Ultra-high Performance Liquid Chromatography Research Articles

UPLC Estimation of an Antidiabetic Drug by Method Development and Validation Followed by Stability Indicating Studies.

The analytical method development and validation of a common antidiabetic drug, Canaglifozin, by taking the aid of ‘Ultra High-Performance Liquid Chromatography’, along with stability indicating method. The canaglifozin drug was eluted from the optimized mobile phase in 70:30 ratio of water and methanol, at 1 ml/ min flow rate, under separation with C18 column of 100 x 2mm size with 1.8 diameters at isocratic mode. The detection was carried at 286 nm wavelength, resulting in 3.80 retention time, as run time being 10 min. The detection limit and quantification limit, was noticed at ‘0.0039 and 0.0119 µg/ ml’ correspondingly. The percentage RSD and recovery percentage was under ICH guidelines only. The stress degradation studies under acid, basic, neutral, and thermal conditions were carried on.

Integrating network pharmacology, molecular docking and non-targeted serum metabolomics to illustrate pharmacodynamic ingredients and pharmacologic mechanism of Haizao Yuhu Decoction in treating hyperthyroidism.

To explore the pharmacodynamic ingredients and pharmacologic mechanism of Haizao Yuhu Decoction (HYD) in treating hyperthyroidism via an analysis integrating network pharmacology, molecular docking, and non-targeted serum metabolomics. Therapeutic targets of hyperthyroidism were searched through multi-array analyses in the Gene Expression Omnibus (GEO) database. Hub genes were subjected to the construction of a protein-protein interaction (PPI) network, and GO and KEGG enrichment analyses. Targets of active pharmaceutical ingredients (APIs) in HYD and those of hyperthyroidism were intersected to yield hub genes, followed by validations via molecular docking and non-targeted serum metabolomics. 112 hub genes were identified by intersecting APIs of HYD and therapeutic targets of hyperthyroidism. Using ultra-high performance liquid chromatography with quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS) in both negative and positive ion polarity modes, 279 compounds of HYD absorbed in the plasma were fingerprinted. Through summarizing data yielded from network pharmacology and non-targeted serum metabolomics, 214 common targets were identified from compounds of HYD absorbed in the plasma and therapeutic targets of hyperthyroidism, including PTPN11, PIK3CD, EGFR, HRAS, PIK3CA, AKT1, SRC, PIK3CB, and PIK3R1. They were mainly enriched in the biological processes of positive regulation of gene expression, positive regulation of MAPK cascade, signal transduction, protein phosphorylation, negative regulation of apoptotic process, positive regulation of protein kinase B signaling and positive regulation of MAP kinase activity; and molecular functions of identical protein binding, protein serine/threonine/tyrosine kinase activity, protein kinase activity, RNA polymerase II transcription factor activity, ligand-activated sequence-specific DNA binding and protein binding. A total of 185 signaling pathways enriched in the 214 common targets were associated with cell proliferation and angiogenesis. HYD exerts a pharmacological effect on hyperthyroidism via inhibiting pathological angiogenesis in the thyroid and rebalancing immunity.

Citrus flavonoid-pectin conjugates with enhanced emulsifying properties

Pectin and flavonoids are important components of citrus fruits, but the effects of flavonoid grafting on the emulsifying properties of citrus pectin remain unclear. In this study, a novel natural citrus flavonoid-pectin conjugate in citrus peel was identified through alkaline hydrolysis combined with ultra-high performance liquid chromatography. Subsequently, two major citrus flavonoids were enzymatically conjugated to citrus pectin at high grafting rates (hesperidin: 20.21%; naringin dihydrochalcone: 16.11%). The flavonoids increased pectin amphipathy while ensuring a tighter conformation and denser network structure. These structural changes reduced the interfacial tension (from 20.6 to 18.4 mN/m) and increased the adsorbed layer thickness (from 11.8 to 37.8 nm), facilitating rapid pectin anchoring at the oil-water interface. These led to smaller droplet size and better emulsion stability. Prepared flavonoid-pectin conjugates with high grafting rates exhibited better emulsifying properties than those with medium grafting rates; hesperidin-pectin conjugates were more effective than naringin dihydrochalcone-pectin conjugates. The prepared flavonoid-pectin conjugates aid in the structural identification of natural flavonoid-pectin conjugates; they also facilitate analyses of the properties of these conjugates and offer new insights into improving the functional properties of pectins used in the food industry.

