反相高效液相色谱法测定人血浆中异丙酚浓度

用柱前衍生化反相高效液相色谱法测定人血浆中异丙酚浓度,为临床药理学研究提供了依据。提取的异丙酚及内标麝香草酚同时以Ｇｉｂｂｓ试剂衍生化,衍生物经色谱柱分离和紫外检测,异丙酚在５０～１５００μｇ／Ｌ浓度范围内呈线性关系（ｒ＝０．９９９１）。方法平均变异系数为６．１％,最低检出浓度为２４．８μｇ／Ｌ,已满意地用于临床药理学研究。     

High Performance Liquid Chromatographic Determination of Pentamidine in Plasma

Abstract

A high performance liquid chromatographic method for quantitating pentamidine in plasma has been developed. Sample clean-up involved precipitating plasma with acetonitrile containing the internal standard, hexamidine. The supernatant was passed through a C₈ Bond Elut column and eluted with a methanolic solution of sodium 1-heptanesulfonate. The eluate was then analyzed on an Altex C₈ column with a mobile phase consisting of 45% CH₈CN, 0.02% detramethylammonium chloride and 0.1% H₃PO₄. Using fluorescence detection (EX: 275 nm and EM: 340 nm), the detection limit was 1.25 ng/ml for 0.5 ml of plasma. The coefficients of variation for interday and intraday were around 10%.     

High Performance Liquid Chromatographic Determination of Oxamniquine in Plasma Samples

Abstract

A sensitive and reliable liquid chromatographic assay procedure for the quantitation of oxamniquine in plasma or urine was developed. Chromatographic separation was achieved on a reversed-phase phenyl colum using U.V. Detection at 254 nm. The eluting solvent was the mixture of 0.05 M acetate buffer pH 5 and acetonitrile (3:7). With this mobile phase the drug and its external standard were well separated from the interference of the blank samples. The average recovery of oxamniquine from 3 or more replicate dog plasma samples of different concentration (0.125 ? 4.00 μg/ml) was 95.5% and its coefficient of variation was 4.17%. The reproducibility of the assay was confirmed by the analysis of variance test for the slopes of the three standard plots obtained from plasma samples at three different occasions (F=4.2, p > 0.01). The detection limit for plasma samples was approximately 20 ng/ml. The method was applied to measure the plasma level vs, time profile of this drug following a single bolus intravenous dose of 16 mg/kg to a dog.     

High Performance Liquid Chromatographic Determination of Triazolam in Human Serum

Abstract

A high performance liquid chromatographic method which utilizes UV-detection has been developed for the sensitive and specific determination of triazolam in human serum. Using 8-chloro-6-phenyl-l-ethoxymethyl-4H-s-triazolo[4, 3-a][1, 4]benzodiazepine as an internal standard, serum samples were buffered with 2 ml of 4M NaOH and extracted twice with 5 ml aliquots of toluene. The combined toluene extracts were evaporated to dryness and the residue dissolved in the chromatographic mobile phase. The samples were chromatography on a microparticulate reverse-phase column using a 0.06M acetic acid:acetonitrile (61:39) mobile phase. Known metabolites of triazolam did not interfere in the analysis. A linear relationship between peak height ratios and concentrations was observed, with the lower limit of detection being approximately 1 ng of triazolam. The utility of the method was demonstrated by administering therapeutic doses of the drug to human volunteers and monitoring serum triazolam concentrations as a function of time.     

High Performance Liquid Chromatographic Determination of Amiodarone in Plasma and Tissues

Abstract

A simple and rapid high-performance liquid chromatographic (HPLC) method for the determination of amiodarone (AD) in plasma and tissues was developed. The method involved deproteinization of plasma or homogenized tissue with acetonitrile containing an internal standard (N-Cetylpyridinium chloride) followed by reversed phase chromatography using μ bondapack C₁₈ column (10μm) with a mobile phase consisting of acetonitrile - methanol - sodium dihydrogen phosphate buffer (70:10:20%, v/v), the pH adjusted to 4.0 and pumped at flow rate of 1.0 ml/min. The column effluent was monitored at 242 nm. A linear relationship was obtained between peak height ratios (drug to internal standard) versus drug levels over the concentration range of 50–750 ng/ml. The detection limit of AD in plasma and tissues by this method was 20 ng/ml.     

Specific and Sensitive High Performance Liquid Chromatographic Determination of Alprazolam or Triazolam

Abstract

A specific and highly sensitive liquid chromatographic procedure has been developed for the rapid determination of intact alprazolam or triazolam in dog serum, using these structurally similar triazolobenzodiazepines as mutual internal standards. The procedure consists of (1) extracting one ml of alkali buffered serum with toluene, (2) evaporating an aliquot of the toluene to dryness, and (3) quantitating the redissolved residue by HPLC using ultraviolet detection (221 nm). Samples were chromatographed on a microparticulate reverse phase column using a mobile phase composed of acetonitrile: isopropanol: water (94:5:1) and a flow rate of 0.75 ml/min. Metabolites of alprazolam and triazolam did not interfere in the assay. The lower limit of detection was approximately 1 ng/ml of serum extracted. The utility of the analytical methodology for the determination of alprazolam or triazolam in pharmacokinetic studies in the dog was demonstrated.     

