A rapid and reliable solid-phase extraction method for high-performance liquid chromatographic analysis of opium alkaloids from papaver plants

A rapid and reliable solid-phase extraction method for HPLC analysis of opium alkaloids from Papaver plants was established. Fifty mg of dried and powdered plant sample was extracted with 5 ml of 5% acetic acid for 30 min under sonication. After centrifugation, 3 ml of the supernatant was loaded on a reversed-phase cation-exchange solid-phase extraction cartridge. After seriate washings with 0.1 M hydrochloric acid and methanol, alkaloids were eluted with a mixture of 28% ammonia and methanol (1:19). The eluate was concentrated under nitrogen stream at 40 degrees C and the residue was dissolved in 50% aqueous methanol for high performance liquid chromatographic analysis. With this solid-phase extraction method, the recovery of morphine, codeine, oripavine, thebaine, papaverine, noscapine and sanguinarine was from 99.94 to 112.18% when the standard alkaloids were added to the plant samples. Opium alkaloids of a variety of genus Papaver plants cultivated in a field and phytotron were analyzed by this method.     

Reversed-Phase High Performance Liquid Chromatography (HPLC) of Nucleosides in Cell Extracts with Special Reference to Deoxythymidine

Abstract

A reversed-phase high performance liquid chromatography (HPLC) system was designed to assay the nucleoside concentrations, especially deoxythymidine, of tissue cultured cells. Concentration and purification of the acid extracted cell samples were achieved by utilizing a Sep-Pak C₁₈ cartridge. After adsorption of the sample, the cartridge was washed with 0.5 ml of water followed by 2 × 0.5 ml of 2.5% methanol. The compounds of interest were subsequently eluted in 2 × 0.5 ml of methanol. The cartridge procedure was found to be fast, inexpensive and showed good recoveries for most tested nucleosides. The nucleosides uridine, deoxythymidine and adenosine could be detected in green monkey kidney cells. Ribonucleotides and deoxyribonucleotides could be separated from each other with the HPLC system used.     

Rapid determination of citrinin in corn by fluorescence liquid chromatography and enzyme immunoassay

A rapid assay procedure was developed for mycotoxin citrinin in corn using liquid-liquid extraction (LLE) cartridges. Ground corn was extracted with methylene chloride and 0.5 N phosphoric acid. The extract was added to an LLE cartridge containing a diatomaceous-earth adsorbant, previously impregnated with sodium bicarbonate solution. After aspiration to dryness, the cartridge was eluted with methanol-water (4 + 1), and aliquots were taken for quantitation by reversed-phase liquid chromatography with fluorescence detection. Recoveries of citrinin added to ground corn at 200-1600 ng/g ranged from 71.2 to 86.3%, with coefficients of variation between 4.1 and 10.6%. An indirect enzyme immunoassay was also evaluated, using sodium carbonate solution for extraction. Recoveries of citrinin added to ground corn at 200-2000 ng/g ranged from 53.2 to 67.2%, but the coefficients of variation varied between 18.4 and 51.5%. The LLE cartridge procedure offers the advantages of low solvent consumption and speed, and is amenable to automation.     

A solid-phase extraction and size-exclusion liquid chromatographic method for polyethylene glycol 25 p-aminobenzoic acid determination in urine: validation for urinary excretion studies of users of sunscreens

No previous publications about percutaneous absorption of polyethylene glycol 25 p-aminobenzoic acid (PEG-25 PABA) have been found in the literature and the expected levels to be found in human urine after sunscreens use are unknown. The method proposed here is suitable to determine PEG-25 PABA in the urine of sunscreens users in order to carry out studies on body accumulation/excretion. It is based on solid-phase extraction (SPE) with size-exclusion liquid chromatography determination. Solid-phase extraction allows the analyte to be retained and subsequently eluted for a clean-up, using a silica-based cartridge. The size-exclusion liquid chromatography of the eluted allows the rest of matrix interferences to be avoided. Fluorescence intensity was measured at λ_em = 350 nm (λ_exc = 300 nm). The sensitivity of the proposed method is in the order of 450 ± 5 mL ng⁻¹ and the detection limit (3 S_y/x/b) in the measured solutions is in the order of 13 ng mL⁻¹, that is 2.6 ng mL⁻¹ in urine samples. The method enables PEG-25 PABA to be determined in both, spiked and unspiked human urine samples. Results obtained for spiked human urine samples (11-100 ng mL⁻¹) demonstrated the accuracy of the method. The mean relative standard deviation of the results was in the order of 3-10%. Three volunteers applied a sunscreen lotion containing a 8% PEG-25 PABA sunscreen cream and their urinary excretion was controlled from the moment of application until the excreted amounts were no longer detectable.     

