You are on page 1of 16

22

Anti-Inflammatory & Anti-Allergy Agents in Medicinal Chemistry, 2009, 8, 22-37

Analytical Procedure for Determination of Cyclooxygenase-2 Inhibitors in Biological Fluids by High Performance Liquid Chromatography: A Review
Giuseppe Carlucci*
Università degli Studi “ G. D’Annunzio” Chieti-Pescara - Facoltà di Farmacia - Dipartimento di Scienze del Farmaco via dei Vestini - 66100 Chieti –Italy
Abstract: High-performance liquid chromatographic methods for the analysis of cyclooxygenase 2 inhibitors (COX-2) in biological fluids are reviewed. In particular, sample preparation and handling procedures, chromatographic conditions and detection methods are discussed. A summary of published high-performance liquid chromatographic assays for individual nabumetone, celecoxib, rofecoxib, nimesulide, etoricoxib, etodolac, deracoxib, lumiracoxib, valdecoxib, and meloxicam is included.

Keywords: Sample preparation, NSAIDs, COX-2 inhibitors, biological fluids, HPLC. 1. INTRODUCTION The anti-inflammatory, analgesic, and antipyretic drugs are a heterogeneous group of compounds, often chemically unrelated (although most of them are organic acids), which nevertheless share certain therapeutic actions and side effects. The prototype is aspirin, hence these compounds are often referred to as aspirin-like drugs; they also are frequently called non-steroidal anti-inflammatory drugs, or NSAIDs, an abbreviation that is used throughout this review to refer to these agents. There has been substantial progress in elucidating the mechanism of action of NSAIDs. Inhibition of cyclooxygenase (COX), the enzyme responsible for the biosynthesis of the prostaglandins and certain related autacoids, generally is thought to be a major facet of the mechanism of NSAIDs. Some of the shared properties of NSAIDs are considered first; the more important drugs are discussed in some detail. Since the principal therapeutic effects of NSAIDs derive from their ability to inhibit prostaglandin production, the enzymatic activities involved in prostaglandin synthesis are described here briefly. The mechanisms by which varying NSAIDs interfere with prostaglandin synthesis then are outlined. The first enzyme in the prostaglandin synthetic patway is prostaglandin endoperoxide synthase, or fatty acid cyclooxygenase. This enzyme converts arachidonic acid to the unstable intermediate s PGG2 and PGH2. It is now appreciated that there are two forms of cyclooxygenase, termed cyclooxygenase-1 (COX1) and cyclooxygenase-2 (COX-2) [1]. COX-1 is a constitutive isoform found in most normal cells and tissues, while COX-2 is induced in settings of inflammation by cytokines and inflammatory mediators [2]. However, COX-2 also is constitutively expressed in certain areas of kidney and brain [3, 4]. Importantly, COX-1, but not
*Address correspondence to this author at the Università degli Studi “ G. D’Annunzio” Chieti-Pescara - Facoltà di Farmacia - Dipartimento di Scienze del Farmaco - via dei Vestini - 66100 Chieti –Italy; E-mail: g.carlucci@unich.it 1871-5230/09 $55.00+.00

COX-2, is constitutively expressed in the stomach. This accounts for the markedly reduced occurrence of gastric toxicity with the use of selective inhibitors of COX-2. 2. NABUMETONE Nabumetone (1) or 4-(6-Methoxynaphthalenyl)-2-butanone, is a nonsteroidal anti-inflammatory drug of the acetic chemical class. Nabumetone is a prodrug that requires conversion to an active metabolite 6-methoxy-2-naphthyacetic acid (6-MNA) for its anti-inflammatory, analgesic, and antipyretic activity. Structures of nabumetone and its metabolite are shown in Fig. (1). Nabumetone is the only NSAID of the acetic acid class with a half-life long enough to support once daily administration. 6-MNA competitively inhibits both cyclooxygenase isoenzymes, COX-1 and COX-2, by blocking arachidonate binding resulting in analgesic, antipyretic, and anti-inflammatory pharmacologic effects [5-8]. The enzymes COX-1 and COX-2 catalyze the conversion of arachidonic acid to prostaglandin G2 (PGG2), the first step of the synthesis of prostaglandins and thromboxanes that are generated in rapid physiological responses [9, 10]. Nabumetone is used in the treatment of osteoarthritis, rheumatoid arthritis and other musculoskeletal disorders. Preliminary studies to quantify this drug in biological fluids involved radioassay scintillation counting of isotopes and gas chromatography

Fig. (1). Chemical structures of nabumetone and 6-methoxy-2naphtyacetic acid (6-MNA).

© 2009 Bentham Science Publishers Ltd.

phase HPLC column. the organic layer was transferred to another glass tube and evaporated to dryness at 45°C under a gentle stream of nitrogen. respectively and evaporated to dryness under a gentle stream of nitrogen. A simple and sensitive HPLC method for the determination of nabumetone in human plasma was developed by Kobyliska et al. and aliquots of 10 μL were injected into the chromatograph. No. 28]. transferred to autosampler vials and 40 μL aliquot was injected onto the HPLC system for analysis. 2009. fitted with a guard column. These authors developed a simple HPLC method for the simultaneous determination of nabumetone.0 mL of plasma or serum.0 mL of diluted methanolic urine samples were applied to pre-conditioned Bond-Elut Certify II cartridges and slowly drawn through the cartridge. Direct injection of diluted urine [17. and centrifuged at 1500 x g for 5 min. acidified using a combination of 0. [22] and Ray et al.. The phosphorescence signals are a consequence of intermolecular protection when analytes are. fluorimetric detection having a higher sensitivity (0. Pharmacokinetics of nabumetone. spinal cord. while Ray et al. The extraction procedure described by AlMomani et al. 20] and liquid-liquid extraction as a sample clean-up procedure [14.0 mL of ethyl acetate was added. Briefly. [26].5 mL of plasma. made use of UV detection at 280 nm. 19. respectively. Both authors then extracted the analyte into an organic phase ether and n-hexane-ethyl acetate 50:50. (2). These n-hexane -ethyl acetate (1:1) solutions were evaporated to dryness in a water bath at 40°C. [16] appears to be loosely based on the method described by Ray et al. COX-2 is induced at inflammation sites [31. were added. Ray et al. back extraction and/or some derivatization techniques induced to increase the sensitivity are often carried out to avoid interference substances [23-25]. While Jang et al. Vol. 25 μL of an internal standard working solution and 0. The lower limits of quantification in human urine were 150 and 16 ng/mL for nabumetone and 6-MNA. 32] but is also found constitutively in brain. and emission at 350 nm. Jang and Al-Momani used approximately 1. After freezing at –70°C for 20 min. For the liquid-liquid extrac-tion procedure. Several high-performance liquid chromatographic methods have been published for the individual determination of nabumetone in biological fluids.1 M HCl and 0. The ability of this method to distinguish intact nabumetone from its metabolite was demonstrated. A selective. 22] are frequently used for the naphthalene derivatives in biological fluids. CELECOXIB Celecoxib or 4-[5-(4-Methylphenyl)-3-(trifluoromethyl)1H-pyrazol-1-yl]benzenesulfonamide is a selective inhibitor of COX-2 for the treatment of rheumatoid arthritis and osteoarthritis [31-35]. used 1. [30]. 6-chloro-2-naphthylacetic acid [15]. Clinical studies indicate that celecoxib is metabolised in the liver via the oxidative pathway to the corresponding alcohol and carboxylic acid and is removed by renal excretion as a glucuronide metabolite [36]. The identity of the nabumetone metabolites in biological samples was confirmed using HPLC-MS experiments. The assay of Nabumetone and 6-MNA and the internal standard utilized a fluorescence detector with excitation at 280 nm. the plasma was briefly mixed with a vortex mixer and 3. while Ray et al. kidney and some other tissues [33-35]. [27. dry filled with Perisorb RP18 pellicular packing. 6MNA was separated from the internal standard and endogenous plasma components using a LiChrospher 100 RP8 stainless-steel column.01 M phosphate buffer pH 10. using naproxen as internal standard.2 mL of 0.313 ng/mL and the calibration curve was linear up to 20 ng/mL. Mikami et al. Naproxen was chosen as the internal standard. In this study the authors describe a procedure that is well suited for rapid sample processing. requiring only a small volume of plasma (200μL).5M citrate buffer (pH 3). simple and sensitive heavy atom induced-room temperature phosphorimetric method for the determination of 6MNA in biological fluids was developed by Pulgarín et al. [14] both described liquidliquid extractions.5 mL of plasma.5 M HCl only. using methyl p toluate as the internal standard. The residues were redissolved in 200 μL of ethanol containing methyl-p toluate. Then. The cartridges were washed with 4. Jang et al. Briefly. The limit of quantification was established as 0.1 μg/mL). 6-MNA and the other metabolites. after which the samples were reconstituted and injected into a normal . and carboxycelecoxib are shown in Fig.. used 0. 6-MNA and 6-HNA (6-hydroxy-2-naphthylacetic acid) in human plasma and minipig plasma was evaluated and compared. both UV for higher concentrations and fluorescence detection for very low concentrations were used. The disposition of nabumetone after a single oral dose administration of this drug to humans and minipigs was investigated by Nobilis et al. [15] describes a reversed-phase HPLC method for the simultaneous quantitation of nabumetone and its metabolite 6-MNA in human urine. Nabumetone and 6-MNA were extracted from the sample matrix using solidphase extraction by Bond-Elut Certify II cartridges containing reversed-phase and anion exchange functionalities for the analysis of these drugs in human urine with fluorimetric detection. De Jager et al. The analytes were eluted with 6 mL of n-hexane-ethyl acetate (1:1). This technique enables us to determine analytes in complex matrices without the need for a tedious separation process. The chemical structures of celecoxib. while using UV detection at 270 nm. 8. 3. 17. centrifuged. serum and urine samples [17-23]. The clinical pharmacokinetics and pharmackodinamics of cele- . The procedure used involves liquid-liquid extraction with ethyl acetate and reversed-phase chromatography with fluorimetrica detection with excitation at 230 nm and emission at 356 nm. aliquots of 1. 1 23 with flame ionization detection and reversed-phase highperformance liquid chromatography using violet or fluorescence detection [11-16]. 0. detected fluorometrically with excitation at 284 nm and emission at 320 nm. following acidification of drug-containing plasma.Analytical Procedure for Determination of Cyclooxygenase-2 Anti-Inflammatory & Anti-Allergy Agents in Medicinal Chemistry.. exclusively. [25] described an HPLC assay for 6-MNA in which samples are prepared by protein precipitation. in the presence of heavy atom salt and sodium sulfite as scavinger to minimize room temperature phosphorescence quenching. hydroxycelecoxib.0 mL of water and dried for five min by aspiration with a moderate to strong vacuum. The sample residue was dissolved in 200 μL freshly prepared mobile phase. however there have been no reports concerning the simultaneous determination of nabumetone and 6-MNA by isocratic HPLC. -naphtol [22] or naproxen [27. Jang et al. The solution was shaken for 5 min. 32] were used as internal standards for the determination in biological samples. [29] and Ray et al. 19.

