反相高效液相色谱法测定注射用胸腺法新含量

目的: 建立反相高效液相色谱法测定注射用胸腺法新含量,本方法区别于中国药典2010年版二部相应品种下的含量测定法。 方法: 选用Alltima C₁₈柱(4.6 mm×250 mm,5 μm);流动相A:乙腈-水[140:860](含0.072%四甲基氢氧化铵,用磷酸调节pH为2.5),流动相B:乙腈-水[250:750](含0.072%四甲基氢氧化铵,用磷酸调节pH为2.5),采用梯度洗脱,使主峰保留时间在17~23 min;流速:1.0 mL·min^-1;紫外检测波长:210 nm;进样量:10 μL;柱温:室温。 结果: 本方法重复性好,稳定性佳;线性范围为0.010~2 0 μg,r=0.9999,最低检测限为4.0 ng;平均回收率为101.4%,RSD为0.60%(n=6)。试验结果表明,本法优化了主峰与其后紧邻杂质峰的分离度,可以将与主峰相对保留时间为1.07的杂质峰分出,使主峰的峰形得以改善,而采用药典方法无法做到这一点,故含量测定所得结果更加准确。 结论: 采用本方法主峰与杂质峰的分离度好,结果准确可靠,可用于注射用胸腺法新含量的测定。

Objective: To establish HPLC method which is different from the method described in Chinese Pharmacopoeia (2010 Edition) method for the determination of Thymalfasin for injection. Method: Alltima C₁₈column(4.6 mm×250 mm,5 μm),with mobile phase A consisted of acetonitrile-water(140∶860) containing 0.072% Tetramethyl ammonium hydroxide,adjust to pH 2.5 by adding phosphoric acid,and mobile phase B consisted of acetonitrile-water(250∶750) containing 0.072% tetramethyl ammonium hydroxide,adjust to pH 2.5 by adding phosphoric acid,gradient elution;the flow rate was 1.0 mL·min;detection wavelength at 210 nm,the injection volume was 10 μL,and the column temperature was room tempreture. Results: The linear range of the method was 0.010-20 μg (r=0.9999),the limit of detection was 4.0 ng,the average recovery was 101.4%,with RSD of 0.60%(n=6). Conclusion: The method has excellent resolution for the related substance,and is sensitive and accurate,can be used for the determination of thymalfasin for injection.

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