We use cookies to improve your experience. By continuing to browse this site, you accept our cookie policy.×
Research Article

Characterization of matrix effects in developing rugged high-throughput LC–MS/MS methods for bioanalysis

    Fumin Li

    *Author for correspondence:

    E-mail Address: fumin.li@ppdi.com

    PPD Laboratories, 3230 Deming Way, Middleton, WI 53562, USA

    ,
    Jun Wang

    PPD Laboratories, 3230 Deming Way, Middleton, WI 53562, USA

    &
    Rand Jenkins

    PPD Laboratories, 2240 Dabney Rd, Richmond, VA 23230, USA

    Published Online:https://doi.org/10.4155/bio-2016-0005

    Aim: There is an ever-increasing demand for high-throughput LC–MS/MS bioanalytical assays to support drug discovery and development. Results: Matrix effects of sofosbuvir (protonated) and paclitaxel (sodiated) were thoroughly evaluated using high-throughput chromatography (defined as having a run time ≤1 min) under 14 elution conditions with extracts from protein precipitation, liquid–liquid extraction and solid-phase extraction. A slight separation, in terms of retention time, between underlying matrix components and sofosbuvir/paclitaxel can greatly alleviate matrix effects. Conclusion: High-throughput chromatography, with proper optimization, can provide rapid and effective chromatographic separation under 1 min to alleviate matrix effects and enhance assay ruggedness for regulated bioanalysis.

    Papers of special note have been highlighted as: • of interest; •• of considerable interest

