HPLC法测定罗氟司特的有关物质
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摘要
目的:建立罗氟司特有关物质测定的HPLC方法。方法:采用十八烷基硅烷键合硅胶为填料的色谱柱,以0.01 mol·L~(-1)磷酸二氢钾溶液(磷酸调节p H至3.5±0.1)为流动相A,乙腈为流动相B,梯度洗脱,流速0.5 m L·min~(-1),柱温为25℃,检测波长为215 nm,对有关物质进行定性、定量分析。结果:罗氟司特和相邻杂质以及各杂质之间的分离度均大于1.5;3-环丙基甲氧基-4-二氟甲氧基苯甲酸(杂质A)、4-氨基-3,5-二氯吡啶(杂质B)、3-(环丙基甲氧基)-N-(3,5-二氯吡啶-4-基)-4-羟基苯甲酰胺(杂质C)、3-羟基-N-(3,5-二氯吡啶-4-基)-4-(二氟甲氧基)苯甲酰胺(杂质D)、3,4-二羟基-N-(3,5-二氯吡啶-4-基)苯甲酰胺(杂质E)、3-(环丙基甲氧基)-N-(吡啶-4-基)-4-(二氟甲氧基)苯甲酰胺(杂质F)、3-(环丙基甲氧基)-N-(3-氯吡啶-4-基)-4-(二氟甲氧基)苯甲酰胺(杂质G)、3-(二氟甲氧基)-N-(3,5-二氯吡啶-4-基)-4-(环丙基甲氧基)苯甲酰胺(杂质H)、3,4-二(环丙基甲氧基)-N-(3,5-二氯吡啶-4-基)苯甲酰胺(杂质I)、3,4-二(二氟甲氧基)-N-(3,5-二氯吡啶-4-基)苯甲酰胺(杂质J)和罗氟司特氮氧化物(杂质K)在各自的线性范围内线性关系良好(r>0.999 0,n=5),相对校正因子分别为1.14、1.00、1.17、0.96、0.70、1.76、1.08、1.37、1.14、1.65和1.22,检测限分别为0.1、0.03、0.1、0.1、0.1、0.03、0.1、0.1、0.1、0.1和0.1 ng;定量限分别为0.3、0.1、0.3、0.2、0.2、0.1、0.2、0.2、0.2、0.3和0.3 ng,低、中、高3种浓度的平均回收率(n=9)分别为102.9%、100.0%、107.6%、107.5%、98.1%、99.9%、98.3%、97.6%、86.9%、100.0%和98.7%。5批样品有关物质测定结果显示,各已知杂质的含量均低于0.1%,其他最大单个杂质的含量均低于0.1%,杂质总量均低于0.5%。结论:本方法可用于罗氟司特的有关物质测定。
Objective:To establish an HPLC method for the determination of related substances in roflumilast raw material.Methods:HPLC was adopted on a C_(18) column with a gradient elution system of acetonitrile-0.01 mol·L~(-1) potassium dihydrogen phosphate solution(p H was adjusted to 3.5±0.1 with phosphate acid)with a flow rate of 0.5 m L·min~(-1),and the column temperature was maintained at 25 ℃,the detection wavelength was set at 215 nm.The related substances were qualified and quantified by the established method.Results:The resolutions between roflumilast and adjacent impurities,and between the known impurities were greater than 1.5.The calibration curves of 11 known impurities were linear in the concentration range(r>0.999 0,n=5).The relative correction factors were 1.14,1.00,1.17,0.96,0.70,1.76,1.08,1.37,1.14,1.65 and 1.22 for 3-(cyclopropylmethoxy)-4-(difluoromethoxy)benzoic acid(impurity A),3,5-dichloropyridin-4-amine(impurity B),3-(cyclopropylmethoxy)-N-(3,5-dichloropyridin-4-yl)-4-hydroxybenzamide(impurity C),N-(3,5-dichloropyridin-4-yl)-4-(difluoromethoxy)-3-hydroxybenzamide(impurity D),N-(3,5-dichloropyridin-4-yl)-3,4-dihydroxybenzamide(impurity E),3-(cyclopropylmethoxy)-4-(difluoromethoxy)-N-(pyridin-4-yl)benzamide(impurity F),N-(3-chloropyridin-4-yl)-3-(cyclopropylmethoxy)-4-(difluoromethoxy)benzamide(impurity G),4-(cyclopropylmethoxy)-N-(3,5-dichloropyridin-4-yl)-3-(difluoromethoxy)benzamide(impurity H),3,4-bis(cyclopropylmethoxy)-N-(3,5-dichloropyridin-4-yl)benzamide(impurity I),N-(3,5-dichloropyridin-4-yl)-3,4-bis(difluoromethoxy)benzamide(impurity J)and roflumilast oxynitride(impurity K),rspectively;The detection limits were 0.1,0.03,0.1,0.1,0.1,0.03,0.1,0.1,0.1,0.1 and 0.1 ng,and the quantitation limits were 0.3,0.1,0.3,0.2,0.2,0.1,0.2,0.2,0.2,0.3 