Performance Liquid Chromatography Research Articles

Pharmacokinetic profile of phenytoin in dried blood spot with high-performance liquid chromatography-photodiode array.

Phenytoin is a first-line antiepileptic drug with narrow therapeutic range and follows non-linear pharmacokinetics. Pharmacokinetics of phenytoin have been studied in plasma matrix before, however, there were several disadvantages. This study aimed to obtain partial validation data of the analytical method and the pharmacokinetic profile of phenytoin in Dried Blood Spot (DBS) of six healthy subjects. DBS has the advantage of only requiring small sample volumes and could be transported more efficiently. Phenytoin along with carbamazepine as the chosen internal standard was analyzed with a reversed-phase high performance-liquid chromatography system and a photodiode array detector at 205nm. The results of partial validation, which evaluated the linearity, within-run accuracy, and precision, were within the criteria acceptance range. The pharmacokinetic profile showed that average AUC0-t was 83.81 ± 37.32μg.h/mL and AUC0-∞ was 83.65 ± 38.89μg.h/mL with an average ratio of 93%. Previous study quantifying phenytoin in the plasma matrix found the average AUC0-t was 39.41 ± 8.57µg.h/mL and AUC0-∞ was 42.94 ± 9.55µg.h/mL. Despite the difference between parameters of phenytoin analyzed in DBS and plasma matrices, the pharmacokinetic profiles obtained from both matrices were similar indicated by comparable concentration-time curves, thus, proving that DBS matrix can be used interchangeably with the plasma matrix as a more comfortable and effective alternative to phenytoin quantification in blood.

Europium Nanoparticle-Based Lateral Flow Strip Biosensors for the Detection of Quinoxaline Antibiotics and Their Main Metabolites in Fish Feeds and Tissues.

Olaquindox (OLA) and quinocetone (QCT) have been prohibited in aquatic products due to their significant toxicity and side effects. In this study, rapid and visual europium nanoparticle (EuNP)-based lateral flow strip biosensors (LFSBs) were developed for the simultaneous quantitative detection of OLA, QCT, and 3-methyl-quinoxaline-2-carboxylic acid (MQCA) in fish feed and tissue. The EuNP-LFSBs enabled sensitive detection for OLA, QCT, and MQCA with a limit of detection of 0.067, 0.017, and 0.099 ng/mL (R2 ≥ 0.9776) within 10 min. The average recovery of the EuNP-LFSBs was 95.13%, and relative standard deviations were below 9.38%. The method was verified by high-performance liquid chromatography (HPLC), and the test results were consistent. Therefore, the proposed LFSBs serve as a powerful tool to monitor quinoxalines in fish feeds and their residues in fish tissues.

Deciphering the TET3 interactome in primary thymic developing T cells

Ten-eleven translocation (TET) proteins are DNA dioxygenases that mediate active DNA demethylation. TET3 is the most highly expressed TET protein in thymic developing Tcells. TET3, either independently or in cooperation with TET1 or TET2, has been implicated in Tcell lineage specification by regulating DNA demethylation. However, TET-deficient mice exhibit complex phenotypes, suggesting that TET3 exerts multifaceted roles, potentially by interacting with other proteins. We performed liquid chromatography with tandem mass spectrometry in primary developing Tcells to identify TET3 interacting partners in endogenous, invivo conditions. We discover TET3 interacting partners. Our data establish that TET3 participates in a plethora of fundamental biological processes, such as transcriptional regulation, RNA polymerase elongation, splicing, DNA repair, and DNA replication. This resource brings in the spotlight emerging functions of TET3 and sets the stage for systematic studies to dissect the precise mechanistic contributions of TET3 in shaping Tcell biology.

