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A simple and sensitive method was developed and validated here for the analysis of thirteen nonsteroidal anti-inflammatory drugs (NSAIDs) in human plasma samples by hydrophilic interaction liquid chromatography (HILIC)-tandem mass spectrometry (MS/MS). A small volume of plasma (20μL) spiked with compounds was diluted with 80μL of 10-mM ammonium acetate followed by a simple protein precipitation with 400μL of acetonitrile. After centrifugation, the clear supernatant extract was directly injected into the HILIC-MS/MS, without any solvent evaporation and reconstitution steps. The chromatographic separation of the NSAIDs was achieved on a Unison UK-Amino HILIC column (50mm×3mm i.d., particle size 3μm) with a linear gradient elution system composed of 10mM ammonium acetate (pH 6.8) and acetonitrile at a flow rate of 0.4mL/min. The mass spectra obtained by HILIC-MS showed base peak ions due to [M+H](+) for indomethacin, oxaprozin, ketoprofen, alminoprofen, zaltoprofen, tiaprofenic acid, pranoprofen, and ketoprofen-d3 and due to [M-H](-) for etodolac, ibuprofen, diclofenac, fenoprofen, loxoprofen, naproxen, and ibuprofen-d3. Recoveries of these thirteen NSAIDs in plasma were 34.8-113% and the lower limits of quantitation were 0.125-1.25μg/mL. The intra- and interday coefficient of variations for all drugs in plasma were less than 14.6%. The data obtained from actual plasma determinations of zaltoprofen, ibuprofen, and diclofenac are also presented.

A rapid and simple microanalysis method using TopTip C18 spin tips, which are new micropipette tip spin columns for solid-phase extraction, was developed and validated for quantitative determination of methamphetamine and amphetamine in human plasma. All the solid-phase extraction procedures, including conditioning, sample loading, washing, and elution, using the TopTip C18 spin tips, were performed by centrifugation. The analytes were extracted within 7–8 min from 0.1 mL of plasma. The methanol-rich eluate was injected directly into a hydrophilic interaction liquid chromatography/tandem mass spectrometer in positive electrospray ionization mode with selected reaction monitoring. The analytes were separated on an Imtakt Unison UK-silica column using a gradient with methanol and 10 mmol/L ammonium acetate buffer as mobile phase at a flow rate of 0.5 mL/min. The recoveries of methamphetamine and amphetamine spiked into plasma were 82–95% and the limit of quantification of methamphetamine and amphetamine wer...

Three tetracyclic antidepressants, mianserin, mirtazapine, and setiptiline, were extracted from human plasma samples using a new micropipette solid-phase extraction tip containing a monolithic silica gel packing material, the MonoTip C18. The plasma samples (0.1 ml), which contained mianserin, mirtazapine, and setiptiline, were mixed with 330 μl of distilled water and 50 μl of 0.5 M glycine–sodium hydroxide buffer solution (pH 8). After centrifugation, the supernatant was extracted onto the C18 phase of the tip (pipette tip volume, 200 μl) by 25 sequential aspirating/dispensing cycles. The three analytes retained in the C18 phase were eluted with methanol by 5 aspirating/dispensing cycles. The eluate was subjected to measurements by gas chromatography (GC) with nitrogen–phosphorus detection (NPD). The recoveries of the three antidepressants were 84.6–99.6% and the limits of detection for each drug were between 2.4 and 15 ng/ml of plasma. The maximum intraday and interday coefficients of variation for the drugs was below 10.6%. Regression equations for the three drugs showed excellent linearity in the range of 50–5000 ng/ml for mianserin and mirtazapine and 50–10 000 ng/ml for setiptiline. The antidepressants were stable for at least 12 h at 4°C, 4 weeks at −80°C, and through three freeze–thaw cycles in plasma. The validated method was successfully used to quantify the plasma concentrations of mirtazapine in a human subject after oral administration of the drug. Although we achieved analyte detection with GC-NPD, the present monolithic micropipette extraction method for tetracyclic antidepressants seems to be very useful for combining it with GC–mass spectrometry (MS) and/or liquid chromatography–tandem MS to secure high simplicity, purification, reproducibility, and sensitivity.

