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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

Dexamethasone, British Pharmacopoeia (BP) Assay Standard
Sigma-Aldrich
Formic acid, ACS reagent, ≥96%
Naproxen, European Pharmacopoeia (EP) Reference Standard
Sigma-Aldrich
Acetonitrile, ReagentPlus®, 99%
Furosemide for peak identification, European Pharmacopoeia (EP) Reference Standard
Supelco
Furosemide, Pharmaceutical Secondary Standard; Certified Reference Material
Furosemide, European Pharmacopoeia (EP) Reference Standard
Supelco
Naproxen, Pharmaceutical Secondary Standard; Certified Reference Material
Sigma-Aldrich
Meldonium dihydrate, ≥98% (HPLC), powder
Sigma-Aldrich
(S)-(+)-6-Methoxy-α-methyl-2-naphthaleneacetic acid, 98%
Supelco
Dexamethasone, VETRANAL®, analytical standard
Sigma-Aldrich
Dexamethasone, tested according to Ph. Eur.
Sigma-Aldrich
Dexamethasone, powder, γ-irradiated, BioXtra, suitable for cell culture, ≥80% (HPLC)
Sigma-Aldrich
Dexamethasone, meets USP testing specifications
Sigma-Aldrich
Milrinone, ≥97% (TLC), powder
Sigma-Aldrich
Dexamethasone, powder, BioReagent, suitable for cell culture, ≥97%
Sigma-Aldrich
Dexamethasone, ≥98% (HPLC), powder
Sigma-Aldrich
Naproxen, meets USP testing specifications
Diclofenac sodium, European Pharmacopoeia (EP) Reference Standard
Dexamethasone, European Pharmacopoeia (EP) Reference Standard
Supelco
Acetaminophen solution, 1.0 mg/mL in methanol, ampule of 1 mL, certified reference material, Cerilliant®
Sigma-Aldrich
Acetonitrile, electronic grade, 99.999% trace metals basis
Supelco
Terbinafine hydrochloride, Pharmaceutical Secondary Standard; Certified Reference Material
Sigma-Aldrich
Formic acid, ACS reagent, ≥88%
Sigma-Aldrich
Furosemide
Sigma-Aldrich
Formic acid solution, BioUltra, 1.0 M in H2O
Sigma-Aldrich
Formic acid, ≥95%, FCC, FG
Sigma-Aldrich
Formic acid, reagent grade, ≥95%
Supelco
Captopril, Pharmaceutical Secondary Standard; Certified Reference Material
Sigma-Aldrich
Captopril, ≥98% (HPLC), powder