Occurrence of Aflatoxin M1 and Estimate of Dietary Exposure in Cheeses from Organic and Conventional Production Systems

This study aimed to compare AFM1 occurrence in different cheese types produced by organic and conventional systems; and to evaluate the risk of food exposure to AFM1. A total of 176 commercial cheeses of 17 types were analyzed, 84 of organic and 92 of conventional production. Determination of AFM1 was performed by high- performance liquid chromatography (HPLC), being detected in 30.5% of samples, with 4.8% of organic cheese samples presenting quantifiable AFM1 values between 0.88 and 1.50 μg/kg. On the other hand, 4.3% of conventional cheese samples with values between 0.79 and 6.70 μg/kg. Two conventional cheese samples were above the limit of AFM1 allowed for cheeses by the Brazilian legislation. No statistical difference were found between organic and conventional cheeses regarding the occurrence (p = 0.1780) and concentration of AFM1 (p = 0.1810), according to the Chi-square and the T test, respectively. Estimated daily intake (EDI) and hazard index (HI) of dietary exposure to AFM1 were 0.26 ng/kg/day and 1.28 ng/kg/day, respectively, for conventional cheese samples, and 0.09 ng/kg/day and 0.47 ng/kg/day for organic samples, with no statistical difference for EDI (p = 0.1729) and HI (p = 0.1802) between the two production systems. Comparison between several cheese types from conventional and organic systems indicated that AFM1 is an obstacle to dairy production. Control and prevention of AFM1 contamination, as well as detoxification methods in the final products, are necessary. In the case of organic products, additional research is needed in order to determine which control and detoxification methods should be allowed in this production system.

A UPLC method development and validation study of Upadacitinib and its impurities in extended – Release oral tablet dosage forms

The primary objective was to develop a concomitant isocratic ultra-performance liquid chromatographic photo-diode array detection method to estimate Upadacitinib and its process-related impurities: impurity-1 and impurity-2. Further validation was conducted and studied for possible degradants under stress environments. All the chemicals and reagents used were of HPLC (acetonitrile, methanol) and analytical grade (trifluoro acetic acid). The ultra-performance liquid chromatography (Agilent 1290 Infinity II LC system) consists of a quaternary pump, a BEH C18 (50×2.1mm, 1.7μ) column, and photo-diode array detector. The method was developed with acetonitrile: methanol: 0.1% v/v trifluoro acetic acid (50:20:30 v/v/v) mobile phase at 0.2mL/min flow rate within a run time of 5.5min The detection was carried at 231.2nm. The respective retention times achieved were 2.289min (Upadacitinib), 0.972min (Upadacitinib impurity-1), and 3.508min (Upadacitinib impurity-2). The optimized method was validated further, and the linearity range was best fit at 15.0-180.0μg/mL for Upadacitinib and 1.0-12.0μg/mL for both Upadacitinib impurity-1 and 2 respectively. The detection and quantification limits were 4.50μg/mL, 15.00μg/mL (Upadacitinib) and 0.30μg/mL, 1.0μg/mL (Upadacitinib impurity-1 and 2). A fast, isocratic, specific, and reproducible ultra-performance liquid chromatographic method was developed and validated for various parameters according to the ICH Q2 (R1) guidelines studies. Stress studies were conducted exposing the sample dilution to various treatments (acid, alkali, peroxide, HPLC water, heat, and UV light). The degradants were well-separated apart from the peaks of the active substance. The stability indicating nature was observed during the degradation. The optimized method can be applied for the separation and estimation of Upadacitinib and its process-related impurities in pharma sector in tablet dosage forms.

Electrochemical determination of salbutamol in aqueous solutions with UiO‐66/rGO‐modified electrodes

AbstractThe synthesis of the UiO‐66/rGO material involved combining UiO‐66 with reduced graphene oxide (rGO). This resulting composite found application in the electrochemical determination of salbutamol (SAL) through modification of a glassy‐carbon electrode (GCE) with UiO‐66/rGO. Differential pulse voltammetry served as the chosen technique, offering a wide linear range spanning 1–160 µm and demonstrating a low limit of detection at 0.84 µm. To ensure the reliability of the electrode, various aspects such as sensitivity, repeatability, reproducibility, and stability were meticulously studied. When applied to the analysis of SAL in swine urine solutions (n = 3), the electrode exhibited good recovery rates. Remarkably, the results obtained through this method correlated well with those acquired using ultra‐high performance liquid chromatography (UPLC).

Identification of RNAi hypoallergic bread wheat lines for wheat-dependent exercise-induced anaphylaxis patients.

