Journal of pharmaceutical and biomedical analysis最新文献

The metabolism process of amino acids is closely related to the growth of normal and cancer cells. It is still not clear how L/D-configuration amino acids participate in the metabolism of colorectal cell. Herein, intra- and extra-cellular metabolic distribution of L/D-amino acids in colorectal cell (HCT116) and human normal intestinal epithelial cell (NCM460) were profiled utilizing HPLC-MS/MS coupled with a chiral probe. The results displayed the differential metabolic portrayal for the two cell lines. Compared with NCM460 cell, 13 kinds of significant differential amino acids were founded in a lower concentration within HCT116 cell, and L-Gln was even not detected for intra-cell; as for extra-cell culture medium, the HCT116 cell consumed more L-Gln, D-Phe and D-Leu, while L-Met was low ingested in HCT116 cell. L-Ala and Gly were excretion in both two cell lines, excepted L-Cit which was uptake in HCT116 and excretion in NCM460 cell. Furthermore, the dynamic changes of chiral amino acids displayed that phenylalanine, tyrosine and tryptophan biosynthesis and arginine biosynthesis is the major pathway for intra-cellular metabolites and extra-cellular metabolites, respectively. Moreover, with additional D-amino acids in culture medium, the results exhibited that high concentration of D-amino acids have no significant effect on the proliferation of NCM460 cell, but could influence the profiling of amino acids metabolites, and further affect the proliferation of HCT116 cell. This present work enhances the understanding of these differential amino acids metabolic network and depicts a dynamic process of metabolic dysregulation of HCT116 and NCM460 cell.

The medicinal fungus Phellinus Igniarius (P. igniarius) has been demonstrated to possess a variety of pharmacological effects, including anti-oxidant, anti-tumor, blood circulation promotion, anti-diarrheal and sedative properties, etc. In order to gain a deeper understanding of the components in P. igniarius extract and its dynamic process in vivo, an ultra-performance liquid chromatography-triple quadrupole mass spectrometry method was developed and validated for the simultaneous determination of 12 major components (nobiletin tangeretin, narirutin, 3,4-dihydroxybenzaldehyde, hesperidin, hispidin, caffeic acid, hispolon, osmundacetone, amygdalin, salvianolic acid B and protocatechuic acid) of P. igniarius extract in rat plasma. The analyses were conducted using an ACQUITY UPLC BEH C18 column with acetonitrile and 0.1 % formic acid (v/v) in aqueous solution as the mobile phases. The intra-day and inter-day precision was less than 12.61 % for all 12 experiments, with a precision range of -11.28-12.25 %. Extraction recovery exhibited a range of 74.03-114.33 %, while the matrix effect demonstrated a range of 83.95-119.28 %. The stability tests demonstrated that the analytes remained stable, with relative standard deviations below 11.65 %. The pharmacokinetic parameters of the 12 compounds in rat plasma after oral administration of P. igniarius extract were successfully determined by the established UPLC-MS/MS method. The findings presented a pivotal foundation for advancing future research on the in vivo processes and mechanisms underlying the effects of P. igniarius extracts.

Organophosphorus (OP) pesticides (e.g., parathion) and nerve agents (e.g., soman) can produce acute and long-term neurological problems. Exposure to OP chemicals is responsible for an estimated 200,000 deaths annually. Pz-1 (N-(5-(tert butyl)isoxazol-3-yl)-2-(4-(5-(1-methyl-1H-pyrazol-4-yl)-1H-benzo[d]imidazol-1-yl)phenyl)acetamide) is a muscle specific kinase (MuSK) inhibitor which has shown potential as a treatment for OP chemical exposure and as a tyrosine kinase inhibitor to impede the growth of cancer cells. While development of this treatment requires the availability of a validated analytical method, no method currently exists for analysis of Pz-1 from biological samples. In this study, an analytical method was developed for Pz-1 from rat (and mouse) plasma. Plasma was prepared by precipitating plasma proteins, isolating the supernatant, evaporating to dryness and reconstituting in 1:1 MeOH:water. Prepared samples were analyzed by reversed-phase liquid chromatography tandem mass-spectrometry (LC-MS/MS). The method produced excellent sensitivity, with a limit of detection of 1 nM (455 ng/L). The calibration range was 3-100 nM and the calibration curve produced excellent linear behavior (R² ≥ 0.99 and PRA ≥ 91 %). The method also showed good accuracy and precision. The validated method was used to detect Pz-1 in mouse plasma following intraperitoneal (IP) treatment with 5 mg/kg Pz-1. In summary, this method shows promise as a simple and sensitive method to analyze Pz-1 in rat plasma to facilitate its continued development as a treatment for OP toxicity.

