High Performance Liquid Chromatography Determination of Pcmx In Blood Plasma Using Electrochemical Detection

Abstract

A rapid and specific high performance liquid chromatographic (HPLC) procedure has been developed for the determination of p-chloro-mxylenol (PCMX) in blood plasma. the method is based on the extraction of PCMX from plasma with benzene in the presence of a known amount of dichloro-m-xylenol (internal standard). the benzene extract is evaporated to dryness and the residue dissolved in 200 μl of mobil phase. the HPLC system consisted of a reversed-phase column and a 65% methanol:35% ammonium carbonate 0.05% solution as a mobile phase. an electrochemical (EC) detector/glassy carbon electrode set a potential of +0.9V versus Ag/AgCl/3M NaCl is used to monitor the drug. the recovery of PCMX is approximately 98%, the limit of quantitation is 2 ng/ml of plasma for the HPLC-EC procedure. the coefficient of variation is 5.1% over the range of 10-1000 ng/ml of plasma. Data are presented to illustrate the practicality of this method for evaluation of PCMX plasma levels after a single intravenous administration of 500 mg dose of PCMX to five mongrel dogs.     

高效液相色谱法测定肾上腺素的光学纯度

用2,6-二甲基-β-环糊精作手性流动相添加剂,C18柱为固定相,建立了D,L-肾上腺素拆分的定量分析方法。用归一化法分析D,L-异构体时引入了定量校正因子。比较了外标法和内标法定量测定L-肾上腺素的回归方程、精密度及回收率。测定了不同储存期的L-肾上腺素滴眼剂中D型异构体的质量分数。     

反相离子对液相色谱－电化学检测器测定生物样本中的儿茶酚胺

讨论了使用反相离子对高效液相色谱－电化学检测器同时分离和测定儿茶酚胺的条件，并对生物样本（脑组织、血液、羊水、尿等）进行了实际测定，结果令人满意。     

Determination of Danazol in Plasma by High Performance Liquid Chromatography

Abstract

An HPLC method has been developed for the determination of danazol in human plasma. the chromatographic conditions consisted of an ODS column 80 × 7.0 mm, 3 μm; a mobile phase of 71:29, methanol:20 mM potassium dihydrogen phosphate. A UV-Visible detector was set at 288 nm to monitor the danazol peak. Danazol was extracted from plasma with acetonitrile which was salted out with potassium carbonate. Prior to salting out, cadmium sulfate was mixed with the acetonitrile-plasma mixture to remove any interfering constituents. the extracted acetonitrile layer was evaporated and the residue was reconstituted in the mobile phase before injection. the method was found to be reproducible with relative standard deviation (%rsd) from 2.3 to 7.2%. A number of clinically important drugs did not interfere with danazol determination.     

高效液相色谱法测定血浆中莲心碱的浓度

采用高效液相色谱法（甲基莲心碱为内标）测定了血浆中莲心碱的浓度。以ＵｌｔｒａｓｐｈｅｒｅＳｉ为固定相、二氯甲烷－异丙醇－二乙胺（７５１５０．２,Ｖ／Ｖ）为流动相,检测波长为２８２ｎｍ。血样用氨－氯化铵缓冲液调ｐＨ１０后用乙醚提取。平均回收率９３．６％,ＲＳＤ为１．９％,最低检测浓度０．０２５ｍｇ／Ｌ。     

Simultaneous Determination of Dopamine,Norepinephrine, Tyramine and Octopamine by Reverse-Phase High Performance Liquid Chromatography with Electrochemical Detection

Abstract

A method is described for the simultaneous separation and estimation of the catecholamines, norepinephrine and dopamine and their monohydroxy-equivalents, octopamine and tyramine. The method employs high-performance liquid chromatographic separation of the compounds on a C18 reverse-phase column with a mobile phase containing methanol as the organic modifier, octane sulphonate as an ion-pair reagent and acetic acid/ammonium hydroxide buffer. The influences of electrode potential and solvent pH on detector response were studied, and the optimal conditions identified as detector potential of 0.95 volts and pH 6.0. The technique of post-column mixing was introduced to provide optimal pH conditions for detector response without the constraint of on-column oxidation of catecholamines. The effects of buffer ionic strength on retention factors and detector response were also investigated and, on the basis of the results obtained, the optimal buffer strength was identified as 0.08–0.09 molar. The described procedure can be used for simultaneous estimation of catecholamines and monohydroxyphenolamines at concentrations between 200–5000 pg.     

