Further studies on the resolution of human mercapt- and nonmercaptalbumin and on human serum albumin in the elderly by high-performance liquid chromatography

High-performance liquid chromatographic (HPLC) analysis of human serum albumin (HSA) on Asahipak GS-520 showed at least two peaks, the principal component corresponding to human mercaptalbumin (HMA) and the secondary one to nonmercaptalbumin (HNA). HPLC analysis of HSA on Asahipak ES-520 N showed three peaks, the principal component corresponding to HMA, the secondary one to HNA having mixed disulfide with cysteine or glutathione and the tertiary one to HNA oxidized higher than mixed disulfide. Two kinds of rapid HPLC for the resolution of HSA into HMA and HNA were developed by the present authors. Using these HPLC, the present authors found a significant decrease in the fraction of HMA in the elderly.     

HPLC-studies on nonmercapt-mercapt conversion of human serum albumin

Human mercaptalbumin (HMA) and nonmercaptalbumin (HNA) could be separated by high-performance liquid chromatography (HPLC) at neutral pH. Using HPLC, the present authors found the nonmercapt-mercapt conversion (HNA ← HMA) during hemodialysis and the mercapt-nonmercapt conversion (HMA ← HNA) after hemodialysis in chronic renal failure, indicating HMA as the covalent carrier protein for sulfur-containing amino acids.     

高效液相色谱法测定人血浆中氟罗沙星浓度

目的：建立HPLC方法测定人血浆中氟罗沙星的浓度。方法：取0.1ml血浆样品,二氯甲烷提取,挥干溶剂,残渣加流动相进样;用C18分析柱,以甲醇－20mmol.L^-1磷酸二氢钾－10mmol.L^-1四丁基溴化铵（20：70：10）为流动相,流速：0.8ml.min^-1,紫外检测波长268nm,用峰面积定量。结果：标准曲线在0.25-8μg.ml^-1范围内线性关系良好（r=0.9999),最低检测浓度为0.1μg.ml^-1;方法回收率为95．93％－101．32％,日内RSD＜4％,日间RSD＜8％。结论：本法准确,精密,简便,快速,取样量少,可满足临床血药浓度测试要求。     

高效液相色谱法测定人血中表阿霉素的浓度

目的:建立人血浆中表阿霉素的HPLC测定方法。方法:采用Hypersil ODS-C18柱为分析柱;乙腈-甲醇-水(32∶50∶18,v/v)为流动相,流速为1.0mL.min-1;荧光检测器的激发波长480nm、发射波长560nm;以柔红霉素作为内标,采用沉淀蛋白-脱水法提取样品。结果:表阿霉素、内标与人血浆中其他成分分离良好,在10~1 000μg.L-1的范围内线性良好,r=0.999 1(n=5),回收率为102.63%,日内及日间RSD均小于5%,最低检测限为5μg.L-1。结论:本法取样量小,操作简便快速,适用于临床表阿霉素的血药浓度监测和药动学研究。     

猪胰岛素与重组人胰岛素的高效液相色谱分析

目的鉴别猪胰岛素与重组人胰岛素。方法采用C8、C1 8及常规孔径 (6～ 1 0nm)与大孔径 (30nm)色谱柱用梯度洗脱和等度洗脱对猪胰岛素与重组人胰岛素的分离方法进行研究。结果用梯度洗脱代替英国药典中的洗脱体系能达到相同的分离效果。结论用常规C1 8柱通过梯度洗脱可分离猪胰岛素与重组人胰岛素     

The determination of metoclopramide in plasma by reversed-phase ion-pair high-performance liquid chromatography

Methodology based on reversed-phase ion-pair high-performance liquid chromatography is described for the determination of metoclopramide in plasma. The chromatography was optimized in terms of the peak shape for the drug and its resolution from endogenous plasma components by investigating the effects of quaternary ammonium (competing) ions and alkylsulphate (pairing) ions in an acidic mobile phase containing acetonitrile (20%) and 20 mM acetic acid. Optimum chromatographic conditions were obtained with an ODS-Hypersil column and a mobile phase containing 20% acetonitrile, 20 mM acetic acid, 0.6 mM sodium octylsulphate and 0.5 mM tetrabutylammonium chloride. A simplified method of sample preparation is described in which only 1 ml of plasma is required. The limit of detection (at 310 nm) was 7 ng/ml and no interference from endogenous plasma components or from any drugs commonly used in the treatment of cancer was observed. Consequently the methodology should be applicable to pharmacokinetic studies on metoclopramide, when used clinically to control the gastro-intestinal side-effects of chemotherapy.     