A fully automated catecholamines analyzer based on cartridge extraction and HPLC separation

Summary A fully automated analyzer is described for the HPLC analysis of catecholamines. Firstly urinary and plasma catecholamines are reacted with diphenylboronic acid giving a complex without requiring a pH-meter step. This complex is strongly retained on a C 18 cartridge and elution with acetic acid solutions shows recoveries higher than 90 %. The catecholamines are eluted also by the mobile phase employed for the HPLC’ analysis. An intelligent autosampler makes the complex and fills a loop with all the solvents necessary for the sample cleanup. It then inverts the flow and pumps the sample and the solvents throught the cartridge. In the elution step the disposable extraction cartridge is positioned on stream with the HPLC analytical column by an automatic sampler. The separation is performed by ion-pair reversed-phase high-performance liquid chromatography, with sodium dodecyl sulphate as pairing ion, a short analytical column and electrochemical detection. The HPLC analysis time is 3 min and the total sample turnover time is 8 min. The recoveries and the precision of the analyzer are given together with correlation with manual HPLC.     

Semi-automated, solid-phase extraction procedure for liquid chromatographic determination of papaverine, diltiazem, desipramine and nicardipine in urine

Summary A solid-phase extraction (SPE) method for sample clean-up followed by a reversed-phase HPLC procedure is reported for the assay of papaverine, diltiazem, desipramine and nicardipine in urine. Disposable extraction cartridges (DECs) filled with C18, C8, C2, CH and PH silica-bonded phases were used. The effect on recovery of sample pH, composition of washing and elution solvents and nature of SPE cartridge were evaluated. The selectivity of SPE was examined using spiked urine samples and the PH cartridge gave rise to the cleanest extracts. Phenyl cartridges were conditioned with methanol and acetic acid-sodium acetate buffer. Urine sample was buffered and then applied to the DEC. The washing step was with acetone-water and subsequently with methanol-acetate buffer. The analytes were eluted with methanol-acetate buffer. The extract was evaporated to dryness, reconstituted in mobile phase, and chromatographed on a reversed-phase C18 column with UV detection at 212 nm. Recoveries of the tested compounds from spiked urine samples using the PH cartridge were in all cases>80%. The within-day and between-day repeatabilities were<5% and 9%, respectively.     

Selective solid-phase extraction of catecholamines by the chemically modified polymeric adsorbents with crown ether

A simple and selective one-step solid-phase extraction procedure using chemically modified polymer resin (Amberlite XAD-4) with crown ether was investigated for the measurement of urinary catecholamines. After loading the urine samples (adjusted to pH 4) on the synthesized adsorbent cartridge, the column was washed with methanol followed by water and then the adsorbed catecholamines were eluted by 1.0 mL of 6.0 M acetic acid. The effectiveness of sample clean-up method was demonstrated by reversed-phase ion-pair high-performance liquid chromatography with electrochemical detection. Under optimal condition, the recoveries of epinephrine, norepinephrine, and dopamine from spiked urine sample were >86% for all catecholamines. The detection limits (n=5) for epinephrine, norepinephrine, and dopamine were 37, 52, and 46 nmol/L, respectively.     

Simultaneous determination of the aromatic retinoids etretin and etretinate and their main metabolites by reversed-phase liquid chromatography

A rapid and simple method is described for the simultaneous determination of methyl, ethyl, isopropyl, n-propyl, isobutyl and n-butyl p-hydroxybenzoic acid esters (parabens) in cosmetics by high-performance liquid chromatography (HPLC). The method involves a single extraction of parabens with diethyl ether and clean-up on a Sep-Pak Florisil cartridge. Fat-soluble excipients in the diethyl ether extracts are removed through the cartridges with hexane-chloroform (75:25). Parabens are then eluted from the cartridges with hexane-ethyl acetate (70:30) and determined by HPLC on a reversed-phase column with water-methanol (50:50) as the mobile phase using sec.-butylpraben as an internal standard. The method was applied to samples with complicated matrices such as cream, milk lotion, lotion and cleansing foam, and the recoveries were 99.0-102.3% with coefficients of variation of 0.3-1.2%.     

