毛细管气相色谱法测定人血浆中5-单硝基异山梨醇酯

目的:建立测定人血浆中5-单硝基异山梨醇酯(IS-5-MN)浓度的气相色谱检测法.方法:分析柱为HP- 5毛细管柱,进样口、色谱柱及检测室温度分别为210、160及200℃,检测器为~(63)Ni电子捕获检测器.采用乙酸乙酯萃取血浆中IS-5-MN及内标 2,4-二硝基甲苯.结果:IS-5-MN血药浓度在10～800ng/ml范围内线性关系良好(r=0.999 8),最低检测浓度为5ng/ml,日内及日间差异均小于10%.结论:本方法适用于IS-5-MN的药物动力学研究.     

胶束液相色谱法测定血清中硝苯地平的含量

目的:建立以胶束液相色谱法测定人血清中硝苯地平含量的方法。方法:色谱柱为C18,流动相为甲醇-水(60∶40,内含25mmol/L的SDS),检测波长为248nm,灵敏度为0.01AUFS,流速为1.0ml/min,记录纸速为2.1mm/min。结果:硝苯地平检测浓度在4～40ng/ml范围内线性关系良好(r=l.0002),平均回收率为98.15%(RSD=1.49%)。结论:本方法操作简便、准确、易行、重现性好,可为硝苯地平的临床血药浓度监测提供一种新的方法。     

在线固相萃取-HPLC法测定比格尔犬血浆中丹参素的含量

目的:建立测定比格尔犬血浆中丹参素含量的在线固相萃取-HPLC法。方法:血浆样品经蛋白质沉淀,采用Lichrospher C18为富集柱,以乙腈-10 mmol/L NaH2 PO4(5∶95)为富集流动相,流速2 ml/min;选择Ultimate XB-C18(50 mm×4.6 mm,5μm)为分析柱,以乙腈-10 mmol/L NaH2 PO4(11∶89)为分析流动相,流速为0.8 ml/min,检测波长285 nm。结果:丹参素在20~2 000 ng/ml浓度范围内线性关系良好,最低定量限为20.0 ng/ml,日内和日间RSD均<10%,平均绝对回收率>80%。结论:本方法简便、快速、灵敏、可靠,适用于比格尔犬血浆中丹参素含量的测定。     

HPLC测定血浆中吡咯地尔浓度

建立了测定吡咯地尔血浆药物浓度的HPLC.采用YWG-C_(18)柱(10μm,15cm×4.6mm I.D.),检测波UV254nm,流动相为甲醇:水:10%三乙胺磷酸缓冲液(pH4.5):二氯甲烷=75:15:10:5,流速1.0ml/min;1.0ml血浆用正己烷:异丁醇(98:2)提取,内标法定量.线性范围10～2000ng/ml,最低检测浓度3ng/ml.平均提取回收率90.02%,平均方法回收率100.33%,日内RSD<4.0%,日间RSD<5.5%.     

用HPLC柱切换法血浆直接进样测定法莫替丁

本文采用HPLC柱切换技术,建立了以YWG-C18(3cm)为预处理柱,乙酸(0.2mol/L)为预处理流动相,血浆直接进样。在线(on-line)固相净化血浆样品;以Shimpack CLC-ODS(15 cm)为分析柱,甲醇-0.2 mol/L乙酸铵缓冲液(14:86)为流动相,267 nm波长检测,采用外标法定量。测定H₂-受体拮抗剂法莫替丁人体血药浓度的方法。该法净化过程简便,为自动净化血浆样品提供了一选择方法。回收率好(81.10～84.75%);法莫替丁的检测限约为2.4 ng,血浆中最低检出浓度约为12 ng/ml。日内变异系数为4.1%,日间变异系数为4.9%;在浓度为15～210 ng/ml血浆范围内呈线性关系。     

阿替洛尔与硝苯地平复方片的HPLC测定

采用高效液相色谱法测定复方阿替洛尔片中阿替洛尔与硝苯地平的含量.色谱柱为Kromasil C18,流动相为甲醇-水-磷酸盐缓冲液(pH3.0)(55:42:3,含6.0 mmol/L辛烷磺酸钠),流速0.8ml/min,检测波长274 nm.阿替洛尔和硝苯地平的线性范围分别为20～80μg/ml和8～32μg/ml,平均回收率分别为100.2%(RSD 0.45%)和100.4%(RSD 0.77%).     