Multi-residue analysis method based on QuEChERS followed by ultra-high performance liquid chromatography coupled with diode-array detector for pesticides in human serum and breast milk.

Background: Maternal fluids play a key role in the risk assessment regarding early life pesticide exposure as the chemicals can transfer to neonate through prenatal exposure and lactation.Aim: A developed UHPLC-DAD and modified QuChERS methods were validated for human serum and breast milk. Matrix effect of the biological samples were evaluated.Methods & results: Serum was extracted by unbuffered QuChERS method while breast milk was extracted by citrate buffered method with addition of hexane. Remaining lipid in breast milk extract was later removed using lipid-removal sorbent. Sample matrices caused huge impacted on low-sensitivity pesticides.Conclusion: The modified QuEChERS methods coupled with UHPLC-DAD were fully validated. Application in paired-serum and breast milk samples revealed 6 detected pesticides.

Metabolomics-based biomarkers of fermented dairy and red meat intake: a randomized controlled trial in healthy adults.

Dietary assessment is usually performed through imprecise tools, leading to error-prone associations between diet and health-related outcomes. Metabolomics has been applied in recent years to develop biomarkers of food intake (BFIs) and to study metabolites in the diet-microbiome crosstalk. Candidate BFIs exist to detect intake of meat and to a lesser extent dairy, but validation and further development of BFIs are needed. Here, we aim to identify biomarkers that differentiate between intakes of red meat and dairy, to validate previously reported BFIs for these foods, and to explore the effect of protein-matched meals on selected microbial metabolites. We conducted a randomized, controlled, cross-over single-meal study comparing a meal with highly fermented yogurt and cheese, and a meal with beef and pork meatballs. Postprandial urine samples from 17 subjects were collected sequentially after each meal up to 24h and analyzed by untargeted metabolomics through ultra-high-performance-liquid chromatography (UHPLC) coupled via electrospray (ESI) source to a qTOF mass spectrometer. Univariate (repeated measures ANOVA) and multivariate (PLSDA, ML-PLSDA) data analyses were used to select BFIs differentiating the two meals. 3-Indoxyl sulfate, p-cresol sulfate, and several other microbial amino acid catabolites were additionally explored within the urine profiles. Thirty-eight markers of meat and dairy intake were selected and are presented along with their excretion kinetics. Carnosine, taurine, and creatine, as well as hydroxyproline-based dipeptides are confirmed as meat BFIs. For dairy, previously reported metabolites such as acyl-glycines are confirmed, while proline-based dipeptides are reported as novel putative BFIs. Microbial metabolites showed only marginal evidence of differential formation after the two meals. This study allowed us to validate the postprandial kinetics of previously suggested biomarkers of meat and dairy intake and to identify new potential biomarkers. The excretion kinetics are useful to ensure that the collection of urine covers the correct time window in future dietary studies. The BFIs add to the existing body of biomarkers and may further be used in combination to provide a more reliable assessment of meat and dairy intake. Proteolytic microbial metabolites should be further investigated to assess the effect of different protein sources on health.