Determination of carbendazim,thiabendazole, and<Emphasis Type="Italic"> o-</Emphasis>phenylphenol residues in lemons by HPLC following sample clean-up by ion-pairing

An efficient analytical method is presented involving effective sample clean-up with solid-phase extraction and HPLC-UV analysis for the simultaneous determination of carbendazim, thiabendazole, and o-phenylphenol residues in lemons. Sample preparation involves extraction with acetonitrile acidified with trifluoroacetic acid and an ethyl acetate/petroleum ether mixture. Purification of the crude extract was carried out with liquid–liquid partitioning after addition of an aqueous ammonia solution. Final clean-up was performed on polymeric reversed-phase cartridges pretreated with sodium dodecyl sulfate. Chromatographic analysis was performed on a reversed-phase HPLC column isocratically eluted with an acetonitrile/water/ammonia mixture and UV detection at 254 nm. The chromatographic method is repeatable, reproducible, and sensitive. Fungicide recoveries from lemon samples fortified at levels of 5 and 1 mg kg^–1 were 81–85% for carbendazim, 96–98% for thiabendazole, and 81–106% for o-phenylphenol with coefficients of variation of 2.5–7.4%. Detection limits for carbendazim, thiabendazole, and o-phenylphenol in lemons were 0.21, 0.27, and 0.51 mg kg^–1, respectively.     

Syringe-cartridge solid-phase extraction method for patulin in apple juice

A syringe-cartridge solid-phase extraction (SPE) method was developed for determination of patulin in apple juice. A 2.5 mL portion of test sample was passed through a conditioned macroporous SPE cartridge and washed with 2 mL 1% sodium bicarbonate followed by 2 mL 1% acetic acid. Patulin was eluted with 1 mL 10% ethyl acetate in ethyl ether and determined by reversed-phase liquid chromatography using a mobile phase consisting of 81% acetonitrile, 9% water, and 10% 0.05M potassium phosphate buffer, pH 2.4. Recoveries averaged 92% and the relative standard deviation was 8.0% in test samples spiked with 50 ng/mL patulin. The method appears to be applicable for monitoring apple juice samples to meet the U.S. Food and Drug Administration compliance action level of 50 microg/kg in an industrial quality assurance laboratory environment.     

多壁碳纳米管固相萃取净化-高效液相色谱法测定猪肉和鸡肉中的磺胺多残留

建立了多壁碳纳米管为吸附剂的固相萃取净化-高效液相色谱-紫外检测测定猪肉和鸡肉中多种磺胺类药物多残留的方法。样品采用乙腈提取,多壁碳纳米管固相萃取净化,NaH₂PO₄缓冲溶液（pH 5.5~6.0）溶解上样,5%（v/v）丙酮-正己烷淋洗,丙酮-二氯甲烷（1：1,v/v）洗脱。色谱分离以50 mmol/L NaH₂PO₄-乙腈（7：3,v/v）为流动相,方法的线性范围为0.01~1.00 mg/L,线性相关系数大于0.998,检出限（LOD）为0.003 mg/L,定量限（LOQ）为0.01 mg/L。在0.02~0.2 mg/kg添加范围内,9种磺胺类药物的回收率高于70%,RSD低于8%,表明多壁碳纳米管对磺胺类药物具有较强的吸附富集能力。该方法简便、准确可用于动物组织及产品中磺胺药物残留的检测。     

Development of a tandem thin-layer chromatography-high-performance liquid chromatography method for the identification and determination of corticosteroids in cosmetic products

A solid-phase extraction clean-up and and a liquid chromatographic method with ultraviolet detection were developed for the analysis of 51 corticosteroids in cosmetic samples in order to screen commercial samples for the presence of undeclared synthetic corticosteroids. A thin-layer chromatographic analysis was carried out on silica gel plates, using different eluants and detection reagents. When such a preliminary chromatographic separation gave some indications about the presence of steroid compounds, the methanol extracts from real samples were applied to a solid-phase extraction C₁₈ cartridge, and the analytes eluted with ethyl ether. The high-performance liquid chromatographic separation was then carried out for the identification and determination of the analytes using a Purospher RP-18 column, an isocratic or a gradient elution with a mixture acetonitrile-water and a photodiode-array detector. The accuracy of the method was determined by spiking experiments on home-made cosmetic samples. The analytical recoveries were satisfactory.     

