LC-MS/MS法测定人血浆中伊伐布雷定及其活性代谢产物的浓度

目的:建立测定人血浆中伊伐布雷定及其活性代谢产物浓度的方法。方法:血样以叔丁基醚提取后,采用液-质联用(LC-MS/MS)法进样测定。色谱柱为Waters C18,流动相为5 mmol/L醋酸铵-甲酸-甲醇(70∶0.1∶30),流速为0.3 ml/min,柱温为30℃,内标为氨溴索;采用电喷雾离子化(ESI)离子源,以多反应监测(MRM)模式检测,伊伐布雷定、去甲伊伐布雷离子对的m/z分别为469.2/177.1、455.2/177.1。结果:伊伐布雷定、去甲伊伐布雷定血药浓度分别在0.48¹20、0.16120、0.16⁴0 ng/ml范围内线性关系良好(r分别为0.999 8和0.999 7);方法回收率分别为96.35%40 ng/ml范围内线性关系良好(r分别为0.999 8和0.999 7);方法回收率分别为96.35%⁹8.33%、96.20%98.33%、96.20%¹02.50%;日内、日间RSD均<7%;血浆样品长期冻存(-20℃冷冻1个月)稳定性良好,反复冻融3次及室温放置4 h条件下,样品浓度均无显著变化。结论:本方法快速、简便、灵敏、准确,可用于伊伐布雷定的药动学研究。     

液相色谱-质谱联用法测定人血浆中氯吡格雷及其3种代谢产物的浓度

目的:建立同时测定人血浆中氯吡格雷(CLP)及其中间代谢产物2-氧-氯吡格雷(2-O-CLP)、非活性代谢产物氯吡格雷羧酸代谢物(CLPCA)和活性代谢产物氯吡格雷硫醇代谢物(CLPTM)浓度的方法。方法:选取陆军军医大学第一附属医院确诊为脑卒中的90名患者,早晨空腹口服1片氯吡格雷片(75 mg/片),于服药2 h后采集血样,将CLPTM经2-溴-3’-甲氧基苯乙酮衍生形成CLPTM-D后与其余3种待测物一起通过乙腈蛋白沉淀提取后,采用液相色谱-串联质谱法(LC-MS/MS)测定其浓度。色谱柱为Agilent poroshell 120 EC-C18,流动相为0.1%甲酸的乙腈溶液-0.1%甲酸水溶液(90∶10,V/V),采用多反应监测模式进行正离子检测,检测离子对分别为CLPCA质荷比(m/z)308.1→198.1、CLP m/z 322.3→212.0、2-O-CLP m/z 338.3→155.0、CLPTM-D m/z504.4→354.1、内标噻氯吡啶m/z 264.0→154.1。结果:CLPCA、CLP、2-O-CLP、CLPTM-D和内标的保留时间分别为2.01、3.32、2.83、2.68、1.87 min,CLPCA、CLP、2-O-CLP、CLPTM-D检测质量浓度的线性范围分别为100～10 000、0.2～20、0.3～30、0.5～50ng/mL(r均≥0.999 5),日内、日间精密度试验的RSD均≤9.5%(n=5),准确度为93.5%～98.9%(n=5),提取回收率为85.4%～95.9%(n=5),基质效应的CV为2.7%～6.2%(n=5)。稳定性(-80℃放置3个月、3次冷冻-解冻循环、4℃放置8 h)试验中,CLPCA、CLP、2-O-CLP和CLPTM-D的RE均≤10.0%(n=5)。结论:建立的LC-MS/MS法特异性强,测定结果准确可靠,可用于检测人血浆中CLP及其3种代谢产物的浓度。     

高效液相-串联质谱联用法测定人血浆中阿昔洛韦的浓度

目的：建立高效液相-串联质谱联用（LC-MS／MS）法测定人血浆中阿昔洛韦的浓度。方法：采用Waters公司SYMMETRY^TM C18（3．9mm×150mm,5gm）谱柱;流动相为甲醇－水（15：85）;流速为0．6mL／min。串联质谱条件：采用多反应离子监测（MRM）检测,每个样品分析时间为2min。结果：阿昔洛韦在20．0～1000．0ng／mL浓度范围内有良好的线性关系;最低检测浓度为20．0ng／mL;日内、目间RSD均〈15％;准确度RE均在±15％范围内。结论：本法简单、快速、灵敏、准确,可用于人血浆中阿昔洛韦浓度的检测。     