UHPLC method for simultaneous assessment of pharmacokinetic parameters of co-administered voriconazole and omeprazole using rat plasma

Voriconazole (VRC) is a first-line therapeutic agent for the treatment of invasive fungal infections, whereas omeprazole (OMZ) is a commonly used acid suppressant; however, the two drugs are often used in combination in clinical practice. The aim of this research was to investigate how the co-administration of OMZ and VRC affects the pharmacokinetic characteristics of VRC in rats. A new ultra high-performance liquid chromatography (UHPLC) analytical method was developed and validated for simultaneous analysis of co-administered drugs using VRC and OMZ. A Shim-pack GIST-HP C18 column with 0.1 M triethylamine:acetonitrile (70:30, v/v) as the mobile phase (flow rate: 0.3 mL/min) was used, and UV detection was performed at 240 nm. Liquid–liquid extraction of plasma samples was carried out using dichloromethane. The bioanalytical method was linear over a range of VRC concentrations (100–2000 ng/mL) and OMZ (50–10,000 ng/mL) concentrations, and exhibited both accuracy and precision within the acceptable respective ranges. The average extraction recoveries for VRC and OMZ in plasma were 94.88 % and 82.76 %, respectively. Additionally, the method was successfully applied in vivo to estimate the pharmacokinetic features of VRC in the plasma of rats receiving gavage with low and high doses of OMZ. In conclusion, using a new UHPLC method, we determined that co-administration of OMZ substantially decreased the bioavailability of VRC in rats. Potentially significant drug interactions should be considered in patients receiving the combination of OMZ and VRC.

Identification and screening of potential anti-pneumonia active ingredients and targets of Qing-Kai-Ling oral liquid via UHPLC-Q-Exactive Orbitrap mass spectrometry based on data post-processing

Qing-Kai-Ling oral liquid is commonly used clinically for the treatment of fever and upper respiratory tract infection. Moreover, studies have shown that Qing-Kai-Ling oral liquid has an anti-pneumonia effect. However, owing to its complex pharmacodynamic material basis, its pharmacological research and clinical application are limited. To address this problem, the chemical constituents of Qing-Kai-Ling oral liquid were identified by ultra-high performance liquid chromatography quadrupole-Exactive Orbitrap mass (UHPLC-Q-Exactive Orbitrap MS) and network pharmacology methods, which were used to predict its potential anti-pneumonia target and signalling pathway. A total of 150 compounds were identified and tentatively characterized, including 35 amino acids and their derivatives, 36 organic acids, 20 terpenoids, 20 alkaloids, 12 glycosides, 7 flavonoids, and 20 others. Among them, 14 compounds were accurately identified by comparing their retention time and mass spectrum data with those of reference substances. Additionally, we performed molecular simulation calculations via Density Function Theory to determine the plausibility of the compound cleavage reactions and further confirm compound structures. Furthermore, 90 key targets were screened through network pharmacology, with the particular focus on the PI3K-AKT, MAPK and TNF signalling pathways. This method achieved the first comprehensive identification of the chemical composition of Qing-Kai-Ling oral liquid and elucidated its potential mechanism of anti-pneumonia. The results provide valuable reference and data support for pharmacodynamic substance research and quality control.

Revealing the sterilization impact on paprika fingerprint: Key markers identified using untargeted metabolomics by liquid chromatography-Orbitrap high-resolution mass spectrometry

Paprika (Capsicum annuum L.) is a popular spice known for its unique properties. Spices are susceptible to microbiological risks arising from harvest factors such as high moisture or environmental contamination. To ensure microbiological safety, post-harvest processing based on heat sterilization, free of chemicals and radiation, is becoming essential in the European market. This study introduces a novel metabolomics approach using ultra-high performance liquid chromatography (UHPLC) coupled with quadrupole-Orbitrap-high-resolution mass spectrometry (HRMS) to assess the sterilization impact on paprika's metabolomic composition. Sterilized and untreated samples were distinguished by OPLS-DA, achieving perfect predictability with high-quality parameters (R2Y = 0.988, Q2 = 0.904). The methodology identified 19 key markers, including fatty acids, amino acids, etc. Sterilization reduced fatty acids such as linoleic acid but increased other metabolites such as DL-malic acid and flazin. This research introduces new metabolomics strategies to ensure paprika quality and other valuable spices, focusing on unexplored sterilization processes.

