硝西泮及其片剂中有关物质的液相色谱电喷雾质谱联用分析

目的: 建立HPLC-MS法测定硝西泮原料及片剂中3种杂质。 方法: 采用Shimadzu VP-ODS柱(250 mm×4.6 mm,5 μm),以乙腈:10 mmol·L^-1磷酸二氢钾(45∶55)为流动相,流速1.0 mL·min^-1,柱后分流;进样量10 μL;DAD检测器,检测波长220 nm。电喷雾电离源(ESI),雾化器干燥气流速8 mL·min^-1,干燥气温度:350 ℃,静电喷雾电压3500 V;质谱扫描质量范围50~1200 m/z。采用全扫描一级质谱和选择离子全扫描二级质谱(MS/MS)2种方式同时测定。 结果: 硝西泮与各杂质峰均能良好的分离,杂质A、B 、C在0.24~30.0 μg·mL^-1浓度范围内与峰面积成良好的线性关系,回归方程分别为Y=3.411×10⁴X+384.1, Y=2.411×10⁴ X-258.4, Y=8.982×10⁴ X-1.378×10³,r均为1.000,最低检测限分别为0.24 ng、0.36 ng、0.24 ng,高、中、低三浓度的平均回收率分别为100.8%,99.4%,101.3%;并对硝西泮的未知降解产物进行结构分析。 结论: 该方法简便、灵敏、专属性好,硝西泮和3种已知杂质有良好的分离。

Objective: To establish an HPLC-MS method for the determination of three impurities in nitrazepam and its tablets. Method: The separation was performed on a Shimadzu VP-ODS(250 mm×4.6 mm,5 μm) column with 10 mmol·L^-1 potassium dihydrogen phosphate solution-acetonitrile(55∶45) as mobile phase at the flow rate of 1.0 mL·min^-1,diode-array detector wavelength was set at 220 nm;ion trap mass spectrometey,spray capillary voltage 3.5 kV,dry gas N₂ 8 mL·min^-1,dry gas temperature 350 ℃,scan range 50-1200 m/z,Sample was analyzed in two modes:MS and MS/MS full scan modes. Results: A good separation for nitrazepam and three impurities was achieved.The linear ranges of impurities A,B and C from 0.24 μg·mL^-1 to 30.0 μg·mL^-1.The average recoveries of impurities A,B and C at three levels were 100.8%,99.4% and 101.3%,respectively. Conclusion: The established HPLC method can be used for the determination of impurities in nitrazepam and its tablets.

1 TANG Xi-lang(唐细兰),SONG Xiang-zhi(宋湘芝),CHEN Zi-he(陈子和),et al.Simultaneous determination of 4 kinds of benzodiazepines in serum by HPLC(高效液相色谱法同时测定血清中4种苯二氮类药物浓度).Guangdong Pham J(广东药学),2001,11(6):39
2 MAO Gui-fu(毛桂福).HPLC determination of diazepam,nitrazepam and clonazepam in human plasma(高效液相色谱法同时测定人血浆中地西泮、硝西泮、氯硝西泮).Chin Hosp Pharm J(中国医院药学杂志),2004,24(11):684
3 LIU Xiang-chen(刘香臣).Determination of Nitrazepam in human serum by RP-HPLC and its clinical applicability(RP-HPLC法测定血清硝基安定及临床适用性).Chin Pham Aff(中国药事),2005,19(1):56
4 TAN Sheng-jian(谭生建),LIU Gang(刘刚),ZHANG Jie(张捷).Determination of phenobarbital chlorphenamine maleate and diazepam in compound benxiaonam in tablets by HPLC(HPLC法测定复方苯硝那敏片中苯巴比妥马来酸氯苯那敏和硝西泮含量).Pham J Chin PLA(解放军药学学报),2007,23(5):384
5 BP.2009.VolⅡ:1457
6 ChP(中国药典).2010.Vol Ⅱ(二部):942
7 JP.2006:925
8 LIU Jing(刘菁),CAI Mei(蔡梅),CAI Mei-Ming(蔡美明),et al.HPLC determination of impurities in nitrazepam and its tablets(HPLC 法测定硝西泮及其片剂中有关物质).Chin J Pharm Anal(药物分析杂质),2010,30(8):1477