犬血浆中氨氯地平和阿托伐他汀的LC-MS／MS测定

建立了LC-MS/MS法同时测定犬血浆中氨氯地平和阿托伐他汀的含量.用甲醇沉淀血样中的蛋白质.采用C18色谱柱,2 mmol/L醋酸胺溶液(含0.5%甲酸).甲醇(32:68)为流动相,以地西泮为内标,电喷雾离子化,正离子多反应监测,检测离子分别为m/z409.2→m/z 238.1(氨氯地平)、m/z 559.2→m/z 440.5(阿托伐他汀)和m/z285.1→m/z 193.1(地西泮).结果氨氯地平和阿托伐他汀在0.1～20ng/ml和0.2～50 ng/ml度范围内线性关系良好,定量限为0.1和0.2 ng/ml,方法回收率均大于94%,日内、日间RSD均小于9%.     

HPLC法快速测定人血浆中酒石酸美托洛尔的浓度

目的:建立快速测定人血浆中酒石酸美托洛尔浓度的方法。方法:血浆样品以3-叔丁基甲醚处理后,采用高效液相色谱法进样测定,色谱柱为Kromasil LC-18DB,流动相为乙腈-10 mmol/L辛烷磺酸钠溶液(p H=2.0)(30∶70),柱温为25℃,流速为1.3ml/min,荧光检测波长为267 nm(激发波长)、290 nm(发射波长),内标为吲哚洛尔。结果:酒石酸美托洛尔血药浓度在2~300 ng/ml范围内线性关系良好(r=0.999 9),最低检测浓度为1 ng/ml。平均方法回收率为(101.13±4.0)%,日内RSD≤2.47%,日间RSD≤4.51%。结论:该方法简便、快速、灵敏、重现性好,适用于酒石酸美托洛尔临床血药浓度监测及人体药动学研究。     

液相色谱串联质谱法测定人血浆中硝苯地平的浓度

目的 建立测定人血浆中硝苯地平浓度的LC-MS/MS方法.方法 血浆样品经液液提取后,以甲醇(含0.1%乙酸)∶水(含0.1%乙酸 2.5 mmol·L-1甲酸胺)=80∶20为流动相,使用Agilent1100 VL型离子阱质谱仪,电喷雾离子源正离子模式,采用多反应离子监测方式测定硝苯地平(MRM,m/z 347→315).结果 血浆中硝苯地平的线性范围为1.0～100.0 ng·mL-1.定量下限为1.0 ng·mL-1.准确度在85%～115%之间,日内、日间精密度(RSD)在±15%之内.结论 该方法准确,特异性强,可用于硝苯地平药动学研究.     

HPLC法测定体液中氨溴索浓度及其药代动力学参数

建立了 HPLC测定人血浆及尿中盐酸氨溴索含量的方法 :Hypersill C18柱 (4.6mm× 2 50 mm,5μm) ,乙腈 -甲醇 - 0 .0 1 mol/L磷酸盐缓冲液 -四氢呋喃 (35∶ 35∶ 2 7.5∶ 2 .5,V/V)为流动相 ,流速 1 .5ml/min,检测波长 2 4 2 nm。结果表明 :最低检测浓度为 5ng/ml,血药浓度在 1 0～ 32 0 ng/ml范围内线性良好 ,尿药浓度在 0 .2 5～ 8.0μg/ml范围内线性良好 ,氨溴索生物半衰期为 (4.2 1± 0 .93) h。     

Reversed-phase high performance liquid chromatographic determination of nifedipine in human plasma

