Abstract

Background: There continues to be consistent pressure for bioanalytical scientists to achieve lower limits of quantitation. The reasons range from smaller sample volumes available for analysis, to more potent analytes and the growth of biologics in drug development. This has led scientists to investigate alternative LC techniques, including microflow and nanoflow. These techniques have been shown to increase sensitivity of electrospray methods and reduce ionization matrix effects. Because high-resolution MS has significant benefits for the analysis of biologics, this type of mass spectrometer is becoming increasingly important in bioanalysis. Results: For microflow analysis, a new ion source and significant extra sample preparation or chromatographic separation are not required. However, increased sensitivity and reduced matrix effects were consistently demonstrated when compared with UHPLC flow rates. The extent of matrix effects observed were compound dependent. Discussion: This paper presents the utility of combining high-resolution/accurate mass with microflow LC from a quantitative standpoint. This includes evaluating the typical quantitative parameters of sensitivity, linearity/dynamic range, precision and accuracy. It also includes the evaluation of changes in signal suppression using microflow LC and microspray ionization. The benefits and disadvantages of using the combination of these two technologies for quantitative bioanalysis are also discussed.

References

1 Baranowska I, Magiera S, Baranowski J. Clinical applications of fast liquid chromatography: a review on the analysis of cardiovascular drugs and their metabolites. J. Chromatogr. B Analyt. Technol. Biomed. Life Sci. doi:10.1016/j.jchromb.02.002 (Epub ahead of print) (2013).
Google Scholar
2 Nováková L, Vlcková H. A review of current trends and advances in modern bio-analytical methods: chromatography and sample preparation. Anal. Chim. Acta656(1–2),8–35 (2009).
Medline CASGoogle Scholar
3 Makarov A, Scigelova M. Coupling liquid chromatography to Orbitrap mass spectrometry. J. Chromatogr. A1217(25),3938–3945 (2010).
Medline CASGoogle Scholar
4 Valaskovic GA, Utley L, Lee MS, Wu JT. Ultra-low flow nanospray for the normalization of conventional liquid chromatography/mass spectrometry through equimolar response: standard-free quantitative estimation of metabolite levels in drug discovery. Rapid Commun. Mass Spectrom.20(7),1087–1096 (2006).
Medline CASGoogle Scholar
5 Chelius D, Zhang T, Wang GH, Shen RF. Global protein identification and quantification technology using two dimensional liquid chromatography nanospray mass spectrometry. Anal. Chem.75(23),6658–6665 (2003).
Medline CASGoogle Scholar
6 Hall TG, Smukste I, Bresciano KR, Wang YX, McKearn D, Savage RE. Chapter 19. Identifying and overcoming matrix effects in drug discovery and development. In: Tandem Mass Spectrometry – Applications and Principles. Prasain J (Ed.). ArQule Inc., MA, USA, 389–420 (2012).
Google Scholar
7 Gangl ET, Annan MM, Spooner N, Vouros P. Reduction of signal suppression effects in ESI-MS using a nanosplitting device. Anal. Chem.73(23),5635–5644 (2002).
Google Scholar
8 Percy AJ, Chambers AG, Yang J, Domanski D, Borchers CH. Comparison of standard- and nano-flow liquid chromatography platforms for MRM-based quantitation of putative plasma biomarker proteins. Anal. Bioanal. Chem.404(4),1089–1101 (2012).
Medline CASGoogle Scholar
9 Chen J, Yang L, Kapron JT et al. Determination of SCH 211803 by nanoelectrospray infusion mass spectrometry: evaluation of matrix effect and comparison with liquid chromatography–tandem mass spectrometry. J. Chromatogr. B Analyt. Technol. Biomed. Life Sci.809(2),205–210 (2004).
