Spe c ial Iss ue : Review Arti c le

Received 12 August 2010, Revised 10 September 2010, Accepted 10 September 2010 Published online in Wiley Online Library: 6 November 2010

(wileyonlinelibrary.com) DOI 10.1002/bmc.1548

Derivatization reagent s in liquid c hromatography/ele c tro s pray ionization tandem ma ss s pe c trometry
Tomofumi Santa *
ABSTRACT: Liquid c hromatography/ele c tro s pray ionization tandem ma ss s pe c trometry (LC/ESI- MS/MS) is one of the mo s t prominent analyti c al te c hnique s owing to it s inherent s ele c tivity and s en sitivity. In LC/ESI- MS/MS, c hemi c al derivatization is often u s ed to enhan c e the dete c tion s en sitivity. Derivatization improve s the c hromatographi c s eparation , and enhan c e s the ma ss s pe c trometri c ionization e c ien c y and MS/MS dete c tability. In this review, an overview of the derivatization reagent s whi c h have been applied to LC/ESI- MS/MS is pre s ented , fo c u sing on the appli c ation s to low mole c ular weight c ompound s. Copyright © 2010 J ohn Wiley & Son s, Ltd . Key word s: derivatization; chromatography; electrospray; tandem mass spectrometry

Introdu c tion
The development of sensitive and selective determination methods for trace level compounds is essential to elucidate their biological roles and functions in living systems. Recently, liquid chromatography/mass spectrometry (LC/MS) has frequently been utilized for this purpose. In particular, liquid chromatography/tandem mass spectrometry (LC/MS/MS) equipped with electrospray ionization (ESI) ion source is the most prominent method, since it requires lower temperature for ionization compared with other ionization methods, and thus it can be used for thermally unstable compounds. Furthermore, the decrease in the noise level by MS/MS detection improves the signal-to-noise ratios and hence enables sensitive detection of the targeted compounds. However, not all compounds can be favorably analyzed by LC/ESI-MS/MS. For example, the ionization efficiencies are often extremely low and such compounds cannot be sensitively detected. An analyte should have the following properties to be sensitively analyzed by LC/ESI-MS/MS. Firstly, it must be in its ionic form in the solution phase or be chargeable through adduct formation in gas-phase reaction (Cech and Enke, 2001). Secondly, it is preferable to have a non-polar region, since hydrophobic compounds can be well separated from salts and interfering compounds possessing suppression effects on ESI (Nordstrom et al., 2004). The non-polar ions prefer the droplet–air interface and reside at the droplet surface. Consequently these ions enter the gas phase more readily than those in the droplet interior and show a higher ESI response (Cech et al., 2001; Cech and Enke, 2000, 2001; Zhou and Cook, 2001). Furthermore, the hydrophobic compounds are eluted by the mobile phase with the higher organic solvent content on the reversed-phase column. The higher organic solvent content is suitable for the generation of charged droplets by electrospray (Cech and Enke, 2001) and thus gives the higher ESI response (Zhou and Hamburger, 1995). Generally, the chargeability and hydrophobicity of the analytes are the critical factors for the ESI response (Ehrmann et al., 2008; Henriksen et al., 2005; Okamoto et al., 1995; Nord-

strom et al., 2004). Thirdly, it is desirable that the target analyte fragments efficiently upon collision-induced dissociation (CID) and generates an intense product ion for the sensitive MS/MS detection.

* Correspondence to: T. Santa, Graduate School of Pharmaceutical Sciences, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, Japan. E-mail: santa@mol.f.u-tokyo.ac.jp Graduate School of Pharmaceutical Sciences, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, Japan Abbreviation s u s ed : 4-APC, 4-[2-(trimethylammonio)ethoxy] benzenaminium halide; APDS, 3-aminopyridyl-N-hydroxysuccimidyl carbamate; 4-APEBA, 4-{2-[(4-bromophenethyl)dimethylammonio] ethoxy}benzenaminium dibromide; APZ, 4-(4-methyl-1-piperazyl)-3nitrobenzoyl azide; AQC, 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate; CID, collision-induced dissociation; DAABD-AE, 4-[2-(N,Ndimethylamino)ethylaminosulfonyl]-7-(2-aminoethylamino)-2,1,3benzoxadiazole; DAABD-MHz, 4-[2-(N,N-dimethylamino)ethylaminosulfonyl ]-7-N-methylhydrazino-2,1,3-benzoxadiazole; DMAP-NCS, p(dimethylamino)phenyl isothiocyanate; DMIS, 1,2-dimethylimidazole-4sulfonyl; DNPH, 2,4-dinitrophenylhydrazine; DMAE, dimethylaminoethyl; Dns-Cl, 5-dimethylamino-1-naphthalenesulfonyl chloride; Dns-Hz, 5dimethylaminonaphthalene-1-sulfonyl hydrazine; ESI, electrospray ionization; FMOC, 9-fluorenylmethyl chloroformate; FPMTS, 2-fluoro-1methylpyridinium p-toluensulfonate; Gir P, 1-(carboxymethyl)pyridium chloride hydrazide; Gir T, (carboxymethyl)trimethylammonium chloride hydrazide; HMP, 2-hydrazino-1-methyl-pyridine; HP, 2-hydrazinopyridine; HTMOB, 4-hydrazino-N,N,N-trimethyl-4-oxobutanaminium iodide; MDMAES, mono-(dimethylaminoethyl) succinyl; NA, isonicotinoyl azide; NBD-F, 4-fluoro-7-nitro-2,1,3-benzoxadiazole; NP-NCS, m-nitrophenyl isothiocyanate; 17-OHP, 17-hydroxyprogesterone; PA, 2-picolylamine; PPZ, 1-(2,4dinitro-5-fluorophenyl)-4-methylpiperazine; PBS, 4-(1H-pyrazol-1-yl) benzenesulfonyl; PS, pyridine-3-sulfonyl; PrCl, propyl chloroformate; Py-NCS, 3-pyridyl isothiocyanate; SPTPP, 5-N-succinimidoxy-5-oxopententyl) triphenylphosphonium bromide; SRM, selected reaction monitoring; TFA, trifluoroacetic acid; TMAE, trimethylaminoethyl; THAS, 4-(trimethylammonium)anilyl-N-hydroxysuccidimidyl carbamate iodide.

1

Biomed. Chromatogr. 2011; 25: 1–10

Copyright © 2010 John Wiley & Sons, Ltd.

the peroxidation products of fatty acids.. A significant increase of detection sensitivity for altrenogest was observed (Lampinen-Salomonsson et al. Gir T was also applied to the analysis of succinylacetone in urine and dried bloodspots (Johnson et al.. Derivatization Reagent s For Ketone s and Aldehyde s Conventional reagent s. 5.. The product ion spectra of the derivatives were rather simple and clear. by the loss of a trimethylamino moiety (59 Da. derived from 1-methylpyrizinoamino moiety (Fig. testosterone in rat serum and brain (Higashi et al.5% trifluoroacetic acid (TFA). 2005a. 93 and 121 were generated by CID. Higashi and Shimada 2004. These compounds gave the common product ion at m/z 216 by CID. 2000). derived from the tri-cyclic structure. Kushnir et al. In the past decade. HMP reacted with carbonyl compounds such as keto-steroids at 60°C within 1 h in ethanol containing 0. Aldehydes were condensed with two molecules of cyclohexanedione in the presence of ammonia to form the tri-cyclic compounds (Hantzsch reaction). Chromatogr. focusing on the applications to low molecular weight compounds.ions to these product ions were used for SRM (selected reaction monitoring). Therefore a chargeable moiety was often introduced to enhance the ionization efficiency. 2006). the product ion spectra of the derivatives with these reagents were not simple and clear. They were efficiently fragmented by CID to generate intense and specific product ions. were observed. The transition of m/z 299 ([M]2+) to m/z 80 was used for SRM (Lai et al. Rashed et al. 2006).MS/MS. (b) HMP. 5a-dihydrotestosterone in prostate and prostatic tissue (Higashi et al. The transition of m/z 462 ([M + H]+) to m/z 170 was used for SRM (Al-Dirbashi et al. 2007a. Santa Chemical derivatization of the analyte is often used to enhance the detection sensitivity in ESI-MS. 2002). a major thymidine lesion generated by reactive oxygen species. Most of these reagents are aimed to enhance the ESI response but they are not designed to generate a particular product ion by CID. including 4-hydroxynonenal and nonanal. 2007. and the product ion of the derivative: (a) Dns-Hz. the fragment patterns of the derivatives depended on the structures of the generated derivatives. derived from the reagent skeleton.4dinitrophenylhydrazine) was used for the determination of aldehydes and ketones in disinfected water (Zwiener et al. Gir T was used for 5-formyl2’-deoxyuridine. Fragment ions at m/z 80.. 2006). 2011. Hydroxylamine was applied to the analysis of testosterone (Borrey et al.. the reagents developed specifically for LC/ESI-MS/MS carried a structure suitable for MS/MS detection. 2006). 2007).. HMP was applied to the analysis of keto-steroids such as testosterone.. Girard’s reagent P [1-(carboxymethyl)pyridium chloride hydrazide. The transitions of [M + H]+ ions of the derivatives to m/z 216 were used for SRM (O’Brien-Coker and Mallet. The transitions of the [M . 1994.. Dns-Hz) was used for the analysis of malondialdehyde (Lord et al.. Reagent s de signed for LC/ESI. 2 View this article online at wileyonlinelibrary. The ionization efficiencies in ESI of these compounds are sometimes low. Derivatization reaction for ketones and aldehydes.. In this review. The transitions of [M + H]+ ions of the derivatives to m/z 273 or 274 were used for SRM.. DNPH (2. Hydroxylamine reacts with ketones to form the corresponding oximes.H]. Honda et al. One of the prominent reagents for carbonyl compounds is HMP (2-hydrazino-1-methylpyridine. 2009) and succinylacetone in dried blood spot specimens (Al-Dirbashi et al... Higashi 2006. The derivatives were detected in the negative-ion mode. 25: 1–10 . Therefore. assigned to the cleavage of dimethyaminonaphtyl moiety originating from the reagent (Fig.. 2007). Johnson 2005.. 2001). 1a). The ionization efficiencies of HMP derivatives were rather high and the generated derivatives gave a product ion at m/z 108 by CID. Conventional reagent s having a s uitable s tru c ture for MS/MS. Similarly. 2001). b). 1998. 2007a. Van Berkel et al. These have summarized in several review papers (Gao et al. Ketones and aldehydes are neutral functional groups. The derivatives gave the common product ions at m/z 273 or 274 by CID (Williams et al. Dansyl hydrazine (5-dimethylaminonaphthalene-1sulfonyl hydrazine. Biomed.5’-dimethyl-1.. Since then. 2006b). Gir T] are reagents that possess a permanent cationic charge. The generated dansyl hydrazone of succinylacetone selectively gave the product ion at m/z 170 by CID. adorenal steroids (Kushnir et al. the marker for congenital adrenal hyperplasia. 2005). Blair 2010). Figure 1 .. 2005.com Copyright © 2010 John Wiley & Sons.3cyclohexanedione was used for biogenic aldehydes. 2005a). Usually. several conventional reagents originally utilized in ultraviolet or fluorescence detection were borrowed for this purpose. On the contrary. Higashi et al. Ltd. The generated derivative gave the product ion at m/z 195.. Lampinen-Salomonsson et al. altrenogest (a steroid used for the control of estrus in horses. The generated derivatives usually gave several product ions dependant on the analyte structure. and succinylacetone (the hallmark of hepatorenal tyrosinemia.T. Santa et al. Cyclohexanedione was used for the analysis of aliphatic aldehydes (C3–C10)... an overview of the derivatization reagents which have been applied to LC/ESI-MS/MS is presented. 1994. Hong and Wang. it was reported that the chemical derivatization of the analyte by the chargeable compounds improved ESI-MS responses of the target analytes (Quirke et al. m/z 152 and m/z 122. Common product ions such as m/z 163. 1b). (c) DAABD-MHz. 2006a). Gir P was used for the derivatization of 17-hydroxyprogesterone (17-OHP). 2005). Gir P] and Girard’s reagent T [(carboxymethyl)trimethylammonium chloride hydrazide. Dansyl moiety is suitable for MS/MS detection.

