Journal of pharmaceutical and biomedical analysis最新文献

Danning tablet (DNT) is a traditional Chinese medicine (TCM) that contains seven herbal ingredients. It has been clinically used to treat liver and gallbladder diseases in humans. However, the complex composition of TCM prescriptions makes it challenging to fully analyze different polar range compounds. The supercritical fluid chromatography (SFC) method has stronger selectivity for weak polarity and low volatility substances. In contrast, ultra-high performance liquid chromatography (UHPLC) has stronger selectivity for compounds with strong polarity and high boiling points, which offsets the disadvantages of SFC. We aimed to establish a complementary and integrated strategy for multicomponent characterization and attribution of DNT based on ultra-performance convergence chromatography (UPCC) and UHPLC combined with quadrupole-time-of-flight mass spectrometry (QTOF-MS) and identify the potential qualitative indicator. The chemical compounds of DNT were analyzed by matching the self-built databases on the UNIFI platform. Network pharmacology was used to verify the reasonableness of the qualitative indicators with the relevant targets and the enrichment pathways related to the treatment of DNT. A total of 247 compounds were characterized. Specifically, the UPCC-QTOF-MS technology individually characterized 73 compounds. The UHPLC-QTOF-MS technology individually characterized 75 compounds. As a result, the study defined 11 compounds as the potential qualitative indicators. The relevant targets and the enrichment pathways related to the treatment of DNT were constructed. This study completed the comprehensive characterization of the full coverage of the polarity of DNT. The potential qualitative indicators can be extended to improve the accuracy of DNT quality evaluation.

Following regulatory pressure, the manufacture of long-chain per- and polyfluoroalkyl substances (PFAS) has been phased out, and alternatives such as short-chain homologs and ether-PFAS have replaced the bioaccumulative long-chain PFAS. However, data are lacking regarding the toxicokinetic (TK) properties of certain PFAS, particularly emergent substitutes for long-chain compounds. Additionally, the existing analytical methods used for TK studies measure a single compound or only a few simultaneously. For this reason, an LC-MS/MS method was developed for the simultaneous quantification in mouse plasma of 11 PFAS representative of some of the most important categories of these compounds, for application in TK studies. The method was successfully validated in the range of 0.5-1000 ng/mL, in accordance with the European Medicines Agency guidelines, and applied to a 24-h pilot TK study conducted in mice. All compounds were monitored over 24 hours in the pilot study. The present method is therefore suitable for the simultaneous quantification of PFAS in plasma samples and can be applied for future TK studies.

Profiling the site-specific transcriptomes of microregions of interest (mROIs) contributes to a complete understanding of multicellular organisms. However, the simple and efficient isolation of mROIs for spatially detecting gene expression remains challenging. Here, we develop an efficient capillary-based microdissection system (CMS) for precisely isolating targeted samples from tissue sections. Optimized sampling procedures reveal that CMS can perform mROI isolation with an efficiency of 97.9 %, and detect a sufficient number of genes for gene expression profiling (CMS-seq). We apply CMS-seq to uncover spatial heterogeneity in the cortex region of the mouse, and the subregions of hippocampus in an Alzheimer's disease (AD) mouse. Results demonstrate that CMS-seq can profile spatial transcriptomes in tissue sections and holds promise for application spatial multi-omics.

Congelex Laxative Granules is an in-house preparation of Hebei Provincial Hospital of Traditional Chinese Medicine. This study aims to establish the HPLC fingerprint of Congelex Laxative Granules and evaluate its quality using chemometric methods. The Agilent Eclipse Plus C18 column and a methanol-water gradient elution system were employed, with detection at 224 nm. The High-performance liquid chromatography (HPLC) analysis of 20 batches of samples successfully established a fingerprint with 17 common peaks and a similarity exceeding 0.95. Seven main active components, including salidroside, echinacoside, acteoside, specnuezhenide, wedelolactone, aurantio-obtusin, and chrysophanol, were quantitatively analyzed. Hierarchical Cluster Analysis (HCA), principal component analysis (PCA), and orthogonal partial least squares-discriminant analysis (OPLS-DA) were used to comprehensively evaluate sample quality. Results indicated that the 20 batches could be divided into two categories, with consistent results from PCA and HCA. The OPLS-DA model was stable and reliable, identifying salidroside, acteoside, and chrysophanol as key differential markers. The conclusion shows that the established fingerprint and content determination method provide an accurate and reliable tool for the quality control and comprehensive evaluation of Congelex Laxative Granules.