    References

    • 1 Ackermann BL, Berna MJ, Murphy AT. Recent advances in use of LC/MS/MS for quantitative high-throughput bioanalytical support of drug discovery. Curr. Top. Med. Chem. 2(1), 53–66 (2002).
    • 2 Balani SK, Miwa GT, Gan LS, Wu JT, Lee FW. Strategy of utilizing in vitro and in vivo ADME tools for lead optimization and drug candidate selection. Curr. Top. Med. Chem. 5(11), 1033–1038 (2005).
    • 3 Jemal M, Xia YQ. LC–MS Development strategies for quantitative bioanalysis. Curr. Drug. Metab. 7(5), 491–502 (2006).
    • 4 Swartz ME. UPLC: an introduction and review. J. Liquid. Chromatogr. Relat. Technol. 28(7–8), 1253–1263 (2005).
    • 5 Churchwell MI, Twaddle NC, Meeker LR, Doerge DR. Improving LC–MS sensitivity through increased in chromatographic performance: comparisons of UPLC-ES/MS/MS to HPLC-ES/MS/MS. J. Chromatogr. B Analyt. Technol. Biomed. Life Sci. 825(2), 134–143 (2005).
    • 6 Yu K, Little D, Plumb R, Smith B. High-throughput quantification for a drug mixture in rat plasma-a comparison of Ultra Performance liquid chromatography/tandem mass spectrometry with high-performance liquid chromatography/tandem mass spectrometry. Rapid Commun. Mass Spectrom. 20(4), 544–552 (2006).
    • 7 Hsieh Y, Duncan CJ, Brisson JM. Fused-core silica column high-performance liquid chromatography/tandem mass spectrometric determination of rimonabant in mouse plasma. Anal. Chem. 79(15), 5668–5673 (2007).
    • 8 Mallett DN, Ramirez-Molina C. The use of partially porous particle columns for the routine, generic analysis of biological samples for pharmacokinetic studies in drug discovery by reversed-phase ultra-high performance liquid chromatography-tandem mass spectrometry. J. Pharm. Biomed. Anal. 49(1), 100–107 (2009). • Utility of high flow monolithic chromatography in high-throughput bioanalysis.
    • 9 Deng Y, Wu JT, Lloyd TL, Chi CL, Olah TV, Unger SE. High-speed liquid chromatography/tandem mass spectrometry using a monolithic column for high-throughput bioanalysis. Rapid Commun. Mass Spectrom. 16(11), 1116–1123 (2002).
    • 10 Huang MQ, Mao Y, Jemal M, Arnold M. Increased productivity in quantitative bioanalysis using a monolithic column coupled with high-flow direct-injection liquid chromatography/tandem mass spectrometry. Rapid Commun. Mass Spectrom. 20(11), 1709–1714 (2006).
    • 11 Xu RN, Fan L, Rieser MJ, El-Shourbagy TA. Recent advances in high-throughput quantitative bioanalysis by LC–MS/MS. J. Pharm. Biomed. Anal. 44(2), 342–345 (2007).
    • 12 Korfmacher WA, Palmer CA, Nardo C et al. Development of an automated mass spectrometry system for the quantitative analysis of liver microsomal incubation samples: a tool for rapid screening of new compounds for metabolic stability. Rapid Commun. Mass Spectrom. 13(10), 901–907 (1999).
    • 13 Jian W, Edom RW, Xu Y, Weng N. Recent advances in application of hydrophilic interaction chromatography for quantitative bioanalysis. J. Sep Sci. 33(6–7), 681–697 (2010). • Bioanalytical applications of ambient desorption ionization mass spectrometry (DART, DESI).
    • 14 Chernetsova ES, Morlock GE. Ambient desorption ionization mass spectrometry (DART, DESI) and its bioanalytical applications. Bioanalyt. Rev. 3(1), 1–9 (2011).
    • 15 Harris GA, Galhena AS, Fernández FM. Ambient sampling/ionization mass spectrometry: applications and current trends. Anal. Chem. 83(12), 4508–4538 (2011).
    • 16 Chen H, Talaty NN, Takáts Z, Cooks RG. Desorption electrospray ionization mass spectrometry for high-throughput analysis of pharmaceutical samples in the ambient environment. Anal. Chem. 77(21), 6915–6927 (2005).
    • 17 Manicke NE, Kistler T, Ifa DR, Cooks RG, Ouyang Z. High-throughput quantitative analysis by desorption electrospray ionization mass spectrometry. J. Am. Soc. Mass Spectrom. 20(2), 321–325 (2009). • Evaluation of laser diode thermal desorption (LDTD)-MS/MS for high-throughput discovery bioanalysis.
    • 18 Beattie I, Smith A, Weston DJ, White P, Szwandt S, Sealey L. Evaluation of laser diode thermal desorption (LDTD) coupled with tandem mass spectrometry (MS/MS) for support of in vitro drug discovery assays: Increasing scope, robustness and throughput of the LDTD technique for use with chemically diverse compound libraries. J. Pharm. Biomed. Anal. 59, 18–28 (2012).
    • 19 Wagner AD, Kolb JM, Ozbal CC et al. Ultrafast mass spectrometry based bioanalytical method for digoxin supporting an in vitro P-glycoprotein (P-gp) inhibition screen. Rapid Commun. Mass Spectrom. 25(9), 1231–1240 (2011). • Evaluation a high-throughput SPE-MS/MS strategy for bioanalysis with analysis time of approximately 10 s/sample.
    • 20 Jian W, Romm MV, Edom RW, Miller VP, LaMarr WA, Weng N. Evaluation of a high-throughput online solid phase extraction–tandem mass spectrometry system for in vivo bioanalytical studies. Anal. Chem. 83(21), 8259–8266 (2011).
    • 21 Ramanathan R, Jemal M, Ramagiri S et al. It is time for a paradigm shift in drug discovery bioanalysis: from SRM to HRMS. J. Mass Spectrom. 46(6), 595–601 (2011).
    • 22 Fung EN, Jemal M, Aubry AF. High-resolution MS in regulated bioanalysis: where are we now and where do we go from here? Bioanalysis 5(10), 1277–1284 (2013).