Alcohols and phenols are neutral and rather hydrophilic compounds... Xia et al. 2008). SRM using the transition of quasimolecular ion of the derivative to m/z 171 or 170 resulted in a sensitive detection of the derivatives. Scarth et al. derived from the neutral loss of 59 Da (trimethylamino moiety). icariin (Gong et al. HP (2-hydrazinopyridine) was used for di-oxo-steroids such as androsterone and progesterone. Similarly. For Al c ohols Conventional reagent s. Xu et al. Beaudry et al. b).. Yamashita et al. 2002.. 4-dimethylaminophenol (Zhuang et al. The derivatives also gave an almost single product ion at m/z 171 by CID. 2010. ethinyl estradiol.. 2b). 2007... respectively (Higashi et al. propofol (veterinary medicine. 2009). Conventional reagent s having a s tru c ture s uitable for MS/MS One of the most widely used derivatization reagents for phenols in LC/ESI-MS/MS is dansyl chloride (5-dimethylamino-1- Figure 2 . 2009a. 2008... 2006). The ESI response was enhanced by the formation of hydrophobic derivatives..Derivatization reagents in LC/ESI-MS/MS 2006).. Li et al. The transition of m/z 506 ([M + H]+) to m/z 383 was used for SRM (Honda et al. 2004. 2008). 2010). 2004.. 2007). deivatization is often used to enhance the ionization efficiency and to increase the hydrophobicity. 2007). (c) NA.87]+. Li et al. HTMOB was applied to the profiling of ketones. To overcome these problems. it was used for determination of steroids having hydroxyl group such as estradiol. since small molecules with a multi-charge are unstable in the gas phase and provided multiple ions.. HP was also applied for the analysis of 17a-hydroxyprogesterone (Shibayama et al. it was inferred that they were formed by the loss of one HP moiety together with a part of the A-ring.5. Ltd. acetyl chloride was used for the derivatization of cholesterol (Liebisch et al. 2008).. 2008.. Santa et al. cytokinins.N-dimethylamino)ethylaminosulfonyl moiety of the reagent (Fig. 2007.. The generated derivatives are easily separated on the reversed-phase column and sensitively detected by ESIMS. 5a-reduced pregnane-type neurosteroids in rat brain and serum (Higashi et al. 2005. ketoacids and ketodiacids in the children’s urine samples by ESI-MS/MS without chromatographic separation (Johnson.. derived from the loss of trimethylammonium moiety and trimethylaminoethyl moiety. The derivatives gave a rather simple product ion spectra. due to the loss of picolinic acid moiety (Fig.. Dns-Cl has a dimethylamino group as an ionization moiety and a hydrophobic aromatic structure. 2008a). View this article online at wileyonlinelibrary... Anari et al. Pedreira et al. 2005.N-trimethyl-4-oxobutanaminium iodide) was a modified Girard’ reagent. Chromatogr. 25: 1–10 Copyright © 2010 John Wiley & Sons. Shou et al. 2007c). The derivative gave the product ion at m/z 383 by CID. Tai and Welch. ribosides and intact nucleotides such as AMP. It contains a bromophenyl group and the isotopic signature of 79Br and 81Br isotope (100:98) provided confirmation of the presence of bromine in the derivatives (Eggink et al. 1-hydroxypyrene (biomarker to monitor exposure to polycyclic aromatic hydrocarbons. 2009. 4-{2-[(4-bromophenethyl)dimethylammonio]ethoxy} benzenaminium dibromide (4-APEBA) was reported. Propionyl and benzoyl anhydride were used for the derivatization of bases. 2004). 2010). 17ahydroxypregnenolone and 17a-hydroxyprogesterone in dried blood spots from low-birthweight infants (Higashi et al. 4-[2-(Trimethylammonio) ethoxy]benzenaminium halide (4-APC) reacted with aldehydes and generated amines in the presence of NaBH3CN. Therefore.. 2010). (d) MDMAES imidazole. testosterone in saliva (Shibayama et al. Picolinic acid was used for the analysis of the steroids having hydroxyl groups (Honda et al. 2-Fluoro-1-methylpyridinium p-toluensulfonate (FPMTS) reacted with hydroxyl group at room temperature for 1 h in dichloromethane–acetonitrile containing 3 Biomed.. Lin et al. Similarly..5% TFA. 2011. naphthalenesulfonyl chloride. 2005a. Although the structures of these ions could not be identified. Farlow et al.. 2008a–c. Picolynic acid was sometimes used for the derivatization of alcohols. known for its unwanted suppression effects on ionization.The generated derivatives gave the product ions at m/z [M . which was assigned to protonated dimethylaminonaphtyl moiety (Fig. Nelson et al.3-benzoxadiazole} reacted with aldehydes at 50°C within 10 min in acetonitrile containing 0. estrone and the related compounds (Anari et al. dehydroepiandrosterone (Higashi et al.. and the derivatives were separated without the need for an ion paring reagent. Simple esterification of the analytes has sometimes been adopted for this purpose. 2007b).N. So far. 4-APC was used for the analysis of aldehydes in biological mixtures (Eggink et al. 2004. Kushnir et al. 2002). 2007a). in 30 min at room temperature in the presence of the condensation reagents in tetrahydrofuran..com .1. 2007b).. Dns-Cl). DAABD-MHz {4-[2-(N. (b) picolynic acid. 2007a–c. 1c. The generated derivatives gave the intense product ions at m/z 322 and 348 for the derivatives of androsterone and progesterone.. a biomarker for bile acid biosynthesis. It reacted with phenols at 60°C within several minutes in acetone and sodium bicarbonate buffer (pH 10. HTMOB (4-hydrazino-N. ADP and AMP (Nordstrom et al. In addition.N-dimethylamino)ethylaminosulfonyl]-7N-methylhydrazino-2. HMP is not effective for increasing the detection responses of di-oxo-steroids. 2005). and neuroactive androgen (Higashi et al. 2a). 2008) and multiple classes of phenols in blood (Chang et al... Derivatization reaction for alcohols and the product ion of the derivative: (a) Dns-Cl. It reacted with 7a-hydroxy-4-cholestene-3-one.. the retention on a reversed-phase column was greatly increased.. derived from the protonated (N. 2009a.. Petucci et al. 2009). 2004.59]+ and [M . The generated derivatives gave a predominant product ion at m/z 151 by CID. Zhang et al. 2005b)..

