反相高效液相色谱法测定注射用胸腺法新含量

RP-HPLC determination of thymalfasin for injection

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,5μm);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:10μL;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 0μg,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： Alhima C 18 column （4. 6 mm × 250 mm ,5 μm）,with mobile phase A consisted of acetonitrilewater（ 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- 1 ; 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.