Liquid Chromatography-electrospray Triple Quadrupole Research Articles

A Qualitative Method to Detect Paraproteins from Serum Using Ultra Performance Liquid Chromatography Electrospray Triple Quadrupole Mass Spectrometry.

Mass spectrometry-based techniques are increasingly reported in the literature for identifying paraproteins due to their improved specificity and sensitivity. The present study demonstrates the capability of ultra performance liquid chromatography (UPLC) electrospray ionization triple quadrupole mass spectrometry for the qualitative analysis of paraproteins. Paraproteins from patient serum (n = 40) were immunopurified using agarose beads coated with camelid antibodies that are specific for various subtypes of immunoglobulins (Igs; G, A, M, and light chains κ, λ). The extracted Igs are reduced to separate light chains from heavy chains in solution. The reduced sample was subjected to UPLC and mass measured using electrospray ionization-mass spectrometry. The mass spectral peaks at specific retention times were deconvoluted after clean-up to obtain the mass of light chains. The interpretation of liquid chromatography peaks and LC-MS data was validated by comparing them with immunofixation electrophoresis (IFE) results. The interpretation from the chromatographic pattern had a 92.5% (37/40) agreement when compared with mass information. The correlation of mass spectrometry data to IFE was 90% (36/40). The high mass of light chains (>25 kDa) was suggestive of glycosylation. Patient sera positive for IgGκ on IFE (n = 15) were analyzed for the interference of tAbs. The mass of Daratumumab observed in a sample was confirmed by the treating physician. A biclonal of same isotype (IgGκ) was identified. The feasibility of using liquid chromatography triple quadrupole mass spectrometry for the identification of the subtype of paraproteins has been demonstrated. The method's applicability to screen for interference from tAbs and identification of biclonals of the same isotype has been highlighted.

Hepatic Metabolomics Analysis of Hybrid Grouper (Epinephelus fuscoguttatus♀×Epinephelus polyphekadion♂) Fed With Quercetin and Sodium Quercetin-5′-Sulfonates

Combining feeding trials and metabolomics analyses of tissues and biofluids could shed light on nutrient effects and changes in feed intake. In the present study, hybrid grouper (F1 hybrid Epinephelus fuscoguttatus♀×Epinephelus polyphekadion♂) was used as the marine fish model to quantify the impacts of quercetin and sodium quercetin-5′-sulfonates on serum biochemistry and the status of hepatic lipid accumulation and the changes of metabolites in the liver using ultra-performance liquid chromatography-electrospray triple quadrupole mass spectrometry (UPLC-QTRAP/MS/MS). The study shows that total protein, albumin, alkaline phosphatase and glucose were not significantly different among the three groups (P > 0.05). Total cholesterol, triglyceride, and alanine aminotransferase of fish fed quercetin, and sodium quercetin-5′-sulfonates were significantly lower than fish fed without additives (P < 0.05). Low-density lipoprotein cholesterol and aspartate aminotransferase of fish fed quercetin were significantly lower than that of fish fed without additives and sodium quercetin-5′-sulfonates (P < 0.05). The high-density lipoprotein cholesterol of fish fed quercetin, and sodium quercetin-5′-sulfonates were significantly higher than that of fish fed without additives (P < 0.05). Lipid accumulation in the quercetin and sodium quercetin-5′-sulfonates groups decreased significantly. Quercetin and sodium quercetin-5′-sulfonates were effective by increasing hypolipidemic and hepatoprotective compounds that are known for reducing blood lipid levels and liver fat accumulation. As a result of this study, we provide international data for metabolic adaptations during the additives feeding using the fish liver as the study model. By understanding the metabolic effects of these feed additives, this study provides a first step toward understanding the molecular mechanisms of these additives and how they function.

Liquid chromatography tandem mass spectrometry determination of chemical markers and principal component analysis of Vitex agnus-castus L. fruits (Verbenaceae) and derived food supplements

A validated analytical method for the quantitative determination of seven chemical markers occurring in a hydroalcoholic extract of Vitex agnus-castus fruits by liquid chromatography electrospray triple quadrupole tandem mass spectrometry (LC/ESI/(QqQ)MSMS) is reported. To carry out a comparative study, five commercial food supplements corresponding to hydroalcoholic extracts of V. agnus-castus fruits were analysed under the same chromatographic conditions of the crude extract. Principal component analysis (PCA), based only on the variation of the amount of the seven chemical markers, was applied in order to find similarities between the hydroalcoholic extract and the food supplements. A second PCA analysis was carried out considering the whole spectroscopic data deriving from liquid chromatography electrospray linear ion trap mass spectrometry (LC/ESI/(LIT)MS) analysis. High similarity between the two PCA was observed, showing the possibility to select one of these two approaches for future applications in the field of comparative analysis of food supplements and quality control procedures.