Development of sensitive and specific analytical techniques for the determination of celecoxib in biological samples is highly required for clinical investigations and pharmacokinetic studies. Vol. No. The assay was linear in the concentration range of 25-2000 ng/mL. the buffered plasma was aspirated through the SPE cartridges using a vacuum pressure. 1. The limits of quantification and detection of celecoxib in plasma were determined by analysing plasma samples spiked with celecoxib at relatively low concentrations (20-50 ng/mL) using the developed LC-MS method under the described conditions. Thereafter compounds were separated on a narrow bore RP-C18 column. pH 3. The method involved extraction of the analytes from plasma samples at pH 5 with ethyl acetate and evaporation of the organic layer. [40]. while for microdialysis samples a modified chromatographic procedure was used. The HPLC method included a column switching procedure. The tubes were vortexed. Microdialysis samples were injected directly into the LC-MSMS system.i. hydroxycelecoxib and carboxycelecoxib. 8. SPE cartridges for plasma were conditioned using sequential washes of 2 mL acetonitrile and 2 mL water. [38] described a method for the determination of celecoxib in human plasma. 30 min).D. Woolf et al. The limit of quantification for celecoxib in plasma was found to be 50 ng/mL (mean . Bräutigam et al. and the solvent was evaporated using nitrogen (50° C. to reduce run-time to 12 min. Extraction was not necessary. respectively. The cartridges were rinsed with 1 mL of water and 1 mL of acetonitrile-water (25:75). Chromatographic separation of extracted plasma samples was performed in isocratic mode with a Nucleosil C18 column (30 x 2.5 ng/mL. highly reliable and reproducible liquid chromatographic-mass spectrometric assay is developed for the determination of celecoxib in human plasma using sulindac as an internal standard by Abdel-Hamid et al. Celecoxib and the internal standard were extracted from plasma by solid-phase extraction with a C18 cartridges. 2009.1). The mobile phase consisted of a mixture of acetonitrile-water-ammonium hydroxide solution 25% (85:15:0. 5μm particle size and 100Å pore size).D. The mass spectrometer was programmed in the positive single-ion monitoring mode to permit the detection and quantification of the molecular ions of celecoxib and sulindac at m/z 382 and 357. (2). Briefly.0 mm I.10 M. The cartridges were allowed to aspirate to dryness after which they were removed from the manifold and suspended into 15 mL centrifuge tubes. The supernatant was decanted into a disposable glass culture tube containing 3 mL of 0.s. Upon transfer of the sample to conditioned cartridge. The samples were chromatographed under normal-phase HPLC conditions using a Nucleosil-NO2 column. 1 Giuseppe Carlucci Fig.0 mL/min. The reconstituted solution of the residue was injected onto a Shim Pack GLC-CN. The residue in the tube was reconstituted using 250 μL of mobile phase and transferred to a low-volume conical glass vial prior to injection onto the HPLC. in which late eluting compounds were diverted to waste. Detection was accomplished using UV absorbance at 260 nm. The tube containing the elution solvent was placed in a Zymark Turbovap LV evaporator. C18 column and chromatographed with a mobile phase comprised of acetonitrile-1% acetic acid solution (4:1) at a flow-rate of 1.. The tubes were capped and centrifuged for 10 min at 2500 g.0. 15 p. Chemical structures of Celecoxib. respectively. The analytes were eluted from the SPE cartridges by drawing two 1 mL aliquots of acetonitrile through each cartridge using centrifugation.0 mm I. A robust.25 ng/mL and in microdialysis samples 0. [39] describes a liquid chromatography-tandem mass spectrometry for the quantification of celecoxib in human plasma and rat microdialysis samples using 4-[5-phenyl-3-(trifluoromethyl)1H-pyrazol-1-yl]benzesulfonamide as the internal standard. coxib have been reported [37]. Four 250 μL aliquots of acetonitrile were added to each tube vortexing for 10 s after each addition. 5μm particle size and 100Å pore size) with a mobile phase acetonitrile-waterammonium hydroxide solution 25% (65:35:0. sodium phosphate buffer and mixed. The buffered samples were poured into individual 10 mm/6mL 3M Empore C18 solid-phase cartridges positioned on a 20-place vacuum manifold equipped with stopcocks at each position.0 mL of plasma was pipetted into a 15 mL polypropylene conical tube. Microdialysates were separated using a Nucleosil C18 column (70 x 2. The detection was by a PE Sciex API 3000 mass spectrometer equipped with a turbo ion spray interface.. The limits of quantitation for celecoxib were in human plasma 0. The limit of quantification for celecoxib was 25 ng/mL..1). A 25 μL aliquot of 10 μg/mL working internal standard solution was pipetted into each of the tubes containing the samples and the previously prepared standards.24 Anti-Inflammatory & Anti-Allergy Agents in Medicinal Chemistry. Tubes containing samples received an additional aliquot of acetonitrile (50 μL) to make these samples equivalent in organic content to the standards. Many chromatographic methods for the assay of celecoxib were reported.

D. The method is rapid. The lower limit of quantification for the celecoxib and rofecoxib is 20 and 10 μg/mL.0 mm I.0 mm I.D. On the other hand. A simplified solid-phase extraction procedure and liquid chromatographic determination of cele- coxib in rat plasma was developed by Guermouche et al. sodium diclofenac and niflumic acid in human serum by Navas et al.1 mm. [47] described a procedure for determination of celecoxib in human plasma and breast milk by HPLC..Analytical Procedure for Determination of Cyclooxygenase-2 Anti-Inflammatory & Anti-Allergy Agents in Medicinal Chemistry. plasma samples are extracted into an organic solvent (1-chlorobutane) and evaporated under an air flow. 2009. connected to an Aqua C18 (4 mm x 3. The samples were chromatographed on a Nova Pak C8 column (150 x 3. in addition to two well-known non-steroidal antiinflammatory drugs. hydroxycelecoxib. after protein precipitation with acetonitrile.5 (50:50) containing 0. Briefly. At all concentrations intra. No.2 mL each of blank. [41]. was taken. the limit of quantification was 12. using flutamide as the internal standard. Each mixture was vortexed for 30 s.01 mg/L.4 with 85% orthophosphoric acid (43:58). 5 μm) analytical column. The 4-n-pentyl-phenylacetic acid was used as the internal standard.5 ng/mL for a sample size of 0. Zhang et al.9 mm. Aliquots of 10 μL were used for HPLC analysis.5 mL of serum. I. and 40 μL of clear supernatant was injected into the HPLC system. The method was applied to the determination of celecoxib for pharmacokinetic studies in man. applying a gradient with acetonitrile and . the samples were separated by gradient reversed-phase HPLC and quantified using UV detection at 254 nm. [42]. The achieved limits of quantification of 10 ng/mL for each analyte allowed the determination of the pharmacokinetic parameters of the analytes after administration of 100 mg celecoxib. the method was used to study the pharmacokinetic profile of celecoxib following administration of a single oral dose (200 mg) in 12 healthy volunteers.and inter-assay variabilities were below 11% with a less than 9% error. The chromatographic separation is achieved using a Zorbax SBCN 5 μm analytical column operated at room temperature and mobile phase consisting of acetonitrile and water containing 0. centrifuged at 15000 x g for 10 min. 5 μm). Use of the analytical column was preceded by that of a direct-connect column prefilter. Jalalizadeh et al. The method is based on liquid-liquid extraction with chloroform.0% expressed as (% deviation) of the nominal concentration. 3 μm particle size) at a flow-rate of 0. The optimal chromatographic conditions for the celecoxib determination were investigated. acetonitrile 0. The method was linear over the concentration range 10-500 ng/mL. 100 mg) for sample preparation. followed by HPLC with ultraviolet detection set at 215 nm. 1 25 predicted concentration 41. The plasma and milk calibration curves of celecoxib were constructed by spiking drug-free human plasma or breast milk with celecoxib working standard solution at concentrations of 10 to 2000 μg/L. A simultaneous determination of celecoxib.1mm I. Chromatographic separation was achieved using a Hypersil ODS (150 x 4.0). The mobile phase consists of acetonitrile-water (60:40). celecoxib and refecoxib. using 1.2% and an accuracy of –9. Briefly.002 mg/L.1% triethylamine. After pilot investigations. For the limit of quantification 0. The limit of quantification of celecoxib was around 10 μg/L in both plasma and milk.D. Briefly.) precolumn. Werner et al.) preceded by a C18 home-made guard column (20 x 4. standard.. The limit of detection with a signal to noise ratio of 3:1 was 4 ng/mL in plasma. The method did not include an internal standard.acetate buffer 0.and inter-day coefficients of variation were < 5%. samples were extracted with chloroform. A simple and sensitive method for the determination of celecoxib in human serum by HPLC with fluorescence detection was developed by Schönberger et al. Briefly. connected to a C18 (5 x 4. Rofecoxib was used as an internal standard. respectively. [43] developed a method for the quantification of celecoxib in human plasma based on reversed-phase high-performance liquid chromatography coupled to atmospheric pressure chemical ionisation (APCI) mass spectrometry after liquidliquid extraction. a poly(divinylbenzene-co-N-divinylpirrolidone) Oasis HLB 60 mg SPE cartridge was used to directly extract celecoxib from rat serum without any supplementary step. ibuprofen was choose as the internal standard. A demethylated analogue of celecoxib was used as internal standard. at which the mean values were within ± 15% of the spiked values and the intra. A reversed-phase HPLC method was developed for the separation and simultaneous determination of two cyclooxygenase inhibitors.) guard column. [49]. [45]. The assay utilizes a Nucleosil CN column and UV spectrophotometer detection at 260 nm. [46] describes a reversed-phase HPLC method for the quantitation of celecoxib in human plasma. Using fluorescence detection with excitation at 240 nm and emission at 380 nm. An innovative reversedphase high-performance liquid chromatographic method is validated for the simultaneous determination of celecoxib and rofecoxib in human plasma by Hamama et al.0 mL of human plasma.D. After validation.35 mL/min using water-acetonitrile (40:60) as the mobile phase.075 M (pH 5. Chow et al. The limit of quantitation was 5 μg/L. respectively.0 mm) using a mobile phase of acetonitrile.5 – 1500 ng/mL and showed good accuracy and reproducibility. Separations were carried out on a Novapak C18 column (150 x 4.25 mL of plasma. The limit of quantification of celecoxib was 40 ng/mL with 0. In this work. [44].5 ng/mL). and carboxycelecoxib in human plasma using gradient reversed-phase liquid chromatography with ultraviolet absorbance detection was developed by Störmer et al. the limit of detection for celecoxib in plasma was 20 ng/mL.01 M phosphate buffer pH 3. This concentration yielded an RSD of 10. Separation was achieved on a Prontosil C18 AQ column (150 x 3 mm I. Vol.0±4. The assay of celecoxib and rofecoxib in human plasma utilized a ultraviolet detector set at 254 nm.1 M potassium dihydrogen orthophosphate buffer adjusted to pH 2. [48] described a method for the determination of celecoxib in human plasma using a solid-phase extraction (BakerBond Octadecyl SPE cartridges. The detection limit at a signal to noise ratio of 3 was 0. following a solid-phase extraction procedure. The assay was linear in the concentration range of 12. The chromatographic separation was achieved in an Aqua C18 5 μm reversed-phase column. defined as the minimum concentration that could be measured with a CV<5% was found to be 10 ng/mL in 500 μL of plasma sample. 8. The residue after extraction was reconstituted in 100 μL of mobile phase. The calibration curves for the two drugs were linear over the range 20 to 2000 μg/mL for celecoxib and 10 to 500 μg/mL for rofecoxib.2 mL was added to 0. The limit of quantification of the method. The mobile phase used was a mixture of acetonitrile and 0.D. sensitive and highly selective. mixed well and 60 μL of the final clear solution was injected into HPLC system. QC or the samples plasma or milk to precipitate the proteins.

Accordingly.45) containing 0.1 M. Detection was made using a diode array detector (DAD). Then 20 μL of supernatant was injected into the liquid chromatography. and injected into the chromatograph for analysis. A simple and rapid HPLC method for determination of celecoxib in human plasma for application in pharmacokinetic studies was developed and validated by Zarghi et al.5 mg/L. Under the chromatographic conditions described. [60] also developed an HPLC method with tandem mass spectrometric detection of rofecoxib in human plasma. 2009. [51] and Matheson et al. and analytical recovery was 100. Recently its use has become more controversial due to cardiovascular side effects on prolonged use. The method is based on high-performance liquid chromatography with atmospheric pressure chemical ionisation tandem mass spectrometric (APCI-MS-MS) detection in negative ionisation mode using a heated nebulizer interface.5 ng/mL. The separation was performed on Chromolith Performance (RP-18e.1 to 100 ng/mL of plasma.1M. the chromatographic elution step is undertaken in a short time (less than 6 min).) was used. In this case a mobile phase was a mixture of acetonitrile-water (35:65).8 mg/L and 2.0% trifluoracetic acid as an organic modifier. 100 x 4. No. [61]. 500 μL of acetonitrile and 100mg NaCl.0 mL of human serum were placed into a 5. to 450 μL of plasma in a glassstoppered 15 mL centrifuge tube were added 50 μL of nefenamic acid as internal standard (5.6 mm I. In many cases the technique has been reported to be highly competitive with HPLC with tandem mass spectrometric detection in terms of speed of analysis. methanol and water (45:10. [53]. The residue was dissolved in 200 μL of water-acetonitrile (50:50). liquid chromatographic assay for the determination rofecoxib in human plasma utilizing liquid-liquid extraction for sample preparation followed by HPLC with post. which has lower separation impedence compared with the particulate packings.D. (3). A C8 SPE plate was used for the extraction of the drug from human plasma while a mixed mode (C8Cation) SPE plate was used to isolate the analytes from human urine. Its chemical structure is shown in Fig. The mobile phase containing 1. The mobile phase used was a mixture of acetonitrile-water (50:50). The analyte and internal standard were chroma- . and the pH of the sample was adjusted with 6 mL of pH 6 phosphate buffer solution 0. semi-automated determination of Rofecoxib in human plasma and urine using solid-phase in the 96-well format and high-performance liquid chromatography with post-column photochemical derivatization-fluorescence detection was developed by Mattews et al. The limit of quantification of the assay. from 15 to 60% acetonitrile. The assay utilizes a reversed-phase BDS-Hypersil C18 analytical column (100 x 4.0 mL plastic tube. The assay enables the measurement of celecoxib for therapeutic drug monitoring with a minimum quantification limit of 10 ng/mL. Vol.0 mL of water was performed and the cartridges were dried by passing air. [50].0 mm I. solid phase extraction was performed using a C18 solid phase extraction 200 mg cartridge fitted with 3. Briefly.column photochemical derivatization and fluorescence detection was developed by Woolf et al.0 mL of pH 6 phosphate buffer solution 0. an Astec Beam Boost photochemical reactor equipped with a 254 lamp and a 10 m reaction coil (0. The cartridges were conditioned with 2.0 mm I. The columns operated at ambient temperature (approximately 22°C). was 0. The use of HPLC-with post-column photochemical derivatization-fluorescence detection has been reported for the determination of various drugs in biological fluids [54-58]. The chromatographic separation was achieved in a YMC ODS AQ (100 x 3. one-step extraction procedure. the mixture was centrifuged for 15 min at 8000 g. Details of its pharmacokinetics. The method involves a simple.D. 8.0 mL of methanol and 2. respectively. A high-throughput.D. After mixing.0 mL reservoir. After blending. The buffered serum samples were passed through the C18 cartridges by gravity. 3 μm) analytical column.0 mL of acetonitrile. The sample injection volume was 150 μL. [52]. The assay of celecoxib in plasma utilized a UV absorbance detector set at 254 nm with a mobile phase consisting of acetonitrile.0 μg/mL). 4.5±1. Rofecoxib and the internal standard were isolated from basified plasma using liquid-liquid extraction.2% acetic acid (pH 3..5). a clean up procedure with 3. The fluorescence detector was set at an excitation wavelength of 250 nm and an emission wavelength of 375 nm.3%. 5 μm) preceded by a threaded guard column (20 x 4 mm) packed with the same material used for the separation. precolumn). The eluted drugs were collected in a glass tube. Chemical structure of Rofecoxib. Owing to the use of a monolithic column. and therefore. 1 Giuseppe Carlucci water. therapeutic efficacy.6 mm) column. celecoxib and the internal standard peaks were well resolved. The eluate was evaporated to dryness with nitrogen. Chavez-Eng et al. it allows easy optimising chromatographic conditions to obtain desirable resolution in a shot time. The analytical procedure in the serum pre-treatment consisting of extraction using a C18 solid-phase extraction cartridges. Before eluting the drugs with 3.3 mm I. A high-performance Fig.D. The assay was validated in the concentration range of 0. (3). with high resolution. Cleaner extracts were obtained when methyl-tert-butyl ether (MBTE) was utilized as an extraction solvent and plasma was basified before extraction. connected to a YMC ODS AQ (20 x 3.26 Anti-Inflammatory & Anti-Allergy Agents in Medicinal Chemistry. Separation was performed on a reversed-phase monolithic column. much faster separations are possible as compared with traditional chromatographic columns packed with porous particles. Isolation of Rofecoxib and the internal standard was achieved by solid-phase extraction (SPE) in the 96well format. Briefly. Briefly. selectivity and sensitivity [59]. ROFECOXIB Rofecoxib or 4-[4-Methylsulfonyl-phenyl]-3-phenyl-2(5H)-furanone is a selective inhibitor of COX-2 for the treatment of rheumatoid arthritis and osteoarthritis. Endogenous plasma components did not give any interfering peaks. 2. and toxicity have been reviewed by Scott et al. The limits of quantification for celecoxib and refecoxib were 1.