高效液相-串联质谱法测定人血浆中头孢拉定的质量浓度

目的:确立高效液相-串联质谱法(LC-MS/MS)测定人血浆中头孢拉定的质量浓度分析方法。方法:采用色谱柱为SymmetryShieldTMRp18(100mm×2.1mm,3.5μm);流动相为乙腈-水(8∶92);流速为0.3mL/min;内标为阿昔洛韦。结果:头孢拉定的血浆质量浓度在0.1～30.0μg/mL范围内线性良好,标准曲线方程为A=1.3ρ-0.00863(r=0.9970,n=8),定量下限为0.1μg/mL,日内RSD<3.90%,日间RSD<14.90%,方法回收率在107.60%～108.70%。结论:本法操作简单,结果准确,专属性强,灵敏度高,适用于头孢拉定药动学及生物等效性研究。     

LC-MS/MS快速测定生物检材中乌头碱的含量

目的 建立快速测定生物检材中乌头碱的LC-MS/MS方法。方法 生物检材经乙醚-二氯甲烷(3∶2)提取后,用Venusil Mp-C₁₈柱进行分离,采用ESI多反应监测(MRM)扫描方式。结果 测定的线性范围为3~5 000 ng&#903;mL^-1,RSD<1.8%,日间和日内精密度分别为2.5%和4.6%,提取回收率为85.61%~89.04%。结论 该方法简便、快捷,适用于生物检材中乌头碱的快速测定。     

Oxidative Metabolism of BDE-99 by Human Liver Microsomes: Predominant Role of CYP2B6

Hydroxylated polybrominated diphenyl ethers (PBDEs) have been found in human serum, suggesting that they are formed by in vivo oxidative metabolism of PBDEs. However, the biotransformation of 2,2',4,4',5-pentabromodiphenyl ether (BDE-99), a major PBDE detected in human tissue and environmental samples, is poorly understood. In the present study, the oxidative metabolism of BDE-99 was assessed using pooled and single-donor human liver microsomes, a panel of human recombinant cytochrome P450 (CYP) enzymes, and CYP-specific antibodies. Hydroxylated metabolites were quantified using a liquid chromatography/tandem mass spectrometry-based method. In total, 10 hydroxylated metabolites of BDE-99 were produced by human liver microsomes. Six metabolites were identified as 2,4,5-tribromophenol (2,4,5-TBP), 4-OH-BDE-90, 5'-OH-BDE-99, 6'-OH-BDE-99, 4'-OH-BDE-101, and 2-OH-BDE-123 using authentic standards. Three monohydroxy- and one dihydroxy-pentabrominated metabolites were unidentified. Rates of formation of the three major metabolites (2,4,5-TBP, 5'-OH-BDE-99, and 4'-OH-BDE-101) by human liver microsomes ranged from 24.4 to 44.8 pmol/min/mg protein. Additional experiments demonstrated that the dihydroxylated metabolite was a primary metabolite of BDE-99 and was not produced by hydroxylation of a monohydroxy metabolite. Among the panel of recombinant CYP enzymes tested, formation of all 10 hydroxylated metabolites was catalyzed solely by CYP2B6. A combined approach using antibodies to CYP2B6 and single-donor liver microsomes expressing a wide range of CYP2B6 levels confirmed that CYP2B6 was responsible for the biotransformation of BDE-99. Collectively, the results show that the oxidative metabolism of BDE-99 by human liver microsomes is catalyzed solely by CYP2B6 and is an important determinant of the toxicity and bioaccumulation of BDE-99 in humans.     

LC-MS/MS法测定人血清中石杉碱甲的浓度

目的:建立以高效液相色谱串联质谱电喷雾检测(LC-MS/MS)法的方法测定人血清中石杉碱甲浓度。方法:以可乐定为内标,待测血清样品经乙酸乙酯液-液萃取,有机相吹干,流动相复溶后进样,色谱柱为Phenomenex Luna CN柱,流动相为甲醇∶0.08%甲酸水溶液(含0.2%乙酸铵)=46∶54。ESI离子源正离子方式检测,选择反应监测(SRM),用于定量的离子分别为石杉碱甲m/z:母离子243.2,子离子210.9,碰撞能30V;内标可乐定m/z:母离子229.9,子离子212.9,碰撞能32V;扫描时间为0.5s。结果:血清中内源性物质不干扰石杉碱甲的检测,石杉碱甲血药浓度在0.08～8ng·mL-1范围内线性关系良好(r=0.9930～0.9960);日内、日间RSD均<15%,低、中、高浓度的平均提取回收率分别为69.9%、63.7%、63.7%。结论:该法操作快速、灵敏、简单、准确,介质用量少,符合生物样品检测要求,可用于临床药动学研究中大批量血样的处理。     