and 0.3 ng,respectively;And the average recovery rates(n=9)were 102.9%,100.0%,107.6%,107.5%,98.1%,99.9%,98.3%,97.6%,86.9%,100.0% and 98.7% at the concentration level of low,medium and high concentrations,respectively.The determination results of the five batches of samples showed that the known impurities were less than 0.1%,the maximum single unknown impurity was less than 0.1%,the total impurities were less than 0.5%.Conclusion:The method is suitable for the quality control of roflumilast.
Objective:To establish an HPLC method for the determination of related substances in roflumilast raw material.Methods:HPLC was adopted on a C_(18) column with a gradient elution system of acetonitrile-0.01 mol·L~(-1) potassium dihydrogen phosphate solution(p H was adjusted to 3.5±0.1 with phosphate acid)with a flow rate of 0.5 m L·min~(-1),and the column temperature was maintained at 25 ℃,the detection wavelength was set at 215 nm.The related substances were qualified and quantified by the established method.Results:The resolutions between roflumilast and adjacent impurities,and between the known impurities were greater than 1.5.The calibration curves of 11 known impurities were linear in the concentration range(r>0.999 0,n=5).The relative correction factors were 1.14,1.00,1.17,0.96,0.70,1.76,1.08,1.37,1.14,1.65 and 1.22 for 3-(cyclopropylmethoxy)-4-(difluoromethoxy)benzoic acid(impurity A),3,5-dichloropyridin-4-amine(impurity B),3-(cyclopropylmethoxy)-N-(3,5-dichloropyridin-4-yl)-4-hydroxybenzamide(impurity C),N-(3,5-dichloropyridin-4-yl)-4-(difluoromethoxy)-3-hydroxybenzamide(impurity D),N-(3,5-dichloropyridin-4-yl)-3,4-dihydroxybenzamide(impurity E),3-(cyclopropylmethoxy)-4-(difluoromethoxy)-N-(pyridin-4-yl)benzamide(impurity F),N-(3-chloropyridin-4-yl)-3-(cyclopropylmethoxy)-4-(difluoromethoxy)benzamide(impurity G),4-(cyclopropylmethoxy)-N-(3,5-dichloropyridin-4-yl)-3-(difluoromethoxy)benzamide(impurity H),3,4-bis(cyclopropylmethoxy)-N-(3,5-dichloropyridin-4-yl)benzamide(impurity I),N-(3,5-dichloropyridin-4-yl)-3,4-bis(difluoromethoxy)benzamide(impurity J)and roflumilast oxynitride(impurity K),rspectively;The detection limits were 0.1,0.03,0.1,0.1,0.1,0.03,0.1,0.1,0.1,0.1 and 0.1 ng,and the quantitation limits were 0.3,0.1,0.3,0.2,0.2,0.1,0.2,0.2,0.2,0.3 and 0.3 ng,respectively;And the average recovery rates(n=9)were 102.9%,100.0%,107.6%,107.5%,98.1%,99.9%,98.3%,97.6%,86.9%,100.0% and 98.7% at the concentration level of low,medium and high concentrations,respectively.The determination results of the five batches of samples showed that the known impurities were less than 0.1%,the maximum single unknown impurity was less than 0.1%,the total impurities were less than 0.5%.Conclusion:The method is suitable for the quality control of roflumilast.
