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Published Online:https://doi.org/10.4155/bio.11.306

Chemical derivatization is the only technique that directly affects the physicochemical property of an analyte in an LC–MS/MS assay platform. On the other side, current MS instruments are extremely sensitive, but still their absolute sensitivity is analyte-dependent. In this review, first, difficulty in analyzing neutral compounds and introducing acidity/basicity and/or proton affinity will be described. Second, the sweet spot of the conventional MS ion source across clogP values is presented. Third, optimization of hydrophilicity/hydrophobicity by derivatization is described. Lastly, development of a new derivatizing reagent specifically designed for LC–MS/MS is described and its significance in pharmacokinetic analysis is discussed.

Papers of special note have been highlighted as: ▪ of interest

References

  • Shah VP, Bansal S. Historical perspective on the development and evolution of bioanalytical guidance and technology. Bioanalysis3(8),823–827 (2011).
  • Johann Jr DJ, Rodriguez-Canales J, Mukherjee S et al. Approaching solid tumor heterogeneity on a cellular basis by tissue proteomics using laser capture microdissection and biological mass spectrometry. J. Proteome. Res.8(5),2310–2318 (2009).
  • Yamane N, Tozuka Z, Kusama M, Maeda K, Ikeda T, Sugiyama Y. Clinical relevance of liquid chromatography–tandem mass spectrometry as an analytical method in microdose clinical studies. Pharm. Res.28(8),1–10 (2011).▪ Example of trace level quantification by LC–MS/MS in a microdose clinical trial.
  • Li Y, Xu LL, Ruan JX, Zhang ZQ. Research progress of enhancing quantitative sensitivity by using LC–MSn with derivatization method in bio-matrices. Yaoxue Xuebao46(6),637–641 (2011).
  • Xu F, Zou L, Liu Y, Zhang Z, Ong CN. Enhancement of the capabilities of liquid chromatography– mass spectrometry with derivatization: general principles and applications. Mass Spectrom. Rev.30(6),1143–1172 (2011).▪ Summarizes derivatization reactions that enhance LC–MS/MS sensitivity.
  • Iwasaki Y, Nakano Y, Mochizuki K et al. A new strategy for ionization enhancement by derivatization for mass spectrometry. J. Chromatogr. B879(17–18),1159–1165 (2011).
  • Wang K, Nano M, Mulligan T, Bush ED, Edom RW. Derivatization of 5-fluorouracil with 4-bromomethyl-7-methoxycoumarin for determination by liquid chromatography–mass spectrometry. J. Am. Soc. Mass Spectrom.9(9),970–976 (1998).
  • Licea-Perez H, Wang S, Bowen C. Development of a sensitive and selective LC–MS/MS method for the determination of α-fluoro-β-alanine, 5-fluorouracil and capecitabine in human plasma. J. Chromatogr. B877(11–12),1040–1046 (2009).▪ Improvement in chromatographic behavior by derivatization.
  • Niwa M, Kawashiro T. Applicability of atmospheric pressure photoionization mass spectrometry to trace analysis of hydrophilic drugs. Yakugaku Zasshi129(8),993–999 (2009).
  • 10  Cech NB, Enke CG. Relating electrospray ionization response to nonpolar character of small peptides. Anal. Chem.72(13),2717–2723 (2000).
  • 11  Cech NB, Krone JR, Enke CG. Predicting electrospray response from chromatographic retention time. Anal. Chem.73(2),208–213 (2001).
  • 12  Ghose AK, Viswanadhan VN, Wendoloski JJ. A knowledge-based approach in designing combinatorial or medicinal chemistry libraries for drug discovery. 1. A qualitative and quantitative characterization of known drug databases. J. Comb. Chem.1(1),55–68 (1999).
  • 13  Lipinski CA, Lombardo F, Dominy BW, Feeney PJ. Experimental and computational approaches to estimate solubility and permeability in drug discovery and development settings. Adv. Drug Deliv. Rev.23(1–3),3–25 (1997).
  • 14  Liu DQ, Hop CECA. Strategies for characterization of drug metabolites using liquid chromatography–tandem mass spectrometry in conjunction with chemical derivatization and on-line H/D exchange approaches. J. Pharm. Biomed. Anal.37(1),1–18 (2005).▪ Summarizes derivatization strategy for metaboite analysis by LC–MS/MS.
  • 15  Prakash C, Shaffer CL, Nedderman A. Analytical strategies for identifying drug metabolites. Mass Spectrom. Rev.26(3),340–369 (2007).▪ Summarizes derivatization of hydrophilic metabolites for LC–MS/MS.
  • 16  Tsume Y, Provoda CJ, Amidon GL. The achievement of mass balance by simultaneous quantification of floxuridine prodrug, floxuridine, 5-fluorouracil, 5-dihydrouracil, α-fluoro-β-ureidopropionate, α-fluoro-β-alanine using LC–MS. J. Chromatogr. B; 879(13–14),915–920 (2011).
  • 17  Liu K, Zhong D, Zou H, Chen X. Determination of tegafur, 5-fluorouracil, gimeracil and oxonic acid in human plasma using liquid chromatography–tandem mass spectrometry. J. Pharm. Biomed. Anal.52(4),550–556 (2010).
  • 18  Higashi T, Yamauchi A, Shimada K. 2-Hydrazino-1-methylpyridine: a highly sensitive derivatization reagent for oxosteroids in liquid chromatography–electrospray ionization-mass spectrometry. J. Chromatogr. B825(2),214–222 (2005).
