Current Pharmaceutical Analysis - Volume 19, Issue 4, 2023

Multi-omics approaches have developed as a profitable technique for plant systems, a popular method in medical and biological sciences underlining the necessity to outline new integrative technology and functions to facilitate the multi-scale depiction of biological systems. Understanding a biological system through various omics layers reveals supplementary sources of variability and probably inferring the sequence of cases leading to a definitive process. Manuscripts and reviews were searched on PubMed with the keywords of multi-omics, data analysis, omics, data analysis, data integration, deep learning multi-omics, and multi-omics integration. Articles that were published after 2010 were prioritized. The authors focused mainly on popular publications developing new approaches. Omics reveal interesting tools to produce behavioral and interactions data in microbial communities, and integrating omics details into microbial risk assessment will have an impact on food safety, and also on relevant spoilage control procedures. Omics datasets, comprehensively characterizing biological cases at a molecular level, are continually increasing in both dimensionality and complexity. Multi-omics data analysis is appropriate for treatment optimization, molecular testing and disease prognosis, and to achieve mechanistic understandings of diseases. New effective solutions for multi-omics data analysis together with well-designed components are recommended for many trials. The goal of this mini-review article is to introduce multi-omics technologies considering different multi-omics analyses.

Background: After the hydrolysis of daptomycin in deuterated hydrochloric acid, the deuterium-substituted kynurenine was found, but the structure of deuterium-substituted kynurenine has not been reported. Introduction: The deuterium-substituted kynurenines were simply synthesized and confirmed to be tri- and tetra-substituted products by high resolution mass spectrum and NMR. In further, the deuterium-substituted kynurenines were used to determine the conformation of kynurenine to be L-type in daptomycin through conformation analysis combined with derivation and high performance liquid chromatography-quadrupole time-of-flight mass spectrometry (HPLCQ/ TOF-MS). Methods: In the present study, a simple synthesis method was developed for deuteriumsubstituted kynurenine, and its structure was confirmed by high resolution mass spectrometry and NMR. L-kynurenine was mixed with the deuterated hydrochloric acid and heated at 110 °C for 7 h. The hydrogen/deuterium exchange products of L-kynurenine were obtained through the hydrogen/deuterium exchange method. After the derivation of deuterium-substituted L-kynurenine by Marfey’s reagent, the conformation of kynurenine in daptomycin was deduced by HPLC-Q/TOF-MS. Results: The deuterium-substituted kynurenines were confirmed to be tri- and tetra-substituted products by high resolution mass spectrum. Further, Hydrogen NMR spectrum indicated that the deuterium-substitution positions were β-position on amino acid and 3’ and 5’ positions on the benzene ring. Thus, the tri-deuterium-substituted product was L-[β, 3’, 5’-²H3] kynurenine-d3, while the tetra-deuterium-substituted product was L-[β, β, 3’, 5’-²H4] kynurenine-d4. Furthermore, the deuterium-substituted kynurenines were used to determine the conformation of kynurenine to be L-type in daptomycin through conformation analysis combined with derivation and HPLC-Q/TOF-MS. Conclusion: The synthesis, structures, and application of tri- or tetra- deuterium-substituted kynurenine were reported in this study.

Background: Daphnoretin, as a known bicoumarin compound that contained various pharmacological activities, was isolated from Wikstroemia indica C.A. Mey (RWI). Objective: The study aims to investigate the pharmacokinetic characteristics of daphnoretin from RWI ethanol extracts in rat plasma and to determine daphnetin in rat plasma and various tissues by a rapid, reliable and sensitive ultra high performance liquid chromatography with tandem mass spectrometry method. Methods: The UPLC-MS/MS method was established. Daphnoretin and IS (buspirone) were chromatographed on an agilent Zorbax XDB-C18 column (2.1 mm × 50 mm, 3.5 μm), and Gradient elution of acetonitrile-0.15% formic acid in aqueous solution. Quantification was performed using electrospray ionization in positive ion multiple reaction monitoring mode of the transitions m/z 353.1→179.1 for daphnoretin and m/z 386.3→122.3 for IS. Results: Good linearity between 5-10000 ng/mL for cyperidin in plasma and tissue samples (r ≥ 0.99) was resulted. The accuracies of plasma and tissue homogenates ranged from-3.31% to 9.00%, and the precision was less than 5.78%. After that, the validated method was successfully applied to the pharmacokinetics and tissue distribution study of daphnoretin after oral administration of ethanol extract from the roots of RWI to rats. Conclusion: Daphnoretin was well absorbed in the systemic circulation after oral administration and was widely distributed in tissues, with the highest concentration in lung tissue. This study is beneficial to the development and utilization of RWI and provides a reasonable reference for its clinical administration.