Identification and quantitation of carbon–carbon double bond isomers of fatty acid in livestock and poultry meat

Livestock and poultry meat is one of the main consumption sources of fatty acids (FAs) in China. FAs are the main component of lipids and essential nutrients for the human body. The location of carbon–carbon double bond (CC) in unsaturated fatty acids (UFAs) significantly affects the nutrition and flavor quality of meat products. This study established an analytical method for identifying the CC positions of UFAs based on the Paternò-Büchi (PB) reaction using ultra-high performance liquid chromatography quadrupole time-of-flight mass spectrometry (LC-QTOF). The FA extract was derivatized with 2-acetylpyridine under ultraviolet light at 254 nm. The derivatized FA was then analyzed using targeted MS/MS mode in positive electrospray ionization. Limits of detection (LOD) and limits of quantitation (LOQ) of 21 UFAs were 0.004–0.680 and 0.013–2.267 μg/L, respectively. Furthermore, for the diagnostic ions at the same functional group terminal fragmented from derivatives of FA CC isomers, the abundance ratio was linear with that of the corresponding content, which could be applied to non-targeted screening of CC positions in FAs. Based on this method, a total of 28 UFAs were screened in five types of livestock and poultry meats in this study. Among them, four isomers of C17:1, four of C18:1, two of C18:2, two of C20:3 and two of C22:5 were identified. The precise identification and quantitation of the double bond positions of fatty acids will be of great significance for the exploration of the nutritional components in livestock and poultry meat.

Method Development and Validation of Alcaftadine by UHPLC in Bulk and Dosage Form

This research developed and validated a new, simple sensitive, suitable, economical, accurate, and robust ultra-high performance liquid chromatography (UHPLC) method for determining Alcaftadine in bulk drug and ophthalmic dosage formulation. The separation was performed using an HPLC method with a UV detector and Open lab EZchrome workstation program, Kromasil C18, 250mm X 4.6mm ID, 2.7μm Acetonitrile: 0.05% OPA (60:40%V/V) with a flow rate of 1.0mL/min and detected at 282nm. The developed UHPLC method yielded a suitable retention time for Alcaftadine of 2.20min, which was optimised using a trial-and-error basis. The linearity of the determined method was found a correlation coefficient (r2) of 0.9998 over the concentration range of 2.0-30.0μg/mL. The percentage RSD for the method's precision was found to be less than 2.0percent. The percentage recovery was discovered within the limit 0.259ug/mL and 0.784ug/mL were found to be the LOD and LOQ, respectively. The developed and validated UHPLC system takes less time and cheaply can be used in the industry for routine quality control/analysis of bulk drugs and marketed Alcaftadine products. In present studies, the retention time was less than previously reported. The developed method was validated in terms of specificity, linearity, accuracy, precision and robustness according to the ICH guidelines.

Multidrug-resistant high-risk clonal Escherichia coli lineages occur along an antibiotic residue gradient in the Baltic Sea

The global spread of multidrug-resistant Escherichia coli threatens human, animal, and environmental health. Despite the recognition of water bodies as potential reservoirs, research on antimicrobial resistance (AMR) and antibiotic residues in the Baltic Sea is limited. In this study, we examined the presence of ESBL-producing E. coli in surface water in northeastern Germany. We then conducted phenotypic and genotypic analyses of the strains, which encompassed AMR and heavy metal/metalloid tolerance, biofilm formation, and detailed plasmid examination. In addition, we used solid-phase extraction with ultra-high performance liquid chromatography and mass spectrometry to quantify antibiotic residues. Our analysis identified 30 ESBL-producing E. coli isolates, some of which were associated with clinically relevant high-risk clonal lineages. We detected antibiotic residues in the environmental samples, but the antibiotic concentrations were significantly lower compared to wastewater samples. These results suggest that the Baltic Sea surface water could serve as a reservoir for ESBL-producing E. coli and clinically relevant antibiotics.

