Determination of non-steroidal anti-inflammatory drugs in environmental samples by chromatographic and electrophoretic techniques

Non-steroidal anti-inflammatory drugs (NSAIDs) are a group of pharmaceutical compounds widely used in human health care and often found in the aquatic environment, with their metabolites. After an introduction that describes the general problem of drug contamination, the properties of NSAIDs, and environmental risk assessment, this review surveys the chromatographic and electrophoretic methods of analysis in use today for monitoring the most important representatives of this pharmaceutical class in different environmental samples.     

Click chemistry-based synthesis of water-dispersible hydrophobic magnetic nanoparticles for use in solid phase extraction of non-steroidal anti-inflammatory drugs

Magnetic nanoparticles (MNPs), prepared via thiol-ene click chemistry and containing both diol and octadecyl groups, are shown to possess both hydrophobic and hydrophilic functionalities. They display excellent dispersibility in water and also are capable of extracting non-steroidal anti-inflammatory drugs (NSAIDs) from water samples. The MNPs can be magnetically separated, and the NSAIDs eluted with acetonitrile-water (9:1, v:v) and submitted to high performance liquid chromatographic analysis. Extraction variables, such as the kind of ion-pairing reagents, amount of MNPs, pH of sample solution, extraction and desorption time, volume of desorption solvent and salt addition, were optimized. Under optimum conditions, the method has a wide analytical range (from 5 to 800 ng∙mL‾¹), good reproducibility with intra-day and inter-day relative standard deviations of <19.2 % (for n = 6), and low detection limits of 0.32 to 1.44 ng∙mL‾¹ for water samples. The results demonstrate that the material possesses good water compatibility, thus warranting ease of operation and good reproducibility.

The water-dispersible C18/diol-Fe₃O₄ MNPs were prepared via “Thiol-ene” click reaction. The material can be used as MSPE sorbent to extract non-steroidal anti-inflammatory drugs from river water. Satisfactory results were obtained with convenient operation and good reproducibility.

     

In situ aqueous derivatization and determination of non-steroidal anti-inflammatory drugs by salting-out-assisted liquid-liquid extraction and gas chromatography-mass spectrometry

A new analytical method for the determination of trace levels of five non-steroidal anti-inflammatory drugs (NSAIDs: clofibric acid, ibuprofen, naproxen, diclofenac and ketoprofen) in water samples is described. The analytical procedure involves in situ aqueous derivatization with N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (EDC) and 2,2,2-trifluoroethylamine hydrochloride (TFEA) and salting-out liquid-liquid extraction (SALLE), followed by gas chromatography-programmed temperature vaporizer-mass spectrometry (GC-PTV-MS). The influence of several parameters on the efficiency of the derivatization (stirring time, reaction time, reagent concentration and pH), and the extraction (solvent, volume, salts and stirring time) and injection steps (liner, injection volume, liner temperature, injection time, venting time and venting flow) was investigated. The detection limits of the method in water varied from 0.042 μg/L for ibuprofen to 1.2 μg/L for ketoprofen. The relative standard deviations (RSD) values were found to be relatively low (<10% for all compounds). The methodology developed was applied to the determination of NSAIDs in several environmental matrices including tap, river, sea and influent and effluent waste water samples. The results obtained show the presence of ibuprofen and naproxen in the influent waste water sample.     

基于金属有机骨架复合气凝胶的分散固相萃取-超高效液相色谱-串联质谱法测定水中5种非甾体类抗炎药

非甾体类抗炎药(NSAIDs)能在环境水体中长期稳定存在,不仅对生物有慢性毒性还能增加病原体的耐药性,开发可靠的测定水样中痕量非甾体抗炎药的分析方法至关重要.该文制备新型金属有机骨架/壳聚糖复合气凝胶材料Co-UiO-67(bpy)/CS分散固相萃取吸附剂,将其用于环境水体中酮洛芬、萘普生、氟比洛芬、双氯芬酸、布洛芬5...     

