RP-HPLC法测定人尿中甲苯磺丁脲及其代谢产物的浓度

目的:建立以反相高效液相色谱法测定人尿中甲苯磺丁脲及其代谢产物羧基甲苯磺丁脲和羟基甲苯磺丁脲浓度的方法。方法:以氯磺丁脲为内标,尿样经酸化后用异丙醚萃取,并进样分析;色谱柱为DiamonsilTM C18,流动相为乙腈-0.05 mol.L-1NaH2PO4(pH4.0,梯度洗脱),流速为1.0 mL.min-1,检测波长为230 nm,柱温为40℃。结果:甲苯磺丁脲、羧基甲苯磺丁脲、羟基甲苯磺丁脲尿药浓度均在0.3～10μg.mL-1范围内线性关系良好(r分别为0.999 9、1.000 0、0.999 9),最低检测浓度均为0.1μg.mL-1。各样本低、中、高浓度的方法回收率在95.00%～105.88%之间,日内、日间RSD均<6%。结论:本方法简便、准确、灵敏、重复性好,适用于细胞色素P4502C9表型分析及甲苯磺丁脲与其代谢产物的人体药动学研究。     

超高效液相串联质谱法测定大鼠格列美脲及其代谢物的血药浓度

目的建立超高效液相串联质谱法快速测定大鼠体内格列美脲及其代谢物羟基格列美脲的浓度。方法用乙腈沉淀蛋白的方法处理血浆,色谱柱为ACQUITY UPLC HSS T3柱(50 mm×2.1 mm,1.8μm);流动相为乙腈-0.1%甲酸水,梯度洗脱;流速为0.4 m L·min-1;用正离子多离子反应监测(MRM)扫描,内标为甲苯磺丁脲。结果血浆中格列美脲和羟基格列美脲的线性范围为10~800μg·L-1和1~80μg·L-1(r=0.999 9和0.999 5),最低定量限为2.00μg·L-1和0.50μg·L-1,回收率均为94.05%~105.33%。两者的日内、日间精密度RSD均<8.20%。结论该方法操作简便、快捷,灵敏度高,适于大鼠体内格列美脲及其代谢物的药动学研究。     

反相高效液相色谱法测定人血浆中甲磺丁脲的含量

目的建立反相高效液相色谱法用于测定人血浆中甲磺丁脲的含量。方法采用Eclipse XDB-C_(18)色谱柱,以乙腈-25 mmol·L~(-1)乙酸钠缓冲液(pH=3.3,32:68)为流动相,流速1.0 mL·min~(-1),检测波长为229 nm,柱温35℃。血样经等体积比的0.6 mol·L~(-1)三氯乙酸处理后,离心,取上清液20μL,进样检测。结果甲磺丁脲血浆药物浓度在0.5～100 mg·L~(-1)内,线性关系良好(r=0.999 6);回收率为93.0%～105.0%;日内RSD≤3.80%,日间RSD≤6.31%。结论本方法简单快速、准确灵敏、回收率高、重现性好,适用于甲磺丁脲的血药浓度测定。     

HPLC法测定甲苯磺丁脲片的含量

目的：建立HPLC法测定甲苯磺丁脲片的含量。方法：色谱柱为VP—ODSC18（250mm×4．6mm,5μm）,甲醇一磷酸二氢铵溶液（pH：3．5）（65：35）为流动相,流速为1．0ml·min^-1,检测波长为254nm,柱温：室温,进样量：20μl。结果：甲苯磺丁脲在0．08—1．20mg·ml^-1浓度范围内与峰面积呈良好的线性（r=0．9998）。平均回收率为99．61％（RSD=0．8％,n=9）。结论：本法定量准确,可用于甲苯磺丁睬片的含量测定。     

高效液相色谱法测定甲苯磺丁脲片的含量

目的 建立了高效液相色谱法测定甲苯磺丁脲片中甲苯磺丁脲含量的方法.方法 主要参数为:色谱柱为KromasilC18(250mm×4.6mm,5μm),流动相为:甲醇-0.02mol/L的乙酸铵溶液-0.1%醋酸溶液(20:20:60),检测波长为235nm,流速为lmL/min.该方法 时甲苯磺丁脲的平均回收率为99.10%.RSD为0.62%(n=5).本法与标准法比较,没有显著性差别.结果 方法 简便,结果 准确,专属性强.结论 可用于含量的测定.     

HPLC法测定荷胆胶囊中牛磺熊去氧胆酸的含量

目的:建立高效液相色谱法测定荷胆胶囊中牛磺熊去氧胆酸的含量.方法:色谱柱:Alltech Alltima C18(250 mm×4.6mm,5μm);流动相∶甲醇-0.03 mol·L-1磷酸二氢钠溶液(65∶ 30)(用磷酸调节pH至4.4);流速:1ml·min-1;检测波长:210 nm.结果:牛磺熊去氧胆酸在4.1～12.2μg范围内线性关系良好(r=0.999 9);平均回收率为98.0％(RSD=1.46％,n=6).结论:该含量测定方法快速,准确,有效,可用于荷胆胶囊中牛磺熊去氧胆酸的含量测定.     