Study on the metabolism of Xiao-Jian-Zhong-Tang in rats with chronic atrophic gastritis coupled with bioinformatics.

Xiao-Jian-Zhong-Tang (XJZT) has the effect of warming the middle and tonifying the deficiency, easing the urgency and relieving pain according to the theory of traditional Chinese medicine (TCM), and is able to treat spleen deficiency type chronic atrophic gastritis (CAG). Metabolites of TCM in cecum contents are common metabolites of intestinal bacteria and hosts, which can reflect the metabolic status in disease states. The present work was performed to study the effect of XJZT against CAG coupled with the cecal metabolites analysis and bioinformatics. A total of nine prototypical components and 144 metabolites were firstly identified in the cecum metabolites of XJZT using ultra-high performance liquid chromatography added to the quadrupole-time of flight mass spectrometry (UHPLC-Q-TOF/MS), which underwent the metabolism of oxidation, reduction, methylation, and glucuronic acid reaction Furthermore, different prototypical compounds might metabolize into identical metabolites in the presence of intestinal flora. Bioinformatics was further used to correlate these metabolites with the disease and intestinal flora. Components and targets were screened by Cytoscape, and molecular docking of key targets and core components showed good binding ability. This study provided important information for exploring the mechanism of TCM formulae.

Plasma Endocannabinoids Are Independently Associated With the Metabolic Function of White Adipose Tissue

Abstract Context Little is known about the link between the endocannabinoid (EC) system and the in vivo metabolic function of white adipose tissue (WAT). Objective We aimed to evaluate whether ECs are linked to postprandial fatty acid metabolism and WAT metabolic function. Methods Men and women, with (IGT, n = 20) or without impaired glucose tolerance (NGT, n = 20) underwent meal testing with oral and intravenous stable isotope palmitate tracers and positron emission tomography with intravenous [11C]-palmitate and oral [18F]-fluoro-thia-heptadecanoic acid to determine systemic and organ-specific dietary fatty acid (DFA) and nonesterified fatty acid (NEFA) metabolism and partitioning. We determined fasting and postprandial plasma levels of EC by ultra-high performance liquid chromatography–tandem mass spectrometry. Results All ECs of the 2-monoacylglycerol (2-MAG) family displayed a progressive postprandial increase up to 360 minutes after meal intake that was more pronounced in women with IGT. N-acylethanolamine (NAE) levels decreased between fasting and 180 minutes, followed by a return to preprandial values at 360 minutes and were also increased in women with IGT. Postprandial area under the curve (AUC) of palmitate appearance rate was significantly and independently associated with postprandial AUC of anandamide (AEA; P = .0003) and total energy expenditure (P = .0009). DFA storage in abdominal subcutaneous adipose tissue was positively predicted by fasting 2-arachidonoylglycerol (2-AG; P < .04). Conclusion EC levels of the NAE family independently follow plasma NEFA metabolism, whereas 2-MAG closely follow the spillover of triglyceride-rich lipoprotein intravascular lipolytic products. Whether these associations are causal requires further investigation.

Assessment of sulfamethoxazole and oxytetracycline uptake and transformation in Eisenia fetida earthworms