Rapid and sensitive determination of phenylurea herbicides in water in the presence of their anilines by extraction with a Carbopack cartridge followed by liquid chromatography

A rapid and sensitive method for determining phenylurea herbicides in environmental aqueous samples in the presence of their anilines is described. The water sample is preconcentrated by passage at a flow-rate of ca. 150 ml/min through a 250-mg graphitized carbon black (Carbopack B) cartridge. After washing with 0.6 ml of methanol, the Carbopack B trap is connected with a cartridge containing a strong cation exchanger. Organics trapped by the Carbopack cartridge are eluted by passage of 6 ml of methylene chloride-methanol (95:5, v/v). Anilines and other basic compounds are quantitatively subtracted from the solvent system while flowing through the cation-exchange cartridge. After evaporation and redissolution, the sample is subjected to reversed-phase gradient elution high-performance liquid chromatography with UV detection at 250 nm. Recoveries of phenylureas added to water at levels between 30 and 3000 ng/l were higher than 92%. The limit of detection was about 1 ng/l, for a 2-1 sample. With respect to an octadecyl (C18)-bonded silica cartridge, the Carbopack B cartridge had a far better extraction efficiency for polar phenylureas.     

A high throughput and selective method for the estimation of valproic acid an antiepileptic drug in human plasma by tandem LC-MS/MS

A high throughput liquid chromatography-tandem mass spectrometric (LC-MS/MS) method for the determination of valproic acid, an antiepileptic drug, in human plasma is described. It is a rapid and sensitive isocratic reversed-phase liquid chromatography-tandem mass spectrometric method equipped with turbo ion spray (TIS) source, operating in the negative ion and pseudo selective reaction monitoring (SRM) acquisition mode to quantify valproic acid. The extraction of valproic acid and hydrochlorothiazide (IS) from the plasma involved sample treatment with phosphoric acid followed by solid-phase extraction using Waters hydrophilic-lipophilic balance (HLB) cartridge giving extracts free from endogenous interferences. Sample preparation by this method yielded very good and consistent mean recoveries of 99.73 and 74.47% for valproic acid and IS, respectively. The method was linear over the dynamic range of 2.0-200.0 μg/ml (covering entire therapeutic range) with a correlation coefficient r ≥ 0.9989. The coefficient of variance (CV, %) was 7.03% at 2.0 μg/ml (LLOQ). This method was fully validated for its accuracy, precision, recovery and matrix effect especially because the pattern of elution of all the analytes may appear as flow injection type. The analyte stability was examined under conditions mimicking the sample storage, handling and analysis procedures. The method was successfully applied for bioequivalence studies in human subject samples after oral administration of 500 mg formulations.     

Separation and concentration of sulfonylurea herbicides in milk by ionic‐liquid‐based foam flotation solid‐phase extraction

The ultrasound‐assisted ionic liquid foam flotation solid‐phase extraction of sulfonylurea herbicides in milk was developed and validated. The proteins and lipids were isolated from the sample matrix by adding salt and adjusting the pH value. The target analytes eluted from the solid‐phase extraction cartridge were determined by high‐performance liquid chromatography. Some experimental parameters, including the pH value of sample solution, amount of NaCl, ionic liquid type, extraction time, flow rate of carrier gas, flotation time, and solid‐phase extraction cartridge type were investigated and optimized. Under the optimized experimental conditions, the limits of detection for metsulfuron, pyrazosulfuron, chlorimuron‐ethyl, and nicosulfuron were 1.3, 0.6, 0.7, and 1.1 μg/L, respectively. When the present method was applied to the analysis of milk samples the recoveries of the analytes ranged from 84.3 to 105.2% and relative standard deviations were >5.7%.     

Determination of Triazines and N-Methylcarbamate Pesticides in Water by High-Performance Liquid Chromatography-Diode Array Detection

A rapid and selective method for the simultaneous determination of triazine herbicides (atrazine, its degradation product desethylatrazine, simazine, prometryn, terbutryn) and N-methylcarbamate insecticides (propoxur, carbaryl and methiocarb) in surface water has been developed. A 0.5 L of the water sample was preconcentrated by passage through a 1 g C₁₈ solid-phase extraction cartridge. The retained compounds were eluted with 5 mL of methanol from the cartridge. The pesticides were separated and quantified by reversed-phase high-performance liquid chromatography with UV diode-array detection. Analytical separation was performed using a concave gradient elution with acetonitrile and water on a C₁₈ column. Prometryn and terbutryn were determined at 240 nm; propoxur, methiocarb at 204 nm and the others at 220 nm. Recoveries varied from 85 to 102% over concentrations at 0.025 and 0.2 µg L^?1. The limits of detection for the compounds investigated are in the range of 0.005-0.012 µg L^?1.     