Methamphetamine and amphetamine were extracted from human whole blood samples using pipette tip solid-phase extraction (SPE) with MonoTip C(18) tips, on which C(18)-bonded monolithic silica gel was fixed. Human whole blood (0.1 mL) containing methamphetamine and amphetamine, with N-methylbenzylamine as an internal standard, was mixed with 0.4 mL of distilled water and 50 microL of 5 M sodium hydroxide solution. After centrifugation, the supernatant was extracted to the C(18) phase of the tip (pipette tip volume, 200 microL) by 25 repeated aspirating/dispensing cycles using a manual micropipettor. Analytes retained in the C(18) phase were eluted with methanol by five repeated aspirating/dispensing cycles. After derivatization with trifluoroacetic anhydride, analytes were measured by gas chromatography - mass spectrometry with selected ion monitoring in the positive-ion electron impact mode. Recoveries of methamphetamine and amphetamine spiked into whole blood were more than 87.6 and 81.7%, respectively. Regression equations for methamphetamine and amphetamine showed excellent linearity in the range of 0.5-100 ng/0.1 mL. The limits of detection for methamphetamine and amphetamine were 0.15 and 0.11 ng/0.1 mL, respectively. Intra- and interday coefficients of variation for both stimulants were not greater than 9.6 and 13.8%, respectively. The determination of methamphetamine and amphetamine in autopsy whole blood samples is presented, and was shown to validate the present methodology.

Mequitazine has been found to be extractable from human plasma samples using MonoTip C 18 tips, inside which C 18 -bonded monolithic silica gel was fixed. Human plasma (0.1 mL) containing mequitazine and cyproheptadine as an internal standard (IS) was mixed with 0.4 mL of distilled water and 25 μL of 1 M potassium phosphate buffer (pH 8.0). After centrifugation of the mixture, the supernatant fraction was extracted to the C 18 phase of the tip by 25 repeated aspirating/dispensing cycles using a manual micropipettor. The analytes retained on the C 18 phase were then eluted with methanol by five repeated aspirating/dispensing cycles. Without evaporation and reconstitution, the eluate was injected into a gas chromatograph injector and detected by a mass spectrometer with selected ion monitoring in the positive-ion electron impact mode. The separation of mequitazine and the IS from each other and from impurities was generally satisfactory using a DB-1MS capillary column (30 m × 0.32 mm i.d., film thickness 0.25 μm). The recoveries of mequitazine and the IS spiked into plasma were more than 90.0%. The regression equation for mequitazine showed excellent linearity in the range of 0.2–200 ng 0.1 mL −1 , and the detection limit was 0.05 ng 0.1 mL −1 of plasma. The intra-day and inter-day coefficients of variation for mequitazine in human plasma were not greater than 8.16 and 9.24%, respectively. Accuracy for the drug was in the range of 90.0–97.4%. The data obtained from determination of mequitazine in human plasma after oral administration of the drug are also presented.

Ten antihistamine drugs, diphenhydramine, orphenadrine, chlorpheniramine, diphenylpyraline, triprolidine, promethazine, homochlorcyclizine, cyproheptadine, cloperastine and clemastine, have been found to be extractable from human plasma samples using MonoTip C18 tips, inside which C18-bonded monolithic silica gel was fixed. Human plasma (0.1 mL) containing the ten antihistamines was mixed with 0.4 mL of distilled water and 25 µL of a 1 M potassium phosphate buffer (pH 8.0). After centrifugation of the mixture, the supernatant fraction was extracted to the C18 phase of the tip by 25 repeated aspirating/dispensing cycles using a manual micropipettor. The analytes retained on the C18 phase were then eluted with methanol by five repeated aspirating/dispensing cycles. The eluate was injected into a gas chromatography (GC) injector without evaporation and reconstitution steps, and was detected by a mass spectrometer with selected ion monitoring in the positive-ion electron impact mode. The separation of the ten drugs from each other and from impurities was generally satisfactory using a DB-1MS column (30 m × 0.32 mm i.d., film thickness 0.25 µm). The recoveries of the ten antihistamines spiked into plasma were 73.8–105%. The regression equations for the ten antihistamines showed excellent linearity with detection limits of 0.02–5.0 ng/0.1 mL. The within-day and day-to-day coefficients of variation for plasma were not greater than 9.9%. The data obtained from determination of diphenhydramine and chlorpheniramine in human plasma after oral administration of the drugs are also presented. Copyright © 2006 John Wiley & Sons, Ltd.

