高效液相色谱法测定硝苯地平血药浓度

目的采用高效液相色谱(HPLC)法测定硝苯地平临床治疗血药浓度.方法选择乙酸乙酯-正己烷(32)混合液为提取溶剂,在Hypersil-BDS(250 mm×4.0 mm, ID 5 μm)分离柱上,以乙腈-20 mmol/L磷酸二氢钠(6040)为流动相,紫外吸收波长350 nm检测临床样本.结果硝苯地平血浆样品在5.0～200.0 ng/ml范围内线性关系良好(r=0.999 9,n=5),回收率>95.0%,日内日间的RSD<8.0%.结论本法简便、准确、重现性好,适合于硝苯地平临床治疗血药浓度的定量监测.     

高效液相色谱法测定人血清中双氯芬酸钠的血药浓度

目的建立人血清中双氯芬酸钠的HPLC测定法.方法取血清0.5 ml,以吲哚美辛为内标,用叔丁基甲醚提取,在50 ℃水浴条件下氮气挥干,残渣用200μl流动相复溶,40 μl进样.色谱条件SP HERI-C18柱(4.6 mm×250 mm,10μm);流动相0.25 mol/L磷酸盐缓冲液(pH7.0)-甲醇(3664 v/v),流速为1.0 ml/min;检测波长263 nm.结果本方法在0.067～17.12μg/ml范围内线性良好,相关系数r=0.999 5(n=5).检测限为8 ng/ml.高、中、低三种浓度的绝对和相对回收率分别为90.08%～92.10%,94.03%～99.88%,日内和日间RSD分别为0.87%～3.03%,1.73%～4.76%.结论本方法灵敏、特异、简便,可用于临床双氯芬酸钠血药浓度检测.     

甲巯咪唑血药浓度的高效液相色谱法快速测定

目的建立适合甲巯咪唑血药浓度快速测定的HPLC法.方法色谱柱Hypersil ODS 柱(150 mm×4.6 mm,5 μm);流动相∶甲醇-0.05%乙酸溶液(1∶99);波长252 nm.结果该方法在含甲巯咪唑血浆浓度0.1～2.0 μg/ml及2.0～50.0 μg/ml范围内定量线性良好(r分别为0.999 4和0.999 9),最低检测限为1 ng.精密度和回收率均良好.结论本方法适用于服用甲巯咪唑及卡比马唑的临床病人的血药浓度测定,以便进行临床药学研究.     

高效液相色谱法测定人血浆中霉酚酸的浓度

目的建立用高效液相色谱测定人血浆中霉酚酸浓度的方法。方法采用SUPELCOSIL C18色谱柱（4.6mm×250 mm,5μm） 流动相：乙腈-0.01 mol.L-1磷酸二氢钾（pH=3.0）溶液（40∶60,v/v） 流速：1.0mL.min-1 检测波长：254 nm。结果霉酚酸在0.4~51.2 mg.L-1线性关系良好（r=0.999 3）,最低检测浓度为0.4 mg.L-1,霉酚酸低、中、高浓度检测的日内、日间RSD分别为2.22%、5.15%、4.13%和1.75%、6.75%、3.35%。结论该法具有操作简便可靠、准确、稳定性高等特点,适合应用于霉酚酸的治疗药物监测和药动学研究。     

反相高效液相色谱法测定水杨酸血药浓度及其应用

目的 :建立反相高效液相色谱测定水杨酸血药浓度的方法 ,并将其用于阿司匹林复方制剂中水杨酸药代动力学及生物利用度的研究。方法 :采用Waters2690高效液相色谱仪 ,以DiamonsilC18 为色谱柱 (4 6mm×250mm ,5μm) ,以甲醇 -乙腈 -0 2 %磷酸 (18∶32∶50)为流动相 ,检测波长为237nm ,流速为1 0ml/min。结果 :水杨酸的浓度在0 40～101 00μg/ml范围内线性关系良好 (r=0 9999)。结论 :本法操作简便、灵敏、快速 ,适用于水杨酸的血药浓度测定及药代动力学研究。     

反相高效液相色谱法测定环孢素A血药浓度

目的采用RP-HPLC法测定环孢素A血药浓度.方法色谱柱为HypersilBDSC18柱(4.6mm×150mm,5μm),乙腈-水(62∶38)为流动相,柱温70℃,流速1.8ml*min-1,检测波长205nm.结果CsA血药浓度在50～1200ng*ml-1范围内,浓度与峰面积比有良好的线性关系,最低检测浓度为10ng*ml-1.平均回收率为(97.5±5.8)%(n=15).日内RSD≤6.8%,日间RSD≤9.6%.结论方法简单、快速、准确,可用于环孢素的临床药动学研究.     