    • 23 Xia YQ, Lau J, Olah T, Jemal M. Targeted quantitative bioanalysis in plasma using liquid chromatography/high-resolution accurate mass spectrometry: an evaluation of global selectivity as a function of mass resolving power and extraction window, with comparison of centroid and profile modes. Rapid Commun. Mass Spectrom. 25(19), 2863–2878 (2011).
    • 24 Ding J, Lund ET, Zulkoski J, Lindsay JP, McKenzie DL. High-throughput bioanalysis of bile acids and their conjugates using UHPLC coupled to HRMS. Bioanalysis 5(20), 2481–2494 (2013).
    • 25 Murphy K, Bennett PK, Duczak N Jr. High-throughput quantitation of large molecules using multiplexed chromatography and high-resolution/accurate mass LC–MS. Bioanalysis 4(9), 1013–1024 (2012).
    • 26 Heudi O. High-throughput quantitative analysis of pharmaceutical compounds in biological matrices. Bioanalysis 3(8), 819–821 (2011).
    • 27 European Medicines Agency. Guideline on bioanalytical method validation. www.ema.europa.eu/docs/en_GB/document_library/Scientific_guideline/2011/08/WC500109686.pdf.
    • 28 US FDA Guidence for Industry. Bioanalytical Method Validation. www.fda.gov/downloads/Drugs/./Guidances/ucm070107.pdf.
    • 29 Bergeron A, Garofolo F. Importance of matrix effects in LC–MS/MS bioanalysis. Bioanalysis 5(19), 2331–2332 (2013). •• Comprehensive review of matrix effects in LC–MS-based bioanalysis.
    • 30 Li F, Ewles M, Pelzer M et al. Case studies: the impact of non-analyte components on LC–MS/MS based bioanalysis – strategies for identifying and overcoming matrix effects. Bioanalysis 5(19), 2409–2441 (2013). •• Useful strategies to reduce matrix effects in LC–MS-based bioanalysis.
    • 31 Chambers E, Wagrowski-Diehl DM, Lu Z, Mazzeo JR. Systematic and comprehensive strategy for reducing matrix effects in LC/MS/MS analyses. J. Chromatogr. B 852(1–2), 22–34 (2007).
    • 32 Xia YQ, Jemal M. Phospholipids in liquid chromatography/mass spectrometry bioanalysis: comparison of three tandem mass spectrometric techniques for monitoring plasma phospholipids, the effect of mobile phase composition on phospholipids elution and the association of phospholipids with matrix effects. Rapid Commun. Mass Spectrom. 23(14), 2125–2138 (2009).
    • 33 Taylor PJ. Matrix effects: the Achilles heel of quantitative high-performance liquid chromatography–electrospray–tandem mass spectrometry. Clin. Biochem. 38(4), 328–334 (2005).
    • 34 Gosetti F, Mazzucco E, Zampieri D, Gennaro MC. Signal suppression/enhancement in high-performance liquid chromatography tandem mass spectrometry. J. Chromatogr. A 1217(25), 3929–3937 (2010).
    • 35 Trufelli H, Palma P, Famiglini G, Cappiello A. An overview of matrix effects in liquid chromatography-mass spectrometry. Mass Spectrom. Rev. 30(3), 491–509 (2011). • Useful information on the causes of ion suppression in LC–MS analysis and strategies to address it.
    • 36 Furey A, Moriarty M, Bane V, Kinsella B, Lehane M. Ion suppression: a critical review on causes, evaluation, prevention and applications. Talanta 115, 104–122 (2013). •• Demonstration of high-throughput (e.g., sub-minute) LC–MS methods to support regulated bioanalysis.
    • 37 Badman ER, Beardsley RL, Liang Z, Bansal S. Accelerating high quality bioanalytical LC/MS/MS assays using fused-core columns. J. Chromatogr. B Analyt. Technol. Biomed. Life Sci. 878(25), 2307–2313 (2010).
    • 38 Ye Z, Tsao H, Gao H, Brummel CL. Minimizing matrix effects while preserving throughput in LC–MS/MS bioanalysis. Bioanalysis 3(14), 1587–1601 (2011).
    • 39 Chromatography Online. Glossary of HPLC/LC Separation Terms. www.chromatographyonline.com/glossary-hplclc-separation-terms.
    • 40 Hawkridge AM. Chapter 1: practical considerations and current limitations. in quantitative mass spectrometry-based proteomics. In: Quantitative Proteomics. Claire E Eyers, Simon Gaskell (Eds). Royal Society of Chemistry, UK (2014).
    • 41 Tang K, Page JS, Smith RD. Charge competition and the linear dynamic range of detection in electrospray ionization mass spectrometry. J. Am. Soc. Mass Spectrom. 15(10), 1416–1423 (2004).
    • 42 Golay MJE. The hetp of an unretained component of fluid flow in a coiled round pipe. J. Chromatogr. A 186, 341–351 (1979).
    • 43 Zheng J, Patel D, Tang Q, Markovich RJ, Rustum AM. Comparison study of porous, fused-core, and monolithic silica-based C18 HPLC columns for Celestoderm-V ointment analysis. J. Pharm. Biomed. Anal. 50(5), 815–822 (2009).
    • 44 Jiang H, Ouyang Z, Zeng J et al. A user-friendly robotic sample preparation program for fully automated biological sample pipetting and dilution to benefit the regulated bioanalysis. J. Lab Autom. 17(3), 211–221 (2012).
    • 45 Yu S, Crawford E, Tice J, Musselman B, Wu JT. Bioanalysis without sample cleanup or chromatography: the evaluation and initial implementation of direct analysis in real time ionization mass spectrometry for the quantification of drugs in biological matrixes. Anal. Chem. 81(1), 193–202 (2009). • Demonstration of high-throughput DESI MS in pharmaceutical analysis.