HPCE与HPLC测定科博肽注射液含量的方法比较

胡亦沁; 李元圆; 徐骏军; 李范珠

HPCE与HPLC测定科博肽注射液含量的方法比较

Comparison of Determination of Cobratide Injection by HPCE and HPLC

摘要

摘要: 目的系统地比较高效毛细管电泳法(HPCE)和高效液相色谱法(HPLC)在测定科博肽注射液含量方面的差异。方法HPCE分离条件为：未涂层毛细管柱(50 μm×40 cm，有效长度30 cm)，运行缓冲液为0.1 mol·L^-1磷酸盐缓冲液(pH 2.5)，分离电压20 kV，柱温25 ℃，检测波长200 nm；HPLC分离条件为：Aeris PEPTIDE XB-C₁₈柱(4.6 mm×150 mm，3.6 μm)，柱温30 ℃，流速0.9 mL·min^-1，检测波长200 nm，流动相A：0.1%三氟乙酸-水、流动相B：0.1%三氟乙酸-乙腈，梯度洗脱。结果科博肽在HPLC和HPCE各自的分离条件下进样浓度分别在17.33~138.60 μg·mL^-11、1.08~138.60 μg·mL^-1内表现出良好的线性关系(r均为0.999 7)，平均加样加收率分别为100.3%和99.3%，RSD 分别为0.7%和1.2%。结论 HPCE与HPLC均可用于科博肽注射液的含量测定，但HPCE在使用成本、分离时间、检出限等方面更具有优势。

Abstract: OBJECTIVE To compare HPCE method and HPLC method for determining the content of Cobratide injection. METHODS In HPCE analysis, an uncoated fused silica capillary(50 μm × 40 cm, effective length 30 cm) was used, with 0.1?mol·L^-1 phosphate buffer(pH=2.5) as the running buffer under the voltage of 20 kV at 25 ℃, and the detection wavelength was 200 nm. In HPLC method, the separation was performed on an Aeris PEPTIDE XB-C₁₈ column(4.6 mm×150 mm, 3.6 μm) under the temperature of 30 ℃ with 0.1% TFA-H2O and 0.1% TFA-CH3CN as the mobile phase at flow rate of 0.9 mL·min^-1. The detection wavelength was set at 200 nm. RESULTS The linear range of HPLC and HPCE were 17.33-138.60 μg·mL^-1　(r=0.999 7) and 1.08-138.60 μg·mL^-1(r=0.999 7), respectively. Their average recoveries were 100.3% and 99.3% with RSD of 0.7% and 1.2%, relatively. CONCLUSION Both methods can be applied to determine the content of Cobratide injection, but HPCE earns more benefits on shorter separation time and lower detection limit and cost.

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