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  • A rapid method for determination of 22 selected drugs in human urine by UHPLC/MS/MS for clinical application.

A rapid method for determination of 22 selected drugs in human urine by UHPLC/MS/MS for clinical application.

Journal of AOAC International (2015-01-30)
Sylwia Magiera, Irena Baranowska
ABSTRACT

A rapid and sensitive ultra-HPLC/MSIMS (UHPLC/MSIMS) assay method for the simultaneous determination in human urine of 22 drugs belonging to different pharmaceutical groups was developed. The drugs were extracted from urine samples and then separated on a Zorbax Rapid Resolution High Definition SB-C18 column. The mobile phase consisted of methanol and water containing formic acid with gradient elution. The chromatographic separation time was 7 min. The MSIMS detector, equipped with an electrospray ionization source, was set up in both positive and negative modes. The lower LOQs for the drugs in this method were between 0.05 and 0.60 ng/mL. Calibration curves in human urine were generated in the range of 0.05-600 ng/mL. Method validation parameters such as intraday and interday precision, accuracy, extraction recovery, stability, selectivity, dilution integrity, and carryover effect for all the compounds were within the acceptable ranges. This simple and fast method was applied successfully to study the pharmacokinetics of four selected drugs in human urine collected from patients. This UHPLC/MS/MS method offers an attractive way forward for the development of a routine rapid analysis for selected substances, particularly given the growing amount of new information about drug properties.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Salicylic acid, ≥99%, FG
Sigma-Aldrich
Methanol, NMR reference standard
Sigma-Aldrich
Acenocoumarol, ≥98% (HPLC)
USP
Nifedipine, United States Pharmacopeia (USP) Reference Standard
Nifedipine, European Pharmacopoeia (EP) Reference Standard
USP
Carbamazepine, United States Pharmacopeia (USP) Reference Standard
Salicylic acid, European Pharmacopoeia (EP) Reference Standard
Supelco
Salicylic acid, 1.0 mg/mL in acetonitrile, ampule of 1 mL, certified reference material, Cerilliant®
Dexamethasone for peak identification, European Pharmacopoeia (EP) Reference Standard
Carbamazepine, European Pharmacopoeia (EP) Reference Standard
USP
Propranolol hydrochloride, United States Pharmacopeia (USP) Reference Standard
Sildenafil citrate, European Pharmacopoeia (EP) Reference Standard
Terbinafine for system suitability, European Pharmacopoeia (EP) Reference Standard
Supelco
Carbamazepine, analytical standard
Diclofenac for system suitability, European Pharmacopoeia (EP) Reference Standard
Propranolol hydrochloride, European Pharmacopoeia (EP) Reference Standard
Ibuprofen, European Pharmacopoeia (EP) Reference Standard
Paracetamol, European Pharmacopoeia (EP) Reference Standard
USP
Ibuprofen, United States Pharmacopeia (USP) Reference Standard
Ibuprofen for peak identification, European Pharmacopoeia (EP) Reference Standard
Sigma-Aldrich
Methanol, Absolute - Acetone free
Sigma-Aldrich
Methanol, ACS reagent, ≥99.8%
Sigma-Aldrich
Methanol, Laboratory Reagent, ≥99.6%
Captopril for system suitability, European Pharmacopoeia (EP) Reference Standard
Sigma-Aldrich
Carbamazepine, powder
Sigma-Aldrich
Carbamazepine, meets USP testing specifications
Sigma-Aldrich
Ser-Phe-Leu-Leu-Arg-Asn-Pro-Asn-Asp-Lys-Tyr-Glu-Pro-Phe, ≥97% (HPLC)
Sigma-Aldrich
Nifedipine, ≥98% (HPLC), powder
Sigma-Aldrich
Salicylic acid, ReagentPlus®, ≥99%
Supelco
Dipyron monohydrate, VETRANAL®, analytical standard