and plasma concentrations vs time profiles are used to calculate the necessary pharmacokinetic parameters to determine the oral bioavailability. the isotopic integrity of the subsequently synthesized [13C7] rofecoxib was maintained. A 1. 5 μm) using a mobile phase consisting of 45% acetonitrile in 10 mM acetate buffer (pH 4). with a mobile phase consisting of acetonitrile-water (1:1) at a flowrate of 0. metabolism. The analyte and an internal standard were extracted from the plasma matrix using solid-phase extraction in the 96-well format with an Empore extraction plate. the analytes were chromatographed on a YMC ODS AQ narrow-bore (100 x 3.The method is based on HPLC with atmospheric pressure chemical ionisation tandem mass spectrometry (APCI-MS-MS) detection in the negative ionisation mode using a heated nebulizer interface.5 μm) connected to a Water Symmetry C18 cartridge guard column with a mobile phase consisting of acetonitrile-water (35:65). Its chemical structure is shown in Fig. The resulting solutions in each of the wells of the plate were mixed. The resulting residue was reconstituted in 140 μL of mobile phase and aliquots of 100 μL injected onto the column. No. Briefly.75 mL of the diluted plasma solutions were transferred into the conditioned SPE plate.3 mL of methanol followed by 0.0 mm I.6 mm I. The analytes are chromatographed on a Waters Symmetry C18 analytical column (50 x 4. was conditioned by passing 0. and excretion of drugs in animal and human subjects. after liquid-liquid extraction of rofecoxib. The assay was validated in the concentration range of 0.. em = 375 nm) following post-column photochemical derivatization. is a non . 100 μL acetonitrile.D. stable isotope labelled analogs of drugs are utilized to determine absolute or relative bioavailability of a test compounds [67-71]. In these bioavailability studies. Each well in this plate contained 300 μL of water. 1. Numerous studies have demonstrated the . The MS-MS detection was performed on a PE Sciex API III Plus tandem mass spectrometer operated in the selective reaction monitoring mode. Chavez-Eng et al. at therapeutic doses. Acetonitrile 150 μL was added to each of the extraction wells. The use of stable isotope lebeled compounds to study the pharmacokinetics of drugs has been reviewed in two review papers [65. the unlabeled and labelled drugs are administered simultaneously by a different route of administration. [64] describes a high-performance liquid chromatographic-tandem mass spectrometric evaluation and determination of stable isotope labeled analogs of rofecoxib in human plasma samples from oral bioavailability studies. distribution. To conduct these studies. (4). [13C7] rofecoxib and internal standard from plasma. The extraction plate was then placed on top of a 96-deep well plate. The plate was sealed with a 96-well storage mat and placed on the 96-well plate compatible autosampler. 5. HPLC was carried out isocratically at ambient temperature using a Nucleosil C8 column. the method was used to study the pharmacokinetic profile of rofecoxib in 12 healthy volunteers after administration of a single oral dose (12. and 75 μL internal standard solution were wortex-mixed for 1 min. The organic layer was transferred to another tube.D.0 mL volume of 10% acetonitrile in water was passed through each of the extraction wells. 3 μm) C18 analytical column connected to a YMC ODS AQ precolumn.6 mL of water through each of the wells.5 mg).Analytical Procedure for Determination of Cyclooxygenase-2 Anti-Inflammatory & Anti-Allergy Agents in Medicinal Chemistry. NIMESULIDE Nimesulide or 4-nitro-2-phenoxymethanesulfonanilide. Werner et al. as expected. [13CD3] rofecoxib was shown to be isotopically unstable in plasma and water containing solvent and an efficient deuterium exchange prevented its use in the study. 3. Afterwards. The method has been validated over a linear range from 1 to 500 μg/L using celecoxib as internal standard.0) mL.5 ng/mL in human plasma using 0. Aliquots of 0. The SPE disk plate/deep round well plate assembly was centrifuged for 5 min at 1500 rpm to elute the analytes. The analytical technique is based on reversed-phase high-performance liquid chromatography coupled to atmospheric pressure chemical ionisation mass spectrometry. in plasma and other solvent systems.. Injection volume was 35 μL for both the human plasma and urine assays. 2009.. The tubes were capped.5 mL of plasma. 5 μm) connected to a Prism-RP analytical column (150 x 4. 8. Analyte detection was via fluorescence following post-column photochemical derivatization. 1 27 tographed on a Keystone Scientific Prism-RP guard column (20 x 4.99) when a 1/y weighed linear regression model was employed. The limit of quantification of the assay was 0.0 mL 0.5 mL/min. markedly inhibits cyclooxygenase (COX)2 with less effect on COX-1 [72].steroidal anti-inflammatory drug (NSAID) that.6 mm I. the analytes were detected by fluorescence (ex = 260 nm. The solutions were draw through the plate wells using negative pressure. Most commonly. After validation. [63]. An improved procedure for the determination of rofecoxib in human plasma involving 96-well solid-phase extraction and fluorescence detection was developed by Mattews et al. Thus. agitated in an overhead shaker for 10 min and centrifuged at 4000 g for 10 min. The results of these experiments clearly demonstrated the need for the careful evaluation of the isotopic integrity of standards in the quantitative analysis of drugs in biological fluids.1 to 100 ng/mL of plasma for both rofecoxib and [13C7] rofecoxib. On the other hand. Vol.1M sodium acetate buffer (pH 5) and 4 mL dichloromethane-hexane (50:50) were added. Analytes were found to form highly fluorescent products after exposure to UV light (254 nm). and the eluent comprised of methanol.water (50:50) and 1% acetic acid at a flow-rate of 200 μL/min.D. Briefly. Briefly. [62] developed a selective and rapid liquid chromatography-mass spectrometry method for the quantification of Rofecoxib in pharmacokinetic studies with humans.6 mm I. The plate was then positioned on the top of an ELISA plate and the extraction plate/ELISA plate assembly was centrifuged for 5 min at 1500 rpm to removal residual solvent. Eight point calibration curves over the concentration range of 5 to 500 ng/mL for human plasma and urine yielded a linear response (R2 > 0. validated quantitative methods are developed for the simultaneous determination of the labelled and an unlabeled drug. Low negative pressure was applied during plate conditioning to prevent drying of the wells. The method was partially automated using a combination of a Packard Multi-Probe liquid handling system and a TomTec Quadra 96 workstation. The labeled compounds have been employed to study absorption. 66]. an aliquot of plasma (1. Two different stable isotope labeled anaogs of rocecoxib were initially evaluated for their use as intravenous markers in the bioavailability study. the outlet coupled to the mass spectrometer’s APCI source. The solvent was evaporated under a stream of nitrogen at 40°C.D. a 96-well disk SPE plate C8-standard density.

[80]. 5 μm) analytical column and the mobile phase consisted of acetonitrilemethanol-15 mM potassium dihydrogenphosphate buffer.6 mm I.3 (30:5:65). The samples were chromatographed on a C18 reversed-phase column (Ultracarb ODS. Only 250 μL of plasma are used for sample preparation and no internal standard is necessary. 1mL. very few HPLC methods are reported in the literature. 5 μm). a simple pre-treatment with acetonitrile was used to deproteinize aqueous humor samples. Being the latest molecule. The limit of quantification was 50 ng/mL. 77]. 20 ng/mL for cerebrospinal fluid and 25 ng/mL for brain tissue.6 mm I. conditioned with 1 x 1 mL methanol and equilibrated with 1 x 1 mL water. Chemical structure of Nimesulide. plasma samples (250 μL) and brain homogenates added with the right amount of internal standard are extracted on C18 disposable cartridges by solid-phase extraction (SPE) while cerebrospinal fluid samples are analysed without any extraction. The cartridges were placed on a 16-place manifold equipped with stopcocks.0 (53:47) as mobile phase. . A rapid and sensitive method for determination of nimesulide in human plasma by high-performance liquid chromatography was developed by Khaksa et al. an aliquot of 200 μL was spiked with 20 μL phenazone solution. Nimesulide was extracted in a single step into dichloromethane. Maltese et al. 1 Giuseppe Carlucci Fig. A simultaneous determination of nimesulide and hydroxynimesulide in rat plasma. and detection at 240 nm. The assay was used for pharmacokinetic studies. Furthermore.D. vortexed and centrifuged at 12000 g.05 to 5. [94]. The method is based on protein precipitation with methanol and reversed-phase chromatography with spectrophotometric detection at 404 nm.. cerebrospinal fluid and brain by liquid chromatography using solid-phase extraction was developed by Ferrario et al. The HPLC method described did not include an internal standard and required 1. 2009. In particular. (5). The separation was performed on a Nucleosil 120-5 C18 (50 x 4.. Ptáek et al. No. The solvent was evaporated under reduced pressure at 45°C with a centrifugal evaporator.4 mL/mim.3’-bipyridine is the newest addition to the group of non-steroidal anti-inflammatory drugs (NSAIDs) known as selective cyclooxygenase-2 inhibitors [90-93].). (4). 8. The cartridges were dried under reduced pressure for 5 min. For all these reasons. some investigators have recently suggested a potential utility of NSAIDs for the treatment of different disorders such as cancer and dementia [76. Th lower limits of quantitation for either nimesulide and hydroxynimesulide are 25 ng/mL for plasma. 75]. Briefly. 150 x 4.2 to 250 ng/mL.5 μm) column with acetonitrile-sodium citrate buffer pH 3.6 mm I. The calibration curve was linear in the concentration range of 0. In this case the mobile phase was a mixture of acetonitrile-water containing 1% triethylamine (TEA) adjusted to pH 3.2 with orthophosphoric acid. the selective COX-2 inhibitors slow the progression of dementia in Alzheimer patients [78. prior to the extraction procedure. [93] developed a LC-MS-MS with atmospheric pressure chemical ionization (APCI) using stable isotope of etoricoxib as an internal standard. The prepared plasma samples were cautiously loaded onto the cartridges and washed with 1 x 1 mL water-methanol (95:5). Briefly. with UV detection at 230 nm the mobile phase consisted of phosphate buffer (5. The validation of a liquid chromatography-tandem mass spectrometry method for the determination of the cyclooxygenase-2 inhibitors etoricoxib in human plasma with phenazone as internal standard is described by Bräutigam et al.0 mL of sample. 3. The separation is performed at room temperature on a Waters Symmetry C18 (150 x 4. Other HPLC methods have been reported for the analysis of nimesulide in biological fluids [84-89].0 mm. [81] developed a rapid and simple HPLC determination of nimesulide in human plasma. The analyte was eluted with 1 x 2 mL acetonitrile-ethyl acetate (50:50). 79].6 mm I.28 Anti-Inflammatory & Anti-Allergy Agents in Medicinal Chemistry.. pH 7.D. I. Vol. The plasma samples were extracted by solid-phase extraction using polymerbased cartridges (Oasis HLB. protected by a guard column ODS ( 100 x 4. analgesic and anti-inflammatory activities of nimesulide in a wide range of clinical conditions [73]. ETORICOXIB Etoricoxib or 5-chloro-6’-methyl-3-[4-(methylsulphonyl) phenyl]-2. 5 μm) analytical column.D. the opportunity to determine nimesulide in human plasma and other biological fluids is of crucial importance to better evaluate the effects of the drug and to understanding the pharmacokinetic-pharmacodynamic relationship in the inhibition of neuroinfiammation exerted by this drug. 30 mg).. in the light of epidemiological findings. Rose et al. Its chemical structure is shown in Fig.D. [83] also developed an HPLC method with ultraviolet detection (300 nm) for the analysis of nimesulide in rabbit aqueous humor.5)-methanol-acetonitrile (50:20:30) at a flowrate of 1.0 μg/L. The calibration curve is linear up to 10000 ng/mL and the limit of quantifiction is 80 ng/mL. (5). The method was validated over the concentration range of 0. Chemical structure of Etoricoxib. The residue was re Fig. good antipyretic. Briefly. and the lower limit of detection was 30 ng/mL. 6.D. [82]. the chromatographic system uses a reversed-phase Zorbax ODS ( 250 x 4. Briefly. In the last few years it has been demonstrated that prostaglandins. play an important role in the modulation of spinal neuron activity and that NSAIDs have a direct action on central processing of peripheral information [74. which are synthesized from arachidonic acid by COX.

