中国药理学通报
中國藥理學通報
중국약이학통보
CHINESE PHARMACOLOGICAL BULLETIN
2014年
4期
578-582
,共5页
脑代谢%微透析技术%LC-MSn%咪达唑仑%1′-羟基咪达唑仑%CYP3A
腦代謝%微透析技術%LC-MSn%咪達唑崙%1′-羥基咪達唑崙%CYP3A
뇌대사%미투석기술%LC-MSn%미체서륜%1′-간기미체서륜%CYP3A
metabolism in brain%microdialysis%LC-MSn%midazolam%1′-hydroxymidazolam%CYP3A
目的：建立灵敏、快速、准确的LC-MSn 方法测定咪达唑仑/1′-羟基咪达唑仑及其在大鼠脑内的药代动力学过程。方法 SD大鼠股静脉注射CYP3A探针药物咪达唑仑(剂量5 mg·kg-1)。采用微透析技术,在2μl·min-1流速下收集脑微透析液样品2.4 h,收集时间间隔为8 min。以LC-MSn法在线检测微透析样品。选用Agilent Eclipse Plus-C18色谱柱(2．1 mm ×50 mm,3．5μm),流动相为2 mmol·L-1乙酸铵水溶液和乙腈,梯度方式洗脱。采用电喷雾电离源( ESI),扫描方式为多反应离子监测模式( MRM ),咪达唑仑 m/z：326.1/291.1；1′-羟基咪达唑仑m/z：342.1/324.1；内标地西泮m/z：285.1/154．0。结果咪达唑仑和1′-羟基咪达唑仑的线性范围分别为0．78~100和0．195~12.5μg·L-1,最低定量下限0．2μg·L-1,批内、批间RSD均<7%, RE值在-1．34~-8%范围内。结论此检测方法快速、灵敏,可用于脑微透析液中咪达唑仑/1′-羟基咪达唑仑的含量测定,结合微透析技术有助于深入进行脑代谢的相关研究。
目的：建立靈敏、快速、準確的LC-MSn 方法測定咪達唑崙/1′-羥基咪達唑崙及其在大鼠腦內的藥代動力學過程。方法 SD大鼠股靜脈註射CYP3A探針藥物咪達唑崙(劑量5 mg·kg-1)。採用微透析技術,在2μl·min-1流速下收集腦微透析液樣品2.4 h,收集時間間隔為8 min。以LC-MSn法在線檢測微透析樣品。選用Agilent Eclipse Plus-C18色譜柱(2．1 mm ×50 mm,3．5μm),流動相為2 mmol·L-1乙痠銨水溶液和乙腈,梯度方式洗脫。採用電噴霧電離源( ESI),掃描方式為多反應離子鑑測模式( MRM ),咪達唑崙 m/z：326.1/291.1；1′-羥基咪達唑崙m/z：342.1/324.1；內標地西泮m/z：285.1/154．0。結果咪達唑崙和1′-羥基咪達唑崙的線性範圍分彆為0．78~100和0．195~12.5μg·L-1,最低定量下限0．2μg·L-1,批內、批間RSD均<7%, RE值在-1．34~-8%範圍內。結論此檢測方法快速、靈敏,可用于腦微透析液中咪達唑崙/1′-羥基咪達唑崙的含量測定,結閤微透析技術有助于深入進行腦代謝的相關研究。
목적：건립령민、쾌속、준학적LC-MSn 방법측정미체서륜/1′-간기미체서륜급기재대서뇌내적약대동역학과정。방법 SD대서고정맥주사CYP3A탐침약물미체서륜(제량5 mg·kg-1)。채용미투석기술,재2μl·min-1류속하수집뇌미투석액양품2.4 h,수집시간간격위8 min。이LC-MSn법재선검측미투석양품。선용Agilent Eclipse Plus-C18색보주(2．1 mm ×50 mm,3．5μm),류동상위2 mmol·L-1을산안수용액화을정,제도방식세탈。채용전분무전리원( ESI),소묘방식위다반응리자감측모식( MRM ),미체서륜 m/z：326.1/291.1；1′-간기미체서륜m/z：342.1/324.1；내표지서반m/z：285.1/154．0。결과미체서륜화1′-간기미체서륜적선성범위분별위0．78~100화0．195~12.5μg·L-1,최저정량하한0．2μg·L-1,비내、비간RSD균<7%, RE치재-1．34~-8%범위내。결론차검측방법쾌속、령민,가용우뇌미투석액중미체서륜/1′-간기미체서륜적함량측정,결합미투석기술유조우심입진행뇌대사적상관연구。
Aim To develop a sensitive, rapid and ac-curate LC-MSn method for determination of midazolam/1′-hydroxymidazolam and their pharmacokinetic char-acteristics in rat brain. Methods SD rats received in-travenous injection of midazolam ( 5 mg · kg-1 ) via femoral vein, a probe drug of cytochrome P450 3A. The microdialysis ( MD ) samples in situ brain were collected every 8 mins at 2. 0μl·min-1 in 2. 4 hours. Analytes in brain dialysate were quantified by the pro-posed LC-MSn method. Gradient elution was performed on an Agilent Eclipse Plus-C18 column ( 2 . 1 × 50 mm, 3. 5 μm). The mobile phase consisted of 2 mmol ·L-1 ammonium acetate and acetonitrile. The analyte was detected using electrospray ionization ( ESI ) in multiple reaction monitoring ( MRM) modes. The reac-tion selected ions were 326 . 1/291 . 1 m/z for midazo-lam, 342. 1/324. 1 m/z for 1′-hydroxymidazolam and 285. 1/154. 0 m/z for diazepam as internal standard. Result The linear ranges of midazolam and 1′-hydroxymidazolam were 0 . 78~100 and 0 . 195 ~12 . 5μg·L-1 respectively. The lower limit of quantification was 0 . 2 μg · L-1 . The RSD of intra- and inter-batch precisions was less than 7 %. The RSD of accuracy was from -1 . 34 to -8 %. Conclusion This sensi-tive and rapid LC-MSn method is suitable for determi-nation of midazolam/1′-hydroxymidazolam in rat brain dialysate. MD combined with LC-MSn method may give assistance to deep and further studies of drug metabo-lism and CYP3A enzyme in brain.