H]. FPMTS was used for the analysis of testosterone.. The generated ester gave the product ion at m/z 98. used isonicotinoyl azide (NA) for the derivatization of di-hydroxysteroids such as estradiol and 5a-androstane-3a. These reagents were used for the analysis of steroidal estrogens (Xu and Spink. The generated derivatives gave the product ion at m/z 110 by CID. Garg and Dasouki. Reagent s de signed for LC/ESI. Carboxylic acids are detectable in the negative ESI-MS. acylcarnitine profile can be obtained by precursor ion scan of m/z 85. 2008). Reagent s de signed for LC/ESI. For Carboxyli c A c id s Con c entional reagent s. Higashi et al. MDMAES imidazole reacted with hydroxyl group at 70°C for 10 min in dichloromethane containing 1% triethylamine. Most a-amino acids butyl esters gave the intense product ions corresponding to the loss of HCOOC4H8 (102 Da) by CID. due to the loss of 2 C4H8. Roschinger et al. 2009). DAABD-AE {4-[2-(N. Biomed. 3-(hydroxymethyl)-pyridine was used for the polar molecules in plant extracts (Kallenbach et al. 2008b). However.. Santa triethylamine. Higashi et al. 2007). Butanolic HCl derivatization of carboxylic acids is sometimes useful in LC/ESI-MS/MS. 2007d).3 [M 47]+. for example. The detection responses in LC/ESI-MS/MS were significantly increased by the conversion to the quaternary amino compounds (Nishio et al. The transition of m/z 231 ([M + H]+) to m/z 119 was used for SRM (Kushnir et al..17bdiol (Higashi et al.. Acylcarnitines have a quaternary ammonium group and a carboxylic group in their structure. 2007c. Chromatogr. Ltd. the product ion spectra of PS derivatives and PBS derivative showed analytespecific fragments. Chace and Kalas. Therefore. 2006. 1994). 2002). NA reacted with two hydroxyl groups of estradiol at 80°C for 30 min in benzene. 2010a).. The generated derivatives of HP gave the strong product ion at m/z 110 and those of PA gave the strong product ion at m/z 109 by CID (Higashi et al. In addition the mobile phases for the carboxylic acids separation are not always compatible with ESI-MS. Carpenter and Wiley. reported the simple and practical method for the analysis of carboxylic acids using HP and 2-picolylamine (PA). derived from protonated 1-methylpiperidine moiety by CID (Fig. methylated PPZ-estradiol gave the product ion at m/z 504. 2003. a brain-specific di-peptide and the marker of heritable defect in GABA pathway (Jansen et al. Magera et al. The generated butyl esters are rather hydrophobic and they often gave the product ion at m/z of ([M + H – 56]+).Ndimethylamino)ethylaminosulfonyl]-7-(2-aminoethylamino)2. Acylcarnitines are the marker metabolites for inherited disorders related to organic acid and fatty acid metabolism (Millington et al. the amino acid profile in biological samples can be obtained by the neutral loss scan of 102 Da. These methods are currently widely used for the analysis of amino acids and acylcarnitines in urine... The generated derivatives were analyzed after the conversion to the quaternary amino compounds with methyl iodide.. Its di-butyl ester gave the product ion at m/z 119. Methylmalonic acid is the marker for a group of metabolic disorders caused by deficiency in methylmalonyl-CoA mutase or a defect in vitamin B12 metabolism. plasma. 2009) and propofol (Thieme et al. Several biological important compounds having carboxylic acids were derivatizaed with these reagents. In contrast. The quaternary amino compounds provided an almost single product ion by CID. introducing two permanently charged groups to the small molecules did not improve the detection responses. The generated butyl esters of amino acids were introduced to ESIMS/MS without chromatographic separation. 2c). 2005. Nishio et al. Several conventional reagents suitable for MS/MS were reported..MS/MS. Their butyl esters were introduced to ESI-MS/MS and gave the common product ion at m/z 85 by CID (Rashed et al. Chace et al. dihydrotestosterone (O’Brien et al. Cholesterol and dehydrocholesterol in dried spots of plasma were analyzed by ESI-MS/MS without chromatographic separation (Johnson et al.T. Allylic isoprenols were derivatized with 3-nitrophtalic anhydride and analyzed by LC/ESI-MS/MS under the negative ion mode. 3a).. HP (2-hydrazinopyridine) was used for the analysis of chenodeoxycholic acid and its glycine-conjugate. 25: 1–10 . Butanolic HCl derivatization is also applied to the analysis of nitrotyrosine. since the multi-charged small molecules are unstable in the gas phase. MDMAES [mono(dimethylaminoethyl) succinyl] imidazole was used for cholesterol and dehydrocholesterol. 2000. The derivatization reaction proceeded at 70°C for 30 min in pyridine. used 1-(2. The generated derivatives gave the product ion at m/z 166 by CID. Schmedes and Brandslund. 2008). 2001.. Similarly. 3b). derived from the protonated 2-hydrazinopyridine moiety (Fig.com Copyright © 2010 John Wiley & Sons.. One of the successful examples for ESI-MS/MS detection of carboxylic acids with butanolic HCl derivatization is the simultaneous analysis of amino acids and acylcarnitines in dried blood spots for newborn screening (Rashed et al. 2007). 2001a). Dns-Cl.4dinitro-5-fluorophenyl)-4-methylpiperazine (PPZ) and 4-(4methyl-1-piperazyl)-3-nitrobenzoyl azide (APZ) for derivatization of steroids having a hydroxyl group. 2011. 2002) and homocarnosine.. and generated MDMAES ester. The transition of [M + H]+ ion to m/z 139 was usable for SRM analysis (Higashi et al. the tyrosine nitration product (Delatour et al. 2010b). due to the neutral loss of MDMAES moiety (189 Da. 2001. 1. 2d). The generated amide derivative gave the product ion at m/z 120 by CID. 2009). 1997). The generated derivative gave the product ion at m/z 139 by CID..to m/z 166 were monitored (Teshima and Kondo. The transition of m/z 277 ([M + H]+) to m/z 120 was used for SRM (Cheng et al. 1995. 2006). Therefore. d. Therefore carboxylic acids were usually transformed to the hydrophobic derivatives. 1994. Conventional reagent s having a s tru c ture s uitable for MS/MS. Xu and Spink compared four derivatization reagents having sulfonyl chloride as a reaction group. pyridine-3-sulfonyl (PS) chloride and 4-(1H-pyrazol-1yl)benzenesulfonyl (PBS) chloride. The transitions of [M . 3-Hydroxy-1-methyl-piperidine was used for the derivatization of malonic acid (Honda et al. Santa et al.. which was assigned to the protonated pyridyl carbamic acid (Fig. 4Dimethylaminobenzylamine was used for the analysis of valproic acid and its metabolite. due to the loss of C4H8 (56 Da) by CID. The transitions to m/z 110 from the [M + H]+ of the derivatives were used for SRM (Higashi et al.1. Fig.. The ester gave the product ion at m/z 369 by CID. However their sensitivity is rather poor because of the high background noise. derived from dimethylamino phenyl moiety of the reagent. the reagent having a high-proton affinity group such as pyridyl group is favorable for the derivatization of di-hydroxysteroids. serum or blood (dried blood spot) (Rashed.3-benzoxadiazole. Although the product ions could not be identified.. Therefore.2-dimethylimidazole-4-sulfonyl (DMIS) chloride. they were specific ions of the derivatives with satisfactory intensity... 2007b} was synthesized and 4 View this article online at wileyonlinelibrary. The product ion spectra of the dansyl and DMIS derivatives were dominated by ions representing derivatization reagent moieties. 2009b). 1990). 2006).MS/MS. 2003...

