色谱
色譜
색보
CHINESE JOURNAL OF CHROMATOGRAPHY
2014年
3期
290-293
,共4页
熊山%李敬来%朱秀清%王晓英%吕圭源%张振清
熊山%李敬來%硃秀清%王曉英%呂圭源%張振清
웅산%리경래%주수청%왕효영%려규원%장진청
液相色谱-串联质谱%莫诺苷%血浆%比格犬%药代动力学
液相色譜-串聯質譜%莫諾苷%血漿%比格犬%藥代動力學
액상색보-천련질보%막낙감%혈장%비격견%약대동역학
liquid chromatography-tandem mass spectrometry ( LC-MS / MS )%morroniside%plasma%beagle%pharmacokinetics
运用高效液相色谱-串联质谱联用技术,建立了简单、快速、灵敏的比格犬灌胃莫诺苷后血药浓度的检测方法。血浆样品采用蛋白质沉淀法处理,以芍药苷作为内标,色谱柱为 Inertsil ODS-SP 色谱柱(50 mm ×2.1 mm,5μm),流动相为水(含1 mmol / L 甲酸钠)-乙腈,梯度洗脱,流速0.4 mL / min。采用电喷雾离子源( ESI),正离子多反应监测(MRM)模式。绘制血药浓度-时间曲线,并采用 DAS 2.0软件计算药代动力学参数。方法学实验结果表明内源性杂质不干扰莫诺苷和内标的测定,线性范围为2~5000μg / L( r =0.9966),定量限为2μg / L。方法精密度、准确度、回收率和基质效应均符合生物样品测定的要求,适合比格犬血浆中莫诺苷浓度的测定,可以应用该方法进行莫诺苷的药代动力学研究。比格犬灌胃莫诺苷3个剂量(5、15、45 mg / kg)后的血药浓度-时间曲线下面积(AUC(0-∞))分别为(1631.20±238.50)、(3984.05±750.38)、(10397.64±3156.34)μg / L·h,与给药剂量之间呈现良好的线性关系。
運用高效液相色譜-串聯質譜聯用技術,建立瞭簡單、快速、靈敏的比格犬灌胃莫諾苷後血藥濃度的檢測方法。血漿樣品採用蛋白質沉澱法處理,以芍藥苷作為內標,色譜柱為 Inertsil ODS-SP 色譜柱(50 mm ×2.1 mm,5μm),流動相為水(含1 mmol / L 甲痠鈉)-乙腈,梯度洗脫,流速0.4 mL / min。採用電噴霧離子源( ESI),正離子多反應鑑測(MRM)模式。繪製血藥濃度-時間麯線,併採用 DAS 2.0軟件計算藥代動力學參數。方法學實驗結果錶明內源性雜質不榦擾莫諾苷和內標的測定,線性範圍為2~5000μg / L( r =0.9966),定量限為2μg / L。方法精密度、準確度、迴收率和基質效應均符閤生物樣品測定的要求,適閤比格犬血漿中莫諾苷濃度的測定,可以應用該方法進行莫諾苷的藥代動力學研究。比格犬灌胃莫諾苷3箇劑量(5、15、45 mg / kg)後的血藥濃度-時間麯線下麵積(AUC(0-∞))分彆為(1631.20±238.50)、(3984.05±750.38)、(10397.64±3156.34)μg / L·h,與給藥劑量之間呈現良好的線性關繫。
운용고효액상색보-천련질보련용기술,건립료간단、쾌속、령민적비격견관위막낙감후혈약농도적검측방법。혈장양품채용단백질침정법처리,이작약감작위내표,색보주위 Inertsil ODS-SP 색보주(50 mm ×2.1 mm,5μm),류동상위수(함1 mmol / L 갑산납)-을정,제도세탈,류속0.4 mL / min。채용전분무리자원( ESI),정리자다반응감측(MRM)모식。회제혈약농도-시간곡선,병채용 DAS 2.0연건계산약대동역학삼수。방법학실험결과표명내원성잡질불간우막낙감화내표적측정,선성범위위2~5000μg / L( r =0.9966),정량한위2μg / L。방법정밀도、준학도、회수솔화기질효응균부합생물양품측정적요구,괄합비격견혈장중막낙감농도적측정,가이응용해방법진행막낙감적약대동역학연구。비격견관위막낙감3개제량(5、15、45 mg / kg)후적혈약농도-시간곡선하면적(AUC(0-∞))분별위(1631.20±238.50)、(3984.05±750.38)、(10397.64±3156.34)μg / L·h,여급약제량지간정현량호적선성관계。
A sensitive,simple and specific high performance liquid chromatography-electros-pray ionization tandem mass spectrometry( LC-MS / MS)method was developed for the deter-mination of morroniside in the plasma of beagles administered via intragastric( ig)doses of morroniside. The method employed paeoniflorin as the internal standard and extracted by sim-ple protein precipitation. The separation was achieved using an Inertsil ODS-SP column(50 mm ×2. 1 mm,5 μm)with mobile phases of 1 mmol / L sodium formate aqueous solution and aceto-nitrile(gradient elution)at a flow rate of 0. 4 mL / min. The detection was accomplished by a mass spectrometer using multiple reaction monitoring( MRM)in positive mode. Pharmacoki-netic parameters were fitted by software DAS 2. 0. The methodological study showed a good lin-ear relationship of 2-5 000 μg / L(r = 0. 996 6)with a sensitivity of 2 μg / L as the limit of quanti-fication. The precision,accuracy,mean recoveries and the matrix effects were satisfied with the requirements of biological sample measurement. The method described above was success-fully applied to the pharmacokinetic study of morroniside in the beagle plasma samples. The area under the plasma concentration-time curves( AUC( 0-∞ )) of morroniside after single ig administration doses of 5,15 and 45 mg / kg were(1 631. 20±238. 50),(3 984. 05±750. 38)and (10 397. 64±3 156. 34)μg / L·h. The relationship between dose and AUC showed a good linearity. The pharmacokinetic property of morroniside was proposed to be linear pharmacokinetics.