Wheat-dependent exercise-induced anaphylaxis (WDEIA) is one of the most severe forms of wheat allergy. It occurs in patients when they exercise after ingesting wheat-containing foods. Nowadays, the only possible alternative for WDEIA patients is to avoid such foods. This study investigated the potential of six RNA of interference (RNAi) wheat lines with low-prolamin content as alternatives for WDEIA patients. For that purpose, a high performance-liquid chromatography (HPLC) analysis was performed to evaluate differences in gluten protein fractions among these lines. Next, western blots were conducted to measure the immunoglobulin E (IgE) reactivity to wheat proteins in sera from five WDEIA patients. Additionally, monoclonal antibodies (moAb) recognition sites and the IgE binding sites were searched in all peptides identified by LC-MS/MS after protein digestion. The results showed a 61.4%-81.2% reduction in the gliadin content of the RNAi lines, accompanied by an increase in their high-molecular weight (HMW) glutenin content compared to the wild type bread wheat line (WT). In all cases, the reduction in gliadin content correlated with a decrease in IgE reactivity observed in the sera of WDEIA patients, highlighting the E82 and H320 lines. These two RNAi lines exhibited a ≤90% reduction in IgE reactivity. This reduction could be attributed to an absence of IgE binding sites associated with α- and ω5-gliadins, which were present in the WT. Overall, these lines offer a potential alternative for foodstuff for individuals with WDEIA.

Four sides to the story: A proteomic comparison of liquid-phase and matrix-bound extracellular vesicles in 2D and 3D cell cultures.

Multipotent mesenchymal stromal cells (MSCs)-derived extracellular vesicles (EVs) play important roles in cellular communication and are extensively studied as promising therapeutic agents. While there is a substantial pool of studies on liquid-phase EVs, data on EVs bound to the extracellular matrix (ECM) is lacking. There is also an emerging trend of accumulating and comparing data on characteristics of EVs obtained in different culturing conditions. Aiming to reveal proteomic signatures of EVs obtained from conditioned media and ECM of MSCs cultured in 2D and 3D conditions, we performed liquid chromatography with tandem mass spectrometry. Bioinformatic analysis revealed common patterns in proteomic composition of liquid-phase EVs and matrix-bound vesicles (MBVs), namely extracellular environment organization, immune, and transport pathways enrichment. However, extracellular environmental organization pathways are more enriched in liquid-phase EVs than in MBVs, while MBVs proteins noticeably enrich enzymatic pathways. Furthermore, each type of EVs from 2D and 3D cultures has a unique differential abundance profile. We have also performed comparative functional assays, namely scratch assay to assess EVs effect on cell migration and tubulogenesis assay to evaluate EVs angiogenic potential. We found that both liquid-phase EVs and MBVs enhance cell migration, while angiogenic potential is higher in MBVs. Results of the present study suggest that while both liquid-phase EVs and MBVs have therapeutic potential, some unique features of each subgroup may determine optimal areas of their application.

Untargeted metabolomics reveals that declined PE and PC in obesity may be associated with prostate hyperplasia.

Recent studies have shown that obesity may contribute to the pathogenesis of benign prostatic hyperplasia (BPH). However, the mechanism of this pathogenesis is not fully understood. A prospective case-control study was conducted with 30 obese and 30 nonobese patients with BPH. Prostate tissues were collected and analyzed using ultra performance liquid chromatography ion mobility coupled with quadrupole time-of-flight mass spectrometry (UPLC-IMS-Q-TOF). A total of 17 differential metabolites (3 upregulated and 14 downregulated) were identified between the obese and nonobese patients with BPH. Topological pathway analysis indicated that glycerophospholipid (GP) metabolism was the most important metabolic pathway involved in BPH pathogenesis. Seven metabolites were enriched in the GP metabolic pathway. lysoPC (P16:0/0:0), PE (20:0/20:0), PE (24:1(15Z)/18:0), PC (24:1(15Z)/14:0), PC (15:0/24:0), PE (24:0/18:0), and PC (16:0/18:3(9Z,12Z,15Z)) were all significantly downregulated in the obesity group, and the area under the curve (AUC) of LysoPC (P-16:0/0/0:0) was 0.9922. The inclusion of the seven differential metabolites in a joint prediction model had an AUC of 0.9956. Thus, both LysoPC (P-16:0/0/0:0) alone and the joint prediction model demonstrated good predictive ability for obesity-induced BPH mechanisms. In conclusion, obese patients with BPH had a unique metabolic profile, and alterations in PE and PC in these patients be associated with the development and progression of BPH.