Medline CASGoogle Scholar
10 He X, Kozak M, Nimkar S. Ultra-sensitive measurements of 11-Nor-δ (9) tetrahydrocannabinol-9-carboxylic acid in oral fluid by microflow liquid chromatography–tandem mass spectrometry using a benchtop quadrupole/orbitrap mass spectrometer. Anal. Chem.84(18),7643–7647 (2012).
Medline CASGoogle Scholar
11 Hatsis P, Brockman AH, Wu JT. Evaluation of high-field asymmetric waveform ion mobility spectrometry coupled to nanoelectrospray ionization for bioanalysis in drug discovery. Rapid Commun. Mass Spectrom.21(14),2295–2300 (2007).
Medline CASGoogle Scholar
12 Chambers E, Wagnowski-Diehl DM, Lu ZL, Mazzeo JR. Systematic and comprehensive strategy for reducing matrix effects in LC–MS/MS analysis. J. Chromatogr. B852(1–2),22–34 (2007).
Medline CASGoogle Scholar
13 Little J, Wempe M, Buchanan C. Liquid chromatography–mass spectrometry/mass spectrometry method development for drug metabolism studies: examining lipid matrix ionization effects in plasma. J. Chromatogr. B833(2),219–230 (2006).
Medline CASGoogle Scholar
14 Bennett P, Meng M. Source for imprecision resulting from ionization suppression from strongly retained phospholipids and dioctyl phthalate. Presented at: 52th ASMS Conference on Mass Spectrometry and Allied Topics. Nashville, TN, USA, 23–27 May 2004.
Google Scholar
15 Bennett P, Van Horne C. Identification of the major endogenous and persistent compounds in plasma, serum, and tissue that cause matrix effects with electrospray LC/MS techniques. Presented at: AAPS Annual Meeting and Exposition. Salt Lake City, UT, USA, 26–30 October 2003.
Google Scholar
16 Mei H, Hsieh Y, Nardo C et al. Investigation of matrix effects in bioanalytical high-performance liquid chromatography/tandem mass spectrometric assays: Application to drug discovery. Rapid Commun. Mass Spectrom17,97–103 (2003).
Medline CASGoogle Scholar
17 Jemal M, Ouyang Z, Yuan-Qing X. Systematic LC–MS/MS bioanalytical method development that incorporates plasma phospholipids risk avoidance, usage of incurred sample and well thought-out chromatography. Biomed. Chromatogr.24,2–19 (2010).
Medline CASGoogle Scholar
18 Beck A, Wagner-Rousset E, Ayoub D, Van Dorsselaer A, Sanglier-Cianférani S. Characterization of therapeutic antibodies and related products. Anal. Chem.85(2),715–736 (2013).
Medline CASGoogle Scholar
19 Schadt S, Chen LZ, Bischoff D. Evaluation of relative LC/MS response of metabolites to parent drug in LC/nanospray ionization mass spectrometry: potential implications in MIST assessment. J. Mass Spectrom.46(12),1281–1286 (2011).
MedlineGoogle Scholar
20 Ruan Q, Ji QC, Arnold ME, Humphreys GW, Zhu MS. Strategy and its implications of protein bioanalysis utilizing high-resolution mass spectrometric detection of intact protein. Anal. Chem.83(23),8937–8944 (2011).
Medline CASGoogle Scholar
21 Peterson AC, Russell JD, Bailey DJ, Westphall MS, Coon JJ. Parallel reaction monitoring for high-resolution and high mass accuracy quantitative, targeted proteomics. Mol. Cell. Proteomics11(11),1475–1488 (2012).
Medline CASGoogle Scholar
22 Gallien S, Duriez E, Demeure K, Domon B. Selectivity of LC–MS/MS analysis: implication for proteomics experiments. J. Proteomics81,148–158 (2013).
Medline CASGoogle Scholar
23 Fung EN, Xia YQ, Aubry AF, Zeng J, Olah T, Jemal M. Full-scan high-resolution accurate mass spectrometry (HRMS) in regulated bioanalysis: LC–HRMS for the quantitation of prednisone and prednisolone in human plasma. J. Chromatogr. B Analyt. Technol. Biomed. Life Sci.879(27),2919–2927 (2011).
Medline CASGoogle Scholar
24 Kaufmann A, Butcher P, Maden K, Walker S, Widmer M. Comprehensive comparison of liquid chromatography selectivity as provided by two types of liquid chromatography detectors (high-resolution mass spectrometry and tandem mass spectrometry): ‘Where is the crossover point?’ Anal. Chim. Acta700(1–2),86–94 (2011).
