HPLC法测定福多司坦原料药及其制剂中有关物质的含量
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摘要
目的:建立福多司坦原料药及其制剂中有关物质含量测定的方法。方法:以国内8家企业生产的福多司坦原料药或制剂为样品。采用高效液相色谱(HPLC)法(外标法)测定杂质A、B、C的含量,色谱柱为MGⅡC_(18),流动相为0.12%己烷磺酸钠溶液(pH 2.0),流速为1.0 mL/min,检测波长为210 nm,柱温为35℃,进样量为20μL。采用HPLC法(加校正因子的主成分自身对照法)测定杂质E、F、G的含量,色谱柱为Altech Altima C_(18),流动相为0.05 mol/L磷酸盐缓冲液-乙腈-水(梯度洗脱),流速为0.5mL/min,检测波长为200 nm,柱温为30℃,进样量为20μL。结果:杂质A、B、C、E、F、G的检测质量浓度线性范围分别为0.446～22.291、0.202～20.158、0.101～12.082、0.111 0～11.100、0.210 4～10.520、0.221 6～11.080μg/mL,检测限分别为5.57、1.01、1.99、2.22、4.21、4.43 ng,定量限分别为11.14、2.02、3.98、4.45、8.42、8.85 ng;杂质E、F、G的校正因子分别为0.91、1.42、1.73,相对保留时间分别为0.88、1.95、3.08;精密度(n=6)、稳定性[杂质A(4 h,n=3),其余各杂质(24 h,n=7)]试验的RSD均小于2.0%,平均加样回收率分别为98.0%、97.3%、102.4%、99.4%、98.9%、96.4%,RSD分别为1.4%、1.5%、1.1%、0.9%、1.2%、0.5%(n=9);8家福多司坦原料药或制剂生产企业中杂质总含量均<1.1%。结论:该方法灵敏度高、专属性好,可用于福多司坦原料药及其制剂中有关物质的定量研究。
OBJECTIVE:To establish the method for content determination of the related substance in fudosteine raw material and its preparations. METHODS:Fudosteine or its preparations produced by 8 domestic enterprises were taken as samples. HPLC method(external standard)was used to determine the contents of impurities A,B and C. The separation was performed on MGⅡC_(18) column with mobile phase consisted of 0.12% sodium hexane sulfonate solution(pH 2.0) at flow rate of 1.0 mL/min. The detection wavelength was set at 210 nm,column temperature was 35 ℃ and sample size was 20 μL. The contents of impurities E,F,G were determined by HPLC method(principal component self-contrast method with correction factor). The separation was performed on Altech Altima C_(18) column with mobile phase consisted of 0.05 mol/L phosphate buffer-acetonitrile-water(gradient elution)at the flow rate of 0.5 m L/min. The detection wavelength was set at 200 nm,and the column temperature was 30 ℃. The sample size was 20 μL. RESULTS:The linear ranges of impurity A,B,C,E,F and G were 0.446-22.291,0.202-20.158,0.101-12.082,0.111 0-11.100,0.210 4-10.520,0.221 6-11.080 μg/mL,respectively. The limits of detection were 5.57,1.01,1.99,2.22,4.21,4.43 ng,respectively. The limits of quantitation were 11.14,2.02,3.98,4.45,8.42,8.85 ng,respectively. The relative correction factors of impurities E,F and G were 0.91,1.42 and 1.73,respectively;their relative retention time were 0.88,1.95 and 3.08. RSDs of precision(n=6)and stability [impurity A(4 h,n=3),other impurities(24 h,n=7)] tests were all lower than 2.0%. The average recoveries were 98.0%,97.3%,102.4%,99.4%,98.9%,96.4%,respectively;RSDs were 1.4%,1.5%,1.1%,0.9%,1.2%,0.5%(n=9),respectively. Total contents of substances in fudosteine raw material or its preparation produced by 8 enterprises were all lower than 1.1%. CONCLUSIONS:Established method is sensitive and specific. The method can be used for the quantitative study on related substances in fudosteine raw material and its preparations.