The scientific community is becoming increasingly concerned about the recent detection of transformation products (TPs) of antimicrobials (AMs) and their presence in the food chain. There are growing concerns about the potential consequences on food safety and the proliferation of antimicrobial resistance. In this work, the transformation process of sulfamethoxazole (SMX) and oxytetracycline (OTC) in soil was thoroughly evaluated. For that purpose, soils were homogeneously contaminated at three concentration levels of SMX and OTC, independently, and samples were analysed after 7 and 14 days by Ultra High-Performance Liquid Chromatography coupled to a triple quadrupole mass spectrometer (UHPLC-MS/MS). The results have demonstrated a remarkable transformation, particularly noteworthy for SMX, as it exhibited an 89 % - 94 % decrease in concentration within the initial 7 days of the experiment. In addition, to assess whether terrestrial organisms would be able to accumulate the AMs, Eisenia fetida (E. fetida) earthworms were exposed to the above-mentioned concentration levels of AMs in soil. Both AMs were accumulated in the organisms after 14 days, but higher bioaccumulation factor values (BCF) were determined for SMX (0.52–17.84) compared to OTC (0.02–0.21) at all tested concentrations. The analyses were extended to search for TPs in earthworms and soils using a suspect screening approach. Concretely, by means of UHPLC-high resolution mass spectrometry (UHPLC-HRMS) three TPs were identified at 2a and 2b of confidence level. To the best of our knowledge, one SMX-TP and one OTC-TP were identified in earthworms and soil, respectively, for the first time in the present work. Earthworms did not experience weight loss or mortality in the presence of these AMs at levels found in the environment, but there was a decrease in riboflavin levels, which is linked to changes in the immune system. This study represents a significant advancement in understanding the impact of AMs in soil and their subsequent entry into the food chain. It also provides valuable insights into the potential effects of AMs and their TPs on organisms.

Comparison of chemical components and quality evaluation of Panax ginseng and its processed products from different habitats in Northeastern China

Ginseng (Panax ginseng C.A. Meyer) is a widely used dietary supplement in Traditional Chinese Medicine due to its numerous health-promoting properties. However, limited research is available on the distinctions in the chemical composition between different ginsengs. Hence, this study explores the chemical composition of different ginsengs obtained from different habitats in Northeastern China. The fresh, red, and white ginsengs were collected from different places in the Jilin province, China. Ginsenoside and non-ginsenoside contents in the ginseng extracts were analyzed by an Ultra-high performance liquid chromatography system & VION® ion mobility hybrid Quadrupole Time-of-Flight. The results show a notable difference in the ginsenoside content among the samples. The qualification ratio of ginsenoside Re, Rg1, and Rb was 46.2%, with Re being the most abundant, followed by Rg1 and Rb1 in all the ginseng analyzed. Furthermore, the study revealed that white and red ginseng had higher total ginsenosides (8.18 ± 0.04 mg/kg) and total polysaccharides (17.4%) content, respectively. The fresh ginseng samples exhibited higher protein content (20.02 ± 0.2 g/100g) and lower fat content (2.0 g/100g) than others. Further, the ginseng samples analyzed in this study met the requirements specified in the Chinese Pharmacopoeia for total ash, heavy metals, and organochlorine pesticide residues. This study offers a comprehensive comparative analysis of the chemical composition of P. ginseng sourced from various habitats in Northeastern China. It underscores the significance of making informed choices when selecting ginseng types for scientific evaluation.

Positive Intervention of Distinct Peptides in Clostridioides difficile Infection in a Mouse Model