引文
[1]BOLAND S,ALEN J,BOURIN A,et al.Novel roflumilastanalogsas soft PDE4 inhibitors[J].Bioorg Med Chem Lett,2014,24(18):4594
[2]闫文娜,陈艳明,倪成亮,等.罗氟司特中有关物质的鉴定和含量测定[J].药物分析杂志,2013,33(10):1726YAN WN,CHEN YM,NI CL,et al.Identification and quantitative analysis of related substances in roflumilast[J].Chin J Pharm Anal,2013,33(10):1726
[3]MILARA J,MORELL A,BALLESTER B,et al.Roflumilast improves corticosteroid resistance COPD bronchial epithelial cells stimulated with toll like receptor 3 agonist[J].Respir Res,2015,16(1):12
[4]马培奇.慢性阻塞性肺病治疗新药罗氟司特[J].上海医药,2011,32(4):201MA PQ.Roflumilast:a novel approach to treating COPD[J].Shanghai Med Pharm J,2011,32(4):201
[5]封宇飞.磷酸二酯酶-4抑制剂罗氟司特的药理与临床研究新进展[J].中国新药杂志,2011,20(14):1257FENG YF.Progress in pharmacological and clinical studies of the phosphodiesterase-4 inhibitor roflumilast[J].Chin J New Drugs,2011,20(14):1257
[6]祝巍,陈萍.磷酸二酯酶抑制药-罗氟司特在慢性阻塞性肺疾病治疗中的应用[J].中国新药与临床杂志,2011,30(11):819ZHU W,CHEN P.Application of phosphodiesterase inhibitorroflumilastin chronic obstructive pulmonary disease[J].Chin J New Drugs Clin Rem,2011,30(11):819
[7]韩伟,周新.慢性阻塞性肺疾病的治疗新选择-罗氟司特[J].世界临床药物,2011,32(7):396HAN W,ZHOU X.Roflumilast,a highly selective PDE-4 inhibitor in the treatment of chronic obstructive pulmonary disease[J].World Clin Drugs,2011,32(7):396
[8]王江玲,梁建华.HPLC法测定罗氟司特有关物质[J].河南大学学报(医学版),2014,33(4):253WANG JL,LIANG JH.Determination of related substances in roflumilast by HPLC[J].J Henan Univ(Med Sci),2014,33(4):253
[9]郑曦孜.HPLC法测定罗氟司特的有关物质[J].光谱实验室,2013,30(6):3122ZHENG XM.Determination of related substances in roflumilast by HPLC[J].Chin J Spectrosc Lab,2013,30(6):3122
[10]张莉,黎其荣,杨伟涵,等.HPLC法测定罗氟司特有关物质方法研究[J].今日药学,2014,24(2):114ZHANG L,LI QR,YANG WH,et al.HPLC determination of roflumilast related substances[J].Pharm Today,2014,24(2):114
[11]赵永娟,付建,赵海桥.HPLC法测定罗氟司特片有关物质[J].中国化工贸易,2013,12(12):321ZHAO YJ,FU J,ZHAO HQ.Determination of related substances in roflumilast tablets by HPLC[J].China Chem Trade,2013,12(12):321
[12]杨丽霞,闫建军,冯彩婷.高效液相色谱法测定罗氟司特药品中罗氟司特和4种杂质[J].理化检测(化学分册),2015,51(5):594YANG LX,YAN JJ,FENG CT.HPLC determination of roflumilast and 4 impurities in roflumilast drug[J].Phys Test Chem Anal Part B(Chem Anal),2015,51(5):594
[13]何凤琴,周磊,秦妮,等.罗氟司特主药和有关物质的HPLC法测定[J].医学美学美容,2013(3):155HE FQ,ZHOU L,QIN N,et al.Determination of roflumilast and related substances by HPLC[J].Med Aesthet Cosmetol,2013(3):155
[14]BARHATE VD,DEOSTHALEE PC.Validation of rapid liquid chromatographic method for the determination of roflumilast[J].JChem Pharm Res,2011,3(2):770
[15]BARHATE VD,DEOSTHALEE P.Rapid liquid chromatographic method for the determination of roflumilast in the presence of degradation products[J].Indian J Pharm Sci,2010,72(3):401(本文于2016年7月4日收到)
