Development of solid phase extraction strategies to minimize the effect of human urine matrix effect on the response of carnitine by UPLC–MS/MS

Abstract

A rapid, selective and sensitive ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method was developed to determine carnitine in human urine. Solid phase extraction approaches based on the use of polymeric and weak cationic exchange cartridges were evaluated and applied to the treatment of urine samples. After optimizing the various stages of SPE, a satisfactory set up for retaining substances interfering on carnitine's response was achieved for both type of cartridges. The UPLC separation was carried out on a reversed phase column. The detection was performed on a triple quadrupole tandem mass spectrometer by multiple reaction monitoring (MRM) mode via electrospray ionization (ESI) source. Residual matrix components were specific to urine samples and interfered on the carnitine signal (a response suppression of 50% was observed). It was then demonstrated that sample treatment by SPE could reduce the effect of the above mentioned interferents, without needing a preliminary derivatization step. The recovery percentage of carnitine obtained after the application of SPE was of approximately 83±7% and, consequently, the matrix effect was minimized. Thus, a sensitive, precise and reliable methodology was developed to determine traces of carnitine in biological fluids.

Function, Detection and Alteration of Acylcarnitine Metabolism in Hepatocellular Carcinoma

“…Despite these shortcomings, SPE is still extremely applied for the enrichment and isolation of acylcarnitines. For example, after purifying human urine using polymeric and weak cationic exchange cartridges, the matrix effect was significantly reduced as the urinary carnitine was analysed by UPLC-MS/MS [ 70 ]. An online SPE with an Oasis mixed-mode cation exchange (MCX) trapping column combined with LC-MS analysis was a rapid sample work-up method for the quantification of acylcarnitines with different polarity.…”

Quantitative Detection of Ambroxol in Human Plasma Using HPLC-APCI-MS/MS: Application to a Pharmacokinetic Study

“…16 These unpredictable effects are a regular problem for biological sample analysis, and greatly affect the reproducibility and accuracy of bioanalytical method. The common strategies, such as liquid-liquid extraction (LLE), solid-phase extraction (SPE) 17,18 and isotope-labeled internal standard normalized matrix factor, 19 were developed to minimize any matrix effects in bioanalytical assays by LC/MS/MS. However, both the LLE and SPE methods suffered from the multiple complicated procedures that are time and cost consuming.…”

Quantification of Eicosanoids and Their Metabolites in Biological Matrices: A Review

“…Literature from last decade revealed that SPE is a method of choice regardless of sample type (i.e., urine, plasma, saliva, etc.) [23][24][25]. SPE works based on the same principles as column chromatography, the majority of sample components are adsorbed to the stationary phase.…”