论文部分内容阅读
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 a HPLC method for the determination of roflumilast related substances. Methods: The chromatographic column was packed with octadecylsilane bonded silica gel. The mobile phase A consisted of 0.01 mol·L -1 potassium dihydrogen phosphate solution (phosphoric acid adjusted p H to 3.5 ± 0.1) and acetonitrile Phase B with gradient elution at a flow rate of 0.5 m L · min -1 .The column temperature was 25 ℃ and the detection wavelength was 215 nm. The related substances were qualitatively and quantitatively analyzed. Results: The separation between roflumilast and adjacent impurities and the impurities were all greater than 1.5; 3-cyclopropylmethoxy-4-difluoromethoxybenzoic acid (impurity A), 4-amino-3 (Impurity B), 3- (cyclopropylmethoxy) -N- (3,5-dichloropyridin-4-yl) -4-hydroxybenzamide Hydroxy-N- (3,5-dichloropyridin-4-yl) -4- (difluoromethoxy) benzamide (Impurity D) 4-yl) benzamide (Impurity E), 3- (cyclopropylmethoxy) -N- (pyridin-4-yl) -4- (difluoromethoxy) benzamide ( Impurity F), 3- (cyclopropylmethoxy) -N- (3-chloropyridin-4- yl) -4- (difluoromethoxy) benzamide (Impurity G), 3- Methoxy) -N- (3,5-dichloropyridin-4-yl) -4- (cyclopropylmethoxy) benzamide (impurity H), 3,4- bis (3,5-dichloropyridin-4-yl) benzamide (Impurity I) 3,4-bis (difluoromethoxy) -N- (3,5-dichloropyridine- (J) and roflumilast oxide (impurity K) showed good linearity (r> 0.999 0, n = 5) in their respective linear ranges with relative correction factors of 1.14, 1.00, 1.17, 0.96, 0.70, 1.76, 1.08, 1.37, 1.14 1.65 and 1.22, 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; the quantitative 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. The average recoveries (n = 9) of low, medium and high concentrations were 102.9%, 100.0%, 107.6%, 107.5%, 98.1%, 99.9%, 98.3%, 97.6% , 86.9%, 100.0% and 98.7% respectively. The results of five batches of samples showed that the contents of all the known impurities were less than 0.1%, the contents of the other largest single impurities were less than 0.1% and the total amount of impurities was less than 0.5%. Conclusion: This method can be used for the determination of roflumilate related substances.