Objective: This study aims to investigate the main metabolites and metabolic pathways of Portulacatone B in rats, which is an alkaloid isolated from Portulaca oleracea L. Methods: Portulacatone B was administered through the tail vein of the rat, and the orbital blood at 10 and 30 min and urine and feces within 24 h were collected. The metabolites and metabolic pathways in the rat were researched by ultra-high performance liquid chromatographyelectrospray coupled with quadrupole-time of flight mass spectrometry (UHPLC-ESI-QTOF/ MS). Results: The research results of the metabolites and metabolic pathways of Portulacatone B showed that after administration through the tail vein of rats, 3 metabolites were found in the plasma sample, 2 metabolites in the urine sample, and one metabolite in the feces sample. The main metabolic pathways were found to be oxidation, hydrolysis, methylation, glucuronidation, and sulfonation. Conclusion: Six metabolites were found in the rat’s plasma, urine, and feces samples, and the metabolic pathways included oxidation, hydrolysis, methylation, glucuronidation, and sulfonation process.

Background: In intensive care units intravenous medicine may be used in simultaneous infusion in the same intravenous site. Sometimes, the physical compatibility and stability of the combined solutions are unknown. Objective: The objective was to develop, optimize and validate a simple, fast and sensitive stability- indicating high-performance liquid chromatography (HPLC) for simultaneous quantification of binary mixtures of norepinephrine, piperacillin + tazobactam, moxifloxacin for intravenous (IV) administration in different diluents and physical compatibility with mannitol. Methods: The HPLC method was performed on a C18LUNA (4.6x250 mm 5-Micron) column, using acetonitrile: methanol: phosphate buffer pH 3.0 (20:30:50) as eluent and validated according to ICH guidelines and applied to mixtures of norepinephrine, moxifloxacin, piperacillin, tazobactam and mannitol at 0, 2, 6, 9 and 24 h. The substances and their mixtures were also evaluated by visual inspection and pH over time. Results: The analytical method developed was specific, linear, precise, accurate and robust. No visual changes were observed in the mixtures over time, maintaining the pH values (except for piperacillin + tazobactam which changed 0.5 in 24 h) and losses of less than 10% of content over the 24 h under analyzed conditions. Conclusion: The proposed method is suitable for simultaneous analysis of norepinephrine, moxifloxacin, piperacillin and tazobactam. All tested mixtures were compatible and stable for up to 24h, which is an important result for increasing patient safety in clinical practice since it has not been reported in the literature yet. The method can be further investigated and used for different concentration and diluent combinations.

Aims: Flavonoids naturally exist in plants as secondary metabolites. In this study, the aim is to determine and compare the theoretical and in vivo chemical activities of 7,8- dihydroxyflavone (7,8-DHF) and 4'dimethylamino-7,8-dihydroxyflavone (4’-DMA-7,8-DHF), tyrosine receptor kinase B (TrkB) receptor agonist flavonoid molecules with reported potent neuroprotective effects. Methods: The density functional theory (DFT) (RB3LYP) method was used for the theoretical chemical analysis. For the in vivo studies, 6-month-old Wistar rats were used in two groups (n=8). 7,8-DHF and 4’-DMA-7,8-DHF (5 mg/kg) were administered intraperitoneally (ip) to each group. Then, plasma samples were collected by carotid catheterization, and brain samples by the microdialysis technique were collected simultaneously for 12 h from awake rats. The level of 7,8-DHF and 4’-DMA-7,8-DHF in blood and brain samples were analyzed and their pharmacokinetics were determined. Results: Theoretical calculations show that 7,8-DHF is slightly more stable than 4’-DMA-7,8- DHF. The in vivo pharmacokinetic results show that the maximum concentration of 7,8-DHF was about 48 ng/mL, whereas it was only 8 ng/mL for 4’-DMA-7,8-DHF. Conclusion: Our results suggest that the 4'-DMA-7,8-DHF is more unstable and is more prone to binding to TrkB than 7,8-DHF. On the other hand, the in vivo pharmacokinetic results show that 7,8-DHF is more stable than 4’-DMA-7,8-DHF when it is applied systemically at therapeutic concentrations.