피부각질세포에서 청심 맥문동 추출물의 케모카인 발현 억제효과

Background/Objectives: This study aimed to develop Cheongsim Liriope platyphylla (CS) as a functional material for anti-atopic dermatitis.
 Methods: After extracting CS and the traditional Liriope platyphylla (CY) variety using 80% and 60% ethanol, we conducted a comparative analysis of the changes in composition via high- performance liquid chromatography (HPLC). We evaluated the inhibitory effects of CS on the expressions of CCL17 and CCL22, representative markers of atopic dermatitis, in human keratinocyte HaCaT cells induced with TNF-a !IFN-¥-induced inflammation. Further, we analyzed the chromatograms of CS extracts from nine farms located in Cheongyang and measured their effects on reducing inflammatory cytokine expression.
 Results: The HPLC comparative analysis between CS and CY ethanol extracts showed an increasing trend in specific time intervals, specifically at 24 32 minutes, 34 36 minutes, and 38 40 minutes in the CS 80% ethanol extract compared to CY. Moreover, when comparing the inhibitory effects on the expression of CCLI 7 and CCL22 in keratinocytes, the CS 60% ethanol extract outperformed CY. On analyzing the chromatograms of CS (CS1-CS9) from the nine farms, we observed a higher peak at 26 minutes in the chromatograms of CS5 and CS6, suggesting differences in composition. Particularly, the CS6 extract, exhibiting vanous compositions, demonstrated excellent suppression of inflammatory cytokines.
 Conclusion/Implications: The results of this study indicate the potential of CS extracts as a functional material for addressing skin disorders. The results can be utilized as fundamental data for analyzing Cheongsim CS from various farms in Cheongyang.

Blood‐based biomarkers for Alzheimer’s Disease in Down Syndrome

AbstractBackgroundPeople with DS are at high risk to develop Alzheimer’s disease dementia (AD) (Lott & Head, 2001). Predicting and monitoring decline and onset of dementia is a diagnostic challenge and of essence in daily care and support for people with DS. Behavioral and Psychological Symptoms of Dementia (BPSD) are an easily accessible tool for the prediction of dementia onset in the DS population. The BPSD‐DS II scale was developed and validated by our team to identify behavioral changes linked to AD between the last six months and pre‐existing behavior in an individual with DS (Dekker et al., 2021). This project is based on a cohort of DS individuals without AD (n = 94), with AD (n = 43), or with questionable AD (qAD) (n = 54) aged >35 years. At inclusion, the BPSD‐DS II was applied to assess AD‐linked behavioral changes, and biofluid sampling was performed as well. Changes in cerebrospinal fluid (CSF) levels of monoaminergic neurotransmitters and metabolites have been associated with BPSD in AD (Dekker et al., 2015). However, CSF sampling procedures remain difficult in DS, illustrating the need for blood biomarkers. BPSD‐DS II results will be correlated with the serum levels of monoaminergic and classical AD biomarkers. Three years post‐inclusion, a follow‐up study was conducted to assess the evolution of possible AD dementia within this group and to determine the predictive values of the BPSD‐DS II scale and the different analytes.MethodWe will use reversed‐phase ultra‐high performance liquid chromatography with electrochemical detection to determine the levels of (nor)epinephrine, dopamine, serotonin and their metabolic products in serum. We will use the Simoaâ platform on the serum of clients with DS without or with (q)AD to detect the levels of Ab40, Ab42, total tau, phosphorylated tau, Glial Fibrillary Acidic Protein and Neurofilament Light chain.ResultWe expect a correlation between the results of the BPSD‐DS II and the levels of the monoamines and classical biomarkers in patients with DS without or with (q)AD.ConclusionThe diagnosis predicted by the results of the BPSD‐DS II correlates with the levels of monoamines and classical biomarkers in individuals with DS without or with (q)AD.

