整体柱高效液相色谱法测定尿中核苷的方法研究

尿中修饰核苷已被研究用作可能的肿瘤标记物,目前最常用的分析方法为反相高效液相色谱法,其不足之处是分析周期太长。为此,建立了应用整体柱的高效液相色谱方法对尿中修饰核苷进行分析,所用缓冲液为25 mmol/L磷酸二氢钾溶液(pH 4.55)和60%(体积分数）的甲醇水溶液,线性梯度洗脱,检测波长为260 nm。12种目标核苷可被完全分离。所建立的方法在分离度、线性、重现性、灵敏度及回收率等指标上与普通反相液相色谱法相近,但分析周期大大缩短,仅为23 min,适合于大量临床样品的分析。     

High Performance Liquid Chromatographic Analysis of Desipramine in Human Plasma Using a Microbore Column Technique

Abstract

A reversed-phase, isocratic HPLC method has been developed for the quantitation of desipramine in human plasma. the method involved the use of cloimpramine as an internal standard. the chromatographic separation was accomplished with a mobile phase comprising acetonitrile-aqueous solution (60:40. v/v) containing 10 mM disodium hydrogenphosphate and 80 mM sodium dodecyl sulfate adjusted to pH 2. the mobile phase was pumped at a flow rate of 0.5 ml/min. the column used was a microbore column (2 mm I.D. × 100 mm) packed with a C18 reversed-phase material (5μm ODS Hypersil). Plasma samples were extracted at basic pH with diethyl ether followed by back-extraction into 0.1 N sulfuric acid. Using UV detection at 250 nm, the lower limit of sensitivity was 10 ng/ml. the inter- and intra-assay coefficients of variation were found to be less than 10%. the assay procedure was applied to a long term oral dosing study in patients to monitor the plasma concentration of desipramine.     

High Performance Liquid Chromatographic Analysis of Pharmaceuticals Using Oil-In-Water Microemulsion Eluent and Monolithic Column

Novel and efficient separations of pharmaceutical substances were achieved using oil-in-water microemulsion eluent and a conventional C18 packing with a flow rate of 1 mL/min⁻¹. Attempts to decrease analysis time was limited due to the high viscosity of the microemulsion which generated relatively high back-pressures. Monolithic columns gave 3-fold lower back-pressures and allowed flow rates of 4 mL/min⁻¹. with the same microemulsion mobile phase which permitted rapid separations to be achieved. Separation of a test-mix of paraben preservatives was achieved in both isocratic and gradient mode in less than 1 min. The monolith-microemulsion combination was applied to rapidly quantitatively analyse two formulated products with excellent linearity, accuracy and repeatability. Quantitative analysis times were under 90 seconds. Successful quantitation of both nicotine lozenges and naprosyn tablets was performed using this approach.     

High Performance Liquid Chromatographic Determination of Nordihydroguaiaretic Acid

Abstract

A high performance liquid chromatography (HPLC) method was developed for the detection and quantitation of nordihydroguaiaretic acid (NDGA). Very low concentrations of NDGA in various extracts are detectable, thus making the method more sensitive than other previously reported analytical techniques. NDGA was extracted from leaves of Larrea divaricata as well as from rodent food containing NDGA. Since rats are fed NDGA in studies that examine the development of renal cystic disease, we modified extraction procedures to permit isolation of NDGA from tissue and serum samples.     

High Performance Liquid Chromatographic Determination of Emetine in Corn

Abstract

A High performance liquid chromatographic (HPLC) method for the quantitative analysis of emetine in corn is described. Corn kernels are removed from the ears, blended, and extracted with methanol. The extract is filtered through a 0.45 um Acrodisc filter and analyzed by HPLC using a LC-18-DB column. For detection, a fluorescence detector set at excitation of 285 and emission at 316 nm is used. The recoveries of samples fortified between 0.1 ppm to 20 ppm with emetine ranged from 95.5% to 103.3%.     

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 Assay of Methoclopramide in Plasma Using a Silica Gel Column and An Aqueous Meobile Phase

Abstract

A high performance liquid chromatographic method for quantitating matoclopramide in plasma is presented. Proteins ara precipitated from the plasma sample with acetonitri la containing the internal standard, procainamida. The treated samples ara than analyzad using an Ultrasphere Si column, an aqueous solution at pH 7 of 65% CH³CN and 5.0 mM (NH₄)₂HPO₄ as a mobile phase, and a fluorescence detector. The retention times for drug and intarna1 standard ara 11.2 and 13.2 min, respectively. The caibration curve is Linear from 0.89 to 44.5 ng/ml. The detection limit is 0.89 ng/ml [signaL/hoisa = 31] for 0.2 ml plasma samples Pracision is measured by intraday and intarday coefficients o f variation, which are less than 10%. This method is currently being used for pharmacokinetic studies of methoclopramide.     

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 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 Phenacemide in Biological Fluids

Abstract

A sensitive and reliable high performance liquid chromatographic procedure has been developed for the quantitation of phenacemide in plasma or urine. After simple extraction of the drug with ethylacetate from alkalinized samples and evaporation to dryness, the reconstituted extract was chromatographed using a C₈ reversed phase analytical column with UV detection at 254 nm. Regression analyses for the calibration plots obtained on 3 different days for the drug concentrations ranging 1–15 mcg/ml indicated excellent linearity (r >0.999) and reproducibility (CV< 4%, p >0.01). The mean recovery of spiked phenacemide in plasma and urine from the lower limit of quantitation (1 mcg/ml) to 15 mcg/ml was 97.9 and 96.3%, respectively and their respective CV was 3.53 and 2.58%. The method was applied to monitor the plasma vs. time profile of the drug following a single bolus IV dose of 12 mg/kg in a dog.     

