Aim: Mass-selective quantitation is a powerful attribute of LC-MS as a platform for bioanalysis. Here, a sensitive LC-MS approach has been validated for an oligonucleotide having chemical modifications (e.g., N-acetylgalactosamine [GalNAc] conjugated), to distinguish between the conjugated and unconjugated forms of the oligonucleotide, thereby enabling a nuanced view of the pharmacokinetic profile. Results: A high-sensitivity methodology for mass-specific measurement of AZD8233, a GalNAc-conjugated 16-mer oligonucleotide, using LLE-SPE with optimized LC conditions and detection of a low-mass fragment ion was successfully validated in the range of 0.20-100 ng/ml in human plasma. Conclusion: The AZD8233 LC-MS methodology adds valuable insight on the GalNAc linker\'s in vivo stability to the program and should be broadly applicable to oligonucleotides requiring high sensitivity and mass-selective measurement for quantitative discrimination from metabolites and endogenous interferences.

Polysaccharides have broad bioactivities and are major components of water decoction of herb formulae. However, the quality control of polysaccharides remains a challenge. Oligosaccharide-fragment approach has been considered in elucidating chemical structures of polysaccharides, but never been used for quantitation. Using reference chemicals and a real sample Danggui Buxue Tang (DBT) in this study, an oligosaccharide-marker approach was established to quantify specific polysaccharides. Firstly, linear relationships between parent polysaccharides and hydrolysis-produced daughter oligosaccharides were verified using reference polysaccharides. Then in case of DBT, two fluorescence-labeled oligosaccharides with high specificity to individual parent polysaccharides were selected as markers. They were easily isolated and identified. Their potential in quantification of parent polysaccharides were satisfactorily validated in terms of linearity (r≥0.99), repeatability (RSD ≤ 8.4 %), and spike recovery (≥80 %). This method could be a promising approach for quality assessment of polysaccharides in herbal formulae.

The study focused on the application of high-resolution mass spectrometry (HRMS) to postmortem toxicological analysis. Fast and simple sample preparation involved precipitation with acetonitrile, removal of phospholipids using special columns and filtration. Qualitative and quantitative analyses were performed using ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry. The method was validated by determining the limit of quantification, precision, recovery and matrix effect. The use of a high-resolution spectrometer allowed us to determine the precise masses of the fragments of interest and to suggest the fragmentation pathway of baclofen. The usefulness, effectiveness and assets of the procedure were confirmed by an authentic case of a 25-year-old woman fatally intoxicated with baclofen who was found dead in her apartment. Toxicological analysis of postmortem blood samples demonstrated that the baclofen concentration was 30.7 μg/mL. In only one published case describing fatal baclofen intoxication were no other xenobiotics (that could interact with baclofen) found. To our knowledge, this is the first report dealing with analysis of baclofen by HRMS.