Daidzin, as one of isoflavone glycosides, has been reported to have multiple activities with few absorbed into body. However, the metabolic behavior of daidzin by intestinal flora has not been researched, that this defect severely constrains its applications. In this study, daidzin and its metabolites were qualitatively and quantitatively analyzed by HPLC and ultra-high performance liquid chromatography coupled to high-resolution mass spectrometry (UHPLC-HRMS) in the fermentation system for daidzin and fecal bacteria. Meanwhile, the alterations of intestinal flora with daidzin were detected by 16S rRNA sequencing technology. Based on the results of intestinal flora, the daidzin and its metabolites transformed by the screened probiotics were quantified and qualified, which the results would corroborate the transformation of daidzin and fecal bacteria. Eventually, daidzin was decreased from 0.30158 mg/mL at 0 h to 0.01176 mg/mL at 48 h, daidzein, as the aglycone of daidzin, was increased from 0.02963 mg/mL at 0 h to 0.04682 mg/mL at 48 h, suggesting the presence of other metabolites. Next, 31 metabolites including the products of ketone removal, Retro-Diels-Alder (RDA) fragmentation, hydroxylation, methylation, C ring cracking and sulfation were identified. The results of 16S rRNA sequencing showed that the intestinal flora, especially Bifidobacterium, was dramatically altered after incubation with daidzin (p < 0.05). Hereby, the fermentation systems of five probiotics (Lactobacillus 3044, Bifidobacterium adolescentis 1.2190, Bifidobacterium longum 25033, Lactobacillus plantarum F1 and Lactobacillus plantarum B2) and daidzin were approved, and these results showed that most metabolites of daidzin were able to be identified with the identical transformation reactions. The study revealed the rationality of daidzin biotransformation at the new perspective, and constructs a new model for fecal metabolites of compounds. These results will also broaden the continued research on daidzin.

E6011 is a monoclonal antibody that is currently under development for the treatment of rheumatoid arthritis. While ligand binding assays (LBAs) are typically employed for the determination of therapeutic antibodies, ultra-performance liquid chromatography with tandem mass spectrometry (UPLC-MS/MS) represents an alternative platform. E6011 in monkey serum was treated with ammonium sulfate to obtain pellets for subsequent processing. The pellets were subjected to denaturalization reduction, alkylation, and tryptic digestion. The resulting signature peptide of E6011, TLADGVPSR, was assayed. The pellet digestion assay was validated in accordance with the established bioanalytical guidelines. E6011 in monkey serum was quantifiable from 3 to 729 µg/mL, with a sample volume of 0.02 mL. The selectivity was confirmed in 12 individual monkey sera. The accuracy and precision were within ± 11.2 % and 15.0 %, respectively. The validated UPLC-MS/MS assay was employed in a pharmacokinetic study in monkeys. After the intravenous dose at 1 mg/kg, E6011 reached the maximum of 27.4 μg/mL, then declined with the half-life of 169 h. The serum E6011 concentrations determined by the UPLC-MS/MS were comparable to those obtained by the LBA with electrochemiluminescence detection. These findings suggest that the established simple UPLC-MS/MS assay is reproducible and can serve as an alternative assay platform.

For the effective treatment of tuberculosis with first-line anti-tubercular drugs, drug concentrations need to be measured at the site of infection to determine drug exposure. To enable the measurement of the anti-tuberculosis drugs isoniazid and pyrazinamide in the nervous system of patients with tuberculous meningitis, an analytical method was developed and validated for the quantification of these drugs in human cerebrospinal fluid. Samples were prepared by solid phase extraction using Strata-X polymeric extraction plates. The analytes were separated by high-performance liquid chromatography on an Atlantis T3, 100 A, 3 µm, 2.1 mm × 100 mm analytical column with gradient elution, employing a mobile phase that consisted of acetonitrile-methanol-formic acid (50:50:0.1, v/v/v), at a flowrate of 0.25 mL/min. The total run time was 4.5 minutes, and the average retention times of isoniazid and pyrazinamide were 1.1 and 1.3 min, respectively. The analytes and their respective deuterated internal standards were detected on a Sciex API4000 triple quadrupole mass spectrometer applying positive electrospray ionization with multiple reaction monitoring as the detection mode. The method was validated according to the FDA and EMA guidelines. The method was demonstrated to be accurate, reproducible, and robust, showing the necessary sensitivity and specificity for the quantification of isoniazid and pyrazinamide in cerebrospinal fluid. The method was successfully applied to analyze clinical samples from the LASER-TBM and TBM-KIDS clinical studies.