was carried out on a short. Then etodolac enantiomers reacted with (S)-(-)--(1-naphthyl)ethylamine) using 1-(3-dimethylaminopropyl)-3-ethyl carbodiimide (EDC) and 1-hydroxybenzotriazole (HOBT) as coupling agents. This limit of quantification was 5 ng/mL. 1 mL volume of plasma was transferred to a glass test tube. sera spiked with etodolac congeners. The isolation of a unknown metabolite of etodolac in urine and its identification a 5-hydroxy etodolac was developed by Strickmann et al. A validate liquid chromatographic ultraviolet method for the quantitation of etoricoxib in human plasma using liquid-liquid extraction was developed by Ramakrishna et al. (6). [100]. Briefly. with a relative standard deviation of less than 20%. Vol. 50 μg/mL) was spiked). The method is applicable to pharmacokinetic studies in humans.4-b] indole-1-acetic acid. In order to increase the lipophilicity of these metabolites the hydroxy groups were acetylated and the carbonilic functions were methylated. Briefly. Jin et al. [105] describes a sensitive high-performance liquid chromatographic method for the determination of etodolac in serum. 7. the enantiomers of the racemic drug etodolac have been resolved by fractional crystallization of the diastereoisomeric salts with optically active 1-phenylethylamine and analized in urine using a simple HPLC method with a conventional reversed-column. The derivatized products were separated on an Agilent Zorbax C18 (250 x 4. [104] developed a stereoselctive RP-HPLC assay to determine the enantiomers of etodolac in human plasma.4 and detected at a wavelength of 280 nm. Chromatography in isocratic mode.5 μg/mL. 5 μm) column with a mixture of methanol-0. Its chemical structure is shown in Fig. Optimisation of separation was evaluated using various concentrations of 2-propanol (doped with TFA) in hexane as the mobile phase. diethyl ether/ dichloromethane (70:30) was added. Detection was set at a wavelength of 284 nm. Briefly. 5 μm) at 30°C temperature.0 mm I. The limit of quantification was 0. 5 μm particle size and 100Å pore size). The injection volume was 15 μL. and the 50 μL of internal standard (zaleplon. Briefly. Detection was achieved by a Sciex API 3000 triple quadrupole mass spectrometer equipped with a turbo ion spray source working in positive ion mode. Next a 5 mL aliquot of extraction solvent. [101]. chiral drug enantiomers were extracted from human plasma with liquid-liquid extraction. A linear range of 5 to 2500 ng/mL was established. Chemical structure of Etodolac. The s-glucuronide of 7-hydroxy-etodolac was preferentially excreted during the period of time observed. An application of a stereospecific highperformance liquid chromatography assay to a pharmacokinetic study of etodolac enantiomers in humans was developed by Jamali et al. The specificity of control sera. 97]. following the addition of internal standard.3.D. the dried extract was reconstituted in 250 μL of water-methanol (50:50) and a 100 μL aliquot was injected into chromatographic system. [106]. Excellent linear relationships were found between the peak area ratios (etodolac/internal standard and the plasma and the urine concentrations (0.4.6 mm I. The detection wavelength of UV detector was set at 278 nm. and sera obtained from rats treated with a variety of other drugs. The mobile phase consisted of acetonitrile-water (90:10). The limit of quantification of the method was 0. [98]. Then.Analytical Procedure for Determination of Cyclooxygenase-2 Anti-Inflammatory & Anti-Allergy Agents in Medicinal Chemistry.1. The analytical method was validated over the concentration range 0. Cosyns et al. is metabolized in humans by hydroxylation and acyl glucuronidation to yield the corresponding 1-O-glucuronides. [99] by HPLC using a RP18 column. Etodolac. The assay was linear from 0. The enantionmers of the etodolac were separed without derivatization on Chiracel OD and Pirkle (R)-DNBPG colums. The diastereoisomers thus formed were extracted and chromatographed on a normal-phase column. ETODOLAC Etodolac or 1. A direct high-performance liquid chromatography separation of etodolac enantiomers using chiral stationary phases was described by Caccamese [102]. The organic layer was evaporated and the drug and internal standard were sequentially derivatized with ethyl chloroformate and (-)-phenyethylamine. The sample was then centrifuged for 3min at 800 g . The sample was vortexmixed for 5 min. (±)-2-(4benzoylphenyl)butyric acid. The etodolac diastereoisomers were separated with a resolution factor of 6.2 to 200 ng/mL.6 mm I.2 280. (6).. narrow bore RP C18 column (30 x 2. administration of 400 mg of etodolac whereas the elimination of the R-glucuronide predominates at longer periods of time.2 μg/mL. 8. The limit of detection was 0. The S-etodolac-glucuronide is mainly excreted during the first 6-hours after Fig. Enantiomeric purity can be determined in less than 10 min.2 and m/z 189.9-tetrahydropyano[3. [95].2). [103] also developed a stereoselective disposition of etodolac enantiomers in synovial fluids. No.D. For the identification elecrospray ionisation mass . 1 29 constituted in 200 μL mobile phase.2 – 20 mg/L). The organic layer was quantitatively transferred to a 6 mL glass tube and evaporated to dryness at 40°C under a stream of nitrogen. Details of its pharmacology have been reviewed by Balfour et al.2 ng/mL. Brocks et al. The respective mass transitions used for quantification of etoricoxib and phenazone were m/z 359. The acylglucuronides of etodolac and one of its hydroxylated metabolites were determined by Berendes et al.5 to 50 μg/mL for each enantiomer.0104.01 M phosphate buffer (pH 4.D.8-diethyl-1. the constituents were extracted from the specimen into a mixture of isooctane-isopropanol (95:5).5) (70:30) as mobile phase.. An evaluation of the stereoselective metabolism of the chiral drug etodolac by high-perormance liquid chromatography was proposed by Becker-Scharfenkamp et al. 2009. with a mobile phase consisting of hexane-ethyl acetateisopropanol (85:15:0. is a non-steroidal anti-inflammatory drug which has been shown to be effective in the treatment of rheumatoid and osteoarthritis and a selective COX-2 inhibitors in a wide range of clinical relevant assays in direct comparisons with other NSAIDs [96. An unknown metabolite of etodolac could be characterized by chemical and enzymatic hydrolysis and by MS and NMR. The analyte and internal standard were separated using a isocratic mobile phase of water-acetonitrile (58:42) on reversed-phase Waters Symmetry C18 column (250 x 4. Briefly.

that confers weakly acidic properties (pKa 4. [109]. accounting for approximately 43% of the AUC. 3 μ m) with 65% acetonitrile and 35% water containing 10 mM ammonium formate (adjusted to pH 3.0 μg/mL. transferred to Eppendorf tubes and used for HPLC analysis. (7).05% trichloracetic acid (35:65). The procedure produced a linear curve over the concentration range 10 .0 mL of mobile phase and a 100 μL injection was analysed. The column was maintained at 30°C. previously frozen plasma samples were vortexed and 1.2% of dose). Fig.8%). Serial blood and complete urine and feces were collected for 168 h post dose. The limit of quantification was 10 ng/mL. The limit of quantification was determined to be 10. The tubes were capped.1 mL internal standard stock solution (10 μg/mL niflumic acid in methanol) to 1. and dose recovery was almost complete (96. The mobile phase consisted of acetonitrile and 0.1 . samples were prepared by adding 500 μL 1M hydrochloric acid and 0. Samples were centrifuged at 1000 x g for 15 mim. There are no other analytical method available for this drug in the literature. 4’-hydroxy. I. 5 μ m) column with an Atlantis D C18 guard column. The absorption. 1 Giuseppe Carlucci spectrometry (ESI-MS) as well as 1NMR and 13C-NMR spectroscopy as been used. Linearity of the calibration cures for lumiracoxib was achieved for concentrations between 10 and 10000 ng/mL. Briefly. Chemical structure of Deracoxib. 200 μg/mL) and 6 mL of isopropilic alcohol -chloroform (80:20). is chemically distinct from the other coxib in that it lacks a sulfur-containing moiety and possesses a carboxylic group Fig. The reconstituted specimens were vortex.D.1 μg/mL with a relative standard deviation of less than 15%. The determination of deracoxib in feline plasma samples using high-performance liquid chromatography was developed and validated by Cox et al. rheumatoid arthritis. VALDECOXIB Vadecoxib or 4-(5-methyl-3-phenyl-4-isoxazolyl)benzenesulfonamide is a anti-inflammatory drug that is highly . mercapturic acid or glutathione conjugates. Its chemical structure is shown in Fig. [111]. Supernatant were transferred to a clean tube and the organic phase evaporated at 30°C with nitrogen. (8). The compounds were separated with an Atlantis D C18 (150 x 4. The major metabolic patways of lumiracoxib is oxidation of the 5methyl group and hydroxylation of the dihaloaromatic ring. Chromatographic separation was performed isocratically using a Capcellpak MGII C18 column ( 50 x 2. The terminal half-life of lumiracoxib in plasma was 6. A validated high-performance liquid chromatographic assay for determination of lumiracoxib in human plasma was developed by Cheremia et al. Lumiracoxib was rapidly adsorbed.7). which has been developed for the treatment of osteoarthritis.0 mm I. (7). with little unchanged drug in urine (3.6 mm. Major plasma metabolites were the 5-carboxy.5 with formic acid). [110]. 8. 10. The organic layer was transferred into a glass tube and evaporated under a stream of nitrogen at room temperature. Lumiracoxib was extensively metabolised before excretion. The analyte was separated using a reversed-phase Nucleosil (120-3 C8) column protected by a C8 precolumn.. Chemical structure of Lumiracoxib. and acute pain. The limit of quantification was 0.D. The calibration curve exhibited good linearity in the concentration range of 0.0 mL placed in a centrifuge tubes followed by 100 μL of tolbutamide (internal standard. unchanged drug was the major circulating component in plasma. The mobile phase used was an isocratic mixture of 10 mM potassium phosphate buffer (pH 4.5 h. 9. Briefly. disposition and mass balance of 14C-lumiracoxib were investigated in healthy male subjects after a single 400 mg oral dose by Mangold et al. although there was no evidence of cysteine. using indomethacin as an internal standard. DERACOXIB Deracoxib or 4-[3-(difluoromethyl)-5-(3-fluoro-4-methoxyphenyl)-1H-pyrazol-1-yl] benzene sulfonamide is a selective cyclooxygenase –2 inhibitor [108]. Vol. Glucuronic acid conjugates of lumiracoxib metabolites (and to a minor extend lumiracoxib itself). No. Were identified. 8. The correlation coefficients were greater than 0.7/%) routes. suggesting a modest first-pass effect. Lee et al.5 h postdose. The residue was dissolved in 150 μL of methanol-water (50:50).99 or better.30 Anti-Inflammatory & Anti-Allergy Agents in Medicinal Chemistry.0 mL plasma followed by the addition of 4 mL hexane-diethyl ether (70:30).1%) and fecal (42. metabolism. agitated and centrifuged at 4000 rpm for 10 min. The calibration curve had to have a correlation coefficient of 0. The injected volume of the samples was 100 μL. 2009. Samples were reconstituted in 1.25. achieving mean plasma concentrations > 1μg/mL within 1 h of postdosing.1500 ng/mL. and 4’hydroxy-5-carboxy derivatives. (8). Its chemical structure is shown in Fig.997. Excretion involved both renal (54.3% of dose) or feces (2. LUMIRACOXIB Lumiracoxib or 2-[2-fluoro-6-chlorophenyl)amino]-5methyl-benzenacetic acid is a distinct cyclooxygenase-2 selective inhibitor.5) and acetonitrile (52:48).0 ng/mL of lumiracoxib in plasma. [107] also developed a simple and specific method using a one-step liquid-liquid extraction with butyl acetate followed by HPLC coupled with positive ion electrospray ionisation tandem mass spectrometry (ESI-MS/MS) detection for the determination of etodolac in human plasma. Unchanged drug plasma accounted for 81 to 91 % of radioactivity up to 2. Uv detection was set at 270 nm. Ultraviolet detection is carried out at 252 nm.