Metabolism of (+)-fenchone by CYP2A6 and CYP2B6 in human liver microsomes

The in vitro metabolism of (+)-fenchone was examined in human liver microsomes and recombinant enzymes. Biotransformation of (+)-fenchone was investigated by gas chromatography-mass spectrometry. (+)-Fenchone was found to be oxidized to 6-exo-hydroxyfenchone, 6-endo-hydroxyfenchone and 10-hydroxyfenchone by human liver microsomal P450 enzymes. The formation of metabolite of (+)-fenchone was determined by relative abundance of mass fragments and retention time with GC. CYP2A6 and CYP2B6 in human liver microsomes were major enzymes involved in the hydroxylation of (+)-fenchone, based on the following lines of evidence. First, of eleven recombinant human P450 enzymes tested, CYP2A6 and CYP2B6 catalyzed oxidation of (+)-fenchone. Second, oxidation of (+)-fenchone was inhibited by thioTEPA, (+)-menthofuran anti-CYP2A6 and anti-CYP2B6 antibodies. Finally, there was a good correlation between CYP2A6, CYP2B6 contents and (+)-fenchone hydroxylation activities in liver microsomes of 8 human samples.     

LC-MS/MS法测定人血浆中羟基红花黄色素A的浓度

目的建立LC-MS/MS法测定人血浆中羟基红花黄色素A(QA)的浓度。方法血浆样品中加入等体积0.2 mol·L-1的乙酸铵后,采用固相萃取技术进行提取QA,洗脱液直接进行LC-MS/MS分析。QA和内标异鼠李素-3-O-新橙皮苷(SLS)通过Agilent ZORBAX SB C18(3.0 mm×100 mm,3.5μm)色谱柱进行等度洗脱分离,流动相组成为0.2 mmol·L-1乙酸铵水溶液/甲醇=30/70。质谱检测采用负离子模式,扫描方式为多反应检测,QA的目标离子对为m/z 611.131/490.900,SLS的目标离子对为m/z 623.032/298.800。结果 QA和SLS的保留时间分别2.7 min和3.9 min左右,空白血浆无干扰影响含量测定;血浆中QA的线性范围为8.57⁴185μg·L-1(r为0.99494185μg·L-1(r为0.9949⁰.9992),定量下限为8.570μg·L-1,日内日间精密度RSD均小于7%;低、中、高3个浓度下的平均介质效应分别为116.62%、119.06%和115.72%(RSD分别为3.27%、3.42%和4.93%),平均提取回收率分别为77.75%、80.90%和80.76%(RSD分别为1.70%、1.78%和4.15%);血浆样本于室温放置4 h、-70℃反复冻融3次及-70℃冰冻保存31d的情况下均稳定;浓度超出定量上限的QA血浆样本经空白血浆稀释6.25倍后可准确测定。结论建立的测定血浆样本中QA浓度的LC-MS/MS分析方法简便、快速、准确灵敏,适用于QA人体药代动力学的系统研究。     

A sensitive LC-MS/MS assay for the determination of dextromethorphan and metabolites in human urine--application for drug interaction studies assessing potential CYP3A and CYP2D6 inhibition

