLC

摘要:

目的: 建立HPLC同时测定百令胶囊中5种核苷类成分尿苷酸、尿苷、腺苷酸、鸟苷和腺苷含量的方法。方法: 采用Ultimate AQ C₁₈色谱柱(4.6 mm×250 mm, 5 μm),以甲醇-0.05 mol·L^-1磷酸二氢钾溶液为流动相进行梯度洗脱,流速1.0 mL·min^-1,检测波260 nm,柱温25 ℃。结果: 在该色谱条件下,尿苷酸、尿苷、腺苷酸、鸟苷和腺苷分离良好,且分别 在35.25~528.8(r=1.000),38.07~571.1(r=1.000),34.96~524.5(r=0.999 8),38.34~575.1(r=1.000),39.84~597.6 ng (r=1.000)线性关系良好。加样回收率分别为101.2%,103.1%,96.7%,101.4%,103.2%,RSD分别为1.9%,1.1%,1.3%,2.7%,1.6%。结论: 该HPLC含量测定方法结果准确、可靠,具有良好的专属性、精密度、重复性和稳定性,可用于百令胶囊的质量控制。

Abstract:

Objective: To establish an HPLC method for simultaneous determination of five nucleosides, uridine 5'-monophosphate, uridine, adenosine 5'-monophosphate monohydrate, guanosine, adenosine in Bailing capsules. Method: The separation was developed on an Ultimate AQ C₁₈ column (4.6 mm×250 mm, 5 μm) by gradient elution with methanol-0.05 mol·L^-1 potassium dihydrogen phosphate solution, with flow rate at 1.0 mL·min^-1 and column temperature at 25 ℃. The detection wavelength was at 260 nm. Result: The baseline separation of uridine 5'-monophosphate, uridine, adenosine 5'-monophosphate monohydrate, guanosine, and adenosine was successfully achieved. The results showed that these five ingredients had good linearity within the test ranges of 35.25-528.8(r=1.000), 38.07-571.1(r=1.000), 34.96-524.5(r=0.999 8), 38.34-575.1(r=1.000) and 39.84-597.6 ng(r=1.000), respectively. The recovery were 101.2% (RSD 1.9%), 103.1% (RSD 1.1%), 96.7% (RSD 1.3%), 101.4% (RSD 2.7%) and 103.2% (RSD 1.6%), respectively. Conclusion: The developed method of the HPLC-fingerprint and quantitative analysis showed good specificity, precision, reproducibility, stability and could be used for the quality control of Bailing capsules.