Abstract: This is the first of a series of reviews on the application of derivatization in mass spectrometry. A description is given of advances in silylation as a powerful tool used for increasing the volatility, thermal and thermo-catalytic stability, and chromatographic mobility of polar and unstable organic compounds. In addition to chemical aspects of silylation, mass spectral properties of silyl derivatives useful for structure determination and quantitation of various organic and biologically-active compounds, mainly by GC/MS, are described. Practically all tested and widely used silylating agents are described. The role of comprehensive libraries containing reference mass spectra for various silyl derivatives and search systems in structure determination is emphasized. Applications of silylation for particular analyses are summarised.

Abstract: A new, fast, sensitive, and solventless extraction technique was developed in order to analyze beer carbonyl compounds. The method was based on solid-phase microextraction with on-fiber derivatization. A derivatization agent, O-(2,3,4,5,6-pentafluorobenzyl)hydroxylamine (PFBOA), was absorbed onto a divinyl benzene/poly(dimethylsiloxane) 65-μm fiber and exposed to the headspace of a vial with a beer sample. Carbonyl compounds selectively reacted with PFBOA, and the oximes formed were desorbed into a gas chromatograph injection port and quantified by mass spectrometry. This method provided very high reproducibility and linearity. When it was used for the analysis of aged beers, nine aldehydes were detected: 2-methylpropanal, 2-methylbutanal, 3-methylbutanal, pentanal, hexanal, furfural, methional, phenylacetaldehyde, and (E)-2-nonenal. Keywords: Aldehydes; beer analysis; derivatization; SPME; GC/MS

Abstract: This paper describes two highly sensitive analytical methods for the unambiguous determination of pharmaceutical residues and related polar contaminants in environmental water samples. Both analytical methods use solid phase extraction (SPE), chemical derivatization, and detection of the analytes by capillary gas chromatography-mass spectrometry (GC-MS) with selected ion monitoring (SIM). Using the two methods, a total of 19 pharmaceuticals and 7 related polar contaminants, most of them pesticides, can be detected and quantified down to the low ng/L range in sewage, surface, ground, and drinking water. Recoveries between 70 and 110% and standard deviations of less than 15% were determined for 21 analytes when applying the analytical method that uses pentafluorobenzyl bromide (PFBBr) for derivatization. The second method, which employs N-(t-butyldimethylsilyl)-N-methyltrifluoroacetamide (MTBSTFA) for derivatization was found to be slightly less sensitive and reproducible but it also permitted the analysis of several other environmentally important compounds such as the antiepileptic drugs carbamazepine or primidone. From spiking experiments, limits of detection (LODs) between less than 1 and 10 ng/L and limits of quantification (LOQs) between 1 and 40 ng/L were calculated.

Abstract: The present review is devoted to acylation as a widely employed derivatization procedure for protection of OH (alcohols, polyols, phenols, enols), SH (thiols) and NH (amines, amides) groups in order to increase volatility, improve chromatographic properties and, if possible, improve mass spectral properties of derivatives. Chemical aspects of derivatization and various acylating agents are characterized. Mass spectral [electron ionization (EI), chemical ionization (CI) and negative-ion (NI) CI] properties of derivatives that are helpful in identification, structure elucidation and quantitative determination of the analyzed compounds are discussed. Some recent analytical applications of the procedure in synthetic organic chemistry, clinical chemistry, environmental chemistry etc. are summarized.

Abstract: To develop new fluorescent derivatization reagents, we investigated the relationship between the chemical structures and the fluorescence quantum yields (Φf) of coumarins, quinoxalinones and benzoxadinones. Forty-six compounds were synthesized and their fluorescence spectra were measured in n-hexane, ethyl acetate, methanol and water. The energy levels of these compounds were calculated by combination of the semi-empirical AM1 and INDO/S (CI = all) methods. The ΔE(Tn(n,π*), S1(π,π*)) (the energy gap between the Tn(n,π*) and S1(π,π*) states) values were well correlated with the Φf values, which enables us to predict the Φf values from their chemical structures. Based on this relationship, 3-phenyl-7-N-piperazinoquinoxalin-2(1H)-one (PQ-Pz) and 7-(3-(S)-aminopyrrolidin-1-yl)-3-phenylquinoxalin-2-(1H)-one (PQ-APy) were developed as fluorescent derivatization reagents for carboxylic acids. The derivatives of the carboxylic acids with PQ-Pz and PQ-APy showed large Φf values even in polar solvents, suggesting that these reagents are suitable for the microanalysis of biologically important carboxylic acids by reversed phase HPLC.