Liquid Chromatographic Determination of Four Purine Bases Using Porous Graphitic Carbon Column

A high performance liquid chromatographic method has been developed for the simultaneous determination of hypoxanthine (Hx), xanthine (Xa), guanine (Gu) and adenine (Ad) in shellfish. The separation of these compounds was performed on a porous graphitic carbon column (100 × 4.6 mm I.D.) using acetonitrile-phosphate buffer (23/77 v/v) containing 0.5% trichloroacetic acid at pH 2.0 as the mobile phase. The flow rate was fixed to 1 mLmin^–1 and the detection was monitored at 254 nm. Calibration curves were found to be linear in the concentration range from 0.3 to 100 gmL^–1 with correlation coefficients (r²) > 0.999. The lowest detectable concentrations of Hx, Xa, Gu and Ad were 0.04, 0.06, 0.08, 0.07 g mL^–1 respectively. Recovery of all purine standards added to sea urchin gonad was above 97%. The proposed method was successfully applied for the determination of Hx, Xa, Gu and Ad in sea urchin gonad extract.     

Determination of Neomycin in Milk by Reversed Phase Ion-Pairing Liquid Chromatography

Abstract

A fluorometric high performance liquid chromatographic (HPLC) method has been developed for the determination of neomycin in milk. Whole or shelf milk was defatted by initial centrifugation at 4°C. The resulting skim milk was deproteinated with trichloroacetic acid and centrifuged again. The neomycin was determined directly in the supernate by HPLC. The HPLC conditions consisted of an ion-pairing mobile phase, a reversed-phase column, post-column derivatization with o-phthalaldehyde (OPA) reagent and fluorescence detection. The overall recovery of neomycin was 94% (coefficient of variation 6.5%), in whole milk spiked at 0.15–10 ppm and 99% (coefficient of variation 6.4%) in shelf milk spiked at 0.15–5 ppm. The method was used to detect neomycin in milk obtained from cows dosed intramuscularly with neomycin (10 mg/kg). The neomycin concentrations in milk at 8 and 24 h after dosing were 0.3 and 0.2 ug/ml, respectively.     

酶水解-高效液相色谱法测定肉类食品及牛奶中11种甾体激素

碎肉用乙腈超声提取,挥干后加入KH2PO4缓冲液(牛奶直接加入KH2PO4缓冲液),再加β-葡萄糖醛酸酐酶,37℃恒温24 h,稀释后过SPE小柱,用乙腈洗脱,线性梯度变化的KH2PO4-CH3CN分离,二极管阵列检测器检测。所测11种激素的检出限为0.009~0.020 mg.kg-1;平均回收率为51.0%~107.0%;相对标准偏差为0.77%~6.42%之间;相关系数为0.999 5~0.999 9之间。酶水解-高效液相色谱法测定肉类食品及牛奶中11种甾体激素是一种简便、快速、灵敏、线性好的检测方法。     

高效液相色谱法测定阿卡波糖水解产物中的Valienamine含量

1引言Valienam ine((1S,2S,3S,4R)-1-氨基-5-(羟甲基)环己-5-烯-2,3,4-三元醇),分子式C7H13NO4,是一种由假单孢菌降解井冈霉素获得的假氨基糖,可通过阿卡波糖或井冈霉素水解来制备。它不仅能够有效治疗糖尿病及高血糖所导致的肥胖症,而且还能治疗艾滋病等其它疾病,所以对其的研究和开发已引起全世界生物医学领域的广泛关注。Valienam ine的检测方法主要有糖化酶检测法、紫外分光光度法、质谱法和薄层扫描法等,这些方法都存在难于定量或定量不准确、检测不方便的缺陷。有人用p-硝基氟苯与Valienam ine反应衍生生成N-p-硝基苯-Valienam …     

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贝类中的软骨藻酸

 介绍了用反相高效液相色谱 (RP HPLC)测定贝类中软骨藻酸的方法。样品以V(甲醇 )∶V(水 ) =1∶1的溶液提取 ,经LC SAX强阴离子柱固相萃取净化 ,用RP HPLC定量分析。方法的最小检出限为 0 2 μg/ g ,在 1 0mg/L～ 2 5 0mg/L范围内有良好的线性关系 ,测定结果准确 ,重现性好 ,回收率大于 96 %。     

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 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.     

High Performance Liquid Chromatographic Determination of Flurbiprofen in Pharmaceutical Dosage Forms

Abstract

A rapid, specific and reliable high performance liquid chromatographic assay of flurbiprofen in dosage forms has been developed. Reversed-phase chromatography was conducted using a mobile phase of 0.05 M ammonium acetate and acetonitrile, (40% v/v) PH 5.2 and detection at λ 247 nm. The recovery and coefficient of variation from six placebo tablets spiked with 100 mg of flurbiprofen were 100.1% and 0.4% respectively. Replicate regression analyses of three standard plots in the concentration range 0.5 - 9 mcg/ml obtained on three different days gave a correlation coefficient (0.99996) and the coefficient of variation of the slopes 0.159%. The assay was precise within day and between days as indicated by ANOVA test. It is suggested that the proposed HPLC method should be used for routine quality control and dosage form assay of flurbiprofen.     

高效液相色谱法测定西红柿中克线磷残留量

高效液相色谱法测定西红柿中克线磷残留量包宏,夏民洲,胡兹苓（南京林业大学南京２１００３７）１前言克线磷（ｆｅｎａｍｉｐｈｏｓ）,商品名Ｎｅｍａｃｕｒ,是防治作物根结、孢囊与自由线虫的杀线虫农药￣［１］。对它的残留分析,有气相色谱法和薄层色谱法等￣［２...