高效液相色谱法分析环境样品中的苯基胂化合物

二苯氯胂和二苯氰胂是刺激性毒剂,在环境中易于降解,其产物苯胂酸、苯胂氧、二苯胂酸、氧联双二苯胂和三苯胂比较稳定,对环境危害大。建立了同时测定这5种含胂产物的反相高效液相色谱方法,选择了最佳色谱条件,提供了各组分的紫外光谱图。5种化合物的线性范围分别为8~30,5~40,20~4000,120~8000,1~60 mg/L,检测限分别为0.1,0.1,0.2,10,0.1 mg/L。对实际环境样品进行了分析,结果较好。     

Application of probe sonication extraction for the determination of linear alkylbenzene sulfonates from sewage sludge. Comparison with other extraction methods

A new method based on probe sonication extraction (USP) prior to high performance liquid chromatography (HPLC) has been developed for the determination of linear alkylbenzene sulfonates (LAS) from sewage sludge. The optimized method was designed to be cost effective compared to existing extraction methods (ultrasonic assisted extraction, Soxhlet or pressurized liquid extraction) which may require large quantities of organic solvents, or costly instrumentation or equipment.The main factors affecting the extraction efficiency (extractant volume, ultrasounds power and extraction time) were optimized using compost sludge. The detection limit of total LAS in the sludge was 10 mg kg^− 1. The extraction of C₁₀-C₁₃ homologues is carried out using an extraction time of 7 min with 10 mL of methanol. Liquid chromatography with fluorescence (FL) detector is used for determination of LAS homologues. A mobile phase acetonitrile-water containing 0.1 M NaClO₄ (65:35) and isocratic elution was used. Compounds were eluted over 6 min at a flow rate of 1 mL/min. Polar interferences are eluted between 0 and 2 min and no purification of the samples is required prior to the final determination by high performance liquid chromatography (HPLC). The recoveries of LAS in spiked sewage sludge were between 84.0% and 97.0%, which reflect the efficiency of the method for extraction of these analytes from sewage sludge. Concentration levels found were between 11,858 mg kg^− 1 for digested sludge and 2379 mg kg^− 1 for compost sludge.     

Analysis of fatty acids in mouse cells using reversed-phase high-performance liquid chromatography

Summary A reversed-phase high-performance liquid chromatography (RP-HPLC) method was developed to analyze various fatty acids in recombinant mouse L cells. These fatty acids were the metabolites of oleic acid. A process was developed to extract fatty acids from the cell samples before RP-HPLC analysis. The samples were first saponified with 0.5 M NaOH in 96% ethanol then extracted with acidified ethyl acetate. After extraction, the sample was dried and dissolved in HPLC-grade methanol. After centrifugation to remove insoluble impurities, the sample was applied to a C₁₈RP-HPLC column using a gradient of acetonitrile (ACN)-H₂O. The eluted fatty acids were monitored by ultraviolet (UV) absorption at 195 nm and identified by retention time and adsorption spectrum comparison. This method successfully resolved various fatty acids and provided a tool for the elucidation of the fatty acid metabolic pathway in the cells.     

Solid-phase extraction followed by high-performance liquid chromatography-ionspray interface-mass spectrometry for monitoring of herbicides in environmental water

In this work we developed a sensitive and specific multiresidue method, based on reversed-phase liquid chromatography-mass spectrometry, with an ionspray interface (LC-ISI-MS), for determining 52 of most representative compounds of herbicides in water samples. The procedure used involved passing 0.5 l of surface water, 2 l of ground water and 4 l of drinking water samples, respectively, through a 0.5 g graphitized carbon black (GCB) extraction cartridge. Base-neutral and acid herbicides were differential eluted from GCB cartridge and follow analyzed by HPLC-ISI-MS apparatus. A conventional 4.6-mm-ID reversed-phase LC C18 column, operating with a mobile phase flow-rate of 1 ml/min, was used to chromatograph the analytes. A flow of 100 microl/min of the column effluent was diverted to the ISI source. The study demonstrates the sensitivity of the technique, with detection limit under 10 ng/l in drinking water samples. Performance data for the method such as recovery and precision are also reported.