Medline CASGoogle Scholar
25 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.25,2863–2878 (2011).
Medline CASGoogle Scholar
26 Bateman KP, Kellmann M, Muenster H, Papp R, Taylor L. Quantitative–qualitative data acquisition using a benchtop Orbitrap mass spectrometer. J. Am. Soc. Mass Spectrom.20(8),1441–1450 (2009).
Medline CASGoogle Scholar
27 Pinhancos R, Maass S, Ramanathan DM. High-resolution mass spectrometry method for the detection,characterization and quantitation of pharmaceuticals in water. J. Mass Spectrom.46(11),1175–1181 (2011).
Medline CASGoogle Scholar
28 Dolan JW. The role of the signal-to-noise ratio in precision and accuracy. LCGC North America23(12),1256 (2005).
CASGoogle Scholar
29 Matuszewski BK, Constanzer ML, Chavez-Eng CM. A systematic approach to reducing matrix effects in LC–MS/MS analyses. Anal. Chem.75,3019–3030 (2003).
Medline CASGoogle Scholar
30 Shen JX, Motyka RJ, Roach JP, Hayes RN. Minimization of ion suppression in LC–MS/MS analysis through the application of strong cation exchange solid-phase extraction (SCX-SPE). J. Pharm. Biomed. Anal.37,359–367 (2005).
Medline CASGoogle Scholar
31 Du L, White RL. Reducing glycerophosphocholine lipid matrix interference effects in biological fluid assays by using high-turbulance liquid chromatography. Rapid Commun. Mass Spectrom.22,3362–3370 (2008).
Medline CASGoogle Scholar
32 Liang HR, Foltz RL, Meng M, Bennett P. Ionization enhancement in atmospheric pressure chemical ionization and suppression in electrospray ionization between target drugs and stable-isotope-labeled internal standards in quantitative liquid chromatography/tandem mass spectrometry. Rapid Commun. Mass Spectrom.17,2815–2821 (2003).
Medline CASGoogle Scholar
33 Shou WZ, Naidong W. Post-column infusion study of the ‘dosing vehicle effect’ in the liquid chromatography/tandem mass spectrometric analysis of discovery pharmacokinetic samples. Rapid Commun. Mass Spectrom.17,589–597 (2003).
Medline CASGoogle Scholar
34 Xu X, Mei H, Wang S et al. A study of common discovery dosing formulation components and their potential for causing time-dependent matrix effects in high-performance liquid chromatography tandem mass spectrometry assays. Rapid Commun. Mass Spectrom.19(18),2643–2650 (2005).
Medline CASGoogle Scholar
35 Bonfiglio R, King RC, Olah TV, Merkle K. The effects of sample preparation methods on the variability of the electrospray ionization response for model drug compounds. Rapid Commun. Mass Spectrom.13(12),1175–1185 (1999).
Medline CASGoogle Scholar

Vol. 5, No. 10

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Published online 30 May 2013

Published in print May 2013

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Financial & competing interests disclosure

The authors have no relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript. This includes employment, consultancies, honoraria, stock ownership or options, expert testimony, grants or patents received or pending, or royalties.

No writing assistance was utilized in the production of this manuscript.

Ethical conduct of research

The authors state that they have obtained appropriate institutional review board approval or have followed the principles outlined in the Declaration of Helsinki for all human or animal experimental investigations. In addition, for investigations involving human subjects, informed consent has been obtained from the participants involved.

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Performance assessment of microflow LC combined with high-resolution MS in bioanalysis

Abstract

References