2003). saves 75% of instrument time and requires one-tenth of the biological sample volume (Al-Dirbashi et al. Conventional reagent s having a s tru c ture s uitable for MS/MS. The major product ion at m/z 118. 2007).com . View this article online at wileyonlinelibrary. The MS/MS spectra of PrCl derivatives showed more product ions.. b. Ethyl bromoacetate was used for the derivatization of trimethylamine. 2008).3-benzoxadiazole) was used for biogenic amines such as tryptamine. was commonly observed.Derivatization reagents in LC/ESI-MS/MS the generated derivatives can be more easily separated from the interfering compounds on the reversed-phase column. FMOC reacted with the amino group. Fatty acids were treated with oxalyl chloride..H]-) to m/z 402 for S1P by CID. Nitrobenzyl chloroformate was used for 3-amino2(S)hydroxypropylmethylphospheric acid. or m/z 119 corresponding to CF3CF2. In case of agmatine. However.. 2003).ion was selected as precursor ion. 1999. Uutela et al.. was observed. derived from protonated dimethylaminonaphtyl moiety of the reagent. The product ions of [M . the transition of m/z 294 ([M + H]+) to m/z 277 was used for SRM (Song et al. 2007). 2011. m/z 297–279 (NBD-Asp). Dns-Cl was used for musimol and ibotenic acid. since the background is generally lower in the higher mass range. pentafluoropropionic acid anhydride were used for diamines. 2008). 2009). Acetyl anhydride was used for polyaromatic amines such as diaminopyrenes and aminonitropyrenes. (c) DAABD-AE. TMAE (trimethylaminoethyl) ester derivatives were used for the analysis of very long chain fatty acids. markers for the exposure to isocyanates. Trietylamine in urine was analyzed by a flow injection ESI-MS/MS. 2005). m/z 269–223 (NBD-Ser) and m/z 311–293 (NBD-Glu) were monitored for quantification (Song et al.ion. The transitions of (M+) ions to m/z 171 were used for SRM (Tsujikawa et al.. resulting in high signal-to-noise ratios. and each fatty acid derivative was detected by SRM. 2001b. NBD-F was applied to the determination of D-amino acids. 2009). The derivatization of amino acids and dipeptides in cerebrospinal fluids with PrCl was also reported (Fonteh et al.H . Compared with standard gas chromatography–mass spectrometric methods routinely used for this purpose. Johnson et al. was used for SRM (Berdyshev et al. Chemical derivatization makes amines more hydrophobic and 5 Biomed. Dns-Cl was also used for the analysis of glutamic acid. in diffusion medium (Timperio et al.2-difluoro-2-deoxyuridine in human plasma (Bowen et al. The [M H].or [M .1. The transition of [M + H]+ ions to m/z 151 was used for SRM. Acetyl anhydride was also used for sphingosine-1-phosphate (S1P) and dihydrosphingosine-1phosphate. histamine. a-fluoro-balanine. The generated derivatives gave the product ion at m/z 171.. an endogenous sphingolipid and the analog. Similarly.. the analysis of amines is often troublesome because of their high polarity. used for the analysis of very long chain fatty acids. reported the comparison of PrCl (propyl chloroformate) and FMOC (9-fluorenylmethyl chloroformate) as the derivatization reagents for amino acids (Uutela et al. The generated derivatives were suitable for MS/MS detection. derived from loss of 28 Da (ethylene). These derivatives gave the product ion by the loss of 59 Da... 2007). 2000a. However. this LC/ESIMS/MS method is simpler. NBD-F (4-fluoro-7-nitro-2. 2009). assigned to Figure 3 . 2004). 5-fluorourasil. PrCl was used for the amino acids analysis in rat brain microdialysates. 25: 1–10 Copyright © 2010 John Wiley & Sons. 3c).Ndimethylamino)ethylaminosulfonyl moiety of the reagent (Fig. followed by the methylation with methyl iodide. producing a protonated amino acids. were monitored (Marand et al. TMAE or DMAE (dimethylaminoethyl) ester derivatization of fatty acids and ESI-MS/MS analysis without chromatographic separation was reported (Johnson. the diagnostic markers for peroxisomal disorders (Johnson et al.. (b) 3-hydroxy-1-methyl-piperidine. capecitabine (Licea-Perez et al.120]. Acetylation improved peak shapes and reproducibility in LC of diaminopyrene. bioactive compounds in mushroom. the three step derivatization reaction was tedious for routine assay. 2005). 3d). The derivative gave the product ion at m/z 152.. GABAB receptor agonist. The compounds having amino group are easily protonated under acidic conditions and suitable for ESI-MS. the excitatory amino acid. 2007). PrCl reacted with both amino and carboxylic groups. DAABD-AE was also used for the analysis of the dicarboxylic acids such as glutaric acid and 3-hydroxyglutarate.. Ltd. due to the neutral loss of one or two CF3CHF2-group. In addition. The characteristic product ions were produced for each derivative by CID. due to the neutral loss of acetic acid. Johnson and Trinh. gemcitabine and 2. (d) TMAE alcohol. Chromatogr. The loss of propanol (60 Da) was common for all the PrCl-derivatized amino acids.H . N-methylsalsolinol and agmatine. The characteristic precursor to product ion transitions. 2007). the marker metabolites for glutaric acidemia type 1 in human urine (Al-Dirbashi et al.240]-... derived from (CH3)3N moiety of the derivatization reagent (Fig. The cleavage of the FMOC moiety (loss of 222 Da). and the generated derivatives gave the product ion at m/z 179 derived from dissociation of FMOC moiety. The generated amide derivative gave the product ion at m/z 151 by CID. 2004). basicity and high water solubility. and can be sensitively detected in ESI-MS. the increase in the molecular weight decreases in the background noise from the matrix. For Amine s Conventional reagent s. The transition of m/z 462 ([M . the marker metabolite of trimethylaminuria (Johnson. derived from the protonated (N. Derivatization for carboxylic acids and the product ion of the derivative: (a) HP. the markers of peroxisomal disorders. human carcinogens (Straube et al.. dimethylaminoethanol.

4d. Their generated derivatives gave the intense product ions at m/z 137. Referen c e s Al-Dirbashi OY. 4a). Reagent s de signed for LC/ESI. Santa T. Amino acids were analyzed with the detection limits of atto-mole level (Shimbo et al.. Con c lu sion The derivatization reagents in LC/ESI-MS/MS applied to the low molecular weight compounds are reviewed. and pro- A c k nowledgement s This work was partially supported by a Grant-in-Aid for Scientific Research from Ministry of Education. 25: 1–10 .. 2000). Shimbo et al. Al-Ahaidib LY. The hydrophobicity of the analytes increased by derivatization and the resolution of the analytes on the reversed-phase column was improved dramatically.. m/z 179 and m/z 137.. NIT (naphtylisothiocyanate) was used for the determination of eighteen kinds of primary and secondary amines in air samples. The derivatization improved the separation efficiency. which was generated by the cleavage of ester bond of the reagent (Fig. Derivatization for amines and the product ion of the derivative: (a) 4-nitrobenzyl chloroformate. Ester. Biomed. was also applicable for amino acid analysis by LC/ESI-MS/MS. respectively (Fig. Jacob M. On the contrary. The derivatives of the primary amines gave the common base peak at m/z 144 and the fragment ion at m/z 127 by CID. Analysis of organic acid markers relevant to inherited metabolic diseases by ultra-performance liquid chromatography/tandem mass 6 View this article online at wileyonlinelibrary. intense product ions. 4c). This method provided the structural information of the analytes.. 3-pyridyl isothiocyanate (Py-NCS). Urea and thiourea moieties were easily cleaved by CID and suitable structure for MS/MS. 2010). The fragment patterns of the derivatives and the efficiencies depended on the structures of the analytes. 2009a). and thus they were suitable for the separation on the reversed phase column. The reagent developed for proteomics. (b) 3-pyridyl isothiocyanate. Jakobs C. Chromatogr.. 4b. The generated derivatives were hydrophobic compared with corresponding THAS derivatives. 2009b. The transition of all the protonated molecular ions to the common fragment at m/z 121 was monitored (Fig. GC-MS and amino acid analyzer was reported (Kaspar et al. iTRAQTM (Ross et al. p-(dimethylamino)phenyl isothiocyanate (DMAP-NCS) and m-nitrophenyl isothiocyanate (NP-NCS) were applied to the derivatization of amines.com Copyright © 2010 John Wiley & Sons.H]-) to m/z 152 was used for SRM (Blum et al. SPTPP [(5-Nsuccinimidoxy-5-oxopententyl)triphenylphosphonium bromide] was used for the analysis of amines and amino acids. SPTPP was used for the analysis of neurotransmitter 4-aminobutanolic acid (GABA) and oxidative stress markers in rat serum (Inagaki et al. Sports. 2010). urea or thiourea. 831: 274–280. AQC was used for the derivatization of b-N-methylamino-L-alanine and 2. Determination of succinylacetone in dried blood spots and liquid chromatography tandem mass spectrometry. Al-Qahtani K. APDS (3-aminopyridyl-N-hydroxysuccimidyl carbamate) was also designed for LC/ESI-MS/MS and used for the analysis of more than 100 compounds with amino group in biological fluid. a potential human neurotoxin and its isomer. (c) THAS. The urea structure of the derivatives was cleaved by CID and gave the product ion at m/z 145 and 171 derived from the AQC moiety were observed (Spacil et al.. 2009). AQC (6-aminoquinolyl-Nhydroxysuccinimidyl carbamate) is the commonly used fluorescence derivatization reagent for amino acids. The transition of m/z 371 ([M . Santa duced characteristic fragment ions at m/z 177.. 2004). The derivatives gave the characteristic cleavage at the urea bond by CID. The product ions derived from the reagent moiety were observed. and was suited for the analysis of complex environmental samples (Claeson et al. Figure 4 . Ltd. THAS [4(trimethylammonium)anilyl-N-hydroxysuccidimidyl carbamate iodide] was the reagent designed for LC/ESI-MS/MS. 4-nitrobenzyl alcohol anion. It reacted with amino acids to form urea compounds. Al-Amoudi M and Rashed MS. whereas those of secondary amines gave the common base peak at m/z 186 and the fragment ion at m/z 128 by CID. The prominent derivatization regents for LC/ESIMS/MS are still desired for the sensitive and selective detection of the various kinds of the compounds in the fields of biomedical analysis. MRC Bioanalytical Science Group for his kind suggestions and valuable discussion. Journal of Chromatography B 2006. hydrazone and alkyl quaternary amine were often easily cleaved by CID and were suitable for MS/MS detection. Rashed MS. and Technology of Japan. Filimban N. (d) APDS. 2011. Al-Hassnan Z and Faqeih E.3-diaminobutylic acid. Al-Dirbashi OY. They were fragmented easily by CID and efficiently generated particular. Culture. The limits of detection of the SPTPP were sub-femtomole levels. Al-Sayed MM.MS/MS. aromatic sulfonyl compound. derived from the reagent moiety (Fig. the reagents designed for LC/ESI-MS/MS have a suitable structure for MS/MS detection. Santa. Science. 2010).T. Conventional isothiocyanates. The author thanks Dr Chang-Kee Lim. ionization efficiency and MS/MS detectability of the analytes. The thiourea moiety of the derivatives was easily cleaved by CID and gave a product ion. A comparison of iTRAQTM-LC-MS/MS. Some conventional regents aimed to enhance the ESI response but they were not designed to generate a particular product ion by CID. 2010). 2004). Ten Brink HJ.