High Pressure Liquid Chromatographic Determination of Isoniazid and Acetylisoniazid in Human Plasma

Abstract

A quantitative high pressure liquid chromatographic (HPLC) assay has been developed for the determination of isoniazid (INH) and acetylisoniazid (ACINH) in human plasma. Plasma samples were taken from a patient after oral administration of INH (with proven tuberculosis infection). A C₁₈ reversed phase radial compression column was used to separate INH and ACINH from other plasma components. The analysis takes 10 minutes per sample and the lower limit of detection for each compound is 0.10 ug/ml plasma.     

Rapid High Performance Liquid Chromatographic Determination of Ampicillin in Human Urine

Abstract

A rapid, sensitive and specific HPLC assay for the determination of ampicillin in human urine is developed.

Ampicillin was directly measured in human urine at 225 nm using a reversed phase column (Synchropack RP-P) and a mobile phase composed of (1:9 methanol-sodium acetate solution, 0.01 M, pH 4). The analysis required no longer than 10 min. Linear correlation between the peak height ratio of ampicillin to cefoxitin sodium (internal standard) and ampicillin concentration in urine over the range 10–100 μg ml^?1 was obtained. The developed method proved to be advantageous as it monitors ampicillin level in urine. Moreover, the urinary excretion of ampicillin in human subjects after an oral administration of 500 mg ampicillin capsules was established using the proposed method.     

High Pressure Liquid Chromatographic Determination of Mecillinam in Human Plasma and Urine

Abstract

A simple, specific, rapid and sensitive method for the analysis of mecillinam in plasma and urine using high pressure liquid chromatography is described. The assay is performed by direct injection of a plasma protein free supernatant or a dilution of urine. A μBondapak phenyl column with an eluting solvent of 16% CH₃CN-0.2% H₃PO₄ was used, with UV detection of the effluent at 220 nm. Desacetyl-cephalothin was used as the internal standard and quantitation was based on peak height ratio of mecillinam to that of the internal standard. The lowest concentration detectable without extraction was 0.25 μg/ml for plasma and 8.9 μg/ml for urine. No interference from plasma and urine was noted.     

High Performance Liquid Chromatographic Determination of (-)-N-Formylnorephedrine in Plasma

Abstract

An HPLC procedure for the detection and quantitative estimation of (-)-N-formylnorephedrine in rabbit plasma had been developed. The procedure involved the extraction of (-)-N-formylnorephedrine from plasma spiked with the internal standard (phenacetin), using ethyl acetate. The ethyl acetate extract is evaporated under nitrogen and the residue is reconstituted in water and injected onto the column. A u-Bondapak-C18 column 30 cm × 3.9 mm ID was used. The mobile phase is 20% acetonitrile in water; at a flow rate of 1.5 ml/min and uv detection at 256 nm. A linear relationship between concentration and peak height ratio (I/internal standard) was obtained (r = 1.00). The reported procedure allows the measurement of (-)-formylnorephedrine in concentrations as low as 150 ng/ml of plasma with total procedure time of about 10 min. The applicability of the procedure to pharmacokinetic studies is illustrated and metabolites are shown not to interfere with the assay procedure.     

高效液相色谱法测定家兔血浆中桂皮酸的浓度及其药代动力学初探

 建立了采用高效液相色谱(HPLC)测定家兔血浆中桂皮酸含量的方法,并应用此法进行了桂皮酸的药代动力学研究。 采用的色谱柱为Kromasil C18柱(250 mm×4.6 mm i.d.,5 μm)；流动相为甲醇-乙腈-水-冰醋酸(体积比为10∶22∶55 ∶0.5),流速0.8 mL/min；检测波长为270 nm;柱温为室温；内标物为苯丙酸。实验结果表明,低、中、高浓度的提取回 收率分别为84.9%,84.4%,87.7%,方法回收率分别为98.4%,99.2%,100.1%,相对标准偏差(RSD)分别为5.5%,3.6%,3.7%, 日内及日间测定值的RSD均小于6%。所建立的HPLC方法灵敏、专一、准确、精密,可作为桂皮酸在家兔体内药代动力学研 究的检测手段。口服冠心苏合丸和冠心苏合胶囊后,桂皮酸在家兔体内的代谢呈一级吸收双室模型。     

免疫法制备液相色谱生物样品的研究——血中沙丁胺醇的分析

用动物免疫法制备了免疫亲和柱纯化水溶性的沙丁胺醇血浆样品。琥珀酸酐交联沙丁胺醇和牛血清白蛋白获得抗原免疫家兔抗沙丁胺醇抗体——免疫球蛋白。琼脂糖Ｓｅｐｈａｒｏｓｅ４Ｂ与抗体交联制成免疫球蛋白亲和柱。对高效液相色谱法测定中的一般提取方法和固相小柱提取法作了比较，后者具有内源性杂质干扰少的优点，是生物样品预处理的一种有效的方法     

High-Performance Liquid Chromatographic Determination of Acetazolamide in Human Plasma