高效液相色谱法测定人体血浆中甲氨蝶呤浓度方法的改进

目的:建立高效液相色谱法测定人体血浆中甲氨蝶呤浓度的方法。方法:血浆样品经处理后,色谱分离采用Zorbax-ODSC18(250mm×4.6mm,5μm)色谱柱,检测波长为302nm,流动相为0.01mol.L-1磷酸盐(用磷酸溶液调pH6.7)-甲醇(80∶20),流速为1.0mL.min-1,柱温为35℃。结果:甲氨蝶呤血药质量浓度在0.2～50mg.L-1(r=0.9997)范围内线性关系良好,最低检测质量浓度为5μg.L-1,提取回收率为96.5%～98.7%,方法回收率为98.2%～101.2%,日内、日间RSD分别为3.62%和4.57%。结论:该方法简单、快速、准确适用于临床甲氨蝶呤浓度监测及药动学研究。     

高压液相色谱法测定人血浆中表阿霉素的含量

目的：建立高压液相色谱(HPLC法)测定人血浆中表阿霉素含量的方法。方法：用甲醇和硫酸钠沉淀蛋白,以高效液相色谱在荧光检测波长λex=450nm,λem=530nm处测定表阿霉素的血药浓度。结果：表阿霉素的线性范围为0．02ug～3ug／mL(r=0．998)。结论：本方法简便、可靠,适用于表阿霉素的血药浓度监测。     

Analysis of 5-OH-indoles in human gut biopsy tissues by reversed-phase high-performance liquid chromatography with fluorimetric detection

A method was devised for the rapid simultaneous determination of major indoles in human gut tissues. Analysis with picomol detection limits was done by HPLC on a C₁₈ reversed-phase column with fluorimetric detection at 276/350 nm. This simple method for which there is no necessity of derivatization or purification was validated for routine analysis of small mucosa samples (less than 4 mg fresh weight) obtainable during endoscopy. A comprehensive list of 5-OH-indole compounds in human gut tissue is presented.     

Determination of fexofenadine enantiomers in human plasma with high-performance liquid chromatography

A simple and sensitive high-performance liquid chromatography (HPLC) method was developed as an assay for fexofenadine enantiomers in human plasma. Fexofenadine enantiomers were separated using a mobile phase of 0.5% KH(2)PO(4)-acetonitrile (65:35, v/v) on a Chiral CD-Ph column at a flow rate of 0.5 ml/min and measurement at 220 nm. Analysis required 400 microl of plasma and involved solid-phase extraction with an Oasis HLB cartridge, which gave recoveries for both enantiomers from 67.4 to 71.8%. The lower limit of quantification was 25 ng/ml for (R)- and (S)-fexofenadine. The linear range of this assay was between 25 and 625 ng/ml (regression line r(2)>0.993). Inter- and intra-day coefficients of variation were less than 13.6% and accuracies were within 8.8% over the linear range for both analytes. This method can be applied effectively to measure fexofenadine enantiomer concentrations in clinical samples.     

HPLC法同时测定健康人血浆中贝那普利及其代谢物贝那普利拉的浓度

目的建立一种灵敏度较高的高效液相色谱法,可同时测定健康人血浆中贝那普利及其代谢物贝那普利拉的浓度。方法流动相用0.02mol·L-1磷酸二氢钾缓冲液-乙腈(7∶3)(pH2.6),用取梯度洗脱法,血浆样本用C8固相小柱萃取,紫外检测波长为237nm。结果贝那普利:在0.06～2μg·mL-1,线性关系良好;最低检测限为0.03μg·mL-1;日内RSD≤6.3%,日间RSD≤4.0%;提取回收率在73%～78.5%(n=5),相对回收率在93.2%～101.3%。贝那普利拉:在0.06～2μg·mL-1,线性关系良好;最低检测限为0.03μg·mL-1;日内RSD≤8.5%,日间RSD≤6.0%;提取回收率在67.7%～74.1%(n=5),相对回收率在95.6%～103.2%。结论本方法灵敏度高、重现性好,可用于贝那普利人体生物利用度和药代动力学研究。     

Atenolol quantification in human plasma by high-performance liquid chromatography: Application to bioequivalence study

An accurate, precise, and sensitive high-performance liquid chromatography (HPLC) assay was developed for the determination of atenolol in human plasma samples to compare the bioavailability of 2 atenolol tablet (50 mg) formulations in 24 volunteers of both sexes. The study had an open, randomized, 2-period crossover design with a 1-week washout period. Plasma samples were obtained over a 24-hour interval. Atenolol concentrations were analyzed by combined reversed phase liquid chromatography and fluorescence detection (λ_EX = 258 nm, λ_EM = 300 nm). From the atenolol plasma concentration versus time curves, the following pharmacokinetic parameters were obtained: AUC_0–24h, AUC_0–∞, and C_max. The geometric mean of test/reference 50-mg tablets individual percent ratio was 102.2% for AUC_0–24h, and 101.6% for C_max. The 90% confidence intervals (CI) were 100.2% to 105.4% and 100.9% to 103.5%, respectively. Since the 90% CI for both C_max and AUC_0–24h were within the 80% to 125% interval proposed by the Food and Drug Administration, it was concluded that atenolol (50-mg tablets) test formulation was bioequivalent to the reference formulation, with regard to both the rate and extent of absorption.     