860: 166–172. Sensitivity enhancement in liquid chromatography/atmospheric pressure ionization mass spectrometry using derivatization and mobile phase additives. 877: 1327–1334. Amino Acids. Farlow D. Esch IJP. 2007. 382: 134– 137. Shibayama Y and Shimada K. Gong Y. and very long chain fatty acid markers of peroxisomal disorders. Bowen C. Giesy JP and Jones PD. Bakhtiar R. Huskey S. Winterborn A. Targeted LC–MS derivatization for aldehydes and carboxylic acids with a new derivatization agent 4-APEBA. Quantitative measurement of endogenous estrogen metabolites. Zhang J. Xu X and Veenstra TD. Hecker M. Cheng H. Zhu B. 49: 1855–1862. Lingeman H. Koh E. 322: 1–10. Nishio T. Nagura Y. Niessen WMA and Irth H. Journal of Chromatography B 2007b. Wijtmans M. Jacob M and Al-Mokhadab M. Studies on neurosteroids XXIV. Gao S. Moerman E. Analytical Chemistry 2000. Henriksen T. Clinical Biochemistry 2005. Kretschmer A. Froestl W and Desrayaud S. Relating electrospray ionization response to nonpolar character of small peptides. Chace DH. Ostin A and Sunesson AL. 49: 1797–1817. 2-Hydrazino-1-methylpyridine: a highly sensitive derivatization reagent for oxosteroids in liquid chromatography–electrospray ionization mass spectrometry.17bdiol. in commercial milk product by LC-MS/MS. Kool J. Use of tandem mass spectrometry for multianalyte screening of dried blood specimens from newborns. Journal of Chromatography B 2007a. Gorshkova IA. 2011. Berdyshev EV. 74: 4136–4144. Krone JR. 846: 195–201. Santa T. Zhang X. Ltd. 54: 1479–1485. Higashi T. Fukushi M and Maeda M. Garcia JGN. Takayama N and Shimada K. 23: 1015–1019. Clinica Chimica Acta 2007. Determination of the GABAB receptor agonist CGP 44532 (3-amino-2hydroxypropylmethylphosphinic acid) in rat plasma after pre-column derivatization by micro-high-performance liquid chromatography combined with negative electrospray tandem mass spectrometry. Journal of Pharmaceutical and Biomedical Analysis 2005. Application of tandem mass spectrometry to biochemical genetics and newborn screening. Blum W. Mass Spectrometry Review 2001. Borrey D. Journal of American Mass Spectrometry 2008. Quantitative analysis of sphingoid base-1-phosphate as biacetylated derivatives by liquid chromatography–tandem mass spectrometry. 19: 719–728. Ramstein P. 850: 206–212. Marier JF and Vachon P. Nishio T. Liu Z. Higashi T. Columnswitching LC-MS/MS analysis for quantitative determination of testosterone in human serum. Simultaneous determination of 17a-hydroxypregnenolone and 17ahydroxyprogesterone in dried blood spots from low birth weight infant using LC-MS/MS. Wijtmans M. 80: 9042–9051. 38: 296–309. Yokoi H. Journal of Chromatography B 2007. Shimada K. Steroids 2010. An improved liquid chromatography–tandem mass spectrometric method for determination of 5a-androstane-3a. 25: 1–10 Copyright © 2010 John Wiley & Sons. Shimada K. in rat brains. Journal of Pharmaceutical and Biomedical Analysis 2008a. and future. Analytical and Bioanalytical Chemistry 2004. 16: 446–455. androsterone 7 Biomed. Eggink M. Carpenter KH and Wiley V. and Enke CG. Uchida S. Quantification of valproic acid and its metabolite 2-propyl4-pentenoic acid in human plasma using HPLC-MS/MS. Anari MR. Thamarai SK. Guenette SA. Natarajan V and Hubbard WC. Alternative procedure for charged derivatization to enhance detection responses of steroids in electrospray ionization-MS. Nishio T and Shimada K. Grever MR. Svensmark B. Analytical and Bioanalytical Chemistry 2004: 378: 875–882. Trace analysis of icariin in human serum with dansyl chloride derivatization after oral administration of Epimedium decoctin by liquid chromatography tandem mass spectrometry. Otomi N and Shimada K. Journal of Chromatography A 2010. Rapid Communications in Mass Spectrometry 2007. Cech N. Ninomiya Y. Guy PA. Yamauchi A and Shimada K. Chedrawi A. Chemical and Pharmaceutical Bulletin 2007c. de Esch IJP. Determination of prostatic androgens in 10 mg of tissue using liquid chromatography–tandem mass spectrometry with charged derivatization. Determination of neuroactive androgens. Higashi T. 71: 609–617. Analytical Biochemistry 2002. Derivatization of ethinylestradiol with dansyl chloride to enhance electrospray ionization: application in trace analysis of ethinylestradiol in rhusus monkey plasma. 21: 1984–1990. 825: 98–110. Yokoi H. 825: 214–222. Higashi T. Cech NB and Enke CG. Expanded newborn screening of inherited metabolic disorders by tandem mass spectrometry. Beaudry F. Development of a sensitive and selective LC-MS/MS method for simultaneous determination of gemcitabine and 2. 378: 932–939. Ekkebus R. Delatour T. Simultaneous quantification of multiple class of phenolic compounds in blood plasma by liquid chromatography– electrospray tandem mass spectrometry. 20: 362–387. Wan Y. Trace determination of steroids causing age-related diseases using LC/MS combined with detection-oriented derivatization. 748: 349–359. Cech NB. Analytical Sciences 2007d. 302: 10–18. Studies on neurosteroids XIX. Development of a selective ESI-MS derivatization reagent: synthesis and optimization for the analysis of aldehydes in biological mixtures. Shimozawa N. Rapid UPLC-MS/MS method for routine analysis of plasma pristanic. Hayashi N and Shimada K. 339: 129–136. Analysis of estrogens in serum and plasma from postmenopausal women: past present. Lam MHW. Journal of Lipid Research 2008. Kuhnol J. Yip SC. Analytical Chemistry 2002. Higashi T. Analytical and Bioanalytical Chemistry 2010. Chromatogr. Juhler RK. Journal of Chromatography B 2005. Derivatization of neutral steroids to enhance their detection characteristics in liquid chromatography. Clinica Chimica Acta 2002. Kalas TA and Naylor EW. Journal of American Mass Spectrometry 2005. The relative influence of acidity and polarity on responsiveness of small organic molecules to analysis with negative ion electrospray ionization mass spectrometry (ESI-MS). Zhang ZP and Karnes HT. 382: 1035–1043. 75: 297–306. Licea-Perez H. Nishio T. Chace DH and Kalas TA. Fonteh AN. Relative importance of basicity in the gas phase and in solution for determining selectivity in electrospray ionization mass spectrometry. Garg U and Dasouki M. 32: 203–212.2-difluoro-2-deoxyuridine in human plasma. Ehrmann BM. Niessenand WMA and Irth H. 55: 662–665. Eggink M. Cockx A.Derivatization reagents in LC/ESI-MS/MS spectrometry as benzofurazan derivatives. Studies on neurosteroids XXI. Analytical and Bioanalytical Chemistry 2005b. Journal of Chromatography B 2000. Naile J. 848: 188–199. Al-Dirbashi OY. Journal of Chromatography B 2009. A biochemical perspective on the use of tandem mass spectrometry for newborn screening and clinical testing. 1217: 506–513. Analytical Biochemistry 2005. phytanic. Quantification of free amino acids and dipeptides using isotope dilution liquid chromatography and electrospray ionization tandem mass spectrometry. Blum W. 48: 177–182. Lee HK and Yong EL. Harrington RJ and Harrington MG. Rashed MS. Marcucci G and Chan KK. Higashi T. Development of a LC-MS/MS method for the analysis of volatile primary and secondary amines as NIT (naphthylisothiocyanate) derivatives. Mizokami A and Namiki M. Analytical Chemistry 2008. Engelrelst V and Langois MR. 877: 2123–2129. Journal of Chromatography B 2005a. 397: 665–675. risk factors for development of breast cancer. 39: 411–417. Steroids 2006. Higashi T. Lingeman H. Determination of salivary dehydroepiandrosterone using liquid chromatography–tandem mass spectrometry combined with charged derivatization. Franklin RB and Evans DC. Higashi T. Chang H. Higashi T and Shimada K. Ninomiya Y and Shimada K. Aichholz R. Analytical Chemistry 2001. Byrd JC. Yamauchi A. Wang S. 3-Nitrotyrosine butyl ester: a novel derivative to assess tyrosine nitration in rat plasma by liquid chromatography–tandem mass spectrometry detection. Chemical and Pharmaceutical Bulletin 2006. 73: 2713– 2723.com . LC-MS analysis of charges in rat brain and serum testosterone levels induced immobilization stress and ethanol administration. Al-Hassnan Z. 39: 315–332. Journal of Chromatography B 2009. Clinical Biochemistry 2006. Journal of Chromatography B 2007. Wiseman S. Blair IA. Yamauchi A. Cech NB and Enke CG. Development of a rapid and sensitive LC-ESI/MS/MS assay for the quantification of propofol using a simple off-line dansyl chloride derivatization reaction to enhance signal intensity. Kool J. Clinical Chemistry 2003. Predicting electrospray response from chromatographic retention time. Iwaki N. Practical implications of some recent studies in electrospray ionization fundamentals. Higashi T. Henriksen T. Studies on neurosteroids XVIII. 73: 208– 213. View this article online at wileyonlinelibrary. Development and validation of liquid chromatography– tandem mass spectrometric method for determination of 5a-reduced pregnane-type neurosteroids in rat brain and serum. Klisovic R. Nagahama A. Claeson AS. Stadler RH and Tursky RJ. Cech NB. Higashi T.