The commonly used antitussive dextromethorphan can be used to simultaneously assess potential cytochrome P450 3A (CYP3A) and CYP2D6 inhibition during drug development. The metabolism of dextromethorphan to dextrorphan and subsequently to 3-hydroxymorphinan are via the 2D6 pathway, while the metabolism of dextromethorphan to 3-methoxymorphinan is via the 3A pathway. A sensitive and specific LC-MS/MS assay has been developed to determine the human urine concentrations of dextromethorphan and three metabolites (dextrorphan, 3-methoxymorphinan and 3-hydroxymorphinan) in support of drug interaction studies. Urine samples (0.5 ml), after enzymatic hydrolysis of the conjugates and containing 3-ethylmorphine as an internal standard, were extracted with chloroform under basic conditions. Following concentration and reconstitution, the samples were analyzed by LC-MS/MS. The assay was linear over the range of 5.00-500 ng/ml for dextromethorphan and 3-methoxymorphinan; and 200-3000 ng/ml for dextrorphan and 3-hydroxymorphinan using a Perkin-Elmer Sciex triple quadrupole mass spectrometer (API 300). The intra- and inter-day relative standard deviation (RSD) across three validation runs over the entire concentration range for all analytes was less than 15%. Accuracy determined at three or four concentrations (9.00, 200, and 400 ng/ml for dextromethorphan and 3-methoxymorphinan; 250, 400, 1300 and 2500 ng/ml for dextrorphan and 3-hydroxymorphinan) ranged between 96.3 and 113.8%. The stability of analytes in urine was demonstrated for 9 months at -20 degrees C, 24 h under ambient conditions and for up to three freeze/thaw cycles. The method described herein is suitable for the rapid and efficient measurement of dextromethorphan and different metabolites to estimate potential CYP3A inhibition by drug candidates and for screening of extensive and poor metabolizers of CYP2D6 in the heterogeneous population. The method has subsequently been validated on a Sciex API 3000 with lower limit of quantitation; 1.00 ng/ml for dextromethorphan and 3-methoxymorphinan; 60.0 ng/ml for dextrorphan and 100 ng/ml for 3-hydroxymorphinan.     

Metabolism of (-)-fenchone by CYP2A6 and CYP2B6 in human liver microsomes

The in vitro metabolism of (-)-fenchone was examined in human liver microsomes and recombinant enzymes. The biotransformation of (-)-fenchone was investigated by gas chromatography-mass spectrometry. (-)-Fenchone was found to be oxidized to 6-exo-hydroxyfenchone, 6-endo-hydroxyfenchone and 10-hydroxyfenchone by human liver microsomal P450 enzymes. The formation of metabolites was determined by the relative abundance of mass fragments and retention times on gas chromatography (GC). CYP2A6 and CYP2B6 were major enzymes involved in the hydroxylation of (-)-fenchone by human liver microsomes, based on the following lines of evidence. First, of 11 recombinant human P450 enzymes tested, CYP2A6 and CYP2B6 catalysed the oxidation of (-)-fenchone. Second, oxidation of (-)-fenchone was inhibited by thioTEPA and (+)-menthofuran. Finally, there was a good correlation between CYP2A6, CYP2B6 contents and (-)-fenchone hydroxylation activities in liver microsomes of 11 human samples. CYP2A6 may be more important than CYP2B6 in human liver microsomes. Kinetic analysis showed that the Vmax/Km values for (-)-fenchone 6-endo-, 6-exo- and 10-hydroxylation catalysed by liver microsomes of human sample HG-03 were 24.3, 44.0 and 1.3nM(-1)min(-1) , respectively. Human recombinant CYP2A6 and CYP2B6 catalysed (-)-fenchone 6-exo-hydroxylation with Vmax values of 2.7 and 12.9 nmol min(-1) nmol(-1) P450 and apparent Km values of 0.18 and 0.15 mM and (-)-fenchone 6-endo-hydroxylation with Vmax values of 1.26 and 5.33nmolmin(-l) nmol(-1) P450 with apparent Km values of 0.29 and 0.26mM. (-)-Fenchone 10-hydroxylation was catalysed by CYP2B6 with Km and Vmax values of 0.2 mM and 10.66 nmol min(-1) nmol(-1) P450, respectively.     

Metabolism of citalopram enantiomers in CYP2C19/CYP2D6 phenotyped panels of healthy Swedes