高效液相色谱–光电二极管阵列法测定虾青素的含量

建立虾青素含量测定的高效液相色谱–光电二极管阵列法。采用Purospher STAR RP 18(4.6 mm×250mm,5μm)色谱柱,以甲醇–水(体积比为95∶5)为流动相,流速1.0 mL/min,检测波长为482 nm,柱温为30℃,进样量为20μL。在所选定的液相色谱条件下,虾青素主峰与其它杂质峰分离良好,虾青素在0.2~16μg/mL范围内线性良好,线性相关系数r=0.999 9,检出限为0.01μg/mL,测定结果的相对标准偏差为0.42%(n=6),平均回收率为100.4%。该法分析快速准确、灵敏度高、重现性好。     

Assay of Yohimbine in Human Plasma Using High Performance Liquid Chromatography with Electrochemical Detection

Abstract

Yohimbine is a selective α₂ adrenoreceptor antagonist used in the study of α₂ adrenoreceptors in man. In order to better improve administration regimens for the study of yohimbine in man, we have developed an assay for the determination of yohimbine in plasma utilizing reverse phase high performance liquid chromatography with electrochemical detection. Using a C₁₈ column and a methanol:acetate (60:40) mobile phase, we detected yohimbine in plasma following a simple chloroform extraction. Reserpiline was used as an internal standard. The assay was linear over a concentration range of 50–250 ng/ml in spiked plasma and had a lower limit of sensitivity of 10 ng/ml. It was used to detect yohimbine in plasma sampled from 4 volunteers during an infusion of the alkaloid.     

Determination of Apomorphine in Tablets Using High Performance Liquid Chromatography with Electrochemical Detection

Abstract

A rapid and selective method using high performance liquid chromatography with electrochemical detection is described for the determination of apomorphine in tablets. Tablet mixes were dissolved in a standard volume of mobile phase containing the internal standard, N-n-propylnorapomorphine. Separation was achieved on a μ-phenyl column using methanol-acetonitrile-0.05M KH₂PO₄ (5:15:80) as mobile phase. The eluted compounds were detected with a sandwich-type electrochemical detector employing a glassy carbon working electrode and operated at 0.5V. Satisfactory accuracy and precision were obtained during analyses of tablets containing apomorphine.     

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。     

使用限进介质色谱柱的柱切换高效液相色谱法直接进样测定大鼠血浆中舒必利

提出一种直接进样测定大鼠血浆中舒必利浓度的高效液相色谱方法,使用限进介质色谱柱作为预柱在线去除血浆蛋白后,将舒必利通过柱切换转移到分析柱中进行分析。限进介质色谱柱为CAPCELLPAKMFSCX阳离子交换柱（20×4．0mmi．d．,5μm）,分析柱为Kromasil C18柱（150×4．6mm i．d．,5μm）,限进介质柱预分离时流动相为PH=6．88的50mmol／L磷酸盐缓冲液乙腈（100：5,V／V）,切换后分析流动相为PH=6．83的50mmol／L磷酸盐缓冲液-乙腈（100：10,V／V）。流速均为1mL／min,检测波长为240nm。该方法检出限为17ng／mL,定量限为50ng／mL。舒必利在50～1400ng／mL之间线性良好（r=0．9997）,高中低浓度的日内、日间相对标准偏差分别为1．5％～4．2％及2．0％～5．2％,方法回收率为98．8％～104．1％．     

Determination of Saccharin in Plasma and Urine by High Performance Liquid Chromatography

Abstract

A sensitive and specific high performance liquid chromatographic (HPLC) assay for the determination of saccharin in plasma and urine was developed. Saccharin is extracted into diethyl ether at acid pH, evaporated, and reconstituted prior to instrumental analysis. Overall recovery of saccharin is 86.9 + 8.6% and the sensitivity limits of detection is 0.15 μg per ml of plasma or urine using a fluorescence detector. The sensitivity limit in plasma can be extended to 20 ng per ml by use of a 2 ml assay volume and detector attenuation. The assay was used for the determination of saccharin in plasma and urine of rats following oral doses of 5 mg/kg.     