A reversed-phase high performance liquid chromatographic method is presented by which the calcium channel blocker, nifedipine (NFP), may be measured in human serum using 17-alpha-ethinylestradiol as an internal standard. A mobile phase of phosphate buffer (0.01 M, pH 6.1)/methanol/acetonitrile (20:35:45) passed through a muBondapak C-18 column at 1.0 ml/min produced symmetrical bands for nifedipine and internal standard. A rapid and simple chloroform extraction of NFP from 1 ml serum samples proved to be efficient and reproducible. Recovery over a concentration range of 5-100 ng/ml was 92.3 +/- 5.1% (mean +/- SD, n = 6). Ultraviolet absorbance detection at 235 nm was sensitive to serum NFP concentrations of 5 ng/ml. Between-day coefficients of variation at 100 and 5 ng/ml were 4.0 and 11.4%, respectively (n = 10). Serum concentration data from NFP-treated subjects are presented.     

国产盐酸西替利嗪片的药代动力学研究

目的 :建立测定国产盐酸西替利嗪 (西可韦 )血药浓度的方法并进行人体药代动力学研究。方法 :采用反相高效液相色谱法 ,以盐酸普罗帕酮为内标 ;色谱柱 :WaterssymmetryC18 不锈钢柱 (3 9mm×150mm ,5μm) ;流动相 :乙腈 -0 02mol/L磷酸二氢钠 -三乙胺 (50∶50∶0 16 ,V/V) ,含十二烷基硫酸钠 (SDS)4 0mmol/L ;流速 :1 0ml/min ;检测波长 :229nm。测定11名健康男性志愿者单剂量口服西可韦片10mg 的血浆中药物浓度。结果 :线性范围为12 5～800ng/ml,最低检测限为5ng/ml,提取回收率>75 %。11名志愿者的血药浓度数据经3p87软件拟合 ,符合血管外给药二室模型 ,其主要药代动力学参数为 :Cmax=(429 00±108 80)ng/ml,Tmax= (0 91±0 40)h ,K10= (0 18±0 04)/h ,以梯形法计算的AUC0～36= (3312 72±682 39)ng/(ml·h)。结论 :本方法结果准确 ,灵敏度高 ,能满足人体药代动力学研究的需要 ;西可韦主要药代动力学参数与国内、外文献报道一致 ,可广泛应用于临床     

A validated method for the determination of verapamil and norverapamil in human plasma

The aim of the study was to develop a simple and sensitive analytical method to determine verapamil (V) and its metabolite norverapamil (N) in human plasma with use of an HPLC isocratic system with fluorescence detection, following fast extraction of the investigated compounds. Extraction recovery was 92.12% and 89.58% for V and N, respectively. Internal standard in HPLC was propranolol. Its recovery was 82.50% on the average. Limit of detection was 0.924 ng/ml and limit of determination was 3,080 ng/ml for V, what corresponds concentration in plasma 1.232 ng/ml. For N limit of detection was 0.030 ng/ml and limit of determination was 1.001 ng/ml what corresponds 0,4 ng/ml in plasma. Parameters of validation prove that precision of the presented method is very good. The method is fast and one chromatogram separation lasts about 8 minutes. 30-40 manual (without autosampler) analyses per day were done. It was used successfully in pharmacokinetic and bioavailability studies of verapamil administration in drug formulations alternative to tablets: buccal and flotation ones.     

反相高效波相色谱法测定华法令钠血药浓度

建立了反相高效液相色谱测定华法令钠血浆中浓度的方法.方法使用Nova-Pak C_(18)柱;流动相为甲醇:0.001 mol/L醋酸钠溶液=55:45,用冰醋酸调PH 6.0;以卡马西平为内标;提取溶媒为30%异丙醇氯仿;紫外检测波长300nm;流动相流速0.8ml/min.方法最低检出限10ng,最低检出浓度50μg/L.华法令钠血药浓度在0.25～40mg/L范围内线性关系良好(r=0.9999).本方法适用于常规监测华法令钠血药浓度工作.     