Clostridioides difficile infection (CDI) is a common healthcare-associated infection and the leading cause of gastroenteritis-related deaths worldwide. To investigate the effects of peptide composition of different protein products on CDI, we analyzed and compared the peptide sequences and compositions from Engraulis japonicus and Glycine max using Ultra High Performance Liquid Chromatography Tandem Mass Spectrometry (UPLC-MS/MS). An animal model of CDI was also established to investigate the potential therapeutic effects of these peptides in vivo. The peptide compositions of E. japonicus and G. max differed, with only 11% of the peptide sequences being identical. Oral administration of the tested peptides could reduce intestinal inflammation, repair the intestinal barrier, increase the proportion of beneficial bacteria, and reduce the proportion of harmful bacteria, providing a therapeutic effect against CDI. However, the peptides may differ considerably in some aspects. E. japonicus peptides were superior to G. max peptides in promoting colon epithelial cell proliferation and repairing tight intestinal cell junctions. Interestingly, the two sources of peptides have different effects on the cecal microbiome. E. japonicus peptides can effectively restore the diversity and richness of intestinal microbiota, while G. max peptides have poor regulatory effects on the intestinal microbiota structure. Overall, E. japonicus peptides showed better results than G. max peptides in treating CDI. This study supports the potential treatment of CDI with natural peptides and promotes the development of specialty foods for CDI enteritis. Clostridioides difficile infection (CDI) is a common healthcare-associated infection and the leading cause of gastroenteritis-related deaths worldwide. To investigate the effects of peptide composition of different protein products on CDI, we analyzed and compared the peptide sequences and compositions from Engraulis japonicus and Glycine max using Ultra High Performance Liquid Chromatography Tandem Mass Spectrometry (UPLC-MS/MS). An animal model of CDI was also established to investigate the potential therapeutic effects of these peptides in vivo. The peptide compositions of E. japonicus and G. max differed, with only 11% of the peptide sequences being identical. Oral administration of the tested peptides could reduce intestinal inflammation, repair the intestinal barrier, increase the proportion of beneficial bacteria, and reduce the proportion of harmful bacteria, providing a therapeutic effect against CDI. However, the peptides may differ considerably in some aspects. E. japonicus peptides were superior to G. max peptides in promoting colon epithelial cell proliferation and repairing tight intestinal cell junctions. Interestingly, the two sources of peptides have different effects on the cecal microbiome. E. japonicus peptides can effectively restore the diversity and richness of intestinal microbiota, while G. max peptides have poor regulatory effects on the intestinal microbiota structure.

Metabolomics and quantitative analysis to determine differences in the geographical origins and species of Chinese dragon's blood.

The aim of this study was to comprehensively analyze the differences in Chinese dragon's blood (CDB), specifically Dracaena cochinchinensis and Dracaena cambodiana, from different geographical origins. Metabolomic analysis of CDB was performed by liquid chromatography-tandem mass spectrometry (LC-MS/MS). A reliable ultrahigh-performance liquid chromatography method with a photodiode array detector (UHPLC-PDA) was developed and applied for the quantitative analysis of 12 phenolic compounds in 51 batches of samples. A total of 1394 metabolites were detected, of which 467 were identified as differentially accumulated metabolites. Multivariate analysis revealed that both origin and species had an effect on the composition of CDB, with greater variation between species. 19 phenolic compounds were selected as quality markers to distinguish D. cochinchinensis (Hdsp) from D. cambodiana (Hdca), and oppositin and spinoflavanone a were identified as quality markers to discriminate D. cochinchinensis samples from Hainan (Hdsp) and Guangxi Provinces (Gdc). Quantitative analysis indicated that four phenolic compounds, including loureirin D, 4H-1-benzopyran-4-one,2,3-dihydro-3,5,7-trihydroxy-3-[(4-methoxyphenyl)methyl]-,(R)-, loureirin B, and pterostilbene, showed significant differences between Gdc and Hdsp. Additionally, five phenolic compounds, namely resveratrol, loureirin D, pinostilbene, 4H-1-benzopyran-4-one,2,3-dihydro-3,5,7-trihydroxy-3-[(4-methoxyphenyl)methyl]-, (R)-, and loureirin B, exhibited significant differences between Hdsp and Hdca. There are significant differences in the quality of CDB from different geographical origins and species, which lays the foundation for the in-depth development and utilization of different sources of CDB.

Rooted in therapeutics: comprehensive analyses of Cannabis sativa root extracts reveals potent antioxidant, anti-inflammatory, and bactericidal properties.