AIMS: To investigate pharmacokinetics of the enantiomers of citalopram (CT) and its metabolites desmethylcitalopram (DCT) and didesmethylcitalopram (DDCT) in Swedish healthy volunteers in relation to CYP2C19 and CYP2D6 geno- and phenotypes. METHODS: Racemic CT was given for seven days to panels with different genotypes and the following mephenytoin (Me) and debrisoquine (De) hydroxylation phenotypes: EMDe/EMMe, PMDe/EMMe, EMDe/PMMe (n = 6 in all groups), and one PMDe/PMMe subject. Blood sampling was carried out during day 7, and all urine was collected for 12 h after the last dose of CT. RESULTS: The AUC of S-CT was significantly higher in the EMDe/PMMe panel compared to the EMDe/EMMe and PMDe/EMMe panels (P < 0.05), whereas the AUC of R-CT did not differ between the panels. Similar differences, although they did not reach statistical significance, were noted for S-DCT and R-DCT. The enantiomers of DDCT were not quantifiable in PMDe, and there was no difference in DDCT enantiomer concentrations between the other two panels. A PMDe/PMMe subject stopped taking CT after five days due to severe adverse effects. Based on two time points, this subject had a very long CT half-life of 95 h. The value of 1.0 for the S/R ratio of the CT trough in this subject was similar to the mean S/R CT trough ratio of the EMDe/PMMe panel, but higher than the S/R CT ratio of the EMDe/EMMe panel (0.56; 95% CI 0.49-0.63) and the PMDe/EMMe panel (0.44; 95% CI 0.31-0.57). Thus the latter two phenotypes eliminated S-CT more rapidly via CYP2C19. An adverse effect described as an 'alcohol hangover' feeling was reported by one subject from each of the three panels. These individuals had the highest concentrations of both CT enantiomers. CONCLUSIONS: The AUC of S-, but not R-(CT) was found to be significantly higher in PM of mephenytoin compared to EMs, PMs may need a lower dosage of CT.     

应用LC-MS／MS 检测中药材中黄曲霉毒素残留量方法研究-

目的：建立LC—MS／MS对中药材中黄曲霉毒素残留量的检测方法。方法：中药材粗粉甲醇提取,经免疫亲和柱净化吸附,甲醇洗脱,以甲醇／乙腈-1．0mmol乙酸铵为流动相,黄曲霉毒素含量用液相色谱。串联三重四极杆质谱测定,用电喷雾正离子模式ESI,多反应（MRM）监测。结果：黄曲霉毒素G2、B2在0．03—3．2ng·ml^-1。范围内线性关系良好、黄曲霉毒素B1,G1在0．1～10ng·ml^-1“范围内线性关系良好。检测限0．1ng·ml^-1。定量限0．2ng·ml^-1。黄曲霉毒素回收率：86．6％～93．2％,RSD为7．1％-11．8％。结论：本法专属性强,灵敏、简便,准确,通过色谱（保留时间）及质谱特征碎片离子定性,能有效排除假阳性干扰。可作为中药中昔曲霪喜素藏留号测定的方法．     

LC-MS/MS测定注射用聚氯乙烯袋中注射液的邻苯二甲酸二异辛酯

目的 建立测定注射用聚氯乙烯(PVC)袋中注射液的邻苯二甲酸二异辛酯(DEHP)含量的方法.方法 以邻苯二甲酸二乙酯(DEP)为内标,采用LC-MS法测定.结果 DEHP和DEP的tR分别为2.86、1.05 min,在19.6～588.0 ng·ml-1有较好线形关系,RSD=3.5%,平均回收率为99.2%,最低检测限为5.88 ng·ml-1.结论 所建方法专属性强,适用于对注射用PVC袋中注射液的DEHP的含量测定.     

LC-MS/MS法测定氨氯地平血药浓度及其片剂的生物等效性研究

目的:建立测定人血浆中氨氯地平血药浓度的方法,并评价2种苯磺酸氨氯地平片的生物等效性。方法:人血浆样本经液-液萃取后,选用ZorbaxSB-C18NarrowBore色谱柱,以甲醇-10mmol/L乙酸铵(90:10,V/V)为流动相,流速为0.3ml/min,选用API3200型三重四极杆串联质谱仪的多重反应监测(MRM)扫描方式进行监测,电喷雾电离(ESI)离子化源,正离子方式,选择监测离子反应分别为m/z409.2→m/z238.2(氨氯地平)和m/z256.2→m/z167.3(苯海拉明,内标)。24名健康受试者随机于2个周期交叉口服相同剂量的苯磺酸氨氯地平片受试制剂(国产)或参比制剂(进口),考察其生物等效性。结果:氨氯地平和苯海拉明的保留时间分别约为2.79、2.43min;氨氯地平的血药浓度在0.10～10.0ng/ml范围内线性关系良好(r=0.9997),定量下限为0.10ng/ml,日内、日间RSD均<15%,相对偏差(RE)均在±15%范围以内,平均提取回收率为(84.7±5.5)%,平均基质效应因子为(76.5±8.4)%。以AUC0-120h计,受试制剂相对于参比制剂的生物利用度为(105.8±20.9)%。结论:该方法快速、灵敏、专属性强、重现性好,适用于人血浆中氨氯地平的测定,可用于苯磺酸氨氯地平片的人体生物等效性研究;2种制剂生物等效。     