A LCMS-MS assay to measure simultaneous valdecoxib. The urine samples were vortexed and placed in the loading modules of a RapidTrace automatic SPE System. An aliquot of plasma sample (1. Werner et al. [120]. 1 31 selective for inhibition of the inducible form of ciclooxygenase (COX-2) [112]. a 1 mL volume of plasma was transferred to a 15 mL glass tube test.D. Chemical structures of valdecoxib. Detection was set at a wavelength of 210 nm. The analytes were detected by mass spectrometry using negative ion electrospray ionisation with MRM mode.4 mL of human plasma using a Zymark RapidTrace automation system. and transferred into autosampler vials. pH 6..2 M sodium acetate buffer (pH 5. 8. osteoarthritis and pain [113-115]. Measurements of human plasma valdecoxib concentrations were determined by HPLC with ultraviolet detection using rofecoxib as the internal standard.D. The lower limit of quantitation for human urine was 1 ng/mL for both valdecoxib. hydroxylated valdecoxib. (9).0. The separation of compounds was made on a YMC ODS-AQ column (250 x 4. [122] utilized a LC-MS for the measurement of both etoricoxib and valdecoxib in human plasma. Briefly. carboxylic acid valdecoxib and the internal standard were carried out on a narrow-bore reversed-phase Prism RP HPLC column (50 x 2.water (50:50). A versatile and sensitive HPLC assay was developed by Ramakrishna et al. Valdecoxib is metabolized primarily by cytochrome P450 2C9 and 3A4 to the pharmacological active hydroxylated metabolite and the carboxylic acid metabolite in humans [116-118]. (9). and carboxylic acid valdecoxib are shown in Fig. and the 50 μL of internal standard was spiked. and carboxylic acid valdecoxib metabolite in human urine by Zhang et al.6 mm. Then. and carboxylic acid valdecoxib. This drug has been approved for the treatment of rheumatoid arthritis. A simple. 1. No. hydroxylated valdecoxib. and 100 μL internal standard solution were vortexed for 1 mim. The solvent was removed under a stream of nitrogen on a TurboVap at room temperature to obtain residues that were reconstituted in 100 μL of the mobile phase of acetonitrile-water (50:50. Valdecoxib and its metabolites and an internal standard were extract on a C18 solid-phase extraction cartridges using a Zymark RapiTrace automation system.0 mL). After extraction. C18 SPE cartridges (100 mg. and 2-200 ng/mL for carboxylic acid valdecoxib metabolite. and hydroxylated valdecoxib. The column effluent was directly introduced into the mass spectrometer using electrospray ionization in negative mode.0) containing 10 mM 4methylmorpholine.methanol (80:20) and 100 μL aliquot was injected into chromatographic system. Then 20 μL of the reconstituted samples was injected onto the LC-MS-MS system for analyses. The organic layer was quantitatively transferred to a 6 mL glass tube and evaporated to dryness. 5 μm). The mobile phase was a mixture of water-methanol (47:53). diethyl ether/dichloromethane (70:30) was added. hydroxylated valdecoxib. the dried extract was reconstituted in 200 μL of water . 1:1) into 0.5 mL of 0. 2009. Afterwards. The sample was vortex-mixed for 5 min.. Enzymatic hydrolysis of glucuronide coniugate of hydroxylated valdecoxib was conducted by adding approximately 100 units of glucuronidase in 0. Chemical structures of Valdecoxib. Aliquots of 500 μL were placed in disposable glass tubes and 500 μL of internal standard solution (100 ng/mL) in 100 mM ammonium acetate buffer (pH 6. Next a 5 mL aliquot of extraction solvent. frozen urine samples were thawed in a water bath in a microwave oven for 60 s on defrost cycle. The LOQ for carboxylic acid valdecoxib metabolite was raised to 2 ng/mL at the concentration of 1 ng/mL did not meet the validation criteria (<±20%). Mass analysis was performed in the positive ion mode.5 mL of human urine. 1 mL 0.Analytical Procedure for Determination of Cyclooxygenase-2 Anti-Inflammatory & Anti-Allergy Agents in Medicinal Chemistry. Vol. The analytical technique is based on reversed-phase high-performance liquid chromatography coupled to atmospheric pressure chemical ionisation mass spectrometry. and hydroxylated valdecoxib.0 mmI. The procedure consisted of an automated C18 solid-phase extraction (SPE) of valdecoxib. The separation of valdecoxib. [119]. 100 μL acetonitrile . was then centrifuged for 5 min at 800 g.8) was added.0) containing 10 mM 4methylmorpholine. The urine samples were loaded onto the cartridges that were washed with 4 mL of water and eluted with 500 μL of acetonitrile. hydroxylated valdecoxib and an internal standard from 0. sensitive and specific automated SPE-LC-MS-MS assay was developed and validated for determination of valdecoxib. pH 6. [121]. Briefly. the samples were injected onto a reversedphase Zorbaz XDB C8 HPLC column for separation. 5 μ m) with a isocratic mobile phase consisting of acetonitrile-water (50:50. Fig. hydroxylated valdecoxib.0 mL) were conditioned with 2 mL of methanol and 2 mL of water. I. Standard curves were linear over the concentration range of 1 – 200 ng/mL for valdecoxib.1 M sodium acetate buffer . in human plasma samples by Zhang et al. hydroxylated valdecoxib.

1 μg/mL. respectively. The standard curve for etoricoxib. 1 Giuseppe Carlucci (pH 5) and 4 mL dichloromethane . 5 μ m) column with methanol and 0. tographic estimation method of meloxicam in biological samples.. The limit of the method is 50 ng/mL.D.1-dioxide is a representative drug belonging to the oxicam derivatives which shown preferential inhibition of cyclooxygenase-2 and prostaglandin synthesis. valdecoxib. 8. Detection was by UV detector at 355 nm.1 . The chromatographic separation was achieved by gradient elution consisting of 0. (10). osteoarthritis. The mobile phase consisted of an aqueous solution of diammonium hydrogenorthophosphate (50 mM). MELOXICAM Meloxicam or 4-hydroxy-2-methyl-N-(5-methyl-2thiazolyl)-2H-1.D. The residue was reconstituted in the mobile phase and injected onto a Kromasil KR 100-C18 column (250 x 4. was 10 and 5 μg/mL. The mobile phase constituents are methanol and aqueous 20 mM Na2HPO4 buffer solution brought to pH 6 with H3PO4 . It has definite activity in treating rheumatoid arthritis. Velpandian et al. The response was linear over a range of 50 – 1500 ng/mL in human plasma. The organic layer was transferred into another tube. The eluent was monitored by a UV- .05% glacial acetic acid (68:32) as mobile phase. reconstituted in the mobile phase and then a volume of 10 μL of the prepared sample was injected in the column. Briefly. 11. The work of Baeyens et al. Lichrocart (125 x 4.6 mm I.3) 170 mM-acetonitrile (62:38). [124]. The chromatographic analysis was performed on Intersil C18 ODS ( 250 x 4. After washing with the buffer the ADS column was backflushed with the mobile phase 0. The eluate was monitored using an ultraviolet detector set at 235 nm.2benzothiazine-3-carboxamide 1. the sample preparation procedure was based on protein precipitation with a misture of methanol-acetonitrile. The supernatant was directly injected to the HPLC. methanol and acetonitrile in the ratio (50:40:10).999) in the concentration range 0. A number of analytical methods for the determination of meloxicam in biological samples were reported. The stepwise gradient elution profile of the chromatographic method allows injection of a high volume of the sample (500 μL) with the focusing of both analytes in a Chromolith Performance RP-18e column.32 Anti-Inflammatory & Anti-Allergy Agents in Medicinal Chemistry. In this work Baeyens. The UV detection was achieved at 364 nm. rofecoxib. Vol. and other joint diseases [125-128]. Meloxicam was quantified in human plasma after a single 15 mg oral dose of the drug was given to 26 healthy volunteers by Wisner et al.05 M phosphate buffer-30% acetonitrile-25 mM tbutylamine at a pH of 7. A diode array detector was used. Briefly. An online elimination of the protein matrix is achieved with a quantitative recovery together with an on-column enrichment. Piroxicam was used as internal standard.water (50:50) and 1% acetic acid. Nageswara Rao et al.05 M phosphate buffer. 5 μm). An Applied Biosystems Sciex API 2000 triple quadrupole mass spectrometer in multiple reaction monitoring (MRM) mode using TurboIonSpray (TIS) in the positive ione mode. [129] describes a new high-performance liquid chroma- Fig. valdecoxib and celecoxib. pH 6. spiked plasma samples were introduced on the ADS precolumn using a 0. [135] was therefore directed towards the determination of meloxicam in human plama by application of an alkyl-diol silica precolumn in a column switching system. Protein precipitation with acetonitrile was followed by C18 reversed-phase liquid chromatography and tandem mass spectrometry. The samples were chromatographed on a C18 reversed-phase column. They include the methods using high-performance liquid chromatography with UV detector (HPLC-UV) [129. Acidified plasma samples were extracted with chloroform.0 mm. 130]. Its chemical structure is shown in Fig. trifluoracetic acid and sodium sulfate solution. The limit of quantitation was around 30 ng/mL. HPLC with diode array detector [1006] and LC-tandem mass spectrometry (LC-MS-MS) [132-134].96 ng/mL. have been developed as special packing used in the liquid chromatographic integrated sample processing of biofluids. [130] also developed an HPLC method with UV detection for the analysis of meloxicam in human plasma. No. [132]. The organic layer was separated and evaporated under a gentle stream of nitrogen at 40°C. Aliquots of 20 μL (etoricoxib) or 100 μL (valdecoxib) injected onto the column. (10). nimesulide and nabumetone) using 4-chloro-2-nitroaniline as internal standard. describes a hand-operated online switching high-performance liquid chromatographic system for the determination of meloxicam. The assay utilized a UV absorbance detector set at 356 nm. thus transferring the analyte to the analytical column.hexane (50:50) were added. [123] describes a reversed-phase HPLC method for the simultaneous separation and quantitation of COX-2 inhibitors (clecoxib. in pharmaceuticals and its application to biological fluids. Briefly. using piroxicam as the internal standard. 5 μ m) was used for analytical separation.50 μg/mL. The group of LiChrospher alkyl-diol silica (ADS) phases that make-up part of the unique family of restricted-access materials.029 μg/mL and the limit of quantification was found to be 0.05 M formic acid (pH 3)-acetonitrile-methanol-water.0. The mean recovery for meloxicam was 92% with a lower limit of quantification of 8. the method employed a simple liquidliquid extraction of the analytes and internal standard from human plasma (500 μL) into acetonitrile. 2009. The limit of detection was found to be 0. A non-extracting procedure for the determination of meloxicam in plasma by HPLC-diode array detection was developed by Medvedovici et al. was used. The solvent was evaporated and the residue reconstituted in 500 μL of mobile phase. I. Another work for the simultaneous determination several analytes in plasma by HPLC with UV detection was developed by Pavan Kumar et al. The limit of quantitation. In this case the mobile phase was a mixture of sodium acetate buffer (pH 3.6 mm I. HPLC was carried out using a Nucleosil C8 column and an eluent comprising methanol . was linear (r2>0. Piroxicam was used as the internal standard. Dasanti et al.0. [131]. Chemical structure of Meloxicam.D. evaporated to dryness. The analytical procedure in the plasma pre-treatment consists of extraction using perchloric acid and acetonitrile mixture.. The tubes were agitated and centrifuged at 4000 g for 10 min.

