Analysis of Biological Samples in a Contemporary Laboratory Practice (Review)

Introduction. In the present publication highlights the key points of the main stages of development of methods for determining trace amounts of drugs and metabolites in biological samples using chromatographic and chromatography-mass spectrometry methods. The main sources of errors are specified. The main attention is paid to chromatography-mass spectrometry, which is the basic method of analysis of small molecules in biological samples. Examples from literary sources and authors' own practice are given.

Text. The review highlights some of the practical issues of preparation of calibration samples, method of increasing the stability of the sample at the stage of sampling and plasma preparation. In particular, the influence of various anticoagulants on the accuracy of the analysis is reflected. Specify the method of reducing back conversion of some metabolites of carboxyl-containing drugs to parent compound to prevent overestimation of the results of quantitative determination. Some methods of sample preparation, which have become widespread recently, are noted. For example, solid supported liquid-liquid extraction, based on the extraction of the component of interest from the water sample into the liquid layer distributed on a solid high-polar carrier, followed by eluting by a system of non-polar solvents that do not mix with this layer. Recommendations on the use of internal standards, the preparation of the mobile phase for HPLC, on chromatographic separation, validation techniques are given. In the section «Mass spectrometric detection» features of preparation of a mobile phase for chromatography-mass spectrometry experiments are given. The questions of carry-over reduction, ion suppression, matrix effect are covered. The phenomenon of cross-talk in the study of drug metabolism by chromatography-mass spectrometry is discussed. It consists in the mutual distortion of the mass spectrometric response, when the same mass fragments are formed from different ions-precursors. Features of development of techniques for high-performance pharmacokinetic screening are given.

Conclusion. The authors hope that the presented material will be useful for scientists and specialists in the field of pharmacokinetics, biomarker discovery and clinical analyses.

1. Pandey S., Pandey P., Tiwari G., Tiwari R. Bioanalysis in drug discovery and development. Pharm Methods. 2010; 1(1): 14–24. DOI: 10.4103/2229-4708.72223.

2. Glahn-Martínez B., Benito-Peña E., Salis F., Descalzo A. B., Orellana G., Moreno-Bondi M. C. Sensitive rapid fluorescence polarization immunoassay for free mycophenolic acid determination in human serum and plasma. Anal. Chem. 2018; 90(8): 5459–5465. DOI: 10.1021/acs.analchem.8b00780.

3. Li J., Zhang L., Cheng X., Zhang L., Shen B., Qing C., Fan G. Determination of d-amphetamine and diphenhydramine in beagle dog plasma by a 96-well formatted liquid-liquid extraction and capillary zone electrophoresis with field-amplified sample stacking. J. Pharm. Biomed. Anal. 2018; 156: 263–271. DOI: 10.1016/j.jpba.2018.04.040.

4. Brandhorst G., Oellerich M., Maine G., Taylor P., Veen G., Wallemacq P. Liquid chromatography–tandem mass spectrometry or automated immunoassays: what are the future trends in therapeutic drug monitoring? Clin. Chem. 2012; 58(5): 821–825. DOI: 10.1373/clinchem.2011.167189.

5. Aubry A. F. LC-MS/MS bioanalytical challenge: ultra-high sensitivity assays. Bioanalysis. 2011; 3(16): 1819–1825. DOI: 10.4155/bio.11.166.

6. Karinen R., Øiestad E. L., Andresen W., Smith-Kiell A., Christophersen A. Comparison of the stability of stock solutions of drugs of abuse and other drugs stored in a freezer, refrigerator, and at ambient temperature for up to one year. J. Anal. Toxicol. 2011; 35(8): 583–590.

7. Miroshnichenko I. I., Sokolova G. B., Mokhireva L. V. Clinical pharmacokinetics of para-aminosalicylic acid tablets. Antibiot. Khimioter. 2009; 54(1-2): 20–24 (in Russ.).

8. Zhou J., Li F., Duggan J. X. LC–MS bioanalysis of acyl glucuronides. In. Handbook of LC-MS Bioanalysis: Best Practices, Experimental Protocols, and Regulations. Li W., Zhang J., Tse F.L.S. (Eds). John Wiley & Sons, Inc., Hokoben, NJ, USA. 2013; 447–460. https://doi.org/10.1002/9781118671276.ch35.

9. Miroshnichenko I. I., Yurchenko N. I. HPLC Analysis for omeprazole and lansoprazole in blood plasma. Pharmaceutical Chemistry Journal. 2002; 36(7): 389–391 (in Russ).

10. Singleton C. Recent advances in bioanalytical sample preparation for LC-MS analysis. Bioanalysis. 2012; 4(9): 1123–1140. DOI: 10.4155/bio.12.73.

11. Musteata F. M. Extraction and microextraction in bioanalysis. Bioanalysis. 2012; 4(19): 2321–2323. DOI: 10.4155/bio.12.206.

12. Sonawane L. V., Poul B. N., Usnale S. V., Waghmare P. V., Surwase L. H. Bioanalytical method validation and its pharmaceutical application – a review. Pharm. Anal. Acta. 2014; 5(3): 288–295.

13. Schellekens R. C., Stellaard F., Woerdenbag H. J., Frijlink H. W., Kosterink J. G. Applications of stable isotopes in clinical pharmacology. Br. J. Clin. Pharmacol. 2011; 72(6): 879–897. DOI: 10.1111/j.1365-2125.2011.04071.x.

14. George R., Haywood A., Khan S., Radovanovic M., Simmonds J., Norris R. Enhancement and suppression of ionization in drug analysis using HPLC-MS/MS in support of therapeutic drug monitoring: a review of current knowledge of its minimization and assessment. Ther. Drug. Monit. 2018; 40(1): 1–8. DOI: 10.1097/FTD.0000000000000471.

15. Zhou W., Yang S., Wang P. G. Matrix effects and application of matrix effect factor. Bioanalysis. 2017; 9(23): 1839–1844. DOI: 10.4155/bio-2017-0214.

16. Larger P. J., Breda M., Fraier D., Hughes H., James C. A. Ion suppression effects in liquid chromatography-tandem mass spectrometry due to a formulation agent, a case study in drug discovery bioanalysis. J. Pharm. Biomed. Anal. 2005; 39(1–2): 206–216. DOI: 10.1016/j.jpba.2005.03.009.

17. Sotgia S., Murphy R. B., Zinellu A., Elliot D., Paliogiannis P., Pinna G. A., Carru C., Mangoni A. A. Development of an LC-tandem mass spectrometry method for the quantitative analysis of hercynine in human whole blood. Molecules. 2018; 23(12): E3326. DOI: 10.3390/molecules23123326.

18. Bateman K. P., Cohen L., Emary B., Pucci V. Standardized workflows for increasing efficiency and productivity in discovery stage bioanalysis. Bioanalysis. 2013; 5(14): 1783–1794. DOI: 10.4155/bio.13.162.

19. Swales J. G., Tucker J. W., Strittmatter N., Nilsson A., Cobice D., Clench M. R., Mackay C. L., Andren P. E., Takáts Z., Webborn P. J., Goodwin R. J. Mass spectrometry imaging of cassette-dosed drugs for higher throughput pharmacokinetic and biodistribution analysis. Anal. Chem. 2014; 86(16): 8473–8080. DOI: 10.1021/ac502217r.

20. Ho S. Best practices for discovery bioanalysis: balancing data quality and productivity. Bioanalysis. 2014; 6(20): 2705–2708. DOI: 10.4155/bio.14.201.