[2]闫文娜,陈艳明,倪成亮,等.罗氟司特中有关物质的鉴定和含量测定[J].药物分析杂志,2013,33(10):1726YAN WN,CHEN YM,NI CL,et al.Identification and quantitative analysis of related substances in roflumilast[J].Chin J Pharm Anal,2013,33(10):1726
[3]MILARA J,MORELL A,BALLESTER B,et al.Roflumilast improves corticosteroid resistance COPD bronchial epithelial cells stimulated with toll like receptor 3 agonist[J].Respir Res,2015,16(1):12
[4]马培奇.慢性阻塞性肺病治疗新药罗氟司特[J].上海医药,2011,32(4):201MA PQ.Roflumilast:a novel approach to treating COPD[J].Shanghai Med Pharm J,2011,32(4):201
[5]封宇飞.磷酸二酯酶-4抑制剂罗氟司特的药理与临床研究新进展[J].中国新药杂志,2011,20(14):1257FENG YF.Progress in pharmacological and clinical studies of the phosphodiesterase-4 inhibitor roflumilast[J].Chin J New Drugs,2011,20(14):1257
[6]祝巍,陈萍.磷酸二酯酶抑制药-罗氟司特在慢性阻塞性肺疾病治疗中的应用[J].中国新药与临床杂志,2011,30(11):819ZHU W,CHEN P.Application of phosphodiesterase inhibitorroflumilastin chronic obstructive pulmonary disease[J].Chin J New Drugs Clin Rem,2011,30(11):819
[7]韩伟,周新.慢性阻塞性肺疾病的治疗新选择-罗氟司特[J].世界临床药物,2011,32(7):396HAN W,ZHOU X.Roflumilast,a highly selective PDE-4 inhibitor in the treatment of chronic obstructive pulmonary disease[J].World Clin Drugs,2011,32(7):396
[8]王江玲,梁建华.HPLC法测定罗氟司特有关物质[J].河南大学学报(医学版),2014,33(4):253WANG JL,LIANG JH.Determination of related substances in roflumilast by HPLC[J].J Henan Univ(Med Sci),2014,33(4):253
[9]郑曦孜.HPLC法测定罗氟司特的有关物质[J].光谱实验室,2013,30(6):3122ZHENG XM.Determination of related substances in roflumilast by HPLC[J].Chin J Spectrosc Lab,2013,30(6):3122
[10]张莉,黎其荣,杨伟涵,等.HPLC法测定罗氟司特有关物质方法研究[J].今日药学,2014,24(2):114ZHANG L,LI QR,YANG WH,et al.HPLC determination of roflumilast related substances[J].Pharm Today,2014,24(2):114
[11]赵永娟,付建,赵海桥.HPLC法测定罗氟司特片有关物质[J].中国化工贸易,2013,12(12):321ZHAO YJ,FU J,ZHAO HQ.Determination of related substances in roflumilast tablets by HPLC[J].China Chem Trade,2013,12(12):321
[12]杨丽霞,闫建军,冯彩婷.高效液相色谱法测定罗氟司特药品中罗氟司特和4种杂质[J].理化检测(化学分册),2015,51(5):594YANG LX,YAN JJ,FENG CT.HPLC determination of roflumilast and 4 impurities in roflumilast drug[J].Phys Test Chem Anal Part B(Chem Anal),2015,51(5):594
[13]何凤琴,周磊,秦妮,等.罗氟司特主药和有关物质的HPLC法测定[J].医学美学美容,2013(3):155HE FQ,ZHOU L,QIN N,et al.Determination of roflumilast and related substances by HPLC[J].Med Aesthet Cosmetol,2013(3):155
[14]BARHATE VD,DEOSTHALEE PC.Validation of rapid liquid chromatographic method for the determination of roflumilast[J].JChem Pharm Res,2011,3(2):770
[15]BARHATE VD,DEOSTHALEE P.Rapid liquid chromatographic method for the determination of roflumilast in the presence of degradation products[J].Indian J Pharm Sci,2010,72(3):401(本文于2016年7月4日收到)