检验医学
檢驗醫學
검험의학
LABORATORY MEDICINE
2014年
11期
1144-1150
,共7页
茶碱%液相色谱-串联质谱法%治疗药物监测%室间质量评价
茶堿%液相色譜-串聯質譜法%治療藥物鑑測%室間質量評價
다감%액상색보-천련질보법%치료약물감측%실간질량평개
Theophylline%Liquid chromatography-tandem mass spectrometry%Therapeutic drug monitoring%External quality assessment
目的：对液相色谱-串联质谱（LC-MS ／MS）检测茶碱进行方法学评价，探讨其在茶碱治疗药物监测（TDM）中的应用。方法在血清添加放射性核素内标茶碱-D6，经蛋白沉淀稀释后采用 LC-MS ／MS 测定。以Capcell C18 MG Ⅲ（100 mm ×2．0 mm，5μm）为分析柱进行反相色谱分离；以0．1％甲酸乙腈-0．1％甲酸水[20∶80（v／v）]为流动相，流速为0．3 mL／min；以电喷雾离子化串联四级杆质谱、正离子多反应监测进行定量检测。用建立的方法从2008年起连续参加卫生部临检中心茶碱 TDM室间质量评价。结果LC-MS ／MS 检测茶碱的线性范围为1～50μg／mL，批内和批间精密度分别为2．26％～6．65％和4．70％～6．84％，准确度分别为94．14％～104．00％。单个样品的监测分析时间为3．5 min。冻融（-30℃室温反复解冻3次）、室温放置24 h、自动进样器放置24 h、长期保存（-30℃放置28 d）的稳定性均良好。LC-MS ／MS 测定结果与室间质量评价靶值偏差为2．75％，斜率为1．04，相关系数（r2）为0．983。结论该 LC-MS ／MS 采用放射性内标稀释，具有简单、快速、特异性和灵敏度较好的特点，连续6年测定结果符合全国室间质量评价要求，可用于茶碱的临床 TDM。
目的：對液相色譜-串聯質譜（LC-MS ／MS）檢測茶堿進行方法學評價，探討其在茶堿治療藥物鑑測（TDM）中的應用。方法在血清添加放射性覈素內標茶堿-D6，經蛋白沉澱稀釋後採用 LC-MS ／MS 測定。以Capcell C18 MG Ⅲ（100 mm ×2．0 mm，5μm）為分析柱進行反相色譜分離；以0．1％甲痠乙腈-0．1％甲痠水[20∶80（v／v）]為流動相，流速為0．3 mL／min；以電噴霧離子化串聯四級桿質譜、正離子多反應鑑測進行定量檢測。用建立的方法從2008年起連續參加衛生部臨檢中心茶堿 TDM室間質量評價。結果LC-MS ／MS 檢測茶堿的線性範圍為1～50μg／mL，批內和批間精密度分彆為2．26％～6．65％和4．70％～6．84％，準確度分彆為94．14％～104．00％。單箇樣品的鑑測分析時間為3．5 min。凍融（-30℃室溫反複解凍3次）、室溫放置24 h、自動進樣器放置24 h、長期保存（-30℃放置28 d）的穩定性均良好。LC-MS ／MS 測定結果與室間質量評價靶值偏差為2．75％，斜率為1．04，相關繫數（r2）為0．983。結論該 LC-MS ／MS 採用放射性內標稀釋，具有簡單、快速、特異性和靈敏度較好的特點，連續6年測定結果符閤全國室間質量評價要求，可用于茶堿的臨床 TDM。
목적：대액상색보-천련질보（LC-MS ／MS）검측다감진행방법학평개，탐토기재다감치료약물감측（TDM）중적응용。방법재혈청첨가방사성핵소내표다감-D6，경단백침정희석후채용 LC-MS ／MS 측정。이Capcell C18 MG Ⅲ（100 mm ×2．0 mm，5μm）위분석주진행반상색보분리；이0．1％갑산을정-0．1％갑산수[20∶80（v／v）]위류동상，류속위0．3 mL／min；이전분무리자화천련사급간질보、정리자다반응감측진행정량검측。용건립적방법종2008년기련속삼가위생부림검중심다감 TDM실간질량평개。결과LC-MS ／MS 검측다감적선성범위위1～50μg／mL，비내화비간정밀도분별위2．26％～6．65％화4．70％～6．84％，준학도분별위94．14％～104．00％。단개양품적감측분석시간위3．5 min。동융（-30℃실온반복해동3차）、실온방치24 h、자동진양기방치24 h、장기보존（-30℃방치28 d）적은정성균량호。LC-MS ／MS 측정결과여실간질량평개파치편차위2．75％，사솔위1．04，상관계수（r2）위0．983。결론해 LC-MS ／MS 채용방사성내표희석，구유간단、쾌속、특이성화령민도교호적특점，련속6년측정결과부합전국실간질량평개요구，가용우다감적림상 TDM。
Objective To perform the methodology evaluation of a liquid chromatography-tandem mass spectrometry (LC-MS /MS)for the determination of theophylline in serum,and to investigate the application significance in the therapeutic drug monitoring (TDM) for theophylline. Methods After being added with radionuclide theophylline-D6 as internal standard,serum samples were treated with protein precipitation and determined by LC-MS /MS.A reverse phase chromatographic separation was performed on Capcell C1 8 MG Ⅲ analytical column (1 00 mm × 2.0 mm,5 μm)by using 0.1 % formic acid in acetonitrile and 0.1 % formic acid in water [20∶80 (v/v)]as mobile phase.The flow rate was 0.3 mL/min.Theophylline and internal standard were monitored by a positive electrospray ion-tandem mass spectrometry system.The proficiency performance of LC-MS /MS was tested by participation to external quality assessment of theophylline TDMfor the National Center for Clinical Laboratory.Results The linear range of LC-MS /MS for the determination of theophylline was 1 -50 μg/mL.The accuracy was 94.1 4%-1 04.00% with within-run and between-run precisions of 2.26%-6.65% and 4.70%-6.84%.The run time was 3.5 min per sample.Stabilities were good under 3 cycles of frozen-thaw for 3 times from -30 ℃ to room temperature,room temperature for 24 h, autosampler for 24 h and -30℃ for 28 d.The external quality assessment proficiency test showed the bias of 2.75%, regression slope of 1 .04 and correlation coefficient (r2 )of 0.983.Conclusions The radionuclide internal standard-dilution LC-MS /MS is simple,rapid,specific and sensitive for the determination of serum theophylline.It is comparable with the external quality assessment proficiency test for consecutive 6 years,and it is suitable for theophylline TDM in clinical practice.
