HILIC-MS/MS法测定早产儿血浆中咖啡因的浓度

Determination of Caffeine in Human Plasma Samples by HPLC-MS/MS

目的:建立HILIC-MS/MS法测定对使用咖啡因治疗呼吸暂停的早产儿血浆中的咖啡因浓度。方法:以13C3-咖啡因为内标;色谱柱:Atlantis^(■)HILIC Silica(2.1 mm×150 mm,3μm);流动相:5 mmol·L^(-1)甲酸铵水溶液(含0.1%甲酸)和乙腈(梯度洗脱);流速:0.3 mL·min^(-1);进样量:1.0μL。采用电喷雾离子化,正离子模式,多反应监测扫描测定。结果:本法测定咖啡因在0.0500~50.0μg·mL^(-1)范围线性关系良好;批内、批间准确度和精密度均<15.0%;提取回收率为89.7%~92.1%;无明显基质效应和残留效应。结论:本方法快速、灵敏,可准确测定早产儿微量血样中的咖啡因浓度。

Objective:An efficient method of high-performance liquid chromatography-tandem mass spectrometry(HPLC-MS/MS)was established to determine the concentrations of caffeine in human plasma samples and to quantitatively analyze caffeine in plasma samples of premature infants with apnea treated with caffeine.Methods:Caffeine^(-1)3C3 was used as the internal standard;the chromatography column was Atlantis^(■)HILIC Silica column(2.1 mm×150 mm,3μm),the mobile phase was 5 mmol·L^(-1)ammonium formate aqueous solution(containing 0.1%formic acid)and acetonitrile,the flow rate was 0.3 mL·min^(-1),the injection volume was 1.0μL,and the gradient elution was lasted for 7.Electrospray ionization,positive ion mode,and multiple reaction monitoring and scanning were applied for MS/MS detection.Results:The established HPLC-MS/MS method for the determination of caffeine had a good linear relationship in the range of 0.0500-50.0μg·mL^(-1);RSD for intra-day and inter-day accuracy and precision were all less than 15.0%;The recoveries were 89.7%-92.1%;There was no obvious matrix or carryover effect of caffeine.Conclusion:A rapid and sensitive HPLC-MS/MS analysis method has been established to determine caffeine in blood samples of premature infants.