Numazawa M. Analytical Sciences 2007a. Higashi T. Ikegami T. 14: 2019–2024. Analytical Chemistry 2007. Highly sensitive quantification of key regulatory oxysterols in biological samples by LC-ESI-MS/MS. 42: 9–16. Gerace R. Rapid Communications in Mass Spectrometry 1999. Clinical Chemistry 2001. Johnson DW. 2011. Jansen EEW. Li AC. a biomarker for polycyclic aromatic hydrocarbons in human urine. Honda A. Measurement of plasma pristanic. Johnson DW and Trinh MU. Beckman E. Shou WZ. 5a-androstane-3a. Daviglus ML. Min JZ. 13: 2388–2392. Rapid Communications in Mass Spectrometry 2000b. Journal of Lipid Research 2001a. Rockwood AL. 38: 351–361. Johnson DW. Journal of Lipid Research 2001b. Numazawa M and Matsuzaki Y. Journal of Lipid Research 2009b. Homma Y. Shi H. Daniels S. Steroids 2010a. Miyazaki H. Higashi T. 24: 1358–1364. Johnson DW. 32: 1636–1638. Bunker AM and Meikle AWM. Kushnir M. American Journal of Clinical Pathology 2008. Johnson DW. Matsuzaki Y and Miyazaki H. Urinary amino acid analysis: A comparison of iTRAQ (R)-LC-MS/MS. 17b-diol. Rapid Communications in Mass Spectrometry 2005b. Yamashita K. Doy M. Johnson DW.17b-diol. Langmann T. Tsai FJ. Rapid Communications in Mass Spectrometry 2007. 5-fluorouracil and capecitabine in human plasma. Rapid Communications in Mass Spectrometry 2001. Rapid monitoring assay of congenital adrenal hyperplasia with microbore high-performance liquid chromatography/electrospray ionization tandem mass spectrometry from dried blood spots. Bonaventure G. plasma. Inagaki S. Miyazaki T. Simultaneous determination of norethindrone and ethinyl estradiol in human plasma by high-performance liquid chromatography with tandem mass spectrometry-experiences on developing a highly selective method using derivatization reagent for enhancing sensitivity. Zhou S. Johnson DW. Trinh M-U and Fingerhut R. Chan Q. in urine and dried bloodspots by flow injection electrospray ionization tandem mass spectrometry. Kushnir M. Ikegami T. Li YH. Zhou S and Naidong W. Clinical Biochemistry 2005. Schifferer R. Naidong W and Lauterbach JH. Tsai CH. The use of chemical derivatization to enhance liquid chromatography/tandem mass spectrometric determination of 1-hydroxypyrene. Analysis of isomeric long-chain hydroxyl fatty acids by tandem mass spectrometry: application to the diagnosis of long-chain 3-hydroxyacyl CoA dehydrogenase deficiency. Yue B. Inagaki S. Shushan B. Ikegami T. Honda A. Biochimica et Biophysica Acta 2006. 52: 120–128. Fitzgerald RL and Meikle AW. 50: 2124–2130. 52: 809–818. Chromatogr. Ikegawa S and Chiba H. Min JZ and Toyo’oka T. Johnson DW. 79: 322–326. Honda A. Pattison EG. Urry FM and Roberts W. 25: 1–10 . Binder M. Xu G. by LC-ESIMS/MS. Journal of Chromatography B 2009. Bunker AM and Meikle AW. Journal of Mass Spectrometry 2008. Journal of Chromatography B 2005a. Highly sensitive and positively charged precolumn derivatization reagent for amines and amino acids in liquid chromatography/electrospray ionization tandem mass spectrometry. Rockwood AL. Highly sensitive quantification of 7a-hydroxy-4cholesten-3-one in human serum by LC-ESI-MS/MS. as a Girard T derivative.T. in rat brain and serum using liquid chromatography–tandem mass spectrometry. 23: 11–15. 48: 458–464. 47: 1993– 2002. by electrospray tandem mass spectrometry. 49: 2063–2072. 1761: 121–128. 798: 159– 162. 19: 3331–3338. Dimethylaminoethyl esters for trace. GC-MS. 50: 350–357. Baldwin I. 5: 17–27 Kaspar H. Licea-Perez H. Toyo’oka T. Yamashita K. and amino acid analyzer. Lampinen-Salomonsson M. Journal of Inherited Metabolic Disease 2000a. quantitative assay for homocarnosine in cerebrospinal fluid using stable-isotope dilution liquid chromatography–tandem mass spectrometry. Honda A. A flow injection electrospray ionization tandem mass spectrometric method for simultaneous measurement of trimethylamine and trimethylamine N-oxide in urine. Derivatization with Gerard reagent T combined with LC-MS/MS for the sensitive detection of 5-formyl-2’-deoxyuridine in cellular DNA. ten Brink HJ and Jakobs C. Numazawa M. Studies on neurosteroids XXXVI. Rapid Communications in Mass Spectrometry 2010. Analysis of dicarboxylic acids by tandem mass spectrometry. Plant methods 2009. 15: 2145–2151. Schulz B and Schmitz G. Honda A. 8 View this article online at wileyonlinelibrary. 52: 1559–1567. 17: 171–175. Kallenbach M. Tanji N. Suzuki Y and Suzuki K. Journal of Lipid Research 2008. 21: 59–63. Roberts WL. 43: 495–499. Journal of Chromatography B 2006. Journal of Chromatography B 2006. Hara T. Nagura Y. Yamashita K. Shibayama Y. Detection of altrenogest and its metabolites in post administration horse urine using liquid chromatography tandem mass spectrometryincreased sensitivity by chemical derivatization of glucuronic acid conjugate. Highly sensitive analysis of sterol profiles in human serum by LC-ESI-MS/MS. Berquist J. Santa and 5a-androstane-2a. Journal of Pharmaceutical and Bioanalytical Analysis 2010b. 22: 1434–1441. Rapid Communications in Mass Spectrometry 2007. phytanic and very long chain fatty acids by liquid chromatography– electrospray tandem mass spectrometry for the diagnosis of peroxisomal disorders. Highly sensitive quantification of serum malonate. Li Y. Simple and practical derivatization procedure for enhanced detection of carboxylic acids in liquid chromatography–electrospray ionization–tandem mass spectrometry. Roberts WL. Shirai M. Biomedical Chromatography 2008b. and urine. Biomed. Journal of Lipid Research 2009a. in plasma and blood spots. Numazawa M and Matsuzaki Y. 23: 475–486. Schmit RC and Jakobs C. A modified Girard derivatizing reagent for universal profiling and trace analysis of aldehydes and ketones by electrospray ionization tandem mass spectrometry. Ranieri E. Ichikawa T. Roberts WL. 42: 1699– 1705. Performance characteristics of a novel tandem mass spectrometry assay for serum testosterone. Fluoxetine-evoked changes in rat brain and serum level of neuroactive androgen. A rapid screening procedure for cholesterol and dehydrocholesterol by electrospray ionization tandem mass spectrometry. Ltd. Komaromy-Hiller G.com Copyright © 2010 John Wiley & Sons. 830: 196– 200. Hara T. Wang S and Bowen C. Li AC. 75: 338–345. Owen WE. Hayashi S. Shimada K. Shimada K and Toyo’oka T. Higashi T. High throughput quantification of cholesterol and cholesteryl ester by electrospray ionization tandem mass spectrometry (ESI-MS/MS). Kushnir M. Gibson KM. Jiang X. Ikegami T. as dimethylaminoethyl esters. 877: 1838–1846. Xu G. Liebisch G. Bondesson U and Hedeland M. ten Brink HJ. Hong W and Wang Y. Rapid and quantitative analysis of unconjugated C27 bile acids in plasma and blood samples by tandem mass spectrometry. Johnson DW. Kushnir NM. Miyazaki T. Hara T and Matsuzaki Y. Ichikawa T. 129: 530–539. Fukushima T and Toyo’oka T. Shirai M. Dettmer K. Tano Y. High-throughput quantitative measurement of methylmalonic acid in serum. A rapid and sensitive method for the simultaneous analysis of a aliphatic and polar molecules containing free carboxyl groups in plant extract by LC-MS/MS. 877: 1040–1046. Development of a sensitive and selective LC-MS/MS method for the determination of a-fluoro-balanine. a possible marker for de novo lipogenesis. Clinical Chemistry 2006a. Development and performance evaluation of a tandem mass spectrometry assay for 4 adrenal steroids. rapid analysis of fatty acids by electrospray tandem mass spectrometry. Journal of Lipid Research 2007b. Jacobs C and Verhoeven NM. Clinical Chemistry 2006b. 833: 245–256. A novel. High-sensitivity tandem mass spectrometry assay for serum estrone and estradiol. Lai CC. Rapid Communications in Mass Spectrometry 2003. Bunker AM. Journal of Chromatography B 2003. Shigematsu Y. 21: 2926–2932. Rockwood AL. Pattison EG. Journal of Chromatography B 2009. Li W. Biological and Pharmaceutical Bulletin 2009. A rapid screening procedure for the diagnosis of peroxisomal disorders: quantification of very long-chain fatty acids. Analysis of succinylacetone. Iwasawa N. Johnson DW. Higashi T. Trinh MU and Oe T. Lee CC and Lin WD. Nimkar S. Salivary chenodeoxycholic acid and its glycine-conjugate: their determination method using LC-MS/MS and varidation of their concentration with increased saliva flow rate. Elliott P and Oefner PJ. Yamashita K. MPAI (mass probes aided ionization) method for total analysis of biomolecules by mass spectrometry. Yamakata Y. Hifumi H. Hirayama T. Alkyldimethylaminoethyl ester iodides for improved analysis of fatty acids by electrospray ionization tandem mass spectrometry. Stamler J. 825: 223–232. Varshavsky M. Johnson DW. Mitamura K. Contemporary clinical usage of LC/MS: analysis of biologically important carboxylic acids.