after chemical hydrolysis. while no methods were reported for tirmacoxib and cimicoxib. Distribution of COX-1 and COX-2 in normal and inflamed tissues. [138]. rofecoxib. 167-170. Final extract was injected in a liquid chromatograph with an ion-trap mass spectrometer with electrospray interface. S. Seibert. A highly sensitive liquid chromatographytandem mass spectrometry method was developed to determine meloxicam at low concentration in human plasma by Yuan et al. and valdecoxib. Rev. Igualada et al.. Zhang. The intrinsic fluorescence of some COX-2 inhibitors makes them good candidates for fluorescence detection.Analytical Procedure for Determination of Cyclooxygenase-2 Anti-Inflammatory & Anti-Allergy Agents in Medicinal Chemistry. The curve was linear within the concentration range. Botting. equipped with electrospray ion source. Briefly.10 ng/mL. 1998. meloxicam and other drugs (piroxicam and tenoxicam) and isoxicam (internal standard) were extracted from human plasma with ethyl acetate at acidic pH and analysed on a Sunfire column with the mobile phase methanol-ammonium formiate (15 mM. The literature compilation has revealed that a variety of methods are available for the first generation of COX-2 inhibitors viz. The lower limit of quantification was 10 ng/mL.10 to 50. The standard curve was linear (r = 1. K. [137]. M.. The analytes were detected using a mass spectrometer.0) (60:40). Bakhle.2). and also gave good precision and a wide range of linear detector response. an organic extraction from homogenised tissue was performed. ABBREVIATION LIST NSAIDs COX 6-MNA HPLC 6-HNA RP APCI DAD TEA MRM LOQ ADS MRM SRM ESI = Non steroidal antinflammatory drugs = Cyclooxygenase = 6-Methoxy-2-naphtylacetic acid = High-performance liquid chromatography = 6-Hydroxy-2-naphtylacetic acid = Reversed . the role of HPLC in the therapeutic monitoring of COX-2 inhibitors may be expected to increase during the coming years.. I have discussed the present state-of-the art of the analytical methods for the determination of not only preferential COX-2 inhibitors viz. 400A. 38. Med.0 ng/mL. Adv. 8.. 2009. 5 μm) with a mobile phase of acetonitrile-20 mM potassium hydrogen phosphate (40:60) (pH 3. I. Y. The method had a lower limit of quantification of 0. K. The instrument was set in multiple reaction monitoring (MRM) mode. Vol. P.. After extraction with ethyl ether. analysis time and sensitivity. This is because of the fact that selective COX-2 inhibitors were introduced in late nineties. [139] described a rapid method for the determination of NSAIDs drugs in animal tissue by liquid chromatography-mass spectrometry with ion-trap detector. The decision limits (CC) and detection capabilities (CC ) were determined and their values were at concentrations near to the maximum residue limits (MRL) for each substance. Y. No. For quantitative purposes flunix-D3 was used as internal standard. Concentrations of meloxicam were analyzed by HPLC with ultraviolet detection by Bae et al. Annu.phase = Atmospheric pressure chemical ionization = Diode array detector = Triethylamine = Multiresolution mode = Limit of quantitation = Alkyl diol silica = Multiple reaction monitoring = Selected reaction monitoring = Electrospray ionization Zorbax SB = Zorbax stable bond REFERENCES [1] [2] Vane. The lower limit of quantification for meloxicam was 0.. For drugs like celecoxib. LC coupled with mass detector (LC-ESI-MS) was used to detect most of the metabolites of COX-2 inhibitors in human plasma and urine. R. Ji et al. . The mobile phase used consisted of acetonitrile-water-formic acid (80:20:0. nimesulide and meloxicam. The method was validated using fortified blank muscle from different animal species according to the European decision criteria. Toxicol. the chromatographic separation of meloxicam was carried out using a reversed-phase Sunfire C18 column (150 x 4..6 mm. In this review.000) over the concentration range of 50 to 200 ng/mL. Biol. Our analysis of the published data revealed that the HPLC was extensively used for estimation of COX-2 inhibitors in biological fluids. Isakson. showing that it is the technique of choice for analysis of COX-2 inhibitors. R. it is obligatory if only small sample volumes are available and for clinical specimens where interferences are likely to be present.. Pharmacol. but also selective agents such as celecoxib. nimesulide and meloxicam. Briefly. [136] described a procedure for the determination of meloxicam and other drugs in human plasma by LC-MS-MS.. Leahy. pH 3. 1 33 detector set at 364 nm..D. CONCLUSIONS A overview of the current state-of art analytical methods for determination of cyclooxygenase COX-2 inhibitors has been presented. detection was performed on a triple quadrupole tandem mass spectrometer by selected reaction monitoring (SRM) mode via electrospray ionisation (ESI) source. Hauser. repeatability. Many methods were published during the period of 1993-2008. Due to the inherent power of HPLC and the impact of better methods and equipment as detailed above. The calibration curve was demonstrated to be linear over the concentration range of 0. Fluorescence detection was highly sensitive and specific. valdecoxib. lumiracoxib and etoricoxib only a limited number of methods were reported. S. In conclusion.50 ng/mL using a 100 μL plasma sample. J. 97-120. 1997.5) and UV detection at a wavelength of 355 nm. The validate method was successfully applied on the determination of meloxicam in human plasma collected up to 180 h after a transdermal administration of 30 mg meloxicam for evaluation of the pharmacokinetics. 10-2400 ng/mL. several methods are based on HPLC.. J. Exp.. Masferrer. meloxicam and the internal standard piroxicam were chromatographed on a Zorbax SB C18 column. Cyclooxygenases 1 and 2. HPLC has been proven in many laboratories to be an effective tool for monitoring of COX-2 inhibitors. After a simple sample preparation procedure by one-step protein precipitation with methanol. Most of the workers used the reversed-phase mode with UV absorbance detection because this provided the best available reliability.

M. Hundt. Roberts. Mishida. Drugs. No.C. Anal..H.. a major metabolite of nabumetone. M. T. Hirai. 2341-48. J. Effect of nabumetone (BRL 14777). Quantitative determination of the beta-blocker labetalol in pharmaceuticals and human urine by high-performance liquid chromatography with amperometric detection. Vol.K. Metabolism of nabumetone (BRL 14777) by various species including man. Undre. F. 40.. 13.C. the active metabolite of nabumetone in horses.. C. 1997. N.. 403-16. 1996. 1982.. 299-05. J. Svoboda. Kopeck . H. J. Alonso. Henschke.J.. J. 375-88. Anal. K.. Swart. B. 251-57. nabumetone. Chabenat. 1993. Clin. C... Pattrick. C. 1990. Kill. Blagbrough. Bacteriocinas: una estrategia de competencia microbiana propuesta como alternativa de antibióticos dirigidos para el futuro humano. 1978. a novel structural class of antiinflammatory compounds. J.. E.C.C. Kvtina. N. Extractionless highperformance liquid chromatographic method for the determination of diclofenac in human plasma and urine. 2485-92.. A simple isocratic high-performance liquid chromatographic (HPLC) determination of naproxen in human plasma using a microbore column technique J. 97-03. S. A.. Eur. Llloyd. Pharmacokinet. Pharm. 30. Karidas. K. J.. H. 32. Langtry. de Jager. 35. 65S-70S.J. P.. S. P. R... C. Res. J. High-performance liquid chromatographic determination of a new anti-inflammatory agent. Pour.. 1995.A. T.. Lafont. 336. Drugs. Clinical pharmacokinetics of nabumetone. A.. Chromatogr..E.D. Med. S. 57.M. 33. Jimenez. 45.K. 2009. R. Anal.34 Anti-Inflammatory & Anti-Allergy Agents in Medicinal Chemistry. 2003.A. H.. B. 4-(6-Methoxy-2-naphthyl)butan-2one and related analogues..L. L.. Malamataris. Barli ska. Maguregui. 131-56. 1260-64. Matsumoto. Thawley. Jun. 18.. Friedel. J. M. A. H.. P. M.E. J. 692. Liq. Miehlke. J. Lake. M.O.. Hermansson. Chem. Smith. Chromatogr. 505-24. 1998.. in human plasma by high-performance liquid chromatography.. M. Chromatogr. A. Haddock. Friedel.M. D. Hundt. T.. 57-61. 14. Lu. A. on human platelet reactivity ex vivo: comparison with naproxen. Pharmacol.C.W..N. Rel.M.. Clin. B..W. Nobilis. Kobyli ska. J. Disposition and excretion of 6-methoxy-2-naphthylacetic acid. J. A. a new anti-inflammatory drug. Fluorometric high performance liquid chromatography for quantitation of naproxen in serum. 119-129.C. 3105-15.. 1990. Such. Jubb. J. 107.. Muth. 36.R. Day.R. 578.. J. J.. 1988.. B. B.... Vet. J. R. Chromatogr. Al-Momani. L. Chromatogr. Pharmacol..C.. K. 1 [3] Brater. G. Chromatogr.. Lett. R... J. Goto. 517-21. Grahm.J.A. Med. Kill. 26. Chamberlain. Ferranti. R... Hol. Ceniceros. J... Kobyli ska M. nabumetone and its major metabolite 6-methoxy-2-naphthylacetic acid in pharmaceuticals and human urine by high-performance liquid chromatography.M.. Penetration of the active metabolite of nabumetone into synovial fluid and adherent tissue of patients undergoing knee joint surgery. P. J.. M. Kendall. The dawn of selective cyclo-oxygenase-2 inhibition? Clin. V.S. Kune . Mikami. 1989.. Chromatog. 2000..R. Effects of nonsteroidal anti-inflammatory drugs on renal function: focus on cyclooxygenase-2-selective inhibition. 234-38. Xenbiotica. 2000. M. 32328.. Avgerinos. Liq. 23. Chellingsworth.. Matsumoto. Schneider. Shaw.M. 718.R. Chellingsworth. B.F. Soma. Extractionless determination of 6-methoxy-2naphthylacetic acid. Anal. N. Simultaneous determination of primidone and its three major metabolites in rat Giuseppe Carlucci [24] [4] [5] [6] [25] [26] [7] [8] [27] [9] [28] [10] [11] [12] [29] [30] [13] [31] [14] [32] [15] [33] [16] [17] [34] [35] [18] [36] [19] [37] [20] [38] [21] [39] [22] [40] [23] urine by high-performance liquid chromatography using solidphase extraction. H. Jeffrey. 57679.. Todd. A.... Chromatogr. Lett. High-performance liquid chromatographic determination of naproxen.. T.. Technol. 21. Hughes.S. Pharm.D. 740. 8.I. M. Buckley. Chromatogr. I. Nabumetone.. M. Kishi. 1984. A pharmacokinetic study of the active metabolite of nabumetone in young healthy subjects and older arthritis patients.. Z. Thawley. J. 1997. Application to single dose pharmacokinetic studies.. P. Ray. P. Kendall. A. 1999. 299-02 Nunn.. 327-37. A. Pharm.E. 199-04. M.. J. Rose. O..M. J. Sörgel. 705.A.D. Giersch.. 36. Wanwimolruk. Menager. 1997. ibuprofen and diclofenac in plasma and synovial fluid in man. Undre. Ohno. Simultaneous analysis of several non-steroidal anti-inflammatory drugs in human urine by highperformance liquid chromatography with normal solid-phase extraction. M. Am. Pharmacol. Münzel. Biomed.. Analysis of nabumetone in human plasma by HPLC. D. 917-25. Determination of drugs by direct injection of plasma intoa biocompatible extraction olumn based on a protein-entrapped hydrophobic phase. A preliminary review of its pharmacodynamic and pharmacokinetic properties. Thawley... 1984. Nabumetone. R.O. J. Drugs. R. S. A reappraisal of its pharmacology and therapeutic use in rheumatic diseases. Gaster. Freeman. F. 247-51... M. Knight. 1992. M.A. 1611-25. 660.. I. I. M..J. H. Daykin. Miller D..J.. Simultaneous analysis of naproxen. 1993.W. A. Am. Doherty. Biomed. D. A pharmacokinetic study of the active metabolite of nabumetone in young healthy subjects and older arthritis patients. 1998..A.F.. 34. Uboh. H. 1989. Jubb.. Davies. 1994. S. and therapeutic efficacy in rheumatic diseases. D. Goudie. and its major metabolite in plasma using fluorimetric detection.

L.. 2003. 32. Pharm.apek. Biomed. Comparative biotransformation and disposition studies of nabumetone in humans and minipigs using high-performance liquid chromatography with ultraviolet. M. J. Hol. M. fluorescence and mass spectrometric detection. 641-56. Nobilis.. Anal. Koráová..

38. Day. E. J. Prostaglandin G/H synthase-2 is a major contributor of brain prostaglandins in the newborn. Murillo Pulgarín. 24615-20. and its major metabolite in plasma using fluorimetric detection. 377-85. B.. Gregory.W. . Woolf.. B.. Kopeck . Bible.. Dispos. D.. A. 753.. McGarity. 1995.Struebe. Oday. 308-14. J. G.G. nabumetone. D..M.P. 1031. Determination of celecoxib in human plasma by normal-phase high-performance liquid chromatography with column switching and ultraviolet absorbance detection.. G. A.. 47. Clin. 336.. S. K.E.J.performance liquid chomatographic determination of a new anti-inflammatory agent.. Invest. R. S. Tegeder. A. Cyclooxygenase-1 and -2 expression in rheumatoid synovial tissues. Clin. Sánchez-Ferrer Robles. K. L. Pharmacokinet.. 2001. Effects of interleukin-1 beta. Davies. Alañón Molina. Z. 2000.. 761. Drug Metab. phorbol ester. Varma. Heinkele.Y. J. L. Res. Pierges. 165-69. 22542. 203-12..J. Chromatogr. Up-regulation of cyclooxygenase-2 mRNA in the rat spinal cord following peripheral inflammation.. 270. J. Peri. A. 234-238. Simple and rapid determination of the active metabolite of nabumetone in biological fluids by heavy atom-induced room temperature phosphorescence. 229-36. FEBS Lett. S. Goppelt-Struebe. Invest.. B. F. Crofford. B. E. Inflamm. M. R.. I. Epps. Hardy. Isakson..L. Remmers. G... S.. 2000. Brune. Beiche.R. McLachlan. 1996. Clinical pharmacokinetics and pharmacodynamics of celecoxib: a selective cyclo-oxygenase-2 inhibitor.A. M. Rose.. Novotny. Liquid chromatographic-mass spectrometric determination of celecoxib in plasma using single-ion monitoring and its use in clinical pharmacokinetics. 738. Geisslinger. J.apek. Matuszewski.. K.... 1984. A. Geisslinger. J. J.C.. M. P. Expression of cyclooxygenase isoforms in the rat spinal cord and their regulation during adjuvant-induced arthritis. Determination of celecoxib in human plasma and rat microdialysis samples by liquid chromatography tandem mass spectrometry. 2000.. Chromatogr. H. Goppelt. Paulson.. N. Williams. Chromatogr... G.E. Anal.. E. 28. K.O. L. Chromatogr..H. 1998. Ray.. A. 2004.J.D..D.. 1996. 2001. Kune .. Hla. Biol. Hribar.. H. Kvtina.. M.F. P. 390. 554. R. 2672-79. 1095-01.. Hauser.R. Clin. Hamza..M. G. J... Ristimaki. S. Geisslinger. 97. 482-87. M. M..K. R. 93. A. Koráová. Vetter. Chromatogr..K. J. Abdel-Hamid.. Chemtob. Identification and determination of phase II nabumetone metabolites by high-performance liquid chromatography with photodiode array and mass spectrometric detection.. High. J. Brune. .L.. J. Chem.. Anderson. Liu. and corticosteroids. J. Scheuerer..J. Bremer. Hajdu. K.. J. H. Acta. Bräutigam. J. Karim. 401-08. Sano. I. Svoboda. Li. Beiche. N.. Metabolism and excretion of [(14)C]celecoxib in healthy male volunteers. 2005. T. 1994. Selective inhibition of cyclooxygenase (COX)-2 reverses inflammation and expression of COX-2 and interleukin 6 in rat adjuvant arthritis. Chim. Wilder. G. L..D. 37-42. F..