Confirmatory analysis of non-steroidal anti-inflammatory drugs in bovine milk by high-performance liquid chromatography with fluorescence detection

HPLC with fluorescence detection is considered for confirmatory analysis of group B veterinary drugs by the European Union legislation. A procedure for confirming the presence of anti-inflammatory non-steroidal drug (NSAID) residues in bovine milk by reversed phase high-performance liquid chromatography with fluorescence detection is herein described. The native fluorescence of nine drugs belonging to different NSAID sub-classes, namely flurbiprofen, carprofen, naproxen, vedaprofen, 5-hydroxy-flunixin, niflumic acid, mefenamic acid, meclofenamic acid and tolfenamic acid, allowed for detection in bovine milk down to 0.25–20.0 μg/kg. Confirmation of the nine NSAIDs is attained by fluorescence detection at characteristic excitation and emission wavelengths. The procedure described is simple and selective. Limits of quantification (LOQs) ranging between 0.25 and 20 μg/kg were measured; satisfactory trueness and within-laboratory reproducibility data were calculated at LOQ spiking levels, apart from 5-hydroxy-flunixin. The procedure developed is used in our laboratory for confirmation of each one of the above mentioned NSAIDs in bovine milk, to support results after HPLC quantitative analysis with UV–vis detection.     

Direct determination of non-steroidal anti-inflammatory drugs by column-switching LC-MS

A method for determination of 16 non-steroidal anti-inflammatory drugs (NSAIDs) in human plasma samples without time-consuming sample pre-treatments was developed. The system consisted of two pumps for mobile phase delivery, a six-port switching valve, a pre-column (Oasis HLB Cartridge Column), and a reversed phase analytical column (COSMOSIL 3C18-MS-II). The analytes were trapped on the precolumn and subsequently separated on the analytical column. The present method allowed on-line sample clean-up and enrichment, leading to improved sensitivity without any tedious sample preparation. The recoveries of NSAIDs from human plasma by column-switching were greater than 72.6%. The total analysis time for a single analytical run was approximately 11 min. The detection limits of NSAIDs were 0.0025 to 0.2 microg/mL using the selected ion monitoring mode.     

Capillary electrochromatographic separation of non-steroidal anti-inflammatory drugs with a histidine bonded phase

An open tubular wall-coated capillary column containing histidine functional groups was prepared and employed for the capillary electrochromatographic separation of non-steroidal anti-inflammatory drugs. The anion exchange along with the hydrogen bonding and hydrophobic properties of the surface coating allowed the separation of analytes with very similar ionic mobility. Selectivity and resolution were studied by changing the pH over the range from 3.5 to 5.0 and the concentration of the buffer from 10 to 25 mM, as well as variation of the organic modifier, such as methanol, ethanol and 1-propanol over the range 7.5 to 20%. The optimum experimental conditions for the separation of a drug mixture, which consisted of indoprofen, ketoprofen, suprofen, naproxen, flurbiprofen, fenoprofen and ibuprofen were using a mixture of acetate buffer (20 mM, pH 5.0)-ethanol (1:5, v/v) as background electrolyte and an applied voltage of -20 kV with UV detection at 220 nm. The separation of these drugs could be achieved with an average plate number of 1.0 x 10(5) m(-1).     

Recent advances on non-steroidal anti-inflammatory drugs, NSAIDs: Organotin complexes of NSAIDs

An overview is given of the results of organotin-NSAIDs interactions. Several organotin complexes with NSAIDs, derivatives of the carboxylic acid family and oxicam family, have been synthesized and characterized by spectroscopy and X-ray crystallography at the University of Ioannina. Results concerning the biological activity of these organotin complexes will be referred.     