Three muscle relaxants, tolperisone, eperisone, and tizanidine, were found to be extractable from human whole blood and urine samples by solid‐phase extraction (SPE) with a Sep‐Pak C18 cartridge. Their determination was made using capillary gas chromatography with nitrogen phosphorus detection. Recoveries of the three drugs were 80.0%–97.2% for whole blood and 82.0%–94.5% for urine. The coefficients of within‐day variation in terms of recovery for all compounds in whole blood and urine samples were 2.2%–9.1% and 6.9%–9.6%, respectively; those of day‐to‐day variation for the compounds in both samples were not greater than 18.1%. The regression equations for the compounds showed excellent linearity with detection limits of 1.6–7.7 ng mL−1 for whole blood and 6.0–8.0 ng mL−1 for urine. The data obtained for actual determination of tolperisone in a subject after ingestion of the drugs are also presented.

Diazepam and its major metabolites, nordazepam, temazepam and oxazepam, in human urine samples, were analyzed by liquid chromatography (LC)/tandem mass spectrometry (MS/MS) using a hydrophilic polymer column (MSpak GF-310 4B), which enables direct injection of crude biological samples. Matrix compounds in urine were eluted first from the column, while the target compounds were retained on the polymer stationary phase. The analytes retained on the column were then eluted into an acetonitrile-rich mobile phase using a gradient separation technique. All compounds showed base-peak ions due to [M+H]+ ions on LC/MS with positive ion electrospray ionization, and product ions were produced from each [M+H]+ ion by LC/MS/MS. Quantification was performed by selected reaction monitoring. All compounds spiked into urine showed method recoveries of 50.1-82.0%. The regression equations for all compounds showed excellent linearity in the range of 0.5-500 ng/mL of urine. The limits of detection and quantification for each compound were 0.1 and 0.5 ng/mL of urine, respectively. The intra- and inter-day coefficients of variation for all compounds in urine were not greater than 9.6%. The data obtained from actual determination of diazepam and its three metabolites, oxazepam, nordazepam and temazepam, in human urine after oral administration of diazepam, are also presented.

Seven phenothiazine derivatives, perazine, perphenazine, prochlorperazine, propericiazine, thioproperazine, trifluoperazine, and flupentixol, have been found to be extractable from human plasma and urine samples using disk solid-phase extraction (SPE) with an Empore C18 cartridge. Human plasma and urine (1 mL each) containing the 7 phenothiazine derivatives were mixed with 2 mL of 0.1M NaOH and 7 mL distilled water and then poured into the disk SPE cartridges. The drugs were eluted with 1 mL chloroform– acetonitrile (8 + 2) and determined by liquid chromatography with ammonium formate/formic acid–acetonitrile gradient elution. The detection was performed by ultraviolet absorption at 250 nm. The separation of the 7 phenothiazine derivatives from each other and from impurities was generally satisfactory using a SymmetryShield RP8 column (150 × 2.1 mm id, 3.5 μm particle size). The recoveries of the 7 phenothiazine derivatives spiked into plasma and urine samples were 64.0–89.9% and 65.1–92.1%, respectively. Regression equations for the 7 phenothiazine derivatives showed excellent linearity, with detection limits of 0.021–0.30 mg/mL for plasma and 0.017–0.30 μg/mL for urine. The within-day and day-to-day coefficients of variation for both samples were commonly below 9.0 and 14.9%, respectively.