title: Simultaneous determination of creatinine and uric acid in urine by liquid chromatography-tandem mass spectrometry with polarity switching electrospray ionization.
abstract: A simple liquid chromatography-tandem mass spectrometric (LC-MS/MS) method was developed and validated to simultaneously determine creatinine (Cr) and uric acid (UA) levels as a confirmatory method for adulteration or dilution of urine. Centrifuged urine samples (10μL) were diluted with 390μL of distilled water. 30μL of internal standard solution (Cr-d(3), 5μg/mL) and 10μL of acetonitrile were added to 20μL aliquots of diluted urine samples and filtered. The samples (1μL) were introduced into LC-MS/MS with no further pretreatment. Cr and UA were separated on a multi-mode ODS column (Scherzo SM-C18, 75mm×2.0mm I.D., 3μm) and quantified by LC-MS/MS with polarity-switching electrospray ionization. Cr requires the positive-ion mode, whereas the negative-ion mode is required for the analysis of UA. The linear ranges were 1.0-300mg/dL for Cr and 0.5-300mg/dL for UA, with good determination coefficients (R(2)≥0.9988). The intra-day and inter-day precision of the analytes was within 13.0% and 14.4%, respectively. The intra-day and inter-day accuracy was -8.8 to 3.7% and -0.3 to 6.6%, respectively. The lower limits of detection (LLODs) were 0.3mg/dL for Cr and 0.07mg/dL for UA. The applicability of the developed method was examined by analyzing urine samples from suspected drug abusers (n=46).
authors.affiliation: Drug Analysis Laboratory, Forensic Science Division, Supreme Prosecutors' Office, 157 Banpodaero, Seocho-gu, Seoul 137-730, Republic of Korea.
authors.forename: Woonyong
authors.initials: W
authors.lastname: Kwon
authors.forename: Jin Young
authors.initials: JY
authors.lastname: Kim
authors.forename: Sungill
authors.initials: S
authors.lastname: Suh
authors.forename: Moon Kyo
authors.initials: MK
authors.lastname: In
mesh_terms: D002853:Chromatography, Liquid; D003404:Creatinine / Q000652:urine*; D006801:Humans; D057230:Limit of Detection; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods; D015813:Substance Abuse Detection; D053719:Tandem Mass Spectrometry; D014527:Uric Acid / Q000652:urine*
chemical_list: D014527:Uric Acid; D003404:Creatinine