Objective: Controlling coccidiosis disease in poultry is significant for healthy breeding, but antibiotic residues can pose a great threat to food safety, the ecological environment, and human health. A special anticoccidial drug called Hainanmycin has been widely used in Asian countries for many years, while a few studies focus on its detection. In this study, we established a hydrophiliclipophilic balance (HLB) purification and ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method to determine the hainanmycin in chicken liver. Methods: It was extracted from the sample with 0.1% formic acid-methanol (10+90, v/v) and then it was purified on a hydrophilic-lipophilic balance (HLB) cartridge. Chromatography separation was performed on an RP-C18 column, and detection of the hainanmycin was done by positive electrospray ionization (ESI.) under the multiple reaction monitoring (MRM) modes. Results: The quantification ion pair was m/z 907.5 and m/z 846.4. The calibration curve showed excellent linearity in the range of 10 to 1000 μg·L^-1, with a correction coefficient of 0.99. When spiked at 5, 100, and 500 μg·kg^-1, the recoveries ranged from 80.82% to 88.09%, with the intra-day and interday coefficients of variation from 10.76% to 13.48% and from 10.80% to 12.71%, respectively. The limit of quantification (LOQ) and detection(LOD) were 5.05 μg·mL^-1 and 1.55 μg·mL^-1, respectively. Conclusion: This method showed high sensitivity and accuracy for determining hainanmycin in chicken liver.

Background: To control the quality of various butyl rubber closures and assess their applicability to cefamandole nafate, HPLC/ GC-MS methods were developed to qualitatively and quantitatively analyze the organic components in various types of butyl rubber closures and drug powder of cefamandole nafate for injection, and migration of these components from rubber closures to drug also were studied. Materials and Methods: The chemical components in the rubber closures and corresponding drug were identified by GC-MS in full scan mode and NIST mass library. A GC-MS method in SIM mode was established to quantitatively analyze siloxane compounds, aromatic hydrocarbons and saturated chain-hydrocarbons in rubber closures and drugs. An HPLC method was established to quantitatively analyze antioxidants and sulfides from rubber closures and drugs. The turbidity values of powder for injection were measured by a turbidimeter. Further, the correlation between the content of organic components in the drug and the clarity of the solution was analyzed. Results: It was found that the chemical components were released from butyl rubber closures and migrated to the drug, which influenced solution clarity of cefamandole nafate for injection and was the main factor to cause solution turbidity of powder for injection. The gradual migration of components occurred over time with the use of common butyl rubber closures, but the coated butyl rubber closures (rubber plated with membrane) were coated with a polytetrafluoroethylene film to block the contact between rubber closures and the drug, and could effectively prevent the migration of the components from rubber closures to drugs. The organic components in the butyl rubber closures were mainly identified as siloxanes, saturated chain-hydrocarbons, and antioxidants. An unknown ingredient was identified as 1-piperidinecarboxaldehyde, and trace amounts of toxic toluene were also detected within limits. Conclusion: It is suggested that rubber plated with membrane or rubber closures of high quality were applicable to cefamandole nafate for injection and should be used in its production. HPLC/ GC-MS methods can be used as an effective method for quality control of butyl rubber closures and compatibility research with drugs to ensure drug safety for the public.

Background: Botulinum neurotoxins (BoNTs) are among the most potent toxins known and are also used for therapeutic and aesthetic applications. Objective: An alternative in vitro cell culture bioassay based on the induction of apoptosis on T- 47D breast cancer cells, after exposure to BoNTA, was developed and validated. Methods: The T-47D cells (ATCC HTB-133) were seeded at a density of 3 × 10⁵ cells mL^-1, and the bioassay was performed with doses of BoNTA, between 3 and 81 U mL^-1. The responses were assessed using 10 μL of Alamar Blue®. The absorbances were read at 570 and 600 nm. Results: The results were compared with those of the in vivo LD50 mouse bioassay, showing a non-significant 1.08% higher, mean difference of the estimated potencies (p>0.05). Besides, the biopharmaceutics is analyzed by the size exclusion and reversed-phase liquid chromatography methods, showing a significant correlation with values 1.15% higher and 0.85% lower, respectively, related to the cell culture bioassay. Conclusion: It is concluded that the validated T-47D cell culture assay represents an advancement toward the establishment of an alternative approach for the potency assessment, in the context of the 3 Rs. Besides, the employment of chromatographic methods in conjunction with the bioassays contributes to assessing the quality attributes of the biopharmaceutical formulations of BoNTA.