特拉唑嗪胶囊的药物动力学及相对生物利用度研究

目的:建立人血浆中特拉唑嗪浓度的HPLC荧光测定方法,研究特拉唑嗪在人体中的药物动力学行为。方法:血浆样品经碱化后,用乙酸乙酯和苯(1:4)提取,色谱柱为 Hypersil-ODS柱,流动相为0.05 mol/L磷酸二氢钾-乙腈-四氢呋喃(70:29.2:0.8),荧光检测器的激发波长为250 nm,发射波长为 370 nm。测定了20名受试者单剂量口眼特拉唑嗪胶囊和市售片剂后的血药浓度。结果:特拉唑嗪浓度在1.5～150.0 ng/ml范围内,线性关系良好(r=0.9994),最低检测限达 1ng/ml,绝对回收率大于 70％,符合生物样品分析要求。受试者口服特拉唑嗪胶囊和片剂 2 mg后,估算的半衰期分别为 10.90±2.24 h和 11.58±2.47 h,达峰时间分别为 1.6±0.6 h和 1.3±0.5 h,峰浓度分别为 36.97±3.53 ng/ml和 38.95±8.31 ng/ml,特拉唑嗪胶囊剂的相对生物利用度为98.0％。结论:特拉唑嗪胶囊剂与市售特拉唑嗪片剂生物等效。     

Determination of oleanolic acid in human plasma and study of its pharmacokinetics in Chinese healthy male volunteers by HPLC tandem mass spectrometry

A highly selective and sensitive HPLC-ESI-MS-MS method was developed for the determination of oleanolic acid in human plasma. The oleanolic acid and glycyrrhetinic acid (internal standard) were recovered from plasma with ethyl acetate liquid-liquid extraction. The organic extracts were dried under a stream of warm nitrogen, reconstituted in mobile phase and injected into a Zorbax-Extend ODS analytical column (150 mm x 4.6 mm i.d., 5 microm), with the mobile phase consisting of methanol-ammonium acetate (32.5 mM) (85:15, v/v) pumped at a flow rate of 1.0 ml/min, and 30% of the eluent was split into a MS system with electrospray ionization tandem mass (ESI-MS-MS) detection in negative ion mode. The tandem mass detection was performed on a Finnigan Surveyor LC-TSQ Quantum Ultra AM tandem mass spectrometer operated in selected reaction monitoring mode. The parent to product ion combinations of m/z 455.4-->455.4 and 469.3-->425.2 at 38 V 1.5 mTorr Ar CID were used to quantify oleanolic acid and glycyrrhetinic acid, respectively. The assay was validated in the concentration range of 0.02-30.0 ng/ml for oleacolic acid when 0.5 ml of plasma was processed. The precision of the assay (expressed as relative standard deviation, R.S.D.%) was less than 15% at all concentrations levels within the tested range and adequate accuracy, and the limit of quantification was 0.02 ng/ml. The established method was applied for the pharmacokinetics study of oleanolic acid capsules in 18 healthy male Chinese volunteers with the mean values of C(max), T(max), AUC(0-48), AUC(0-infinity), t(1/2,) CL/F, and V/F of oleanolic acid after p.o. a single 40 mg dose obtained were 12.12 +/- 6.84 ng/ml, 5.2 +/- 2.9h, 114.34 +/- 74.87 ng h/ml, 124.29 +/- 106.77 ng h/ml, 8.73 +/- 6.11 h, 555.3 +/- 347.7 L/h, and 3371.1 +/- 1,990.1 L, respectively.     