Abstract: d-Lactic and l-lactic acids were simultaneously determined by means of a column-switching high-performance liquid chromatography (HPLC) with fluorescence detection. As a fluorescence reagent, 4-nitro-7-piperazino-2,1,3-benzoxadiazole (NBD-PZ) was employed for the fluorescence derivatization of lactic acid. The proposed HPLC system adopted both octylsilica (Cadenza CD-C8) and amylose-based chiral columns (CHIRALPAK AD-RH), which proved to give a sufficient enantiomeric separation of the lactic acid derivatives with a separation factor (α) of 1.32 and a resolution (Rs) of 1.98. Moreover, the features of the first elution of d-lactic acid peak in the proposed HPLC were convenient for the determination of trace amount of serum d-lactic acid, which is known to increase under diabetes. Intra-day and inter-day accuracies were in the range of 90.5–101.2 and 89.0–100.7%, and the intra-day and inter-day precisions were 0.3–1.2 and 0.4–4.8%, respectively. The proposed method was applied to determine d-lactic and l-lactic acids in human serum of normal subjects and diabetic patients, showing that both d-lactic and l-lactic acid concentrations were significantly increased in the serum of diabetic patients (n=31) as compared with normal subjects (n=21). This fact was found for the first time owing to the development of the proposed HPLC method which is able to determine d-lactic and l-lactic acid simultaneously. Finally, serum d-lactic acid concentrations determined by the proposed HPLC method were compared with those from a reported enzymatic assay, and the smaller p value between normal subjects and diabetic patients was shown by the proposed HPLC method.

Abstract: A protocol combining immobilized metal ion affinity chromatography and β-elimination with concurrent Michael addition has been developed for enhanced analysis of protein phosphorylation. Immobilized metal ion affinity chromatography was initially used to enrich for phosphorylated peptides. β-Elimination, with or without concurrent Michael addition, was then subsequently used to simultaneously elute and derivatize phosphopeptides bound to the chromatography resin. Derivatization of the phosphate facilitated the precise determination of phosphorylation sites by MALDI-PSD/LIFT tandem mass spectrometry, avoiding complications due to ion suppression and phosphate lability in mass spectrometric analysis of phosphopeptides. Complementary use of immobilized metal ion affinity chromatography and β-elimination with concurrent Michael addition in this manner circumvented several inherent disadvantages of the individual methods. In particular, (i) the protocol discriminated O-linked glycosylated peptides from phospho...

Abstract: Acrylamide (AA) is a reactive compound widely used as an industrial chemical. It is also, as recently shown, present in heated foodstuffs. AA is known to cause tumors in rodents and is classified as probably carcinogenic to humans. The metabolite glycidamide (GA) is assumed to be the predominant genotoxic agent in AA exposure. Therefore, knowledge about in vivo doses of GA is essential for cancer risk assessment of exposure to AA. The in vivo dose of GA could be inferred from the level of the adduct formed by GA with N-terminal valine (GA-Val) in hemoglobin (Hb), detached as a pentafluorophenylthiohydantoin (PFPTH) and measured by gas chromatography/tandem mass spectrometric (GC/MS/MS) analysis. However, due to the highly polar character of the GA-Val-PFPTH derivative, it was found necessary to modify the method through further derivatization. This paper presents an evaluation of acetonization for derivatization of the adjacent bond;OH and bond;NH(2) groups in the adduct formed from GA. Good reproducibility was obtained. Also, acetonization improves the response and thus increases the sensitivity of the GC/MS/MS analysis of the PFPTH derivative of GA-Val. The sensitivity obtained is sufficient for studies of background adduct levels of GA in animals and in humans. Acetonization as a method for derivatization is robust and simple.