Rosenfield J. and amines. Dalene M and Skarping G. Nordstrom A. Macarthur R and Kay J. O’Brien Z. Quantification of D-amino acids in the central nervous system of Aplysia carifornica by liquid chromatography/ 9 Biomed. Alamoudi M. Alwattar M and Ozand PT. Higashi T. Sensitive detection and structural characterization of trimethyl(p-aminophenyl)-ammonium-derivatized oligosaccharides by electrospray ionization–mass spectrometry and tandem mass spectrometry. Doi T. Shimbo K. Enhancing capillary liquid chromatography/tandem mass spectrometry to biogenic amines by pre-column derivatization with 7-fluoro-4-nitrobenzoxadiazole. Rashed MS. Jacobson A and Pappin DJ. Tandem mass spectrometric assay of succinylacetone in urine for the diagnosis of hepatorenal tyrosinemia. Adams CL. 66: 1302–1315. Nishio T. Validation and application of a liquid chromatography–tandem mass spectrometric method for the simultaneous determination of testosterone and dihydrotestosterone in rat prostic tissue using a 96-well format. Analytical Chemistry 2004: 76: 2869– 2877. Kashimada-K. Shimada K. Trace LC/MS/MS quantitation of 17b-estradiol as a biomarker for selective estrogen receptor modulator activity in rat brain. Luo R and Kohout TA. Santa T. Pillai S. Marchese JN. Takahashi K. Al-Dirbashi OY. Nakazawa M and Miyano H. Harris HA. Al-Ahaidib LY. 867: 49–56. 8: 129–133. alcohols. Hattan S. Journal of Chromatography B 2001. Ozawa S. 23: 1483–1492. Volovich V. thiols. European Journal of Pediatrics 2003. 877: 3515–3521. Analysis of methylmalonic acid in plasma by liquid chromatography–tandem mass spectrometry. Tanaka J and Shimada K. 44: 786–795. and Imai K. Liquid chromatography–tandem mass spectrometric method for determination of salivary 17a-hydroxyprogesterone: a noninvasive tool for evaluating efficacy of hormone replacement therapy in congenital adrenal hyperplasia. Clinical Chemistry 2004. Rahbeeni Z. Daniels S. Teale P. Liquid chromatography–electrospray ionization–mass spectrometry method in multiple reaction monitoring mode to determine 17aethynylestradiol residues in cattle hair without previous digestion. Jiang X and Naidong W. He F. Funatsu T and Imai K. 81: 5172–5179. Clarke A. Ai-Sayed MMA. Tarkowski P. Analytical Chemistry 2009a. Chennathukuzhi VM. Santa T. Bucknall MP. Nakamura M. 38: 324–331. Quan Z. Analytical Biochemistry 2005. Journal of Pharmaceutical and Biomedical Analysis 2007. Simultaneous determination of salivary testosterone and dehydroepiandrosterone using LC-MS/MS: method development and evaluation of applicability for diagnosis and medication for lateonset hypogonadism. Rashed MS. Vazquez BI. 21: 1207–1213. Dolezal K. Chen CY and Wang GS. Tarkowska D. Yamazaki J. 877: 1292–1298. Shimbo K. Konaka H. Chromatogr. Chromatographia 2010. Harada Y. Determination of amines as pentafluoropropionic anhydride derivatives in biological samples using liquid chromatography and tandem mass spectrometry. Automated precolumn derivatization system for physiological amino acids by liquid chromatography/mass spectrometry. Derivatization for LC–electrospray ionization–MS: a tool for improving reversed-phase separation and ESI response of bases. 339: 310–317. Agricultural and Food Chemistry 2007. Sandberg G and Moritz T. Lloyd T. Analyst 2004. 46: 1804– 1810. Oonuki T. Ozand PT. Ichibangase T. Clinical Chemistry 2000. Brown M. Oonuki T. Astot C. Song Y. Development and application of electrospray–active derivatization reagents for hydroxysteroids. Higashi T. Little D. Hirayama K. Odani A. 877: 2615– 2623. Shimbo K. Shou WZ. 1: 108–118. Rashed MS. Olgemoller R. Santa T. Kodo N. Journal of Chromatography B 2009. View this article online at wileyonlinelibrary. Hirayama K. Santa T. Rapid Communications in Mass Spectrometry 2001. Tandem mass spectrometry: a new method for acylcarnitine profiling with potential for neonatal screening for inborn errors of metabolism. Khainovski N. Determination of malondialdehyde in human plasma by fully automated solid phase analytical derivatization. 18: 414–421.Derivatization reagents in LC/ESI-MS/MS Lin YH. 758: 27–48. Ando T. Diagnosis of inborn errors of metabolism from blood spots by acylcarnitines and amino acids profiling using automated electrospray tandem mass spectrometry. Methylmalonic acid measured in plasma and urine by stable-isotope dilution and electrospray tandem mass spectrometry. Helgeson JK. 129: 522–528. Analysis of steroid estrogens in water using liquid chromatography/tandem mass spectrometry with chemical derivatizations. Rashed MS. Shibayama Y. Fukushima T. Pedreira D. Mekonnen B and Cai Y. 71: 241–252. Biomedical Chromatography 2010. Coon T. Juhasz P. Kubo S. Journal of Chromatography B 2009. Kumbhare D and Parkinson B. Liebl B and Roscher AA. Mizokami A. Lord HL. Alkyl halides. oestrogen and progestagen abuse in the bovine. Miura J and Miyano H. Multifunctional and highly sensitive precolumn reagents for amino acids in liquid chromatography/tandem mass spectrometry. Magera MJ. Dey S. 162: S67–S76. Amao M. ribosides. Nelson RE. Al-Dirbashi OY. Karisson D. Shibayama Y. 43: 1129–1141. Precolumn derivatization reagents for high-speed analysis of amines and amino acids in biological fluid using liquid chromatography/electrospray ionization tandem mass spectrometry. Rapid Communications in Mass Spectrometry 2009b. Byrd EA and Liu YM. 2011. Van Berkel GJ. Rapid Communications in Mass Spectrometry 1995. Franco CM. Biomedical Chromatography 2004. Matern D and Rinaldo P. Martin S. Rapid Communications in Mass Spectrometry 2004. phenols. Advances in analytical mass spectrometry to improve screening for inherited metabolic diseases. Millington DS. Ono M. Validation of a quantitative multi-residue urinary assay for the detection of androgen. Rashed MS. Miyai K and Mizutani S. Williamson B. Purkayastha S. Higashi T. 22: 115–118. Rapid Communications in Mass Spectrometry 2007. Rashed MS. O’Kane DJ. Singh RJ. Okamoto K. Liquid chromatography– tandem mass spectrometry assay for simultaneous measurement of estradiol and estrone in human plasma. Ichibangase T. Ozand PT. Hands J. Biomedical Chromatography 2008. 25: 1–10 Copyright © 2010 John Wiley & Sons. Al-Dirbashi OY and Fukushima T. Screening blood spots for inborn errors of metabolism by electrospray tandem mass spectrometry with a microplate batch process and a computer algorithm for automated flagging of abnormal profiles. Clinical applications of tandem mass spectrometry: ten years of diagnosis and screening for inherited metabolic diseases. Liang F and Liu YM. Journal of Mass Spectrometry 2010. Quirke JME. Song Y. Huang YLN. Post N. Al-Dbaas A. Pediatric Research 1995. Harrison ME. Funaishi A.com . Al-Hassnan Z. Multiplexed protein quantitation in Saccharomyces cerevisiae using amine-reactive isobaric tagging reagents. Journal of Inherited Metabolic Disease 1990. Petucci C. Parker K. Koh E and Namiki M. 3: 1154– 1169. Grebe SK. Rapid Communications in Mass Spectrometry 1994. Fukushima T. Scarth J. Electrospray tandem mass spectrometry in the diagnosis of organic icidemias. Chemical derivatization for electrospray ionization mass spectrometry. Derivatization reagents in liquid chromatography/electrospray ionization tandem mass spectrometry for biomedical analysis. Schmedes A and Brandslund I. Zhang X. Watkins PJF and Evans S. Roschinger W. O’Brien-Coker IC and Mallet GPAI. Yoshida H. Bartlet-Jones M. Evans L. Al-Owain M and Luanaigh MN. 9: 641–643. 18: 989–994. Fingerhut R. Bucknall MP and Little D. Development and validation of a highsensitivity liquid chromatography/tandem mass spectrometry (LC/ MS/MS) method with chemical derivatization for the determination of ethinylestradiol in human plasma. Yahashi A. Hirayama K and Miyano H. Molecular Cellular Proteomics 2004. 15: 920–928.N-dimethylamino)ethylaminosulfonyl]-7-Nmethylhydrazino-2. Al Amoudi M. 50: 373–384. Norwood DL and Roe CR. Kageyama N. Ltd. 915–918. Lolo M. Synthesis of benzofurazan derivatization reagents for carboxylic acids in liquid chromatography/electrospray ionization– tandem mass spectrometry (LC/ESI-MS/MS). Drug Discovery and Therapeutics 2007a. Onishi T. Aldehyde analysis by high performance liquid chromatography/tandem mass spectrometry. Ceprda A and Fente C. Yokokura T. Rashed MS. Synthesis of 4-[2-(N. Isothiocyanates as derivatization reagents for amines in liquid chromatography/electrospray ionization–tandem mass spectrometry (LC/ESI-MS/MS).3-benzoxadiazole (DAABD-MHz) as a derivatization reagent for aldehydes in liquid chromatography/electrospray ionization–tandem mass spectrometry. Journal of Chromatography 2009. Marand A. Analytical Chemistry 1994. 1. Al-Dirbashi OY. Ross PL. Awad A. Biomedical Chromatography 2010 24. Journal of Chromatography B 2008. Jacob M. Shimada K.1. Biomedical Chromatography 2007b. Yahashi A. Clinical Chemistry 2006. Madan A. 55: 9325–9329. Clinical Chemistry 1997. 45: 65–71. 52: 754–757. 21: 1973–1983. and intact nucleotides. 24: 683–691. 13: 321–324.