高效液相色谱法测定人血浆中酮洛芬浓度

采用高效液相色谱法测定人血浆中酮洛分浓度。方法：用石油醚－乙醚（１：１）提取血浆样品中酮洛芬及萘普生在ＳｐｈｅｒｉｓｏｒｂＣ１８柱上,甲醇－５０ｍｍｏｌ．Ｌ＾－１磷酸二氢钠（６３：３７）为流动相进行分离于２５５ｎｍ检测。结论本法简便,准确,重现性好,用于测定１２名健康示愿者口服酮洛芬胶囊后血药浓度取得良好的效果。     

高效液相色谱法测定人血浆中头孢拉定浓度

目的 探讨用HPLC法测定人血浆中头孢拉定浓度可行性。方法 用乙腈-1mol·L-1盐酸（95:5）混合沉淀血浆蛋白,在Spherisorb C18柱上,以水-甲醇-3.86％醋酸钠溶液-4％酸醋溶液（800:240:15:3）为流动相进行分离,于254mm检测。结果 线性范围为1.0～128.0μg·mL-1,加样回收率为97.8％～100.3％,日内RSD为0.93％～1.92％,日间RSD为1.65％～4.83％。结论 本法简便,准确,重现性好,可用于患者临床血药浓度检测。     

兔血浆中霉酚酸酯的活性代谢物霉酚酸的HPLC测定及其药代动力学研究

目的建立霉酚酸兔血药浓度测定方法 ,并进行了灌胃后药代动力学研究。方法采用HPLC 二极管阵列检测器测定血药浓度。血样采用乙醚萃取 ;色谱柱 :HypersilODS2 (2 0 0mm×4 6mm ,5 μm) ;流动相 :0 0 5mol·L-1KH2 PO4 CH3 CN(5 5∶45 ,pH 2 5 ) ;检测波长 :2 1 5nm ;流速 :1 0mL·min-1。结果霉酚酸血药浓度线性范围为 0 1 6～ 1 9 98μmol·L-1(r =0 9996,n =8) ,血浆最低检测浓度为 0 0 3 μmol·L-1,高、中、低 3种浓度平均回收率分别为 :(1 0 3 2 5± 6 3 2 ) %、(99 63± 2 75 ) %、(1 0 2 47± 3 1 8) % ,兔灌胃后主要的药代动力学参数分别为 :达峰浓度cmax为(8 2 7± 6 1 2 ) μmol·L-1,达峰时间tmax为 (0 49± 0 1 7)h ,消除半衰期t1/ 2 为 (3 1 2± 0 94)h ,药时曲线下面积AUC0 -t为 (1 3 0 8± 9 0 8) μmol·h·L-1,AUC0 -∞ 为 (1 3 80± 8 93 )mg·h·L-1。结论为霉酚酸的血药浓度监测及药动学研究提供了实验方法。     

用HPLC-MS/MS法同时测定Caco-2细胞单层转运介质中吗替麦考酚酯和麦考酚酸的浓度

目的:建立HPLC-MS/MS法同时测定Caco-2细胞单层转运介质中吗替麦考酚酯(mycophenolate mofetil,MMF)和麦考酚酸(mycophenolic acid,MPA)的浓度,用于药物转运实验研究。方法:采用Zorbax Eclipse XDB-C18色谱柱(50mm×2.1mm,3.5μm);流动相A相:甲醇(含0.05%甲酸),B相:水(含0.05%甲酸);梯度洗脱:0～3.60min,40%A,流速:0.3ml/min,3.61～7.00min,65%A,流速:0.6ml/min,7.01～10.00min,40%A,流速:0.6ml/min;柱温:30℃,以吲哚美辛为内标。质谱检测方式:多种反应监测(MRM),选择监测的离子为:m/z433.8→194.8(MMF),m/z319.1→191.0(MPA)和m/z356.0→312.1(吲哚美辛)。结果:MMF和MPA检测的线性范围均为0.01～20μmol/L,线性回归方程分别为MMF:As/Ai=-0.035 2+0.394 5 c(r=0.999 1);MPA:As/Ai=0.005 5+0.050 8 c(r=0.999 5)。低、中、高浓度(0.1、1.5、15μmol/L)MMF的回收率分别为(103.2±7.5)%、(97.3±3.1)%和(96.0±3.4)%,日内、日间RSD<7.27%;低、中、高浓度(0.1、1.5、15μmol/L)MPA的回收率分别为(106.9±1.0)%、(106.0±5.7)%和(108.4±3.2)%,日内、日间RSD<9.25%(n=5)。结论:本研究建立的测定方法简便、准确、灵敏度高,适用于MMF和MPA体外转运实验研究。     