  • 19  Inoda H, Nishiyama T, Yoshikado T, Suwanai Y, Santa T. Compounds having thiourea moiety as derivatization reagents in liquid chromatography/electrospray ionization-tandem mass spectrometry (LC–ESI-MS/MS): synthesis of derivatization reagents for carboxylic acids. Biomed. Chromatogr.25(6),635–640 (2011).
  • 20  Kalhorn TF, Page ST, Howald WN, Mostaghel EA, Nelson PS. Analysis of testosterone and dihydrotestosterone from biological fluids as the oxime derivatives using high-performance liquid chromatography/tandem mass spectrometry. Rapid Commun. Mass Spectrom.21(19),3200–3206 (2007).
  • 21  Santa T. Derivatization reagents in liquid chromatography/electrospray ionization tandem mass spectrometry. Biomed. Chromatogr.25(1–2),1–10 (2011).
  • 22  Higashi T. Trace determination of steroids causing age-related diseases using LC–MS combined with detection-oriented derivatization. Chem. Pharm. Bull.54(11),1479–1485 (2006).
  • 23  Higashi T, Ninomiya Y, Shimada K. Studies on neurosteroids XX. Liquid chromatography–tandem mass spectrometric method for simultaneous determination of testosterone and 5α-dihydrotestosterone in rat brain and serum. J. Chromatogr. Sci.46(7),653–658 (2008).
  • 24  Yamashita K, Yamazaki K, Komatsu S, Numazawa M. Fusaric acid as a novel proton-affinitive derivatizing reagent for highly sensitive quantification of hydroxysteroids by LC–ESI-MS/MS. J. Am. Soc. Mass Spectrom.21(2),249–253 (2010).
  • 25  Eggink M, Wijtmans M, Kretschmer A et al. Targeted LC–MS derivatization for aldehydes and carboxylic acids with a new derivatization agent 4-APEBA. Anal. Bioanal. Chem.397(2),665–675 (2010).
  • 26  Cech NB, Enke CG. Effect of affinity for droplet surfaces on the fraction of analyte molecules charged during electrospray droplet fission. Anal. Chem.73(19),4632–4639 (2001).
  • 27  Zhou S, Cook KD. A mechanistic study of electrospray mass spectrometry: charge gradients within electrospray droplets and their influence on ion response. J. Am. Soc. Mass Spectrom.12(2),206–214 (2001).
  • 28  Center for Drug Evaluation and Research, Food and Drug Administration. Guidance for Industry: safety testing of drug metabolites. (2008).
  • 29  Shibata Y, Ito K, Suzuki K et al. Changes in the endocrine environment of the human prostate transition zone with aging: simultaneous quantitative analysis of prostatic sex steroids and comparison with human prostatic histological composition. Prostate42(1),45–55 (2000).
  • 30  Mitamura K, Shimada K. Derivatization in liquid chromatography/mass spectrometric analysis of neurosteroids. Chromatography22(1),11–15 (2001).
  • 31  Xu X, Ziegler RG, Waterhouse DJ, Saavedra JE, Keefer LK. Stable isotope dilution high-performance liquid chromatography–electrospray ionization mass spectrometry method for endogenous 2- and 4-hydroxyestrones in human urine. J. Chromatogr. B780(2),315–330 (2002).
  • 32  Yamashita K, Miyashiro Y, Maekubo H et al. Development of highly sensitive quantification method for testosterone and dihydrotestosterone in human serum and prostate tissue by liquid chromatography–electrospray ionization tandem mass spectrometry. Steroids74(12),920–926 (2009).▪ Relations between MS response and basicity, proton affinity and hydrophobicity.
  • 33  Higashi T, Ichikawa T, Inagaki S, Min JZ, Fukushima T, Toyo’oka T. Simple and practical derivatization procedure for enhanced detection of carboxylic acids in liquid chromatography–electrospray ionization-tandem mass spectrometry. J. Pharm. Biomed. Anal.52(5),809–818 (2010).
  • 34  Nakagawa Y, Hashimoto Y. Polar Derivatization of 5α-dihydrotestosterone and sensitive analysis by semimicro-LC/ESI-MS. J. Mass Spectrom. Soc. Jpn.50(6),330–336 (2002).
  • 35  Nishio T, Higashi T, Funaishi A, Tanaka J, Shimada K. Development and application of electrospray-active derivatization reagents for hydroxysteroids. J. Pharm. Biomed. Anal.44(3),786–795 (2007).
  • 36  Anari MR, Bakhtiar R, Zhu B, Huskey S, Franklin RB, Evans DC. Derivatization of ethinylestradiol with dansyl chloride to enhance electrospray ionization: application in trace analysis of ethinylestradiol in rhesus monkey plasma. Anal. Chem.74(16),4136–4144 (2002).
  • 37  Niwa M, Watanabe N, Ochiai H, Yamashita K. Determination of testosterone concentrations in rat plasma using liquid chromatography–atmospheric pressure chemical ionization mass spectrometry combined with ethyl oxime and acetyl ester derivatization. J. Chromatogr. B824(1–2),258–266 (2005).
  • 38  You J, Shi Y, Zhao X et al. Enhancement of atmospheric pressure chemical ionization for the determination of free and glycineconjugated bile acids in human serum. J. Sep. Sci.29(18),2837–2846 (2006).
  • 39  Carvalho VM, Kok F. Determination of serum methylmalonic acid by alkylative extraction and liquid chromatography coupled to tandem mass spectrometry. Anal. Biochem.381(1),67–73 (2008).