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  • Fast targeted analysis of 132 acidic and neutral drugs and poisons in whole blood using LC-MS/MS.

Fast targeted analysis of 132 acidic and neutral drugs and poisons in whole blood using LC-MS/MS.

Forensic science international (2014-05-06)
Matthew Di Rago, Eva Saar, Luke N Rodda, Sophie Turfus, Alex Kotsos, Dimitri Gerostamoulos, Olaf H Drummer
ABSTRACT

The aim of this study was to develop an LC-MS/MS based screening technique that covers a broad range of acidic and neutral drugs and poisons by combining a small sample volume and efficient extraction technique with simple automated data processing. After protein precipitation of 100μL of whole blood, 132 common acidic and neutral drugs and poisons including non-steroidal anti-inflammatory drugs, barbiturates, anticonvulsants, antidiabetics, muscle relaxants, diuretics and superwarfarin rodenticides (47 quantitated, 85 reported as detected) were separated using a Shimadzu Prominence HPLC system with a C18 separation column (Kinetex XB-C18, 4.6mm×150mm, 5μm), using gradient elution with a mobile phase of 25mM ammonium acetate buffer (pH 7.5)/acetonitrile. The drugs were detected using an ABSciex(®) API 2000 LC-MS/MS system (ESI+ and -, MRM mode, two transitions per analyte). The method was fully validated in accordance with international guidelines. Quantification data obtained using one-point calibration compared favorably to that using multiple calibrants. The presented LC-MS/MS assay has proven to be applicable for determination of the analytes in blood. The fast and reliable extraction method combined with automated processing gives the opportunity for high throughput and fast turnaround times for forensic and clinical toxicology.

MATERIALS
Product Number
Brand
Product Description

Supelco
Valproic acid, Pharmaceutical Secondary Standard; Certified Reference Material
Supelco
Acetaminophen, Pharmaceutical Secondary Standard; Certified Reference Material
Supelco
Acetaminophen solution, 1.0 mg/mL in methanol, ampule of 1 mL, certified reference material, Cerilliant®
Supelco
Dimethyl sulfoxide, for inorganic trace analysis, ≥99.99995% (metals basis)
USP
Acetaminophen, United States Pharmacopeia (USP) Reference Standard
USP
Amoxicillin, United States Pharmacopeia (USP) Reference Standard
USP
Valproic acid, United States Pharmacopeia (USP) Reference Standard
Sigma-Aldrich
Methanol solution, NMR reference standard, 4% in methanol-d4 (99.8 atom % D), NMR tube size 3 mm × 8 in.
Sigma-Aldrich
Ammonia, anhydrous, ≥99.98%
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Sodium hydroxide, anhydrous, free-flowing, Redi-Dri, reagent grade, ≥98%, pellets
Supelco
Metronidazole, analytical standard
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Prednisolone, ≥99%
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Trimethoprim, ≥98.5%
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Metronidazole, BioXtra
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Papaverine hydrochloride, powder
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Chlorothiazide, thiazide diuretic
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Cyclobenzaprine hydrochloride
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Phenylbutazone
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2-Propylpentanoic acid
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Salicylic acid, suitable for plant cell culture
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Acetazolamide, ≥99%, powder
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Salicylic acid, BioXtra, ≥99.0%
Supelco
Metronidazole, VETRANAL®, analytical standard
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Ammonia solution, 4 M in methanol
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Ammonia-14N, 99.99 atom % 14N
Salicylic acid, European Pharmacopoeia (EP) Reference Standard
Supelco
Salicylic acid, 1.0 mg/mL in acetonitrile, ampule of 1 mL, certified reference material, Cerilliant®
Sigma-Aldrich
Methanol, NMR reference standard
Acetazolamide, European Pharmacopoeia (EP) Reference Standard