In silico screening and characterization, inhibition mechanism on ACE, and stability of antihypertensive peptides with Zn-chelating capacity identified from millet bran albumin hydrolysates

To obtain Angiotensin-I-Converting Enzyme (ACE) inhibition peptides with Zn-chelating capacity, millet bran albumin hydrolysates (MBAH) were subjected to Sephadex G-15 gel and reverse phase-high liquid performance chromatography, and UPLC-ESI-MS/MS analysis. Five oligopeptides including Val-Gly-Ser-Asp-Gly-Leu (VGSDGL), Thr-Gly-His-Gly-Val-Ala (TGHGVA), Gly-Cys-His-His-Tyr (GCHHY), Leu-Val-Glu-Ala-Val-Pro-Arg-Gly (LVEAVPRG) and Val-Leu-Ala-Gln-Glu-Glu (VLAQEE) were identified in MBAH. Of these, only pentapeptide GCHHY showed both ACE-inhibitory activity (IC50: 134.78 μmol/L) and Zn-chelating capacity (14.16 mg/g). Molecular docking and inhibition kinetics showed that GCHHY was a competitive inhibitor of ACE, because it could bind with Lys511 of ACE which belongs to the central S2 pocket. Moreover, GCHHY can affect zinc tetrahedral coordination in ACE through binding with Glu411 and His383. The sulfhydryl, amino and carboxyl groups of GCHHY can bind with zinc ions. Under the gastrointestinal digestion, the ACE-inhibitory activity of GCHHY was relatively stable, and the zinc solubility of GCHHY-Zn complexes was more stable than zinc sulfate (P < 0.05). In addition, both GCHHY and GCHHY-Zn complexes lowered the diastolic blood pressure and systolic blood pressure of spontaneous hypertensive rats at dose of 50–200 mg/kg body weight. These results suggested the potential applications of millet albumin peptides as ingredients for antihypertension or zinc fortification.

Effects of Zhuang medicine compound Xiancao Granule on diabetic kidney disease: A multi-omics analysis

Ethnopharmacological relevanceDiabetic kidney disease (DKD) poses a severe threat to human health. Compound Xiancao Granule (CXCG), a classic Zhuang medicinal formula, is reported as highly effective in treating DKD. However, the mechanisms underlying the action of CXCG in DKD remain unclear. Aim of the studyThis study aimed to investigate the mechanisms of action of CXCG against DKD using multi-omics analysis, including 16s rRNA sequencing, metabolomics, and transcriptomics. Materials and methodsThe chemical compounds of CXCG were identified using ultra-high- performance liquid chromatography quadrupole/electrostatic field orbital trap high-resolution mass spectrometry analysis. A rat model of DKD was established by combining nephrectomy of the left kidney, high-fat diet, and streptozotocin. The therapeutic effects of CXCG on DKD were assessed based on body weight, blood glucose level, renal function, inflammatory cytokine levels, and histological staining. Subsequently, 16s rRNA sequencing, liquid chromatography-tandem mass spectrometry untargeted metabolomic profiling, and RNA sequencing analysis were used to investigate the mechanisms of action of CXCG in DKD. Spearman's correlation analysis was performed to elucidate the correlations between efficacy indicators, gut microbiota, metabolites, and inflammation-related genes. ResultsA total of 118 compounds were identified in CXCG. CXCG significantly ameliorated glucose metabolism disorders, improved renal function, attenuated inflammation, and delayed renal pathological changes in DKD rats. CXCG modulated gut microbiota dysbiosis, including Alloprevotella, Oscillibacter, Anaeroplasma, Anaerotruncus, and Faecalibacterium. In addition, metabolic disruption in DKD rats was regulated by CXCG, which is involved in the metabolism of carbohydrates and amino acids. Transcriptome analysis showed that CXCG affected DKD mainly by regulating inflammation-related genes and pathways, such as the PI3K/Akt and MAPK signaling pathways. Furthermore, there were significant correlations between efficacy indicators, gut microbiota, metabolites, and genes. ConclusionThis multi-omics association study provides novel insights into the effects of CXCG on DKD by remodeling the gut microbiota structure and restoring the metabolic homeostasis through the regulation of carbohydrate metabolism, amino acid metabolism, and inflammation-related pathways, highlighting a potential therapeutic strategy for DKD.

Disruption of the Contents of Endogenous Hormones Cause Pollen Development Obstruction and Abortion in Male-Sterile Hybrid Lily Populations.