Accurate determination of plasma protein binding (PPB) is crucial in understanding the pharmacokinetics and pharmacodynamics of drugs, particularly for highly bound compounds where traditional methods may fall short. In this study, we present a pioneering approach for the precise determination of PPB that takes advantage of the lipophilicity of highly bound compounds. Twenty four highly bound compounds (with a fraction unbound (f_u) from 10^-1 to 10^-6) were tested with the most commonly used method, i.e., the rapid equilibrium dialysis (RED) method, along with its modifications adapted especially for strongly bound compounds, including dilution, presaturation, competition, and flux-dialysis methods. The results of these methods were compared to data obtained with the modification of RED coupled with extraction to organic phase, and the correlations between them were presented. Comparison studies demonstrate the accuracy of our approach across a set of highly bound compounds within a twofold range. Moreover, PPB values were determined for venetoclax, amiodarone, montelukast, and fulvestrant, for which, to the best of our knowledge and after extensive review of the literature, it has not been possible with the standard RED method until now. In conclusion, our new method can be a big step forward in finding the PPB of strongly plasma protein-bound compounds. It gives us a better understanding of how drugs and proteins interact, which helps us make safer and more effective medicines.

Artificial sweeteners (ASs) and dyes are widely used in foods, beverages and pharmaceutical and are recognized as emerging environmental contaminants due to their persistence and widespread occurrence. These substances often pass through the human body unchanged and resist wastewater treatment processes, leading to continuous introduction into aquatic environments and potential long-lasting term environmental effects. This study investigated, for the first time, the biodegradability of nine commercial dietary supplements, both natural and those containing ASs and synthetic dyes, using the Organisation for Economic Cooperation and Development (OECD) 301 F ready biodegradation test (RBT), which is a respirometry-manometric method. While the products showed good biodegradability, those containing ASs and dyes were further studied to determine their fate at the end of the RBT. The study involved developing and validating a chromatographic method to quantitatively determine the presence of Acesulfame K (1), Sucralose (2), Tartrazine (3) and Carmoisine (4) in the RBT mineral medium, using ultra-high performance liquid chromatography (UHPLC) coupled with two detectors: a high-resolution mass-spectrometer with quadrupole time-of-flight (qToF) and a UV-Vis diode array detector (DAD). Results indicated that these additives were not readily biodegraded, highlighting a potential significant environmental concern. This issue extends beyond dietary supplements to all Pharmaceutical and Personal Care Products (PPCP) including drugs and medical devices. The findings underscore the importance of raising cultural awareness about the environmental impact of persistent substances, encouraging the healthcare chain and patients to make informed choices. From a One Health perspective, reducing environmental contamination can lead to positive outcomes for human health.

Huo-Xiang-Zheng-Qi Mixture is a renowned traditional Chinese medicine formula used to treat ailments associated with dampness pathogens. This study employed ultra-high-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry to perform a comprehensive qualitative and quantitative analysis of the chemical compounds in Huo-Xiang-Zheng-Qi Mixture. A total of 155 compounds were identified, including 61 flavonoids and their glycosides, 36 phenylethanoid glycosides, 23 saponins, 14 coumarins, 9 organic acids, 1 amino acid, 2 nucleosides and purines, and 9 additional compounds. For the first time, a practical method based on inductive effects and hydrogen bonding was developed to determine the elution order of PhGs isomers. The relative quantification of 9 isomers and the absolute quantification of 10 compounds in Huo-Xiang-Zheng-Qi Mixture were determined, primarily derived from tangerine peel, licorice and Magnolia officinalis. Notably, 18 β - glycyrrhetinic acid and 9 Phenylethanoid glycosides isomers were quantified for the first time in the Huo-Xiang-Zheng-Qi prescription. These findings were compared with corresponding values in Huo-Xiang-Zheng-Qi oral liquid. The research revealed relatively low levels of 18 β - glycyrrhetinic acid in the mixture and significant differences in the content of four key compounds: magnolol, honokiol, glycyrrhizic acid and hesperidin. This study offers valuable insights into the chemical composition of Huo-Xiang-Zheng-Qi Mixture and provides a foundation for optimizing preparation processes, improving therapeutic efficacy, and establishing quality standards.