高效液相色谱法测定血浆中尼扎替丁浓度

目的:建立测定人血浆中尼扎替丁的高效液相色谱方法.方法:采用Diamonsil C18色谱柱(200 mm×4.6mm,5μm),流动相为0.1 mol·L-1醋酸铵缓冲液-甲醇(60:40),检测波长为320nm.血浆样品加盐析溶液碱化后以氯仿提取,雷尼替丁为内标.结果:尼扎替丁血药浓度线性范围为20～6 000l·L-1(r=0.999 9,n=6),最低检测浓度为10μg·L-1(S/N=3),方法回收率在96.84%～101.39%(n=5),日内和日间RSD均小于4%.结论:本法简便,快速,重现性好,适于尼扎替丁的药动学研究.     

高效液相色谱法测定人血浆中芬太尼浓度

目的:建立高效液相色谱测定人血浆中芬太尼浓度的方法.方法:采用外标法,以Kromasil-C18(4.6 mm×250 mm,5μm)为固定相,舍0.02 mol·L-1磷酸二氢钠水溶液-乙腈(65:35)为流动相,流速1 mL·min-1,检测波长为220 nm.结果:血浆芬太尼浓度在6～300μg·L-1范围内与峰面积呈良好线性关系(r=0.999 6),最低检测浓度为5 μg·L-1,方法回收率为(92.6±1.5)%,提取回收率为(84.9±1.4)%,日内RSD为1.8%,日间RSD为2.6%.结论:本方法具有较高的准确度,线性范围宽,方法灵敏,专一性好,操作简便,适用于临床芬太尼血药浓度的测定及临床药动学研究的要求.     

餐时血糖调节剂米格列奈钙的RP-HPLC法人肝微粒体药物浓度测定

目的:建立人肝微粒体中米格列奈的高效液相色谱测定法.方法:采用Diamonsil-C18色谱柱(200 mm×4.6 mm,5μm);流动相为乙腈-0.02 mol·L-1KH2PO4缓冲液(pH 4.0)(43:57);流速为1.0 mL·min-1;紫外检测波长为210 nm;进样量为20 mL.结果:米格列奈最低检测限为2 μmol·L-1;线性范围为5～1 000 μmol·L-1(r=0.999 9);最低定量浓度为5μmol·L-1(RSD<5%,n=5);低、中、高3种浓度日内、日间RSD(n=5)分别为1.9%,1.6%,1.4%和3.8%,4.0%,3.8%;绝对回收率平均为94.4%,相对回收率平均为101.2%.结论:本实验建立的人肝微粒体中米格列奈的高效液相测定方法操作简便、结果准确.     

SPE-HPLC法测定止嗽立效丸中吗啡的含量

目的:建立HPLC法测定止嗽立效丸中吗啡的含量。方法:预处理应用固相萃取技术;液相色谱采用Shim-pack C18柱(6.0 mm×150 mm,5μm),流动相为乙腈-0.05 mol.L-1磷酸二氢钾-0.005 mol.L-1庚烷磺酸钠(10∶45∶45),流速1 mL.min-1,柱温25℃,检测波长210 nm。结果:吗啡进样量在0.026×10～2.58μg范围内线性关系良好(r=0.999 9),平均回收率为101.3%,RSD为1.2%(n=9)。结论:方法简便、准确,重复性好。     

Quantitative determination of tolbutamide and its metabolites in human plasma and urine by high-performance liquid chromatography and UV detection

An isocratic, high-performance liquid chromatography method has been developed for simultaneous determination of the oral antidiabetic tolbutamide and two of its metabolites, 4-hydroxytolbutamide and carboxytolbutamide, in human plasma and urine. The method was based on simple one-step liquid-liquid extraction with tertiary-butyl methyl ether as extraction solvent. The chromatographic eluent was 23:77 (v/v) methanol: 0.01 M aqueous sodium acetate buffer pH 3.0, and the UV detection was performed at a wavelength of 230 nm. The limit of detection was 0.1 microM for tolbutamide in plasma and 1.5 microM, 0.5 microM, and 0.75 microM for carboxytolbutamide, 4-hydroxytolbutamide, and tolbutamide, respectively, in urine. The limit of quantitation was 0.5 micro for tolbutamide in plasma and 2 microM, 0.75 microM, and 1.25 microM for carboxytolbutamide, 4-hydroxytolbutamide, and tolbutamide, respectively, in urine. The overall mean recoveries ranged from 91% to 109% for tolbutamide in plasma and from 80% to 98% in urine for all three compounds.     