Lilies are well-known flowers with large anthers and a high quantity of pollen that easily contaminates clothing and tepals. The anthers need to be artificially removed, leading to production problems. Cultivating male-sterile or pollen-free lilies could solve these problems. The key period of male sterility in a specific male-sterile hybrid lily population was determined through cytological observation. The contents of hormones, soluble sugar, soluble protein, and proline were determined by high-performance liquid chromatography, tandem mass spectrometry and colorimetry. Transcriptome sequencing was used to identify the genes with altered expression. The key period of male sterility was determined to be the microspore mother and tetrad stages. The hormone contents were abnormal in the sterile line compared with the fertile line. The indole-3-acetic acid (IAA) content was higher in the sterile line than in the fertile line at all stages, while the gibberellic acid 4 (GA4) content showed the opposite result. Abscisic acid (ABA) accumulated in the sterile line in both the microspore mother and tetrad stages, and the zeatin riboside (ZR) content in the sterile line increased at the microspore mother stage but decreased at the tetrad stage. The contents of soluble sugar, soluble protein and proline were higher in the fertile line than in the sterile line. Genes involved in auxin and ABA synthesis and signalling pathways were highly expressed in the male-sterile line. Our data suggested that abnormal contents of hormones in the microspore mother and tetrad stages resulted in pollen abortion in a male-sterile hybrid lily population, which indicated that the hormone balance in specific stages plays critical functions in pollen development in lilies.

Unlocking the genetic potential of Ethiopian durum wheat landraces with high protein quality: Sources to be used in future breeding for pasta production

AbstractThe content and composition of the grain storage proteins in wheat determine to a high extent its end‐use quality for pasta and bread production. This study aimed to evaluate the content and composition of the grain storage proteins in Ethiopian landraces and cultivars to contribute to future breeding toward improved pasta quality. Thus, 116 landraces and 34 cultivars originating from Ethiopia were grown in three locations, and the protein parameters were analyzed using size exclusion‐high performance liquid chromatography (SE‐HPLC). A considerable variation in the amount of the analyzed protein parameters was found. The genotypes, environments, and interactions contributed significantly (p &lt; 0.001) to the differences obtained. The broad‐sense heritability was high (0.75–0.98) for all protein parameters except for unextractable small monomeric protein (uSMP). Using the principal component analysis (PCA) to evaluate the impact of protein parameters and using either PCA or unweighted pair group method with arithmetic mean (UPGMA) to assess the impact of the genetic composition, the cultivar group was found to form a separate cluster. This indicates that durum wheat improvement in Ethiopia has relied on exotic materials, which might result from a narrow genetic base. Unlike most landraces, most released cultivars showed a high and stable gluten strength across environments. Two landraces, G057 and G107, were found genetically distinct from the released cultivars but with high and stable gluten. The two selected landraces might be of extremely high value for future use in durum wheat breeding programs, as they might be adapted to wide‐ranging Ethiopian growing conditions, they might carry genes of relevance to withstand abiotic and biotic stresses, and they seem to hold essential protein properties, which might result in high‐quality grains for industrial processes.

Ultra-high Performance Liquid Chromatography-Ion Mobility-High-Resolution Mass Spectrometry to Evaluate the Metabolomic Response of Durum Wheat to Sustainable Treatments.

Sustainable agriculture aims at achieving a healthy food production while reducing the use of fertilizers and greenhouse gas emissions using biostimulants and soil amendments. Untargeted metabolomics by ultra-high performance liquid chromatography-ion mobility-high-resolution mass spectrometry, operating in a high-definition MSE mode, was applied to investigate the metabolome of durum wheat in response to sustainable treatments, i.e., the addition of biochar, commercial plant growth promoting microbes, and their combination. Partial least squares-discriminant analysis provided a good discrimination among treatments with sensitivity, specificity, and a non-error rate close to 1. A total of 88 and 45 discriminant compounds having biological, nutritional, and technological implications were tentatively identified in samples grown in 2020 and 2021. The addition of biochar-biostimulants produced the highest up-regulation of lipids and flavonoids, with the glycolipid desaturation being the most impacted pathway, whereas carbohydrates were mostly down-regulated. The findings achieved suggest the safe use of the combined biochar-biostimulant treatment for sustainable wheat cultivation.