.L. 21. Ziaee.. Mattews. 949. K. A. B.. 730. 89-94. A. Sci. B. Dobrinska. S. 2004.. Simultaneous measurement of ethanol and ethyl-d5 alcohol by stable isotope gas chromatography-mass spectrometry.. Matuszewski. 16. Gharbi.B. K. 207-12. 760. 8. J. 2001. N. Shafiee. J. A. Simple and rapid high-performance liquid chromatographic method for determination of celecoxib in plasma using UV detection: application in pharmacokinetic studies.. The role of anti-inflammatory drugs in the prevention and treatment of Alzheimer's disease. Werner. Ray. 61. Woolf.. Anal. Anal. Am.. Rofecoxib. Woolf. J. B. Determination of celecoxib in human plasma and breast milk by high-performance liquid chromatographic assay.J. Simplified solid-phase extraction procedure and liquid chromatographic determination of celecoxib in rat serum.K. Delinsky. S.H.D. T. Rofecoxib: a review of its use in the management of osteoarthritis.. Werner. Morris. Dumanual. Selective and rapid liquid chromatography-mass spectrometry method for the quantification of rofecoxib in pharmacokinetic studies with humans. 86.J. 351-54. H. A. 56-60. Siluveru. R.. 2004. A.J. 1995.. Matheson. 100-04. Determination of rofecoxib (MK-0966).D. J.. 2001.. Mitchell.. p. Chromatogr. V. 835. B... 60.. 667. Figgitt. B. L. I. Khoddam. P.Analytical Procedure for Determination of Cyclooxygenase-2 [41] Anti-Inflammatory & Anti-Allergy Agents in Medicinal Chemistry... Biomed. Med. D. Scott. Brenner.. T.. 1033-37.D. Matuszewski. Matuszewski. 52. Constanzer. A. Kohno. 682. I..L.. Nonsteroid drug selectivities for cyclo-oxygenase1 rather than cyclo-oxygenase-2 are associated with human gastrointestinal toxicity: a full in vitro analysis. 47.... Mattews. Lipsky. Cambridge... T. D. J. 31-39. Olah. J. Moore.. No. Chromatogr.R. J.V. 42. Chromatogr. 1996. D. acute pain and rheumatoid arthritis. Chem. J..K. acute pain and rheumatoid arthritis. D.. 61. Foroutan. A. 2004. J. The use of stable isotope labeling and liquid chromatography/tandem mass spectrometry techniques to study the pharmacokinetics and bioavailability of the antimigraine drug. 401-11. 2006. Chromatogr. 1994. T.V.. 167-74.K. in human plasma by high-performance liquid chromatography with post-column photo derivatisation and fluorescence detection. Drugs. 1750-56. MK0462 (rizatriptan) in dogs. 748. Lawrence. Navas. 665-70. The use of stable isotope labeling and liquid chromatographytandem mass spectrometry techniques to simultaneously determine the oral and ophthalmic bioavailability of timolol in dogs. 410.P. B. Werner. [61] [62] [63] [64] [65] [66] [67] [68] [69] [70] [71] [72] [73] [74] [75] [76] [77] Chavez-Eng. Warner. 1997. J. 2002. Pharm. Chromatographia. Heining. C. 81-132.... Chromatogr. J.. J. Bortnick. A. K. Chromatogr. Chromatogr. 1999.. Mürdter. Krauss. Hsieh. 1 [60] 35 [42] [43] [44] [45] [46] [47] [48] [49] [50] [51] [52] [53] [54] [55] [56] [57] [58] [59] Schönberger. oncology. J. Bauer.. Lin. N. Safa.. J. 67..T. N. Roots. Royal Society of Chemistry.. Zhang.C. 237-46. J. E.. 34. Thomasson. Poole. Rev. semi-automated determination of a cyclooxygenase II inhibitor in human plasma and urine using solid-phase extraction in the 96-well format and high-performance liquid chromatography with post-column photochemical derivatization-fluorescence detection.M. C. W. Woolf. 367-72.J..T. Direct activation of rat spinal dorsal horn neurons by prostaglandin E2. Klotz. B.. 16. 661. Gilbert. rofecoxib. 1996... Fang. Stewart. Sci.K. 689. J. W. J. Begg.. Determination of rofecoxib.. Matuszewski.. Nature.Z. J.. sodium diclofenac and niflumic acid in human serum samples by HPLC with DAD detection. Moore. 2001. I. Chromatogr. Ro. Matheson. Investigation of the pharmacokinetics of celecoxib by liquid chromatography-mass spectrometry. L. 471-75.. Simultaneous determination of unlabeled and deuterium-labeled indinavir in human plasma by high-performance liquid chromatography with tandem mass spectrometric detection. 2000. G.A. 1996. R. Stewart J.. a cyclooxygenase-2 inhibitor. J. M. M. 2003. High-throughput.. 1996. A. Acad..L. Rofecoxib: a review of its use in the management of osteoarthritis. 1997. J. Drugs. 2000. B.. M. a novel leukotriene D4 antagonist. W. Z. 36. Boos.C. Drugs. M. 2006. M..F. D.M. A. 221-27. 783. T. 553-59. Barrish. Lamb.A. Salazar... Ureña. Mundkowski. B. Simultaneous determination of rofecoxib and celecoxib in human plasma by highperformance liquid chromatography. Pharmacol. Bukasa. 2530. Figgitt. Nimesulide: some pharmaceutical and pharmacological aspects--an update.. Chromatogr.. Vol. B. Chromatogr. E.A. Clin. R.A. R. 55-61. 1981. Determination of celecoxib in human plasma using solid-phase extraction and high-performance liquid chromatography. A. Gilbert. 2001. 26.. Westerlund. J.. J. Determination of BAY x 7195. T. U. Chromatographia. B... Biomed.. J.C. K.A. Chromatogr.A. 767. D. Pharm. D. D. 2001. Biomed. Anal. B.. B. 83-90.. 2008. Werner. J. Woolf.E. Zarghi. H. Moore. H. Improved procedure for the the determination of rofecoxib in human plasma involving 96-well solid-phase extraction and fluorescence detection. M.. 2002. E. Application of negative-ion chemical ionization isotope dilution gas chromatography--mass spectrometry to single-dose bioavailability studies of mefloquine. 1995.. B. J. 10. 263-69.. S. Bachman. Vol. Brockmöller. Proc. Pahl. 2009. Giuliano. Matuszewski. R. Singla.. D. Michel.G. J. Brune. Guermouche. Determination of celecoxib. Determination of methotrexate and its metabolite 7-hydroxymethotrexate by direct injection of human plasma into a column-switching liquid chromatographic system using post-column photochemical reaction with fluorimetric detection.A. 1998.. Kuhlmann.E. Stewart J. A.Z.J. J. Crocheted ETFE-reactor for on-line post-column photoderivatization of diclofenac in high-performance liquid chromatography.K.. J.K. R. Matuszewski. Brune.J..K.F.. Highperformance liquid chromatographic-tandem mass spectrometric evaluation and determination of stable isotope labeled analogs of rofecoxib in human plasma samples from oral bioavailability studies. 1999. S. M. 833-65.. Frank. Baba. Yu. Rapid Commun. Vojnovic.J. Pharmacol. H. Hamama. S.H. Bonventre.K. 7563-68. Amini. 673.. 499-05. Biomed. 255-60.... Olah. B. F. Baillie. M. B. Determination of fenbufen and its metabolites in serum by reversed-phase high-performance liquid chromatography using solid-phase extraction and on-line postcolumn ultraviolet irradiation and fluorescence detection. Interleukin-1beta-mediated induction of Cox-2 in the CNS contributes to inflammatory pain hypersensitivity. Matuszewski. 58.. Determination of celecoxib in human plasma by highperformance liquid chromatography. Dean.A. Allchorne.M. K.T. C. D. Brunner. Rheumatol. Ch 6... U. Kirchheiner. 2002. Rev. 379-59.J. U. J. 768. Chawla. Natl... and beyond: where is the science headed? J. 35.. Pharm. 833-65. Chromatogr. O. a novel leukotriene D4 antagonist. 83-89. Jalalizadeh... 163-68. Mass Spectrom. Specific COX-2 inhibitors in arthritis. M. 33. Sci. Gillich. Vane.. Chromatogr. S.. in human plasma using highperformance liquid chromatography with post-column photochemical derivatization and fluorescence detection J. Chromatogr. Chavez-Eng. G. Venkateshwaran. U... G. Shafaati.. 2002. M. 341-45. Brien.. J. J. Kunze. C. Alberts.J.N. J. 1997. 24548. B. 751.. Determination of sulindac and its metabolites in human serum by reversed-phase high-performance liquid chromatography using on-line post-column ultraviolet irradiation and fluorescence detection.. Siluveru. B. 193-98. Pharm. Howald. a cyclooxygenase-2 specific inhibitor.. Störmer.. Mundkowski.. Simpson. Gatto. Simple and sensitive method for the determination of celecoxib in human serum by high-performance liquid chromatography with fluorescence detection. T. G. Billet. Gardiner. Farsam. Amann.H.. J. M. T. B. Ha..Reid (Ed). C. 1999. A. 830. A.K. 25. B. 117-29. The use of stable isotopes in pharmacological research. A..R.. H. Neurosci.. K.S. Determination of BAY x 7195.V. . . Heinkele.. 2005.. W. Neal.. A. R. Hassan. F.K.. Woolf. Singh. Woolf. T. 9196. Methodological Surveys in Bioanalysis of Drugs. USA. A.J. in human plasma by high-performance liquid chromatography with tandem mass spectrometric detection. Fermin-Vallvey. J. Gillich. Chromatogr. Anavy. Sapirstein.. C. K. M.P. B.J. Sci. Pharm. in E..J. 1998. A. Li. S. E. Breitner. Chromatogr. J. Samad. Chow. 96.170. Trager.M. Chromatogr.. 567-71. D. E. Fu.. Siluveru. J. L. Hofmann. Constanzer. 2001. 1999.M.. 43. in human plasma by high-performance liquid chromatography with post-column photo derivatisation and fluorescence detection.. Chromatogr. 137-47. M.R.. E.