OBJECTIVE:To establish the method for content determination of the related substance in fudosteine raw material and its preparations. METHODS:Fudosteine or its preparations produced by 8 domestic enterprises were taken as samples. HPLC method(external standard)was used to determine the contents of impurities A,B and C. The separation was performed on MGⅡC_(18) column with mobile phase consisted of 0.12% sodium hexane sulfonate solution(pH 2.0) at flow rate of 1.0 mL/min. The detection wavelength was set at 210 nm,column temperature was 35 ℃ and sample size was 20 μL. The contents of impurities E,F,G were determined by HPLC method(principal component self-contrast method with correction factor). The separation was performed on Altech Altima C_(18) column with mobile phase consisted of 0.05 mol/L phosphate buffer-acetonitrile-water(gradient elution)at the flow rate of 0.5 m L/min. The detection wavelength was set at 200 nm,and the column temperature was 30 ℃. The sample size was 20 μL. RESULTS:The linear ranges of impurity A,B,C,E,F and G were 0.446-22.291,0.202-20.158,0.101-12.082,0.111 0-11.100,0.210 4-10.520,0.221 6-11.080 μg/mL,respectively. The limits of detection were 5.57,1.01,1.99,2.22,4.21,4.43 ng,respectively. The limits of quantitation were 11.14,2.02,3.98,4.45,8.42,8.85 ng,respectively. The relative correction factors of impurities E,F and G were 0.91,1.42 and 1.73,respectively;their relative retention time were 0.88,1.95 and 3.08. RSDs of precision(n=6)and stability [impurity A(4 h,n=3),other impurities(24 h,n=7)] tests were all lower than 2.0%. The average recoveries were 98.0%,97.3%,102.4%,99.4%,98.9%,96.4%,respectively;RSDs were 1.4%,1.5%,1.1%,0.9%,1.2%,0.5%(n=9),respectively. Total contents of substances in fudosteine raw material or its preparation produced by 8 enterprises were all lower than 1.1%. CONCLUSIONS:Established method is sensitive and specific. The method can be used for the quantitative study on related substances in fudosteine raw material and its preparations.
引文
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[2]高橋規行,小北俊夫,矢野憲一,など.SS320A第Ⅰ相臨床試験(第1報)単回経口投与試験[J].臨床医薬,2002,18(1):7-14.
[3]陈光.福多司坦的药理及临床试验[J].泰州职业技术学院学报,2009,9(6):35-38.
[4]章云峰,陈辉,姚晓梅.新型黏痰溶解剂对比氨溴索治疗慢性呼吸道疾病的祛痰效果的系统评价[J].中国药房,2012,23(32):3041-3044.
[5]徐桂清,毛润泽,孙青斌,等.福多司坦合成工艺研究[J].河南师范大学学报,2011,39(3):103-104.
[6]YOSHIKUNI ITOH.Expectorant comprising hydroxyalkyl cysteine derivative:USA,5047428[P].1991-09-10.
[7]国家食品药品监督管理局.福多司坦YBH02592008[S].2008.
[8]国家食品药品监督管理局.福多司坦片YBH02782008[S].2008.
[9]国家食品药品监督管理局.福多司坦胶囊YBH02752008[S].2008.
[10]国家药品审评中心.化学药物杂质研究技术指导原则[EB/OL].(2007-08-23)[2019-01-05].http://www.cde.org.cn/zdyz.do?method=largePage&id=2060.
[11]苏平.HPLC法测定福多司坦口服液的含量和有关物质[J].中国药师,2010,13(6):824-826.
[12]胡士高,高效液相色谱法测定福多司坦胶囊的有关物质[J].中国实用医药,2007,22(8):26-27.
[13]顾霄,罗英,陈悦,等.福多司坦有关物质的亲水作用色谱-质谱结构鉴定[J].药学学报,2017,52(8):1313-1317.
[14]GUPTA PB,RAO BV,SANASI PD,et al.Identification,isolation and characterization of process and degradation impurities of fudosteine and its stability indicating HPLCmethod validation[J].Int J Adv Res,2016,4(7):453-461.
[15]余振喜,庾莉菊,黄海伟,等.浅谈HPLC法测定有关物质时已知杂质的计算方法[J].中国药品标准,2010,11(4):278-282.