A novel incubation direct injection LC/MS/MS technique for in vitro drug metabolism screening studies involving the CYP 2D6 and the CYP 3A4 isozymes

A direct injection LC/MS/MS method involving a novel incubation technique was developed for the inhibition screening of CYP 2D6 and CYP 3A4 isoenzymes using dextromethorphan and midazolam as probe substrates. Both assays were performed using an electrospray ionization source in the positive ion mode. Direct injection was possible by using a short C 18, LC column (2 mm x 20 mm) with large particle diameter packing (10 microm). Analytical characteristics of the direct injection technique were studied by examining matrix effects, which showed suppression of the ESI signal between 0.20 and 0.65 min. The retention times for analytes were adjusted to approximately 0.8 min (k'>3), resulting in no matrix effect. Column lifetime was evaluated and determined to be approximately 160 direct injections of the matrix. The precision and accuracy of the control samples for the quantitation of dextromethorphan was between -0.53 and -12.80, and 3.73 and 6.69% respectively. Unlike conventional incubation techniques, incubations were carried out in an autosampler equipped with a heating accessory. This novel incubation method, which involved no stirring of the incubation mixture, estimated the Cl(int in vitro) for dextromethorphan and midazolam in human liver microsomes to be 1.65+/-0.22 ml/(hmg) and 0.861 ml/(min mg) respectively. The autosampler tray maintained uniform temperature and was sensitive to changes in temperature between 33 and 41 degrees C. High-throughput screening was performed using known inhibitors of the CYP 2D6 isozyme, and the system was evaluated for its ability to differentiate between these inhibitors. The strong inhibitor quinidine resulted in a 25.6% increase in t(1/2), the medium potency inhibitor chlorpromazine resulted in an increase of 6.14% and the weak inhibitor primaquine had no significant effect on half-life. This technique involves no sample preparation, demonstrated run times of 2 min per injection and can be fully automated. The method should therefore prove to be a valuable tool in the drug discovery process.     

LC-MS/MS法测定藏药十三味红花丸中羟基红花黄色素A的含量

目的:建立测定藏药十三味红花丸中羟基红花黄色素A含量的方法。方法:样品经甲醇-水(80∶20,V/V)提取,0.22μm微孔滤膜滤过后采用液相色谱-串联质谱(LC-MS/MS)法测定其中羟基红花黄色素A的含量。色谱柱为AgilentZorbaxSB-C18(50mm×2.1mmI.D.,3.5μm),流动相为水(含0.1%甲酸)-乙腈(含0.1%甲酸),梯度洗脱;离子源为电喷雾离子化源(ESI源),离子源温度为500℃,以多反应监测(MRM)方式分别监测离子对m/z611.2→491.2(羟基红花黄色素A)和m/z268.9→159.0(内标染料木素)。结果:羟基红花黄色素A的检测浓度在10～1000ng·mL-1范围内同其与内标的峰面积比值呈良好线性关系(r=0.9989);平均加样回收率在93.8%～106.2%范围内,RSD均<4.90%(n=9)。结论:该方法快速、灵敏,结果准确,适用于藏药十三味红花丸的质量控制。     

LC-MS/MS法测定人血浆中西布曲明代谢物(M2)

西布曲明代谢物M2(N-双脱甲基西布曲明)是减肥药西布曲明(N-[1-[1-(4-氯苯基)环丁基]-3-甲基丁基]-N,N-二甲基胺,sibutramine)的主要活性代谢产物,其体内浓度较低,难以用常规的HPLC-UV法进行测定.国内至今未有西布曲明血药浓度测定的相关报道,为了解西布曲明制剂的体内过程,我们参考有关文献[1],采用液相色谱-质谱/质谱(LC-MS/MS)法建立了西布曲明主要活性代谢产物M2的血药浓度分析方法,可较好地满足研究工作的需要. 1 材料和方法 1.1 仪器 Waters 2690高效液相色谱仪(美国Waters公司);QuattroLC质谱仪(英国质谱公司,micromass).XW-80A旋涡混合器(上海医科大学仪器厂);CQ50超声波清洗器(上海超声波仪器厂);80-2型离心机(上海手术器械厂);LNG-T83台式快速离心浓缩干燥器(江苏太仓医用器械厂);SHB-Ⅲ循环多用水泵(郑州长城仪器厂);TGL.16G台式高速离心机(上海医用分析仪器厂).