Understanding the pharmacodynamics of ritonavir through metabolomics offers insights into its side effects and helps in the development of safer therapies. This study aimed to investigate the effects of ritonavir treatment on the metabolic profiles of rabbits via a metabolomics approach, with the objective of elucidating its impact on various biochemical pathways and identifying relevant biomarkers. The rabbits were divided into control and ritonavir-treated groups, and their plasma samples were analyzed via ultra-performance liquid chromatography/quadrupole time-of-flight mass spectrometry (UPLC/Q-TOF/MS/MS). Metabolites were identified on the basis of the masscharge ratio (m/z) and validated via XCMS software. Metabolites with a fold change ≥ 1.5 and P ≤ 0.01 were analyzed via principal component analysis (PCA) and orthogonal partial least squares discrimination analysis (OPLS-DA) to distinguish between the groups. MetaboAnalyst 6.0 was used for pathway analysis to identify metabolic pathways affected by ritonavir. The PCA and OPLS-DA models revealed clear separation between the control and ritonavir-treated groups, with high R² and Q² values indicating robust model performance. Pathway analysis revealed that ritonavir treatment significantly affected several metabolic pathways, including those related to ether lipid, phenylalanine, sphingolipid, and glycerophospholipid metabolism. Particularly significant changes were observed in metabolites related to lipid metabolism, oxidative stress responses and cellular signaling. Ritonavir significantly impacts metabolic pathways, particularly those involved in lipid metabolism, and oxidative stress responses, which may influence immune responses and drug interactions. This study also highlights the potential of integrating metabolomics with personalized medicine approaches to optimize ritonavir treatment strategies and reduce adverse effects. These findings indicate that ritonavir significantly influences cellular homeostasis and metabolic processes in addition to its antiviral properties. This highlights the necessity of comprehending the metabolic effects of ritonavir to enhance its clinical application, especially in the management of COVID-19. Further research is warranted to explore these alterations and their implications for therapeutic strategies.

This review article brings together two of the current hot-spots in the field of analytical chemistry, and more specifically in the sample preparation stage: the use of dispersive microextraction techniques, and the analysis of saliva. Due to saliva collection is minimally invasive, it is increasingly being considered in bioanalysis. Moreover, bioanalysis is routine and agglutinates a high number of samples demanding for fast results, thus high-throughput assays are highly required. On the other hand, if something characterizes biological matrices, including saliva, is their complex composition. To adapt the matrix to the analytical method to be applied and to avoid as far as possible the matrix effect, an efficient sample preparation stage is required. To this regard dispersive microextraction techniques, as rapid, efficient and sustainable sample preparation approaches, play a crucial role. In the first part of the review, different workflows for the collection and pretreatment will be briefly described, placing special emphasis on advice to follow. Then, a compilation of the different applications of dispersive techniques for the analysis of saliva is presented, in which the trends observed in both specific analytes and microextraction approaches used are discussed.

In response to the urgent need for effective treatments during the rapid spread and high mortality rate of COVID-19, existing drugs were repurposed for potential antiviral effects, including favipiravir, originally designed as an RNA-dependent RNA polymerase inhibitor for influenza. Despite limited antiviral effectiveness against COVID-19, favipiravir has been reported to cause several adverse drug events (ADEs) in the body. Recent studies have shown that favipiravir can damage various tissues in rats. However, a detailed analysis of its effects on the metabolomics profile of tissues using high-resolution spectroscopic technologies has not yet been conducted. In this study, it was aimed to analyze the metabolomic changes in rat kidney tissues induced by favipiravir, using high-resolution nuclear magnetic resonance (NMR) spectroscopy. Sixty male Wistar Albino rats were randomly divided into three groups: control, low-dose favipiravir (200 mg/kg), and high-dose favipiravir (300 mg/kg), with 20 rats per group. Each group received its respective treatment via oral gavage. After the treatment period, kidney tissue samples were collected and subjected to ¹H NMR analysis. Bioinformatics analysis of the obtained ¹H NMR spectra suggests that favipiravir induces dose-dependent metabolic changes in kidney tissue, with higher doses causing more profound disruptions in several pathways.

Infections with the dengue virus affect more than 100 million people every year. The infected can present a mild form of the disease or a severe form, which can, eventually, lead to death. Dengue prevails in tropical and subtropical regions, although increased incidence has been observed in the last years in tempered climates. Vaccines are available but testing for previous infection is often required prior to application. Commercially available ELISA and rapid tests for the diagnosis of dengue IgG do not fulfill individually the performance required by control agencies. In this context, rapid, simple and decentralized point-of-care testing (POCT) is highly desirable. However, POCT approaches available usually offer expensive solutions, often due to the complex complementary hardware required. In this article, an equipmentless system based on a commercial ELISA kit and a smartphone is developed for POCT of dengue antibodies. A customized app provides guiding, optical reading, result reporting and connectivity. The reading method employes an algorithm which requires no external information, other than the available on the digital images from the smartphone camera, to classify samples into positives, negatives or indeterminates. The full system operation, from sample extraction to result reporting, was tested in a low resource medical facility with real patients (n = 26). After comparison with an ELISA reader, a Cohen's κ coefficient of 0.92 was obtained, showing very good agreement between both methods. These results show that it is possible to perform ELISA with no specific equipment, bringing massive testing at low resource facilities one step closer.