Strategy for successful urine sample preparation for LC-MS/MS device at drug verification laboratory

Year 2022, Volume: 3 Issue: 2, 91 - 97, 16.09.2022

Esin Eren , Asuman Göncü Sürü , Necat Yılmaz , Cemile Öz

https://doi.org/10.56766/ntms.823790

Cited By: 1

Abstract

Objectives
The use of liquid chromatography with tandem mass spectrometry (LC-MS/MS) device has increased significantly in toxicology validation laboratories in recent years. The maintenance cost of this expensive equipment is high as well as the cost of consumables. Reduction of the matrix effect and preparation more clear samples are very important for the validation of methods in clinical laboratories.
The primary goal is to create a cleaner urine sample preparation technique to reduce the cost of maintenance of the LC-MS/MS device without affecting test results.
Methods
We prepared the patients’ urine in two different ways; routine urine preparation method and used our centrifuged method (14000 rpm, 10 minutes) for routine illicit substance use. The standard material used to determine whether there was a statistical difference in the urine sample with both different methods was added to both urine samples.
Results
Our findings showed that there was no statistical difference between the results of both methods for detection of illicit substance use. There was no difference between the high and low quantities of the 14 illicit substances measured and the centrifuged method and routine urine preparation methods (p<0.05). However, the urine sample obtained by our newly developed centrifuged method was cleaner, lucid and homogeneous.
Conclusion
Preliminary studies show that although the new centrifuged method is time-consuming, it can be reliable. The long-term use of this centrifugal method can reduce the costs of device maintenance, repair and consumption. So initial findings suggest that column costs and replacement processes may be positively affected.

Keywords

LC-MS/MS, Urine, Toxicology, Morphine, Validation

References

• Verplaetse R, Henion J. Quantitative determination of opioids in whole blood using fully automated dried blood spot desorption coupled to on-line SPE-LC-MS/MS. Drug Test Anal 2016;8:30-8.
• Verplaetse R, Tytgat J. Development and validation of a sensitive ultra-performance liquid chromatography tandem mass spectrometry method for the analysis of fentanyl and its major metabolite norfentanyl in urine and whole blood in forensic context. J Chromatogr B Analyt Technol Biomed Life Sci 2010;878:1987-96.
• Mareck U, Haenelt N, Geyer H, Guddat S, Kamber M, Brenneisen R, Thevis M, Schänzer W. Temporal indication of cannabis use by means of THC glucuronide determination. Drug Test Anal 2009;1:505-10.
• Henion J, Brewer E, Rule G. Peer Reviewed: Sample Preparation for LC/MS/MS: Analyzing Biological and Environmental Samples. Analytical Chemistry 1998;70:650A-656A.
• Felli M, Martello S, Chiarotti M. LC-MS-MS method for simultaneous determination of THCCOOH and THCCOOH-glucuronide in urine: Application to workplace confirmation tests. Forensic Sci Int 2011;204:67-73.
• Andersson M, Scheidweiler KB, Sempio C, Barnes AJ, Huestis MA. Simultaneous quantification of 11 cannabinoids and metabolites in human urine by liquid chromatography tandem mass spectrometry using WAX-S tips. Anal Bioanal Chem 2016;408:6461-71.
• Andersson M, Scheidweiler KB, Sempio C, Barnes AJ, Huestis MA. Simultaneous quantification of 11 cannabinoids and metabolites in human urine by liquid chromatography tandem mass spectrometry using WAX-S tips. Anal Bioanal Chem 2016;408:6461-71.
• Raćkowska E, Bobrowska-Korczak B, Giebułtowicz J. Development and validation of a rapid LC-MS/MS method for determination of methylated nucleosides and nucleobases in urine. J Chromatogr B Analyt Technol Biomed Life Sci 2019;1128:121775.
• Gaunitz F, Kieliba T, Thevis M, Mercer-Chalmers-Bender K. Solid-phase extraction-liquid chromatography-tandem mass spectrometry method for the qualitative analysis of 61 synthetic cannabinoid metabolites in urine. Drug Test Anal 2020;12:27-40.
• Yanes EG, Lovett DP. High-throughput bioanalytical method for analysis of synthetic cannabinoid metabolites in urine using salting-out sample preparation and LC-MS/MS. J Chromatogr B Analyt Technol Biomed Life Sci 2012;909:42-50.
• Court M, Garin J, Masselon CD. Urine sample preparation and fractionation for global proteome profiling by LC-MS. Methods Mol Biol 2015;1243:175-86.
• Smith G, Barratt D, Rowlinson R, Nickson J, Tonge R. Development of a high-throughput method for preparing human urine for two-dimensional electrophoresis. Proteomics 2005;5:2315-8.
• Staeheli SN, Veloso VP, Bovens M, Bissig C, Kraemer T, Poetzsch M. Liquid chromatography-tandem mass spectrometry screening method using information-dependent acquisition of enhanced product ion mass spectra for synthetic cannabinoids including metabolites in urine. Drug Test Anal. 2019 Jun 20. doi: 10.1002/dta.2664.