Abstract

A simple, rapid and specific HPLC method has been developed to determine acetazolamide concentrations in human plasma. The assay procedure requires only 250 μl of sample with direct injection of the organic supernatant after protein precipitation with acetonitrile. Chlorothiazide was used as an internal standard. A reversed-phase C₁₈ μBondapak column was employed for the chromatographic separation. The eluent was monitored at 265 nm using a UV variable wavelength detector. The retention times for acetazolamide (ACZ) and chlorothiazide (CTZ) were 6 and 8 min respectively. A linear relationship (r).995) was obtained over the 1-20 μg/ml concentration range. The limit of sensitivity for ACZ was 0.5 μg/ml, with greater than 85% recovery of ACZ and internal standard. The method was applied to human plasma samples obtained after administration of a 250 mg acetazolamide tablet.     

A Sensitive Gas Chromatographic Method for the Determination of Metoprolol in Human Plasma

Abstract

A sensitive method for the determination of metoprolol in plasma has been developed. The procedure is based on gas chromatographic measurements of derivatized metoprolol, using 9-bromophenanthrene as internal standard. Metoprolol is derivatized with pentafluoropropionic anhydride. The resulting derivative gives a four-fold increase in sensitivity compared to the published methods where trifluoroacetic anhydride was used for derivatization.     

高效液相色谱法测定血浆中舒必利浓度

讨论了测定舒必利血药浓度的反相高效液相色谱方法。选用美国Ｂｉｏ－Ｒａｄ７００型高效液相色谱仪,ＲＰ－３１８色谱柱,ＵＶ检测波长２９０ｎｍ,流动相为甲醇－水－冰醋酸（６０３０１）,内标物为胃复安。回收率为９７．９５％～９９．９６％,ＲＳＤ为２．６％～５．１％,最低检出浓度１．０ｍｇ／Ｌ,线性范围５～１００ｍｇ／Ｌ。测定３０例服药病人的血药浓度,得到了满意的结果。     

高效液相色谱法测定人血浆中的芦氟沙星

 建立了测定人血浆中芦氟沙星质量浓度的高效液相色谱法 ,血浆用二氯甲烷提取 3次 ,以 UltrasphereODS(4.6mm i.d.× 2 5 0 mm)为色谱柱 ,流动相为甲醇 -1 0 mmol/L 溴化四丁铵 -三乙胺 (体积比为 3 2∶ 68∶0 .5 ) ,用磷酸调 p H2 .8,检测波长 2 95 nm,流速为 1 .2 m L/min,以培氟沙星为内标。血浆中芦氟沙星的线性范围为 0 .1～ 1 0 mg/L ,最低检测质量浓度为 0 .0 5 mg/L ,回收率为 99.7% ,日内、日间 RSD分别为 2 .3 3 %和3 .83 %。方法简便、快速、准确 ,适用于人血浆中芦氟沙星质量浓度的测定。     

高效液相色谱法测定人血浆中的辛伐他汀

 建立了高效液相色谱法监测人口服辛伐他汀药物后的血药浓度。血样用环己烷-二氯甲烷(体积比为3.5∶1)提取,以洛伐他汀为内标,在237nm波长下检测;色谱柱:LichrospherC18(200mm×4.6mmi.d.,5μm),流动相:乙腈-水(体积比为70∶30);流速:1.2mL/min。血药浓度在0.25～50.0μg/L范围内与峰面积和内标峰面积的比值之间线性关系良好,日内及日间相对标准偏差(n=5)分别低于7.94%和8.58%,回收率高于93.3%。方法灵敏、准确、快速,适用于药物动力学和药效学研究。     

高效液相色谱法测定血浆中米非司酮

建立了测定血浆中米非司酮浓度的高效液相色谱分析方法。血浆样品以乙醚提取,常温下负压挥发至干,所得残渣用甲醇浸溶后进行测定。Spherisorb C18(250 mm×4.6 mmi.d.,5μm)为色谱柱,流动相为甲醇-乙腈-水(50 25 25)溶液,检测波长为302 nm,用外标法定量。结果表明,在0.05~10.00 mg.L-1范围内呈线性,检出限为0.01 mg.L-1;平均回收率为98.2%,日内、日间平均精密度分别为7.0%,8.3%。米非司酮血浆样品的萃后残渣于-20℃可保存至少7d。     

A Liquid Chromatographic Method for the Determination of Atenolol in Human Plasma

Abstract

A reliable, highly reproducible, accurate and time-efficient high performance liquid chromatographic (HPLC) method to measure atenolol concentration in human plasma was developed and validated. Sample clean-up consists of simple and efficient liquid-liquid extraction (mean recovery 103%) which allows a high sample throughput. Chromatography on a CN-propyl column yields symmetrical and well resolved peaks for atenolol and for the internal standard (metoprolol) without any interference from endogenous plasma components. Using 1 ml plasma samples the method has a limit of detection of 12.6 ng/ml (calculated at a 99.9% confidence level) with %CV (precision) ≥ 8.8% and bias (accuracy) ≥ 3.8% for concentrations in the range of 10 – 1000 ng/ml. We now routinely use this method in human pharmacokinetic studies of atenolol dosage forms.