A screening test for slow metabolisers of tolbutamide.

1. Six subjects participated in a detailed pharmacokinetic study of tolbutamide (pilot study). Using parameters based on these data, sixty-three non-diabetic volunteers underwent a simple screening test designed to identify slow metabolisers of tolbutamide. 2. The screening test was an estimate of tolbutamide plasma elimination half-life from plasma concentrations at 8 and 24 h after 500 mg tolbutamide orally, and urinary recovery of the hydroxy- and carboxytolbutamide metabolites over the 4-8 h post-dose period. 3. The mean tolbutamide half-life for 61 of the screened subjects was 7.5 +/- 1.5 h (range 5.2-12.2 h). Two subjects had half-lives of 21.6 and 16.1 h. Their urinary metabolite recoveries were within the range of those in the screening test but lower than those in the pilot study. 4. The subject with the 21.6 h half-life was restudied with intensive serial sampling for 72 h post-dose. She was confirmed as a 'slow' metaboliser of tolbutamide since her terminal half-life was 25.9 h but plasma Cmax and tmax were within the range of those in the detailed study. This subject's 24 h urinary recoveries of both hydroxytolbutamide and carboxytolbutamide were clearly different from the mean values for the pilot study subjects implicating hydroxylation of tolbutamide as the metabolic defect. 5. The two point plasma half-life is therefore a discriminatory screening test but a 4-8 h urinary recovery is not. 6. A partial family study did not provide conclusive evidence of the inheritance of slow tolbutamide metabolism but the screening test should allow simple identification of slow metabolisers for further study.     

Method for the quantitative analysis of cilostazol and its metabolites in human plasma using LC/MS/MS

An LC/MS/MS method for the simultaneous determination of cilostazol, a quinolinone derivative, and three active metabolites, OPC-13015, OPC-13213, and OPC-13217, in human plasma was developed and validated. Cilostazol, its metabolites, and the internal standard, OPC-3930 were extracted from human plasma by liquid-liquid partitioning followed by solid-phase extraction (SPE) on a Sep-Pak silica column. The eluent from the SPE column was then evaporated and the residue reconstituted in a mixture of methanol/ammonium acetate buffer (pH 6.5) (2:8 v/v). The analytes in the reconstituted solution were resolved using reversed-phase chromatography on a Supelcosil LC-18-DB HPLC column by an 17.5-min gradient elution. Cilostazol, its metabolites, and the internal standard were detected by tandem mass spectrometry with a Turbo Ionspray interface in the positive ion mode. The method was validated over a linear range of 5.0-1200.0 ng/ml for all the analytes. This method was demonstrated to be specific for the analytes of interest with no interference from endogenous substances in human plasma or from several potential concomitant medications. For cilostazol and its metabolites, the accuracy (relative recovery) of this method was between 92.1 and 106.4%, and the precision (%CV) was between 4.6 and 6.5%. During the validation, standard curve correlation coefficients equalled or exceeded 0.999 for cilostazol and its metabolites. These data demonstrate the reliability and precision of the method. The method was successfully cross-validated with an established HPLC method.     

HPLC法测人血浆中丹皮酚浓度及其药代动力学研究

目的：建立HPLC丹皮酚胶囊血浓度测定方法，评价丹皮酚胶囊的药代动力学特点。方法：24名健康志愿者单剂量口服160mg丹皮酚胶囊，按设定时间采集肘静脉血经乙睛萃取处理，以XB—C18（250mm×4．6mm，5μm）色谱柱为固定相，四氢呋喃-甲醇-水-磷酸（6：60：34：0．1）为流动相测定丹皮酚血浆浓度。采用DAS2．0计算丹皮酚主要药动学参数。结果：丹皮酚线性范围10—500mg／mL，最低检出浓度为10ng／mL。主要药代动力学参数：Cmax为（116±46）ng／mL，tmax为（1．02±0．13）h，t1/2为1．03h。结论：建立的HPLC方法特异性强、灵敏度高，可用于丹皮酚血药浓度测定和人体药动学研究。     

Determination of ketobemidone and its metabolites in plasma and urine using solid-phase extraction and liquid chromatography-mass spectrometry

The authors developed a sensitive, specific, and rapid liquid chromatography--mass spectrometry (LC-MS) method for determining ketobemidone and its major metabolites in plasma and urine. The method involves a solid-phase extraction, high-performance liquid chromatography (HPLC), and electrospray mass spectrometry. The limit of quantification for ketobemidone and norketobemidone was 3 nmol/L. Recovery rates for ketobemidone and norketobemidone were 84.8% and 81.1%, respectively. Coefficients of variation (CV) ranged from 2.8 % to 9.5%. The method was used to determine ketobemidone and its major metabolites in clinical samples from relevant patient groups.     

