LC-MS法同时测定人血浆中对乙酰氨基酚和金刚烷胺的浓度及其制剂的人体生物等效性研究

Simultaneous Determination of Paracetamol and Amantadine in Human Plasma and Bioequivalence of Its Preparations in Healthy Volunteers by LC-MS

目的:建立液-质联用(LC-MS)法同时测定人血浆中对乙酰氨基酚和金刚烷胺的浓度,并应用于小儿氨酚烷胺泡腾片的人体生物等效性研究。方法:人血浆样本以甲醇沉淀蛋白后进样测定,色谱柱为Shim-packVP-ODS,流动相为甲醇-5mmol·L-1乙酸铵测(,含电喷0.5雾%离甲子酸化)(源45,∶正55)离,子流方速式为,0选.3择mL监·测mi离n-子1;选反用应分AP别I3为20m0/型z1三52重.2四→极11杆0.1(串对联乙质酰谱氨仪基的酚多)重、m反/z应15监2.测3(→M13R5M.2()金扫刚描烷方胺式)进和行m监/z166.1→148.1(伪麻黄碱,内标)。结果:乙酰氨基酚和金刚烷胺血药浓度分别在0.0800～12.0mg·L-(1r=0.9964)、0.0080～1.20mg·L-(1r=0.9995)范围内线性关系良好;日内、日间相对标准偏差(RSD)均<6%;平均方法回收率分别为99.23%、101.70%;平均提取回收率分别为(97.0±2.0)%、(92.4±2.7)%。结论:本方法高效、灵敏、准确、专属性强、快速,适用于对人血浆中乙酰氨基酚和金刚烷胺的测定,可应用于小儿氨酚烷胺泡腾片的人体生物等效性研究。

OBJECTIVE： To develop an LC-MS method for the determination of paracetamol and amantadine in human plasma, and to evaluate their bioequivalence in healthy volunteers after given Pediatric paracetamol and amantadine hydrochloride effervescent tables. METHODS： After protein-precipitated with methanol, blood sample was separated on Shim-pack VP-ODS column using the mobile phase consisted of methanol-5 mmol·L-1 ammonium acetate （containing 0.5% formic acid）（45∶55） at a flow rate of 0.3 mL·min-1. Detection was carried out by API 3200 electrospray positive ionization mass spectrometry in the multiple reaction monitoring （MRM） mode. The MRM transitions of m/z 152.2→110.1, m/z 152.3→135.2, m/z 166.1→148.1 were used to quantify paracetamol, amantadine and pseudoephedrine（internal standard）, respectively. RESULTS： The linear ranges were 0.080 0～12.0 mg·L-1 for paracetamol （r=0.996 4） and 0.008 0～1.20 mg·L-1 for amantadine （r=0.999 5）, respectively. The RSDs ofintra-day and inter-day were both less than 6%, and the average method recoveries were 99.23% and 101.70%. The mean extraction recoveries were （97.0±2.0）% and （92.4±2.7）% for paracetamol and amantadine, respectively. CONCLUSION： The method is effective, sensitive, accurate, specific, rapid, and it is suitable for the determination of paracetamol and amantadine in human plasma. The method can be used for bioequivalence study of Pediatric paracetamol and amantadine hydrochloride effervescent tables.