Fabric phase sorptive extraction: A new sorptive microextraction technique for the determination of non-steroidal anti-inflammatory drugs from environmental water samples

Fabric phase sorptive extraction (FPSE) is a new, yet very promising member of the sorbent-based sorptive microextraction family. It has simultaneously improved both the extraction sensitivity and the speed of the extraction by incorporating high volume of sol–gel hybrid inorganic–organic sorbents into permeable fabric substrates. The advantages of FPSE have been investigated for the determination of four non-steroidal anti-inflammatory drugs, ibuprofen, naproxen, ketoprofen and diclofenac, in environmental water samples in combination with gas chromatography–mass spectrometry. Initially, the significance of several parameters affecting FPSE: sorbent chemistry, matrix pH and ionic strength were investigated using a mixed level factorial design (3¹ × 2²). Then, other important parameters e.g., sample volume, extraction kinetics, desorption time and volume were also carefully studied and optimized. Due to the high sorbent loading on the FPSE substrate in the form of ultra-thin coating and the open geometry of the microextraction device, higher mass transfer of the target analytes occurs at a faster rate, leading to high enrichment factors in a relatively short period of time (equilibrium times: 45–100 min). Under optimal operational conditions, the limits of detection (S/N = 3) were found to be in the range of 0.8 ng L⁻¹ to 5 ng L⁻¹. The enrichment factors ranged from 162 to 418 with absolute extraction efficiencies varied from 27 to 70%, and a good trueness (82–116% relative recoveries) indicating that the proposed method can be readily deployed to routine environmental pollution monitoring. The proposed method was successfully applied to the analysis of target analytes in two influent and effluent samples from a wastewater treatment plant and two river water samples in Spain.     

Determination of anti-inflammatory drugs in water samples, by in situ derivatization, solid phase microextraction and gas chromatography-mass spectrometry

In this work, the reactions of various copper ions with 1,3,3-trimethyl-2-[5-(1,3,3-trimethyl-1,3-dihydro-indol-2-ylidene)-penta-1,3-dienyl]-3H-indolium--more commonly known as dimethylindodicarbocyanine polymethyne dye (DIDC)--as well as the application of the results obtained for the development of a spectrophotometric method for the determination of Cu(I), Cu(II) and Cu(III) are described. Cu(I) and Cu(II) in the presence of chloride ions and DIDC reagent are extractable by a variety of organic solvents. It is important to emphasize that Cu(I) was extracted under considerably different experimental conditions than Cu(II). The optimum conditions for the extraction of the Cu ion associates with DIDC by amyl acetate and the determination of Cu(I) and Cu(II) were found to be: pH 3-5 and pH 3-6 and chloride concentrations of 0.5-0.8 mol L(-1) and 3-6 mol L(-1) for Cu(I) and Cu(II), respectively. The molar absorptivities for Cu(I) and Cu(II) are 1.8x10(5) L mol(-1) cm(-1) and 1.2x10(5) L mol(-1) cm(-1), respectively. A reaction mechanism is suggested. Cu(III) does not extract in the presence of chloride ions. However, Cu(III) is a strong oxidative agent which can cause the decolourisation of the DIDC reagent. The optimum conditions for Cu(III) determination were found to be: 2x10(-5) mol L(-1) DIDC; pH 8; water:acetone 4:1 medium. The developed procedures were tested for the determination of Cu(I), Cu(II) and Cu(III) in semiconductor samples.     

Determination of fourteen non-steroidal anti-inflammatory drugs in animal serum and plasma by liquid chromatography/mass spectrometry

The European Union has regulated the use of non-steroidal anti-inflammatory drugs (NSAIDs) in animal production and requires its member states to detect their residues in different matrices. In this work, a detailed MS and MS/MS study by ion-trap mass spectrometry of fourteen NSAIDs is described. Two multi-residue reversed-phase LC/ESI-MS/MS methods were developed, one for the determination of salicylic acid, naproxen, carprofen, flurbiprofen, ibuprofen, niflumic acid and meclofenamic acid in the negative ion mode, and the other for the determination of ketoprofen, suxibutazone, diclofenac, mefenamic acid, tolfenamic acid, phenylbutazone and its metabolite oxyphenbutazone in the positive ion mode. It was thus possible to confirm up to 14 different NSAID residues in serum and plasma samples of farmed animals, after chromatographic separation by a linear gradient. These substances were chosen as representative of different chemical subclasses of NSAIDs. The two methods were also validated in-house at three contamination levels, evaluating specificity and calculating mean recoveries, repeatability and within-laboratory reproducibility. The MS/MS product ion spectra were successfully used for the qualitative identification of all the drugs tested. All the NSAIDs, apart from salicylic acid, were recovered in high amounts, ranging between 71.6% and 100.9%.     