Valorized pineapple waste by conventional and energy-saving ohmic extraction: potentially toxic elements and mycotoxin contamination

Pineapple peel is a food waste with environmental concerns. It can be valorized by extracting its bioactive compound through conventional and emerging technologies such as ohmic heating. However, mycotoxins and heavy metals contaminations could be concerns of such a valorization process. This study assessed the effects of conventional and ohmic heating on selected aflatoxins and metal ions in pineapple peel extract using high- performance liquid chromatography (HPLC) and inductively coupled plasma mass spectrometry (ICP-MS). Also, energy consumption and environmental impacts of the ohmic process were analyzed. According to the results, the ohmic-based valorization system reduced energy consumption and greenhouse gas emissions by more than 83%, contributing to net-zero and sustainable development goals (SDGs). Aflatoxins B1, B2, G1, and G2 were below the detection limit (B1, G1: 0.2µg/kg; B2, G2: 0.1µg/kg). However, ohmic extracts contained higher concentrations of Cr, Pb, Ni, and Fe because of electrochemical reactions and electrode corrosion. Despite the capability of ohmic valorization systems, inspecting food waste and valorized products for hazardous chemicals and investing in durable electrodes (e.g., titanium) are among the considerations for ohmic systems’ industrial implementation.

Anticancer Effects of Date Seed Oil- Loaded Niosomes and Gemcitabine Combination in MCF-7 Human Breast Cancer Cells

Gemcitabine (GEM) is one of the most widely used chemotherapeutic medications for treating various solid tumuors. Niosomes are a novel drug delivery system, self-assembled vesicular nanocarriers and composed of several surfactants and various lipids. Date seed Oil (DSO) contain a high percentage of phenolic and flavonoid compounds that making them important for food and pharmaceutical formulations. In this study, we prepared GEM loaded into anionic niosomes (GEM-niosomes) to target breast tumor cells. The niosomes were prepared with cholesterol (CHOL), span 60, date seed oil and chloroform using the thin film hydration method. GEM-niosomes were fully characterized for their physiochemical properties and evaluated for their cytotoxicity. The targeted niosomes were 100±10nm, the loaded niosomes were in the range of 125±15. GEM-fully loaded DSO-niosomes with drug entrapment percentage (EE%) 82% using high- performance liquid chromatography (HPLC) were developed. The prepared targeted GEM-niosomes showed good stability over 4 weeks at -7ºC and better drug loading. Our targeted GEM-niosomes showed stronger activity against MCF-7 cell by approximately 10 folds compared with the free drug. Taken together, the combination of GEM and “DSO- loaded niosomes” may be of great importance for developing new treatments following in vivo investigations with breast cancer animal models. Future research should explore the in vitro and the in vivo cytotoxicity of this combination on more aggressive breast cancer ( BC).

Supercritical fluid extraction tandem responsive polarity modifier separation of hydroxytyrosol rich phenolic raffinates from olive oil

SummaryThis research includes recovery of hydroxytyrosol (HT) rich bioactive raffinates from olive oil through supercritical fluid extraction (SFE) using CO2 and polarity modifiers including ethanol (EtOH) and methanol (MeOH) at 6% each. The raffinates left after 120 min of SFE (&lt;1%) at 40 °C and 150 bar were subjected to minor phenolic analysis through Reversed Phase‐High Performance Liquid Chromatography (RP‐HPLC). HT being the most consistent phenolic compound, the retention, total phenolic content (TPC) and 1,1‐diphenyl‐2‐picrylhydrazyl (DPPH) radical scavenging activity decreased in the raffinates as Supercritical carbon dioxide (ScCO2) + (MeOH) &gt; ScCO2 + (EtOH) &gt; ScCO2. Multivariate analysis revealed high positive correlations among the individual phenolic compounds and antioxidant potential. Principal components analysis explained 90.5% variability through two principal components. The MeOH, ScCO2, ScCO2 + MeOH and ScCO2 + EtOH extracts were clearly separated in the score plot with positive loadings of parameters on the loading plot. Hierarchical cluster analysis separated the MeOH extraction and SFE in two clusters. In conclusion, ScCO2 + MeOH and ScCO2 + EtOH could be employed for the recovery of HT from olive oil.

Rapid Differentiation of Tea through High - Performance Liquid Chromatography and Chemometric Analysis

Rapid Differentiation of Tea through High - Performance Liquid Chromatography and Chemometric Analysis