Stability of mycophenolic acid in plasma samples from patients during mycophenolate mofetil therapy

The aim of this study was to investigate the stability of mycophenolic acid (MPA) in human plasma from patients receiving mycophenolate mofetil (MMF) therapy. MPA was measured using a validated HPLC method. MPA concentrations were determined immediately after receiving the samples in the laboratory, and after 24 h, 72 h, 96 h, 7, 14, 21 days of storage at two different temperatures at -20 degrees C and 4 degrees C. The number of samples used in different conditions of temperature was limited by plasma volume in patients specimens. Monitoring human plasma samples stored at -20 degrees C showed that MPA was stable at this temperature for at least 3 weeks. MPA concentrations were found to be stable also up to 96 hours when stored at 4 degrees C.     

HPLC法同时测定人血浆中甘草酸和苦参碱的质量浓度

目的建立以HPLC法同时测定人血浆中甘草酸和苦参碱质量浓度的方法。方法 Ultimate AQ-C18柱(250mm×4.6mm,5μm),柱温为室温(25℃),流动相为乙腈-三乙胺调pH至7.5的0.05mol·L-1磷酸二氢钾(22.5∶77.5),流速为1.0mL·min-1,检测波长为220nm(检测苦参碱)和250nm(检测甘草酸),进样量50μL。结果甘草酸单铵盐质量浓度在2.0²00.0μg·mL-1范围内线性良好,苦参碱质量浓度在2.0200.0μg·mL-1范围内线性良好,苦参碱质量浓度在2.0²00.0μg·mL-1范围内线性良好。平均回收率均大于95%,日内、日间RSD均小于4%。结论该方法简便、快速、灵敏、可靠,可用于人血浆中甘草酸和苦参碱质量浓度的监测。     

HPLC determination of mycophenolic acid and mycophenolic acid glucuronide in human plasma with hybrid material

Mycophenolic acid (MPA), the active metabolite of the prodrug mycophenolate mofetil is an immunosuppressive agent which inhibits inosine monophosphate dehydrogenase. MPA is metabolised to phenolic glucuronide (MPAG) that may be hydrolysed in vivo to form free MPA. Drug monitoring is required in patients with multi-organ failure. Here, we report a HPLC method with organic/inorganic hybrid material for the simultaneous analysis of MPA and MPAG in human plasma. MPA and MPAG and carboxy butoxy ether mycophenolic acid (MPAC) used as internal standard were analysed on a bonded X-Terra column with a linear gradient elution mode using orthophosphoric acid and acetonitrile as eluents. Sample treatment procedure consists of deproteinisation with acetonitrile. Analytical recoveries were higher than 98 and 89% at concentrations ranging from 1 to 25 and 20 to 200mg/L for MPA and MPAG, respectively. Calibration curves fitted by plotting the peak area ratio (compound of interest/internal standard) versus concentration were linear in the range 0.2-50mg/L for MPA and in the range 1-500mg/L for MPAG. The quantification limit was 0.2mg/L for MPA and 1mg/L for MPAG with a coefficient of variation less than 20% for a 500microL sample volume. Intra- and inter-assay coefficient of variation was lower than 7% for all compounds. Detection was performed at 215nm. Peak identity was confirmed through library matching by comparison with reference spectra. The X-Terra column provides good peak shape and may be used at low pH with a long life-time column. This HPLC method using a simple sample treatment procedure appears suitable for therapeutic drug monitoring in organ-transplant patients. The method is sensitive enough for monitoring MPA and MPAG during pharmacokinetic studies.     