固相萃取-反相高效液相色谱法测定环境水体中卡马西平

建立了固相萃取-反相高效液相色谱法测定水中痕量卡马西平的方法.以C18固相萃取柱对水样固相萃取,再经Agilent ZORBAX Exlipse XDB-C18色谱柱分离,在紫外检测器上检测.考察了洗脱液、流动相比例的影响.确定洗脱液选用甲醇(17+3)溶液,流动相选择甲醇(11+9)溶液.结果线性关系良好(在0.2～...     

Nano polypyrrole-coated magnetic solid phase extraction followed by dispersive liquid phase microextraction for trace determination of megestrol acetate and levonorgestrel

The aim of the present work is combination of the advantages of magnetic solid phase extraction (MSPE) and dispersive liquid phase microextraction (DLLME) followed by filtration-based phase separation. A new pretreatment method was developed for trace determination of megestrol acetate and levonorgestrel by liquid chromatography/ultraviolet detection in biological and wastewater samples. After magnetic solid phase extraction, the eluent of MSPE was used as the disperser solvent for DLLME. Emulsion resulted from DLLME procedure was passed through the in-line filter for phase separation. Finally the retained analytes in the filter was washed with mobile phase of liquid chromatography and transferred to the column for separation. This approach offers the preconcentration factors of 3680 and 3750 for megestrol acetate and levonorgestrel, respectively. This guarantees determination of the organic compounds at trace levels. The important parameters influencing the extraction efficiency were studied and optimized. Under the optimal extraction conditions, a linear range of 0.05–50 ng mL⁻¹ (R² > 0.998) and limit of detection of 0.03 ng mL⁻¹ were obtained for megestrol acetate and levonorgestrel. Under optimal conditions, the method was successfully applied for determination of target analytes in urine and wastewater samples and satisfactory results were obtained (RSDs < 6.8%).     

A fast-screening approach for the tentative identification of drug-related metabolites from three non-steroidal anti-inflammatory drugs in hydroponically grown edible plants by HPLC-drift-tube-ion-mobility quadrupole time-of-flight mass spectrometry

The (tentative) identification of unknown drug-related phase II metabolites in plants upon drug uptake remains a challenging task despite improved analytical instrument performance. To broaden the knowledge of possible drug metabolization, a fast-screening approach for the tentative identification of drug-related phase II metabolites is presented in this work. Therefore, an in silico database for the three non-steroidal anti-inflammatory drugs (ketoprofen, mefenamic acid, and naproxen) and a sub-group of their theoretical phase II metabolites (based on combinations with glucose, glucuronic acid, and malonic acid) was created. Next, the theoretical exact masses (protonated species and ammonia adducts) were calculated and used as precursor ions in an autoMS/MS measurement method. The applicability of this workflow was tested on the example of eleven edible plants, which were hydroponically grown in solutions containing the respective drug at a concentration level of 20 mg/L. For the three drugs investigated this led to the tentative identification of 41 metabolites (some of them so far not described in this context), such as combinations of hydroxylated mefenamic acid with up to four glucose units or hydroxylated mefenamic acid with two glucose and three malonic acid units.     