Determination of Plasma Pseudo-Ephedrine by Fluorescence Detection and High Performance Liquid Chromatography

Abstract

A sensitive and specific high-performance liquid chromatographic method has been developed for the determination of pseudoephedrine in plasma. The assay was based on the production of a highly fluorescent derivative of pseudoephedrine using 4-chloro-7-nitrobenzo-2, 1, 3-oxadiazole (NBD-Cl) as the derivatization agent. The fluorescent derivatives were separated using normal phase liquid chromatography after an automated, column-switching, sample clean-up procedure. Pseudoephedrine determination was accurate and precise at concentrations as low as 10 nanograms per milliliter of plasma. The chromatographic step separated derivatives of pseudoephedrine from those of norpseudoephedrine and several other amines. Measurement of pseudoephedrine concentrations in plasma following a single 120 milligram oral dose is illustrated.     

Determination of Chlormezanone in Plasma and Urine by High Performance Liquid Chromatography

Abstract

A reversed-phase high-performance liquid chromatographic (HPLC) method with UV detection was developed for quantifying chlormezanone in plasma and urine. an extraction step was needed to avoid endogenous interferences especially in urines and led to an average recovery of 90%. the sensitivity limit was 20 ng/ml in both plasma and urine. the method was reproducible with intra- and inter-assay coefficients of variations below 5%. This method was applied to the determination of plasma and urine levels during a pharmacokinetic study in the elderly. It was found suitable to follow the concentrations as long as 120 h after a single oral administration of 400 mg chlormezanone.     

Determination of Amphotericin-B in Human Plasma by Reversed-Phase High Performance Liquid Chromatography Using a Short Octyl Column

Abstract

Amphotericin-B is a polyene antifungal antibiotic used for the treatment of severe systemic fungal infections. For effective treatment of urinary fungaria and the prevention of significant adverse-effects, monitoring the concentration of Amphotericin-B in biological samples of humans (ingesting the drug) is required. In this experiment, Amphotericin-B was isolated from plasma endogenous substances by adding 200 μL of acetonitrile in 800 μL of plasma. This mixture was vortex mixed, 20 mg of zinc sulfate and 10 mg of monobasic potassium phosphate was added to the mixture. This mixture was again vortex mixed and followed by centrifugation. The supernatant was filtered through a 0.45 μm membrane and a 100 μL aliquot of this solution was injected onto the chromatographic system. A short column of 60 mm × 4.6 mm packed with 3 μm octyl particles was used with an isocratic elution of 50/50, acetonitrile/0.01M KH₂PO₄ (v/v). The pH of the mobile phase mixture was adjusted to 3.5 with H₃PO₄. The intact drug molecule (parent drug) was monitored by a W-visible detector at 410 nm and 0.10-0.005 A.U.F.S. The limits of detection of the method were 0.03 μg/mL for 100 μl injection volume at signal-to-noise ratio of 3.     

高效液相色谱法测定血浆中左亚叶酸钙的含量

建立了测定血浆中左亚叶酸钙含量的RPHPLC方法。采用DiamonsilC18色谱柱(150mm×4.6mm,5μm),乙腈50mmol/L磷酸氢二钾/磷酸二氢钾缓冲溶液(10∶90,V/V,用磷酸调pH至4.0)为流动相,用前经0.45μm的过滤膜,超声脱气,流速为1.0mL/min,紫外检测波长为290nm,柱温为35℃,进样量20μL。该方法最低检出限为0.2ng;线性范围为0.05～10mg/L,方法平均回收率在97.0%以上;日内及日间精密度的相对标准偏差分别小于4%和6%。该方法简单,快速,灵敏度高,重复性好。     

高效液相色谱法测定大鼠血浆中的原儿茶酸

建立了大鼠血浆中原儿茶酸含量测定的高效液相色谱方法。采用的色谱柱为DiamondsilTM C18 柱(150 mm×4.6 mm,5 μm);流动相为乙腈-水(体积比为9∶91,用H3PO4 调pH至2.5);流速1.2 mL/min;检测波长260 nm;内标为对羟基苯甲酸。原儿茶酸的线性范围为0.050~3.20 mg/L,线性相关系数为0.9978，最低定量限为0.050 mg/L,日内和日间测定的精密度（以相对标准偏差表示）均低于7.0%,准确度（以相对误差表示）为-1.4%～2.6%；在0.050,0.40,3.20 mg/L低、中、高3个添加浓度水平下，血浆样品的提取回收率分别为83.4%,87.3%,91.1%。该方法简便,灵敏,准确,适用于大鼠体内原儿茶酸的药物动力学研究。