机械通气患儿血浆中咪达唑仑及其代谢产物的测定

目的 建立测定人血浆中咪达唑仑及其代谢产物浓度的固相萃取和HPLC方法.方法 采用HPLC法分别测定29例ICU机械通气患儿给药24 h后血浆中咪达唑仑及其代谢产物浓度.色谱条件为：ZORBAX Eclipse XDB-C18色谱柱（4.6×250 mm,5 μm）;流动相为7.56 mmol/L硫酸铵溶液-乙腈,梯度洗脱,柱温25 ℃,流速1.0 ml/mmin,紫外检测波长254 nm.结果 4-羟基咪达唑仑、1＇-羟基咪达唑仑和咪达唑仑的保留时间分别为8.14、9.09和11.18 min.咪达唑仑及其羟基代谢产物的相对回收率98.88%～100.01%,RSD＜4%.29例患者血浆中咪达唑仑、1＇-羟基咪达唑仑和4-羟基咪达唑仑的平均浓度为0.68,0.28和0.08mg/L.结论 该方法适用于咪达唑仑及其代谢产物血药浓度的常规监测.     

Determination of cilostazol and its metabolites in human urine by high performance liquid chromatography

A high performance liquid chromatography (HPLC) method with ultraviolet detection for the simultaneous quantification of cilostazol, and its known metabolites in human urine was developed and validated. Cilostazol, its metabolites and the internal standard OPC-3930 (structural analogue of cilostazol) were extracted from human urine using liquid-liquid extraction with chloroform. The organic extract was then evaporated and the residue was reconstituted in 8% acetonitrile in ammonium acetate buffer (pH 6.5). The reconstituted solution was injected onto an HPLC system and was subjected to reverse-phase HPLC on a 5-microm ODS column. A gradient mobile phase with different percentages of acetonitrile in acetate buffer (pH 6.5) was used for the resolution of analytes. Cilostazol, its metabolites and the internal standard were well resolved at baseline with adequate resolution from constituents of human urine. The lower limit of quantification was 100 ng/ml for cilostazol and all metabolites. The method was validated for a linear range of 100-3000 ng/ml for all the metabolites and cilostazol. The overall accuracy (% relative recovery) of this method ranged from 86.1 to 116.8% for all the analytes with overall precision (%CV) being 0.8-19.7%. The long-term stability of clinical urine samples was established for at least 3 months at -20 degrees C in a storage freezer. During validation, calibration curves had correlation coefficients greater than or equal to 0.995 for cilostazol and the seven tested metabolites. The method was successfully used for the analysis of cilostazol and its metabolites in urine samples from clinical studies, demonstrating the reliability and robustness of the method.     

A high-performance liquid chromatographic method for the determination of encainide and its major metabolites in urine and serum using solid-phase extraction

A high-performance liquid chromatography (HPLC) procedure used to quantitate encainide and two of its active metabolites, O-desmethylencainide (ODE) and 3-methoxy-O-desmethylencainide (MODE), is described. All three compounds were simultaneously extracted from urine and serum using an octyl (C-8) solid-phase extraction column. The compounds were then separated by reverse-phase HPLC on a cyanopropylsilane column using ultraviolet detection at 260 nm. Serum samples were quantified over a concentration range of 25-400 ng/ml and urine over a range of 150-10,000 ng/ml. Total run time for the assay was less than 12 min. Within-day and between-day precision and relative error were less than 10% in serum and less than 13% in urine for all three compounds. The lower limit of quantitation was 10 ng/ml for encainide and ODE and 15 ng/ml for MODE. This HPLC procedure represents a quick and reliable method of measuring encainide and its major metabolites in both urine and serum, making the assay applicable as an aid for therapeutic drug monitoring of patients receiving encainide therapy.     

HPLC method for the determination of rosiglitazone in human plasma and its application in a clinical pharmacokinetic study

Rosiglitazone (CAS 155141-29-0, Avandia) is a novel insulin sensitizer used in the treatment of type 2 diabetes. A sensitive high performance liquid chromatography (HPLC) method for its determination in human plasma using fluorescence detection (excitation: 247 nm, emission: 367 nm) with a suitable internal standard (I. S.) is described. Ethyl acetate was used as extraction solvent. A mobile phase consisting of phosphate buffer, acetonitrile and methanol was used at a flow rate of 1.0 ml/min on a C18 column. The absolute recovery was > 90% and the lower limit of quantitation was 5 ng/ml. The intra- and inter-day relative standard deviations ranged from 0.58-6.69% and 0.82-6.63%, respectively. The method described is simple, economical, precise and accurate and has been successfully applied in a pharmacokinetic study conducted in healthy human volunteers.