基质固相分散-反相液相色谱法测定蔬菜中二甲戊乐灵农药残留

应用基质固相分散-反相液相色谱技术建立了提取、检测蔬菜中二甲戊乐灵农药残留的分析方法。研究了提取及测定条件对检测二甲戊乐灵的影响,确定了最佳提取条件:萃取吸附剂为弗罗里硅土,样品与弗罗里硅土吸附剂的比例为1∶4,洗脱剂为20 mL乙酸乙酯。二甲戊乐灵的测定线性范围为0.02~2.0 mg/L、r0.999。应用此方法测定了某些蔬菜样品中二甲戊乐灵农药残留,测定相对标准偏差为2.2%~5.2%。在几种蔬菜中的加标回收率为85%~93%,当样品为0.5 g时,方法检出限在5.5~10 ng范围内。     

Simultaneous determination of non-steroidal anti-inflammatory drugs in river water by gas chromatography-mass spectrometry

The gas chromatography/mass spectrometric assay method was developed for the determination of 13 non‐steroidal anti‐inflammatory drugs (NSAIDs) in river water. Extraction was achieved by a liquid‐phase extraction procedure using methylene chloride. The extract was reacted for 30 min at 80°C based on the formation of methyl ester with 1.0 M HCl in methanol and extraction of the derivative with ethyl acetate, which was then measured by gas chromatography‐mass spectrometry. The limit of quantification of NSAIDs was 1.0–60 ng/L and the calibration curve showed linearity being greater than r=0.997. The method was used to analyze ten river water samples from various regions in Korea. Diclofenac, indoprofen, ketoprofen and loxoprofen were detected at concentration of up to 1.29 μg/L in river water. The developed method may prove valuable for use in the national monitoring project of NSAIDs in surface water.     

In-tube solid phase microextraction using a beta-cyclodextrin coated capillary coupled to high performance liquid chromatography for determination of non-steroidal anti-inflammatory drugs in urine samples

A configuration of in-tube solid-phase microextraction (SPME) coupled to HPLC was constructed by using a pump and a six-port valve combined with a PEEK tube as the pre-extraction segment. The extraction capillary was fixed directly on the HPLC six-port valve to substitute for the sample loop. The whole system could be handled easily to perform accurate on-line extraction, and the possible inaccurate quantification caused by sample/mobile phase mixing when using an autosampler could be eliminated.A β-cyclodextrin coated capillary, prepared by sol-gel method, was used as the extraction capillary for in-tube SPME. Three non-steroidal anti-inflammatory drugs, ketoprofen, fenbufen and ibuprofen, were employed to evaluate the extraction performance of the capillary. After optimizing the extraction conditions, satisfactory extraction efficiency was obtained and detection limits for ketoprofen, fenbufen and ibuprofen in diluted urine samples were 38, 18 and 28 ng/mL, respectively. The extraction reproducibility was evaluated with intra-day and inter-day precision, and the R.S.D.s obtained were lower than 4.9 and 6.9%, respectively. The capillary was proved to be reusable and the extraction efficiency did not decrease after 250 extractions.     

Capillary zone electrophoresis for simultaneous determination of seven nonsteroidal anti-inflammatory drugs in pharmaceuticals

A simple capillary zone electrophoresis (CZE) method has been developed for analyzing seven nonsteroidal anti-inflammatory drugs (NSAIDs)—sulindac (SU), ketoprofen (KE), indomethacin (IN), piroxicam (PI), nimesulide (NI), ibuprofen (IB), and naproxen (NA). The separation was run using borate buffer (60 mmol L^–1, pH 8.5) containing 13% (v/v) methanol at 20 kV, and detected at 200 nm. Several conditions were studied, including concentration and pH of borate buffer, methanol percentage, and separation voltage. In method validation, the calibration plots were linear over the range 40.0–500.0 mol L^–1. In intra-day and inter-day analysis, relative standard deviations (RSD) and relative errors (RE) were all less than 5%. The limits of detection were 10 mol L^–1 for SU, IN, PI, and 20 mol L^–1 for KE, NI, IB, NA (S/N = 3, sampling 6 s by pressure). All recoveries were greater than 95%. This method was applied to the quality control of six NSAIDs in pharmaceuticals using NI as internal standard (IS). The assay results were within the labeled amount required by USP 25.