Clinically heterogeneous spectrum and molecular phenotypes of inflammatory bowel disease (IBD) remain to be comprehensively elucidated. This exploratory multi-omics study investigated the serum molecular profiles of Crohn's disease (CD) and ulcerative colitis (UC), in association with elevated fecal calprotectin and disease activity states. The serum proteome, metabolome, and lipidome of 75 treated IBD patients were profiled. Single- and multi-omic data analysis was performed to determine differential analytes and integrative biosignatures for biological interpretations. We found that chronic inflammation, phosphatidylcholines and bile acid homeostasis disturbances underlined the differences between CD and UC. Besides, elevated calprotectin was associated with higher levels of inflammatory proteins and sphingomyelins (SM) and lower levels of bile acids, amino acids, and triacylglycerols (TG). Relative to the remission disease state, the active form was characterized by decreased abundances of SMs and increased abundances of inflammatory proteins and TGs. We also observed that molecular changes upon treatment escalation were putatively related to altered levels of inflammatory response proteins, amino acids, and TGs. ISM1, ANGPTL4, chenodeoxycholate, Cer(18:1;2 O/24:1), and TG were identified as candidates subject to further investigation. Altogether, our study revealed that disturbances in immune response, bile acid homeostasis, amino acids, and lipids potentially underlie the clinically heterogeneous spectrum of IBD.

Synthetic cannabinoids (SCs) are an evolving class of new psychoactive substances (NPS) with structurally various compounds that are increasing over the past few years. Therefore, they are initially hard to identify because of the lack of analytical information. Moreover, there is little to no information regarding the pharmacology of these compounds despite human abuse. In the present study, gas chromatography-mass spectrometry (GC-MS), ultrahigh-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF MS), and nuclear magnetic resonance (NMR) spectroscopy were used to identify the structure of three compounds obtained from seized materials. The pharmacological properties of these compounds were evaluated by subsequent behavioral testing, including von Frey and cold allodynia tests. The results indicated that these compounds were determined to be 2-(1H-indazole-3-carboxamido)-3,3-dimethylbutanoate (MDMB-INACA), N-(1-amino-3,3-dimethyl-1-oxobutan-2-yl)-1H-indazole-3-carboxamide (ADB-INACA), and N-(1-amino-3,3-dimethyl-1-oxobutan-2-yl)-1-hexyl-1H-indazole-3-carboxamide (ADB-HINACA) via GC-MS, UPLC-Q-TOF MS and NMR analysis, and they can attenuate mechanical and cold allodynia induced by paclitaxel in rats with peripheral neuropathy. Compared with MDMB-INACA and ADB-HINACA, ADB-INACA showed better analgesic effects on paclitaxel-induced peripheral neuropathy (PIPN) in rats, and its effect was similar to that of the positive drug N'-(1-hexyl-2-oxoindolin-3-ylidene) benzohydrazide (MDA-19).

The analysis of biomolecular interactions is important in characterizing and understanding many fundamental processes that occur in the body and biological systems. A variety of methods are available for studying the extent and rate of binding of these interactions. Some of these techniques are homogeneous methods, with all interacting components being present in the solution-phase, while others are heterogeneous, such as involving both solution-phase and solid-phase components. LC and HPLC have often been used to study biomolecular processes. Although these chromatographic methods make use of both a liquid phase (i.e., the mobile phase and applied samples) and a solid phase (the stationary phase and support), they can be used to study solution-phase interactions. This review examines several strategies that have been developed and employed to use LC and HPLC for this purpose. These strategies include the Hummel-Dreyer method, solution-phase frontal analysis, and the use of physical entrapment for a soluble component of a biomolecular interaction. Other strategies that are discussed are those in which the stationary phase of the column is used as a secondary component or capture agent when studying a solution-phase interaction, as occurs in normal-role affinity chromatography and ultrafast affinity extraction. The general principles for each of these strategies will be considered, along with their advantages, potential limitations, and applications.