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Distribution of drugs into tissues is an important determinant of the overall PK and PD profile. Thus, bioanalysis of drugs and their metabolites in tissues can play an important role in understanding the pharmacological and toxicological properties of new drug candidates. Unlike liquid matrices, bioanalysis in tissues offers unique challenges such as proper tissue sampling, appropriate tissue sample preparation, efficient extraction of the analytes from the tissue homogenates, and demonstration of stability and recovery of analytes in intact tissues. This article provides a systematic review of tissue sample analysis for small molecules using LC–MS/MS. The authors provide rationale for tissue sample analysis, and discuss strategies for method development, method qualification or validation, and sample analysis. Unique aspects of method development and qualification/validation are highlighted based on authors’ direct experiences and literature summary. Analysis using intact tissue samples such as MALDI imaging is also briefly discussed.

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The positive- and negative-ion laser desorption ionization Fourier transform ion cyclotron resonance mass spectra of two known, underivatized, linear oligosaccharides obtained by enzymatic hydrolysis with bacterial viruses of the corresponding native bacterial capsular polysaccharides were analyzed. The high-resolution spectra, estimated to be from less than 1 nmol of sample, showed prominent pseudo-molecular ions, as well as fragment ions derived from glycosidic bond and ring cleavages. For some of these frequently observed ring fragments, structures are proposed by analogy to known even-electron gas phase fragmentations and even-electron solution chemistry. The structures of the capsular biopolymers were delineated from these fragmentations, confirming that Klebsiella pneumoniae serotype K3 capsular polysaccharide is made up of a pentasaccharide repeating unit, consisting of four hexoses, one hexuronic acid and one pyruvic acid acetal; and confirming that the tetrasaccharide repeating unit of Escherichia coli serotype K9 is composed of two hexoses, one acetamido hexose and one N -acetyl neuraminic acid. The analysis of oligosaccharides containing N -acetylated sugars was complicated by the fact that an amino hexose attached to a −C 2 H 3 O 2 ring fragment shares the same elemental formula as an acetamido hexose. Nevertheless, other less ambiguous structural ions could be used for sequencing. Furthermore, analysis of the ring fragments also provided some indication of the positions of linkage of certain monosaccharide residues.

The vitamin K dependent carboxylase of liver microsomes is involved in the posttranslational modification of certain serine protease zymogens which are critical components of the blood clotting cascade. During coupled carboxylation/oxygenation this carboxylase converts glutamate residues, dihydrovitamin K, CO2, and O2 to a gamma-carboxyglutamyl (Gla) residue, vitamin K (2R,3S)-epoxide, and H2O with a stoichiometry of 1:1 for all substrates and products. In this paper we investigate the role of molecular oxygen in the reaction by following the course of the oxygen atoms using 18O2. Two different mass spectroscopic techniques, electron ionization positive ion mass spectrometry and supercritical fluid chromatography-negative ion chemical ionization mass spectrometry, were used to quantitate the amount of 18O incorporation into the various oxygens of the vitamin K epoxide product. We found that 0.95 mol atoms of oxygen were incorporated into the epoxide oxygen, 0.05 mol atoms of oxygen were incorporated into the quinone oxygen of vitamin K epoxide, and the remaining ca. 1.0 mol atoms of oxygen were incorporated into H2O. No incorporation of oxygen into vitamin K epoxide from 50% H2(18)O was observed. Thus, the carboxylase operates as a dioxygenase 5% of the time during carboxylation/oxygenation. The relevance of these findings with respect to the nonenzymic "basicity enhancement" model proposed by Ham and Dowd [(1990) J. Am. Chem. Soc. 112, 1660-1661] is discussed.