HPLC法测定吉非替尼原料药有关物质

目的:建立加校正因子的主成分自身对照法测定吉非替尼原料药中有关物质的含量。方法:采用Inertsil ODS-3(3.0 mm×100 mm,3μm)色谱柱,以乙腈-0.05 mol·L^-1的醋酸盐(用冰醋酸调节pH至4.5)为流动相进行梯度洗脱,流速0.5 mL·min^-1,柱温50℃,检测波长248 nm,进样量5μL。测定吉非替尼和杂质SM1、I、XV、SM2、XII、D和C的线性方程,以斜率计算杂质相对于吉非替尼的校正因子,并进行方法学验证。结果:在吉非替尼与相邻杂质(杂质SM2和杂质XII)及各已知杂质之间的分离度均大于1.5。杂质SM1、I、XV、SM2、XII、D和C在各自的线性范围内线性关系良好(r > 0.9990,n=6),相对校正因子分别为0.8、2.3、1.2、4.0、1.1、0.8和1.1,检测限分别为0.02、0.27、0.04、0.31、0.05、0.10和0.05 ng,定量限分别为0.15、0.06、0.54、0.13、0.62、0.15、0.31和0.15 ng,低、中、高3种浓度的平均回收率(n=9)分别为99.1%、93.5%、96.8%、100.3%、102.8%、103.4%和97.9%,RSD分别为1.8%、5.4%、4.3%、2.3%、3.3%、3.1%和3.8%。结论:经验证,该方法的专属性、线性等良好,可采用加校正因子的主成分自身对照法对本品的有关物质进行控制。

Objective: To establish a self-control method of principle component using the correction factor for the determination of related substances in gefitinib raw material.Methods: HPLC was adopted on an Inertsil ODS-3 column(3.0 mm×100 mm, 3 μm) with a gradient elution system of acetonitrile-0.05 mol·L^-1 acetic acid buffer solution(pH was adjusted to 4.5 with glacial acetic acid) with a flow rate of 0.5 mL·min^-1, while the column temperature was maintained at 50℃, the detection wavelength was set at 248 nm and the injection volume was 5 μL.The slope of linear equation was used to determine the correction factor between impurities SM1, I, XV, SM2, XII, D, C and gefitinib.The developed method was validated.Results: The resolutions between gefitinib and adjacent impurities(impurities SM2 and XII), and between the known impurities were greater than 1.5.The calibration curves of seven known impurities were linear in the self-concentration range(r > 0.999 0, n=6).The relative correction factors were 0.8, 2.3, 1.2, 4.0, 1.1, 0.8 and 1.1 for impurities SM1, I, XV, SM2, XII, D and C; the detection limits were 0.02, 0.27, 0.04, 0.31, 0.05, 0.10 and 0.05;the quantitation limits were 0.15, 0.06, 0.54, 0.13, 0.62, 0.15, 0.31 and 0.15 ng;and the average recovery rates(n=9) were 99.1%, 93.5%, 96.8%, 100.3%, 102.8%, 103.4% and 97.9% while RSDs were 1.8%, 5.4%, 4.3%, 2.3%, 3.3%, 3.1% and 3.8% at low, medium, and high concentrations, respeetively.Conclusion: The method is proved that it has good specificity and linearity. The proposed main component self-control method with correction factors is suitable for the quality control of this drug.