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。     

反相高效液相色谱法测定硝苯地平的血药浓度

目的：建立血浆中硝苯地平的高效液相色谱测定方法。方法：取血浆1.0ml，以尼群地平为内樯，用乙酸乙酯及环己烷（3：1）提取两次，有机相在55℃水浴中氮气吹干，残渣用流动相复溶后进样分析。结果：本法在5－200μg.L^-1范围内线性良好，相关系数r=0.9999(n=6)。高、中、低三种浓度硝苯地平的回收率为90．13％－99．43％，日内精密度为1．43％－6．61％，日间精密度为3．85％－8．79％，硝苯地平和内标的平均绝对回收率分别为90．58％和91．59％，内源性物质对测定无干扰。结论：本法灵敏度及特异性高，操作简便，适用于硝苯地平的临床药代动力学研究。     

高效液相色谱法测定盐酸二甲双胍血药浓度

目的:建立一种快速、灵敏、准确的高效液相色谱法用于测定人血浆中盐酸二甲双胍浓度.方法:取血浆样品0.5 mL,用1.0 mL乙腈沉淀蛋白,取上清液10 μL进样,流动相为甲醇-磷酸盐(50 mmol·L-1用氢氧化钠调节pH 6.7),色谱柱为Silica柱(4.6 mm×250 mm,5 μm),紫外检测器,检测波长为233 nm.结果:盐酸二甲双胍在20～4 000 μg·L-1范围内线性关系良好,最低检测限为20μg·L-1.日内、日间精密度(RSD)小于4%.结论:该法操作简单,灵敏度高,适用于临床药动学研究.     

高效液相色谱法测定人血浆中伏立康唑浓度

目的：建立一种简单高效的高效液相色谱（HPLC）法用来检测人体中伏立康唑的血药浓度，并应用于临床中伏立康唑用药监测，以促进其个体化用药。方法：色谱柱：Kromasil C₁₈（4.6 mm×150 mm，5 μm），柱温：35℃，流速：1.0 mL·min^-1，流动相：甲醇-水（60：40），检测波长：257 nm，内标：酮康唑。对该方法进行方法学验证。结果：该方法专属性良好，血浆中伏立康唑在0.1~20.0 μg·mL^-1范围内线性良好（r=0.999 6），定量下限为0.1 μg·mL^-1。高、中、低3个浓度提取回收率分别为（90.68±10.32）%、（92.82±8.26）%、（97.47±4.58）%；日内精密度RSD分别为5.87%、7.85%、4.10%；日间精密度RSD分别为5.64%、3.30%、2.74%。对某院20例（男12例，女8例）使用伏立康唑抗真菌治疗的患者运用该方法进行了监测，结果显示浓度范围在0.71~13.51 μg·mL^-1之间。结论：本方法专属性高，操作简便，结果准确，可用于临床上伏立康唑血药浓度的检测，从而促进其个体化用药的推广。     

反相高效液相色谱法测定人体血浆中表阿霉素的含量

采用反相高效液相色谱法(以柔红霉素为内标)测定血浆中表阿霉素的含量,流动相为5mmol/L H3PO4-甲醇-异丙醇-乙晴(6∶9∶7∶2,pH2.9),YWG C18不锈钢色谱柱,荧光检测波长:λex=450nm,λem=530nm.血浆中样品经氯仿-甲醇-异丙醇=5∶2∶3抽提液抽提处理后进样测定.血浆中表阿霉素的浓度在0.03～2.4mg/L范围内峰面积与柔红霉素的峰面积之比呈线性关系(r=0.9998),方法对表阿霉素的平均回收率为89%～90.7%.最低检测浓度为0.01mg/L.本法灵敏度高,线性范围大,符合临床用药的血药浓度监测和药代动力学研究的分析方法要求,并用于40例已服用表阿霉素的癌症病人的血浆药物含量的测定.     

高效液相色谱法测定人血浆中左布比卡因的浓度

目的建立高效液相色谱法测定人血浆中左布比卡因的浓度。方法以罗哌卡因为内标,色谱柱为Hypersil C_(18)柱(200 mm×4.6 mm,5μm),流动相采用0.01 mol·L~(-1)磷酸二氢钾-乙腈(87:13,V:V,pH=3.4),流速2.0 mL·min~(-1),检测波长为210 nm,柱温40℃。结果标准曲线方程为Y=0.310 1 X+ 0.008 9,r=0.999 7,左布比卡因的质量浓度在0.012 5～2 mg·L~(-1)范围内呈良好的线性关系,最低检测限为0.01 mg·L~(-1);方法回收率在96%～105%之间;日间、日内精密度RSD均<5%。结论本方法可用于临床上左布比卡因血药浓度的监测和药动学研究。     

Determination of ibuprofen in human plasma by solid phase extraction and reversed-phase high-performance liquid chromatography

A new, sensitive and simple method for the rapid quantitative determination of ibuprofen in human plasma has been developed. This method involves the use of a solid phase extraction on "Baker" C-18 disposable extraction columns for sample clean-up and uses mefenamic acid as an internal standard. Separation and quantitation are performed by reversed-phase liquid chromatography using a Nucleosil C18 column and methanol-0.04 M phosphoric acid (80:20, v/v), as the mobile phase. Detection was achieved by UV-absorbance measurements at 229 nm.