Liquid chromatographic assay of nifedipine in human plasma and its application to pharmacokinetic studies

A highly sensitive, selective and reproducible reversed-phase high-performance liquid chromatographic method has been developed for the determination of nifedipine in human plasma with minimum sample preparation. The method is sensitive to 3 ng/ml in plasma, with acceptable within- and between-day reproducibilities and linearity (r2 > 0.99) over a concentration range from 10-200 ng/ml. Acidified plasma samples were extracted using diethyether containing diazepam as internal standard and chromatographic separation was accomplished on C18 column using a mobile phase consisting of acetonitrile, methanol and water (35:17:48, v/v). The within-day precision ranged from 2.22 to 4.64% and accuracy ranged from 102.4-106.4%. The day-to-day precision ranged from 2.34-7.07% and accuracy from 95.1-100.1%. The relative recoveries of nifedipine from plasma ranged from 91.0-107.3% whereas extraction recoveries were 88.6-93.3%. Following eight 6-week freeze-thaw cycles, nifedipine in plasma samples proved to be stable with accuracy ranging from 0.64 to 3.0% and precision ranging from 3.6 to 4.15%. Nifedipine was also found to be photostable for at least 120 min in plasma, 30 min in blood and for 60 min in aqueous solutions after exposure to light. The method is sensitive and reliable for pharmacokinetic studies and therapeutic drug monitoring of nifedipine in humans after the oral administration of immediate-release capsules and sustained-release tablets to five healthy subjects.     

高效液相色谱法测定乙吗噻嗪在人的血浆浓度及药代动力学

建立了反相高效液相色谱法测定人血浆中乙吗噻嗪浓度。色谱柱采用SpherisotbC18柱(25cm×4.6mm,5μm),流动相为甲醇—水—三乙胺(70:30:0.4,pH6.5),检测波长268nm。用乙腈沉淀蛋白后,吹干浓缩进样。血药浓度在20～4000ng/ml范围内呈线性关系,相关系数0.9994,血浆最低检测浓度3ng/ml。方法回收率90～103%,日内、日间RSD2.4～10.2%。应用该法研究了8名志愿者口服乙吗噻嗪片后的药代动力学,用一室模型拟合,消除相半衰期为1.75±0.45h。本法简便、回收率和灵敏度高、重复性好,适于临床药代动力学和药效学的研究。     

高效液相色谱法测定人体5-氟尿嘧啶血药浓度

目的 :以高效液相色谱法测定人血浆中5 -氟尿嘧啶的浓度。方法 :用硫酸铵作为蛋白沉淀剂 ,血浆样品用乙酸乙酯 -异丙醇(85∶15 ,V/V )提取 ,氮气吹干 ,残留物用流动相溶解后进样。色谱柱为DiamonsilC18 柱 (4 6mm×150mm ,5μm ) ,流动相为0 01mol/LKH2PO4(pH5 5) ,流速1 5ml/min ,紫外检测波长267nm ,5 -溴尿嘧啶为内标。结果 :本法最低检测浓度为0 025μg/ml ,线性范围为0 3～10μg/ml ,日内RSD≤5 0 % (n=5) ,日间RSD≤11 5 % (n=5)。结论 :本法简便、灵敏、经济 ,可作为5 -氟尿嘧啶药代动力学研究时血药浓度的测定方法。     

A model study of bioequivalence of dissolved nifedipine from soft gelatin capsules

Oral absorption of nifedipine determined from plasma concentrations was studied in 20 healthy subjects aged 19 to 40 years following application of a single soft gelatin capsule as a generic preparation (P) and a reference preparation (R) containing 10 mg nifedipine. Nifedipine was measurable in plasma 15 min after application and maximal concentrations occurred after 0.44 and 0.64 h (mean), respectively. Maximal concentrations Cmax varied between 46.4 and 251.0 ng/ml (median 100.7, mean 112.6) after P and between 35.5 and 279.7 ng.h/ml (median 115.8, mean 125.2) after R. Mean areas under the curves (AUC0-infinity) were 173.6 (median 151.4 P) and 188.6 ng.h/ml (median 163.1, R). The minor differences in the AUC values and Cmax values were not statistically significant. The shorter tmax after the generic preparation (p less than 0.05) is clinically unimportant. Since the bioavailability of P is 97% the two preparations are bioequivalent.