Following the legalization of recreational Cannabis in Canada in 2018, the associated waste, including Cannabis roots, has significantly increased. Cannabis roots, comprising 30%-50% of the total plant, are often discarded despite their historical use in Ayurvedic medicine for treating inflammatory and infectious disorders. This study evaluates the phytochemical and therapeutic properties of Cannabis root extracts from a high tetrahydrocannabinolic acid, low cannabidiolic acid cultivar (variety Alien Gorilla Glue). We performed ultra high-performance liquid chromatography coupled with mass spectrometry (UPLC-QTOF-MS) to identify the chemical components of the Cannabis roots. Extracts using water, ethanol and acid-base solvents were tested for antioxidant activity through free radical scavenging, metal chelation, and lipoperoxidation inhibition assays. Mitochondrial membrane protection was assessed using flow cytometry with the MitoPerOx probe in THP-1 monocytic leukemia cells. Anti-inflammatory potential was evaluated by measuring interleukin-6 levels in lipopolysaccharide-stimulated THP-1 cells. Bactericidal/fungicidal efficacy against Escherichia coli, Staphylococcus aureus, and Candida albicans was determined using the p-iodonitrophenyltetrazolium assay. Additionally, we investigated the anticholinesterase activity of Cannabis root extracts, given the potential role of plant alkaloids in inhibiting cholinesterase, an enzyme targeted in Alzheimer's disease treatments. UPLC-QTOF-MS analysis suggested the presence of several phenolic compounds, cannabinoids, terpenoids, amino acids, and nitrogen-containing compounds. Our results indicated significant antioxidant, bactericidal, and anticholinesterase properties of Cannabis root extracts from both soil and hydroponic cultivation. Extracts showed strong antioxidant activity across multiple assays, protected mitochondrial membrane in THP-1 cells, and exhibited anti-inflammatory and bactericidal/fungicidal efficacy. Notably, soil-cultivated roots displayed superior anti-inflammatory effects. These findings demonstrate the remarkable antioxidant, anti-inflammatory, and anti-microbial activities of Cannabis roots, supporting their traditional uses and challenging their perception as mere waste. This study highlights the therapeutic potential of Cannabis roots extracts and suggests avenues for further research and application.

Ultrahigh-Performance Liquid Chromatography–Tandem Mass Spectrometry-Based Untargeted Metabolomics to Differentiate Guizhou Glutinous Sorghum and Hongyingzi Sorghum

Ultrahigh-Performance Liquid Chromatography–Tandem Mass Spectrometry-Based Untargeted Metabolomics to Differentiate Guizhou Glutinous Sorghum and Hongyingzi Sorghum

Separation and identification of oligonucleotides impurities and degradation products by reversed phase ultra-high performance liquid chromatography using phenyl-bonded stationary phases without ion pairs - A step towards sustainability

This manuscript discusses the development of a reversed-phase ultra high-performance liquid chromatography (RP UHPLC) method based on phenyl-bonded stationary phases without ion-pairs for the separation and identification of oligonucleotides. The elimination of ion-pair reagents makes the proposed protocol as more compliant to the principles of green chemistry, compared to the traditional ion-pair reversed-phase liquid chromatography methods (IP RP LC). In detail, three phenyl-based stationary phases were tested, namely a C18/AR (a C18 stationary phase with the addition of aromatic groups), a Phenyl-hexyl, and a Diphenyl. Generally, the retention of oligonucleotides increases with the increase of salt concentration and the decrease of the pH, thus confirming the significant impact of van der Waals interactions, salting-out effect, and π-electrons interactions in the retention mechanism. The highest retention and best peak symmetry were observed for the C18/AR stationary phase, while the lowest retention for the Phenyl-hexyl, with retention influenced by the type of salt in the mobile phase. The obtained methods using C18/AR stationary phases allow for the effective separations of positional isomers and for identifying impurities and degradation products using RP UHPLC Q-TOF-MS in a comparatively short time. The application of RP UHPLC Q-TOF-MS provides reasonable selectivity for the resolution of 33 impurities and two degradation products. Both groups of compounds are mainly 3′N and 5′N-shortmers, but in the case of impurities, modifications of cyclic phosphate and phosphate groups were also identified. Nevertheless, Diphenyl and Phenyl-Hexyl may be applied to separate modified oligonucleotides with higher salt concentrations. The proposed separation methods without ion-pair reagents contribute to a more sustainable approach in oligonucleotide analysis.