Morimoto R. Zhang F. Ketola RA. Okuyama M. Biomed. Zhou S and Cook KD. Steroids 2007a. Analytical Chemistry 2005. Van Berkel GJ and Asano KG. Covey TR. Kobayashi S and Numazawa M. Saavedra JE. Nakagawa R. Watanabe Y. 633: 223–231. Measuring fifteen endogenous estrogens simultaneously in human urine by high-performance liquid chromatography–mass spectrometry. Journal of Chromatography A 2008b. 72: 50–59. Development of highly sensitive quantification method for testosterone and dihydrotestosterone in human serum and postrate tissue by liquid chromatography–electrospray ionization tandem mass spectrometry. Honma S. Analytica Chimica Acta 2009. Inoue H. Takahashi M and Numazawa M. 77: 6359–6363. 66: 2096–2102. Microdetermination of catechol estrogen by liquid chromatography–electrospray ionization tandem mass spectrometry combined with picolinyl derivatization. Analytical and Bioanalytical Chemistry 2002. Zwiener C. Sachs H. Takahashi M and Numazawa M. specificity and reproducibility of its detection in blood by liquid chromatography–mass spectrometry. Okuyama M. Determination of muscimol and ibotenic acid in Amanita mushrooms by high-performance liquid chromatography and liquid chromatography–tandem mass spectrometry. Ito S. Analytical Chemistry 2005. Wang XY. 2011. Formation of the N-methylpyridinium ether deribvative of propofol to improve sensitivity. 852: 430–435. Van Berkel GJ. Zhang ZQ and Ruan JX. Journal of Chromatography A 2007c. 77: 6646–6654. Yamashita K. Biomedical Chromatography 2007. 74: 920– 926. Uutela P. Rapid Communications in Mass Spectrometry 2000. 1200: 114–121. 135: 127–132. Journal of Pharmaceutical and Biomedical Analysis 2008. Journal of Mass Spectrometry Society of Japan 2009a. Use of novel picolinoyl derivatization for simultaneous quantification of six corticosteroids by liquid chromatography– electrospray ionization tandem mass spectrometry. Yamashita K. A mechanistic study of elecrospray mass spectrometry: charge gradients within electrospray droplets and their influence on ion response. Ilag LL and Bergman B. Mitsunobu O and Honma S. Analytical Chemistry 1998. Tadokoro Y. Analysis of derivatized biogenic aldehydes by LC tandem mass spectrometry. 3. Aromatic and highly conjugated molecules. Straube E. Analytical Biochemistry 2008. Honma S. 70: 1544–1554. Quirke JME. 21: 1069–1076. Alkenes and alkynes. Quantification of 17a-ethinylestradiol in aquatic samples using liquid– liquid phase extraction. Rapid Communications in Mass Spectrometry 1995. 72: 819–827. Yoshida T and Kishi T. Development of sensitive derivatization method for aldosterone in liquid chromatography– electrospray ionization tandem mass spectrometry of corticosteroids. Yamashita K. Analytical method for determination of allylic isoprenols in rat tissues by liquid chromatography/tandem mass spectrometry following chemical derivatization with 3-nitrophtalic anhydride. Steroids 2009b. Kawahata T. Takahashi M. Yamashita K. Tsujikawa K. Chang SW. Comparison of different amino acid derivatives and analysis of rat brain microdialysates by liquid chromatography tandem mass spectrometry. 18: 2739–2742. Analytical protocol for identification of BMAA and DAB in biological samples. Honma S. Journal of American Society of Mass Spectrometry 2001. Journal of Chromatography B 2007. Tsukamoto S and Numazawa M. 14: 849–858. 73: 727–737. Numazawa M. Kuwayama K. 877: 4055–4058. Xu X. Schlling G and Hornuss C. Satoh F. Yuan M. Zhang ZW. Steroids 2007b.com Copyright © 2010 John Wiley & Sons. Thieme D. Yamashita K. Journal of Chromatography B 2009. Piepponen P and Kostianinen R. Method optimization for the determination of carbonyl compounds in disinfected water by DNPH derivatization and LC-ESI-MS-MS. 375: 105–114. Development and evaluation of a reference measurement procedure for the determination of estradiol-17b in human serum using isotope-dilution liquid chromatography–tandem mass spectrometry. Highly sensitive determination of estrone and estradiol in human serum by liquid chromatography–electrospray ionization tandem mass spectrometry. 9: 1516–1521 (1995). Tsukamoto S. Zhuang XM. dansylderivatization. Timperio AM.T. Glauner T and Frimmel FH. 57: 75–80. Enhanced sensitivity for the determination of amibiphilic polyaromatic amines by LC-MS/MS after acetylation. 21: 73–77. Xia YQ. Spacil Z. 47: 560–566. Fox SD. Miyashiro Y. Adams CL. Veenstra TD. Lovell MA and Lynn BC. 876: 76–82. Patel S. Bakhtiar R. Takahashi M and Numazawa M. Chromatogr. 18: 1621–1628. Okuyama M. Derivatization for electrospray ionization mass spectrometry. 1173: 120–128. McClymont EL and Woodburn KB. Effects of solvent composition on molecular ion response in electrospray mass spectrometry: investigation of the ionization process. Yamashita K. Trace level quantification of deuterated 17b-estradiol and estrone in ovariectomized mouse plasma and brain using liquid chromatography /tandem mass spectrometry following dansylation reaction. Tigani RA. 56: 873– 877. Tai AAC and Welch MJ. 10 View this article online at wileyonlinelibrary. Analytical Chemistry 1994. Determination of 4-dimethylaminophenol concentrations in dog blood using LC-ESI/MS/MS combined with precolumn derivatization. Grandinetti F and Zolla L. Electrochemically ionizable derivatives. Bartels MJ. Quirke JME. Kobayashi S. Rapid Communications in Mass Spectrometry 2004. Rapid Communications in Mass Spectrometry 2004. Van Berkel GJ. 2. 1067: 181–190. Ltd. Dilley AS. Teshima K and Kondo T. Maekubo H. Analysis od steroidal estrogens as pyridine-3-sulfonyl derivatives by liquid chromatography electrospray tandem mass spectrometry. Iwata Y. Xu L and Spink DC. 77: 3383–3389. Chemical derivatization for electrospray ionization mass spectrometry. Brodeur JC. Takahashi M and Numazawa M. Eriksson J. 25: 1–10 . Williams TI. Analytical Chemistry 2005. Yamashita K. Chemical and Pharmaceutical Bulletin 2008a. Rasmussen U. Miyaguchi H. 4. Dekant W and Volkel W. Derivatization for electrospray ionization mass spectrometry. Santa tandem mass spectrometry. Honasson S. Chemically enhanced liquid chromatography/tandem mass spectrometry determination of glutamic acid in the diffusion medium of retinal cells. Simultaneous determination of tetrahydrocortisol. 12: 206–211. Fagioni M. Falk R. Synthesis of pyridine-carboxylate derivatives of hydroxysteroids for liquid chromatography–electrospray ionization–mass spectrometry. Journal of Chromatography B 2008. Preparation and structural elucidation of the pocolinyl ester of aldosterone for liquid chromatography–electrospray ionization tandem mass spectrometry. allotetrahydrocortisol and tetrahydrocortisone in human urine by liquid chromatography–electrospray ionization tandem mass spectrometry. Steroids 2008c. Zhou S and Hamburger M. Analyst 2010. Journal of Chromatography A 2005. 372: 615–621. Yamashita K. Issaq HJ. Roman JM. Keefer LK and Ziegler R. Rapid Communications in Mass Spectrometry 2007. Honma S. Kanamori T. Karanam B and Evans DC. and liquid chromatography/positive electrospray tandem mass spectrometry. Nakagawa R. Takahashi M and Numazawa M. Okuyama M.

Sign up to vote on this title
UsefulNot useful