S.K. Jaworowicz Jr. 215-21. Vishwottam. Douglas. on the pharmacokinetics and pharmacodynamics of propofol. J. Seibert. Cox... R. J. 2002..... Pharmacokinetic profile and in vitro selective cyclooxygenase-2 inhibition by nimesulide in the dog. A. Effect of deracoxib. Curr. D.. 802. K. 275-77.T. No. Simultaneous determination of unlabeled and carbon-13-labeled etoricoxib. A validated high-performance liquid chromatographic assay for determination of lumiracoxib in human plasma.. 2005.. Fricke...H. H... D. Chromatogr. Am. J.. Cho. 33-45..N. Chromatogr. 293-99. 8. Mehvar. J. Cheremina. 788. 723. M. Laurent. A. B. A. E. D.. Opin. 2003.. J. 1 [78] Stewart.. Ramakrishna. Kraml.. J.. Eradiri. assessor-blind study of the pharmacokinetics of parenteral nimesulide versus placebo in healthy indian volunteers. S.D. Carrasco-Portugal. Makarowski.. A.. J.. 1999. B. Hinz. 183-88. 405-12.. Granados-Soto. J.E. Ober. R. S. Caccamese. Koboldt. J. 1983. Rel. 1700-04. Biomed. 20. Chromatogr. B.J.. Bucalo. J. Sci. Ther. 164 . E. Bible. S.44. 2002. Clin.W. J. Rogers. 2002. a new cyclooxygenase-2 inhibitor. S. Flores-Murrieta.T.. Jamali..M.. N. Chromatogr. E. M.F... G. Rapid and simple highperformance liquid chromatographic determination of nimesulide in human plasma. Chromatogr. Capone. Ther. A.. Miller. J.. 2001. 566-71. Wishu. 241-44.S.B. M. 23. 977-84. J.C. K. 621. Clin..J. Varkalis...U. 10. Anal. Sensitive high-performance liquid chromatographic method for the determination of etodolac in serum..W. 227-36.. Pasinetti. Cosyns. Gassel. 1013-21. Shaffer.M. W. 2004. B. ketoprofen and etodolac enantiomers by pre-column derivatization RPHPLC and application to drug-protein binding in human plasma. Evaluation of the stereoselective metabolism of the chiral analgesic drug etodolac by highperformance liquid chromatography.L. J. Toutain. C. S.V. 2003. Frolich. Brune. Fast. Ther. 2002. D. M. Pharmacol. M..S. G. Biomed.. 290-96. W. Jamali.. Liquid Chromatogr. Application of a stereospecific high-performance liquid chromatography assay to a pharmacokinetic study of etodolac enantiomers in humans.. 123-34. 2005. S.. Kuss. 30.. J. M. S. G. M.. S. Forester. Risk of Alzheimer's disease and duration of NSAID use..E.. 87. 66. Dickson. Rordorf. Lee.Y. Gotta.. V.W. Drug Invest. Dispos.. 1993. Blaschke. Pharmacokinetics and metabolism of lumiracoxib in healthy male subjects. Tofanetti. Choi. J.. 2001. Valdecoxib is more efficacious than rofecoxib in relieving pain associated with oral surgery. Lemko.. Jones.. Effects of parecoxib. O. Ji. 72. Richards. R.. 24. K. Zwillich. in human plasma using HPLCMS/MS. Karim. Quantitation of Valdecoxib in human plasma by highperformance liquid chromatography with ultraviolet absorbance detection using liquid–liquid extraction. P.. B. Findlay.) parecoxib sodium in normal subjects. 48.Y.M. Corrada. S. 9. .A. Millis.. Determination of etoricoxib in human plasma by liquid chromatography–tandem mass spectrometry with electrospray ionisation.. Y. cerebrospinal fluid and brain by liquid chromatography using solid-phase extraction. L. Lee. Ptáek. W. Walson. Mhatre. J. J.Y. 819. J.. T. Bonner. Isolation of an unknown metabolite of the non-steroidal anti-inflammatory drug etodolac and its identification as 5-hydroxy etodolac. S. . 158-62. P.. Determination of an antiinflammatory methanesulfonanilide in plasma by high-speed liquid chromatography.. 46. Zweifel. Filipowski. Woolf. 4. Balfour. D.P.. L. Mangold.J.. P. Kharasch.S. 2002. [100] Giuseppe Carlucci [79] [80] [101] [102] [81] [82] [103] [104] [83] [105] [84] [85] [106] [86] [107] [87] [108] [88] [109] [89] [110] [111] [90] [91] [112] [92] [113] [114] [93] [94] [115] [95] [116] [117] [96] [118] [97] [98] [119] [99] Becker-Scharfenkamp.L. Macek.. Drugs.C. A. 1977..M.S. J. Contin. 2004. 2006. M. 4-[5-Methyl-3phenylisoxazol-4-yl]. Ryu. Compend. Pharm. 7. J. Chromatogr. D.. P. Clin.K. Rose. Carte... I..P. 3 . 271-75. 727. 425. Chromatogr. Haak.36 Anti-Inflammatory & Anti-Allergy Agents in Medicinal Chemistry. Chromatogr. 199-07.M. A simple method for determination of nimesulide in rat blood samples by highperformance liquid chromatography. O. A. Kshirsagar.. N. G.S.. G. Boje. B. .benzenesulfonamide. Perkins. A pharmacokinetic study of intramuscular (i. Koteshwara. J. Boni. N. Roark.. J.S. 319-24. Recker. Med. Desai.. J.. N. Talley. 1988. 88-95. Tobias. Spain. Khaksa. Tang. B.. crossover. J. 2003. Dispos. Pharm. Stichtenoth. J. Drugs. Y. Cyclooxygenase-2 expression is increased in frontal cortex of Alzheimer's disease brain. a new COX-2 inhibitor. Simultaneous determination of nimesulide and hydroxynimesulide in human plasma and urine by high-performance liquid chromatography. Tacconelli. Kang. Inflammopharmacol. Dalvi.A.J. 91. Korth-Braddley. Enantiomer.S.. D.A. Ferrario. A. Lee. Wishu. Maltese.B. Bräutigam. Moyers. A. B. F. J. 2000.E. Slater.S. 25.. J.W.. J. Sci. B. Zhao. Yang.. Ibrahim. 2000. 1993. Rennebohm. Masferrer. B. Sci.C. N. D. J. G. 240-45.. J.J. 2237. 441-43. 77. K. Improved high-performance liquid chromatographic assay for nimesulide. 1715-24. 2008. Feldman.. Chromatogr. Simcoe.. Aisen. Buonomo.. 2003. B. J. C. Monzani. 77577. Castoldi. 89-97. J. Biomed. R. P.. Metge.... Determination of valdecoxib and its metabolites in human urine by automated solidphase extraction–liquid chromatography–tandem mass spectrometry.S. Pharmacother. J. Zhang.. 2009. 2000. T. U. 413-8. 1.. Buckley. 1999. Park. 96.R. J. 1999.. Analysis of flurbiprofen. 269-75. 2002.O. J..H. The second generation of COX-2 inhibitors: what advantages do the newest offer? Drugs. Direct high-performance liquid chromatography (HPLC) separation of etodolac enantiomers using chiral stationary phases. Koteshwara.. D. Neuroscience. S. Simultaneous determination of the phase II metabolites of the non steroidal anti-inflammatory drug etodolac in human urine.F.. Brown. M.P. Zeng. Qian.C. 181-84. K. Vol. G.. J. 785. K. Hubbard. valdecoxib. 1997.. C. Gupta. T. J. J. Strickmann..X. Anal. Y.. Rapid determination of nimesulide in rabbit aqueous humor by liquid chromatography.. Patrignani... Breu. 785. Osteoarthr. R. 963-66..M. Perez-Urizar... C.. 953-58. 1988. . D. Pharmacol. M... J. 5. Disposition of a specific cyclooxygenase-2 inhibitor. Yuan.. 274-99. Geisslinger. B.F.. A reappraisal of its pharmacology and therapeutic use in rheumatic diseases and pain states. Chromatogr.W. Educ. 7-10. Drug Metab. J. R. S. Adler. Etodolac: An overview of a selective COX-2 inhibitor. L.. S.C... A.. F. 2004. Berendes. J. Determination of deracoxib in feline plasma samples using high performance liquid chromatography. J. Chromatogr.H. Clin. Agrawal.. Chang. N. Pharm.C. Ramakrishna. S. 32. Russell. Zhang..E. B. 1991... 1999. Graneto. K. Lee. Neurol. 816. 63. Zhang. Metter. Technol. 43. Karim. Rapid and sensitive method for determination of nimesulide in human plasma by high-performance liquid chromatography.. Jin. Vishwottam.K.A.. 741-46. 265-84. Pharmacokinetics of etodolac in patients with stable juvenile rheumatoid arthritis. O.m. J. Bianchi. Nefflen. H. J. M. 41... D. Klíma.L. placebo-controlled comparison with naproxen.A. M. J. 1033-37. J. Pharm.J. 804. M. V.. D.K..V. 2002. 1998. Etodolac. Rodriguez. 2008.. 863. Y. P.C. M. K. H. U. Brocks.. J.J. Development and validation of a high performance liquid chromatography-tandem mass spectrometry for the determination of etodolac in human plasma. 1111 –19.. 42. Blaschke. M.67.M..J. 626-32. Martin.. Pharmacol. 35-42. Weigel. a parenteral COX-2-specific inhibitor.. F.. Gogtay. Stereoselective disposition of etodolac enantiomers in synovial fluid. B.J. Chromatogr.J. Bevirt.N. on the prevention of lameness induced by chemical synovitis in dogs.. Camacho-Vieyra. C. Vet. P.... Sci. 2001. 21... Chromatogr. Vet. Blaschke. J. Pharm.C. 309-15. 22..L.. R. J. Chirality.. A. Udupa.L. J. Recker. Pharm. Matuszewski. CrosbySessoms. Verburg. Expert Opin. 2003. A. Pract. Clinical pharmacology of etoricoxib: a novel selective COX2 inhibitor.D.P.. Chem.. Kawas...Y. 2002. Efficacy and safety of the COX-2 specific inhibitor valdecoxib in the management of osteoarthritis of the hip: a randomized. B.. Seo.M.. Karim. F. 3. double-blind. Validated liquid chromatographic ultraviolet method for the quantitation of Etoricoxib in human plasma using liquid–liquid extraction.. Valdecoxib (Pharmacia). W. Cartil.. 415-22.. Drug Metab. Simultaneous determination of nimesulide and hydroxynimesulide in rat plasma. Invest.L. 17-23. L. E. Jeong. R. Gu. A Randomised. 1991. in human. Valdecoxib: A Potent and Selective Inhibitor of COX-2. Chromatogr. F. Maugeri. M.S. A. 758.F. Anesthesiology. 31. 1996.P. Cester.

Distel. R.M...P. D. 432-39... 2005. Jaiswal. M. Simultaneous determination of piroxicam. Nageswara Rao. Pharm.S.. Huskisson. No. salicylic acid. G.C. 2007. 28... Ghozlan. Srinivas. 11521. Biomed. F.W. Degner... valdecoxib. Y. Zhong. Br. Br. Chen. J.. Chromatogr. 36. 2003. 2004. ketoprofen. Wiesner. 2008 Revised: July 28. Clin. de Jager.L. Chromatogr. F. Development and validation of an automated SPE-LC-MS/MS assay for valdecoxib and its hydroxylated metabolite in human plasma... J. Sensitive and rapid liquid chromatography-tandem mass spectrometry method for the determination of meloxicam in human plasma. J... Chim. J. Bhardwaj. Chromatogr. Ji... Biomed. C. 586. Y.M. S. A non-extracting procedure for the determination of meloxicam in plasma samples by HPLC-diode array detection. 489-98. double-blind comparison with diclofenac sodium. Clinical pharmacokinetics of meloxicam. Vercauteren. . W. Bae. Meloxicam in osteoarthritis: a 6month. A. Y. Swart. Breu. 839-46.M. 2003. N. 362-68.... Anal. A long-term study to evaluate the safety and efficacy of meloxicam therapy in patients with rheumatoid arthritis. CarcíaCampaña. Davies. 852. Lambrecht. B. J.R. 1996. 802. Chromatogr. Br.C. 55. A liquid chromatography-mass spectrometry method for the quantification of both etoricoxib and valdecoxib in human plasma..K. D. 2006.G. J. H. K. David. Medvedovici. Borges. nimesulide and celecoxib in plasma by highperformance liquid chromatography with UV detection. Pharm. 39-43. Yuan. V.Y.W.. 19.. 2006.Y. Rheumatol.F. Anal. Meena. Biomed. Quantitation of Valdecoxib in human plasma by highperformance liquid chromatography with ultraviolet absorbance detection using liquid–liquid extraction.A.K. Lee. J.. Chromatogr.. Simultaneous quantitation of etoricoxib.. Hundt. Hinz.. Anal. J. Rheumatol. Kurthen. . J. C.. A. Development and validation of a new high-performance liquid chromatographic estimation method of meloxicam in biological samples. 2008 . 852. Chromatogr. Chromatogr. M. 271-75.. J. 19. 20. J. 2005. Bluhmki. LC determination and pharmacokinetics of meloxicam. J. Received: June 24.S. 431-36. Zhong.. R.C. J. Georgita. K.H. N.. N. Anal. H. Yuan. E. D’haenick. B. J. Mullangi.. A. Biomed. 35.. Saroj. M. M.. Sutherland. Van der Weken. Jang. 8. A double-blind study to compare the efficacy and safety of meloxicam 15 mg with piroxicam 20 mg in patients with osteoarthritis of the hip. Albu. 826. E.. Baeyens. Vankeirsbilck. S. Pharmacokinet. 738... Vinu. E. Pavan Kumar. B. Ther. Ji. Mendes. Determination of meloxicam in human plasma by liquid chromatography-tandem mass spectrometry following transdermal administration. 2003. V.R. B. C. R. 1996. Wishu. J. 1996. N. 125-32.G.. G.. Shivaprakash. 32. Bluhmki. 1999. B.. [131] [132] [133] [134] [135] [136] [137] [138] [139] Dasandi.. Hosie..J. Bluhmki. Wishwottam. H. 785. S. Biomed. 2007.. Int.. 2005.Analytical Procedure for Determination of Cyclooxygenase-2 [120] Anti-Inflammatory & Anti-Allergy Agents in Medicinal Chemistry. Chen. 35. A. B. K. Moreno.. Acta.C.L.. H. A. Distel.. Chromatogr. Rigato.. X. Fast.. F. Velpandian.. H. Biomed. 2007.. 69-73. Pharm.A. B. Pitarch. Moragues. Vol. A.D. 2008 Accepted: August 05. D. Linden. Arzneimittelforschung. 35-38. R. X. Jeong. Brune. R.V. Clin. M... Deprez.. Meloxicam determination in human plasma by high-performance liquid chromatography coupled with tandem mass spectrometry (LC-MSMS) in Brazilian bioequivalence studies. 2005.. 859. 214-19. Rheumatol. Els.M. Mircioiu. Hundt... Ramakrishna. D.D. Development and validation of a reversed-phase liquid chromatographic method for separation and simultaneous determination of COX-2 inhibitors in pharmaceuticals and its application to biological fluids.M. 115-26. 33. J. T. Werner. U.N...W. 2002. Werner. C. 35.. N. A.. Application of an alkyl-diol silica precolumn in a columnswitching system for the determination of meloxicam in plasma. D. J. Lee. A cyclo-oxygenase-2 preferential nonsteroidal antiinflammatory drug. 61-72. 44.F.K... Kim. Raghu Ram Rao. Bhat. 2000. Igualada. meloxicam and tenoxicam in human plasma by liquid chromatography with tandem mass spectrometry. Pharm. H.. 2007. 113-18.Y.V. R. E. 650-54.. S. Chromatogr. Chromatogr. 29-34. Determination of meloxicam in human plasma using a HPLC method with UV detection and its application to a pharmacokinetic study.. C. 999-04.. B. Rapid method for the determination of non-steroidal anti-inflammatory drugs in animal tissue by liquid chromatography-mass spectrometry with ion-trap detector.. K. Koteshwara. Kim. B.J. T. F. P. E. Gupta. J. Determination of meloxicam in human plasma by liquid chromatography-tandem mass spectrometry following transdermal administration. 1 [130] 37 [121] [122] [123] [124] [125] [126] [127] [128] [129] Zhang. B. J.. Skjodt.. 650-54. 326-31. J.. Lee. 2009.