Relationship between mycophenolate mofetil side effects and mycophenolic acid plasma trough levels in renal transplant patients

There has been limited experience with routine therapeutic monitoring of mycophenolic acid (MPA; CAS 24280-93-1), which is the active metabolite of the new immunosuppressive prodrug mycophenolate mofetil (MMF; CAS 115007-34-6). MMF was introduced with recommendation for fixed oral dosing (1 g twice daily) in combination with cyclosporin A (CSA) and a glucocorticoid for the prevention of renal allograft rejection. In the course of routine CSA monitoring a MPA monitoring was performed in adult renal transplant patients receiving MMF in combination with CSA and methylprednisolone (MEP). For 30 consecutive patients with 234 plasma samples the relationship of MMF doses used and MPA plasma through levels estimated at steady state (C88 min) to clinical outcome was evaluated retrospectively. The MPA concentrations were determined with the enzyme-multiplied immunoassay technique (EMIT mycophenolic acid assay) on a Cobas Mira Plus analyzer. The within-run (n = 10) and between-run (n = 10) coefficients of variation were 3.6%, 3.5%, 3.1% and 3.6%, 5.1%, 6.7% analysing three MPA level plasma controls (1.25 mg/l, 7.5 mg/l, 12.5 mg/l), respectively. The data analysis of the MPA plasma trough levels resulted in a high variability between patients (0.3 to 3.4 mg/l) received the recommended fixed MMF dose (2 g/day). There was a higher incidence of adverse reactions with increasing MPA plasma trough levels (2.13 +/- 1.35 mg/l in 13 patients with side effects versus 1.53 +/- 0.67 mg/l in 17 patients without side effects; p < 0.001), regardless of reduction of MMF dose (1.77 +/- 0.3 g/day versus 1.89 +/- 0.2 g/day; NS), respectively. No acute rejection episodes occured under MMF administration in combination with CSA and MEP. The study shows that the to date recommended MMF dose resulted in individual, quite different MPA plasma trough levels, which were associated with incidence of side effects rather than the MMF doses. Therefore, monitoring of plasma MPA trough levels and individual dose adjustment could be helpful to reduce the incidence of adverse reactions and to increase the safety of MMF therapy.     

HPLC法同时测定人尿液中霉酚酸及其代谢物葡糖苷酸结合物的浓度

目的用HPLC测定人尿液中霉酚酸(MPA)及其代谢物葡糖苷酸结合物(MPAG)的浓度。方法分析柱为Zorbax Eclipse XDB-C8(150 mm×4．6mm,5μm);流动相为甲醇-0．1％三氟乙酸溶液(55:45,V／V);尿稀释5倍后直接进样。MPAG采用紫外检测,检测波长为250nm;MPA采用柱后添加0．2 mol·L-1 NaOH溶液进行荧光检测,荧光激发波长325 nm,发射波长435 nm。结果MPA和MPAG的线性范围分别为0．1-50 mg·L-1和10-500 mg·L-1,MPA、MPAG日内和日间RSD均<10％。结论此法能简便、灵敏、准确地测定人体尿液中MPA与MPAG的浓度,可用于临床药动学研究和治疗药物浓度监测。     

高效液相色谱法测定人血浆中霉酚酸及其代谢产物的浓度

目的建立高效液相色谱法建立测定人血浆中霉酚酸(MPA)、霉酚酸葡萄糖苷(MPAG)、霉酚酸酰基葡萄糖苷(Ac MPAG)的浓度。方法血浆处理采用乙腈蛋白沉淀法。色谱柱为Gemini C18(250mm×4.6 mm,5μm),柱温为40℃。Ac MPAG、MPA采用荧光检测,流动相:甲醇-10 mmol·L-1KH2PO4(43:57,p H=8.0),荧光激发波长343 nm,发射波长425 nm。MPAG采用紫外检测,流动相:甲醇-10 mmol·L-1 KH2PO4(45:55,p H=3.0),检测波长254 nm。结果 MPA、MPAG、Ac MPAG血药浓度分别在0.2~20μg·m L-1、4.75~475μg·m L-1、0.02~2μg·m L-1内与峰面积线性关系良好。MPA、MPAG、Ac MPAG的相对回收率分别为88.1%~104.3%、95.7%~99.6%、94.2%~96.0%。日内及日间RSD均<10%。结论该方法测定人血浆中MPA、MPAG、Ac MPAG的浓度,操作简便,方法灵敏度高,适合用于MPA及其代谢产物的药动学研究及血药浓度监测。