High-pressure Liquid Chromatography Research Articles

Comparison of Capillary Zone Electrophoresis with High-pressure Liquid Chromatography in the Evaluation of Hemoglobinopathies

The capillary zone electrophoresis (CZE) and high-performance liquid chromatography (HPLC) methods were compared in terms of HbA2 measurement for the assessment of hemoglobinopathies. CZE was compared with HPLC for the evaluation of patients without hemoglobinopathy (n=321), with β-thalassemia trait (n=113), and with common (HbD-Punjab, E, C, S/A, and S/S) and rare (HbS/D, O-Arap, Lepore, G-Coushata, Setif, Hamadan, Q-Iran, and H) variants (n=21). The reference range for HbA2 was determined by CZE. Among patients without hemoglobinopathy, the median (2.5th-97.5th percentiles) values were 97.4% (97.0-98.0%) and 97.5% (96.6-98.4%) for HbA (p=0.060) and 2.4% (1.6-3.0%) and 2.5% (1.6-3.1%) for HbA2 (p<0.001) by HPLC and CZE, respectively. The reference range for HbA2 was 1.6-3.1% by CZE. In the comparison of methods for HbA2, there was a constant error of 0.255 (confidence interval: 0.062-0.448) and bias of 0.10% (limit of agreement: 0.33-0.53), and higher values were obtained with CZE. A strong correlation was observed between the methods (r=0.782). Interrater agreement was almost perfect for clinical diagnosis (ϰ=0.911). The two methods detected and identified the common variants similarly. All rare variants, except HbH by HPLC and HbS/D by CZE, were detected as separate peaks by both methods. The two methods were in agreement regarding the preliminary identification of β-thalassemia patients. Different Hb variants were detected by both methods but with possible methodological interference for HbA2 measurements. CZE is a reliable and simple alternative for the evaluation of hemoglobinopathies. The standardization of HbA2 measurements should be prioritized as more techniques become available in routine laboratory practice.

Sesquiterpenoids from the stem bark of Aglaia pachyphylla Miq (Meliaceae) and cytotoxic activity against MCF-7 Cancer Cell Line

Sesquiterpenoids are terpenoid-derived compounds formed from three isoprene units with diverse pharmacological activities. Sesquiterpenoids can be obtained from higher plants, such as the genus Aglaia from the Meliaceae family. This study aims to isolate and characterize the structure of sesquiterpenoidsfrom the n-hexane extract of Aglaia pachyphylla Miq stem bark and to determine their cytotoxic activity against MCF-7 breast cancer cells. The n-hexane extract was separated and purified by various chromatography techniques such as vacuum liquid chromatography, normal-phase chromatography, and reversed-phase chromatography to obtain three sesquiterpenoids. The chemical structures of sesquiterpenoids were identified by various spectroscopic analyses such as IR, MS, 1D-NMR, and 2D-NMR and compared with previously reported spectrum data. Three sesquiterpenoids were identified as β-caryophyllene oxide (1), 1β-Hydroxy-4(15),5-eudesmadiene (2), and spathulenol (3). The three compounds were tested against MCF-7 breast cancer cells using the PrestoBlue method. Compound 2 showed the highest cytotoxic activity with an IC50 value of 262,25 µM.

New insights into the dyes of Central Asian ikat textiles

Central Asian ikat textiles are characterised by their bold and large abstract patterns, made up of vibrant colours with a characteristic “blurred edge” effect, which makes them some of the most recognisable fabrics worldwide. Eleven ikats from the collection of the National Museum of Asian Art, Smithsonian Institution (Washington, DC, USA) and six ikats from the collection of the British Museum (London, UK) were investigated by high pressure liquid chromatography coupled to diode array detector and high-resolution tandem mass spectrometry (HPLC-DAD-HRMS/MS) to identify the dyes and expand on recent research focused on the integration of synthetic dyes in ikat dyeing during the second half of the 19th century. The results confirmed that the palette of natural dyes includes cochineal (probably mostly from a local species native to central Asia) for red, larkspur (Delphinium semibarbatum) for yellow, and indigo for blue as the main dye sources. In some of the textiles, madder (Rubia tinctorum) for red and pagoda tree (Sophora japonica) for yellow were also detected as minor components mixed with cochineal and larkspur, respectively. Synthetic dyes, such as scarlet reds (C.I. 16150 and 16155), rose bengale (C.I. 45435), eosin (C.I. 45380), erythrosine (C.I. 45430), fuchsin (C.I. 42510/42520), methyl violet (C.I. 42535/42555), malachite green (C.I. 42000) and brilliant green (C.I. 42040) were detected in eight textiles. In three textiles, natural dyes were identified in the ikat-dyed silk warp threads and synthetic dyes in the red cotton weft threads, highlighting interesting mixtures of materials and textile-making practices. Natural yellow dyes were used in all textiles, even those for which all the other colours are synthetic. These results provide significant new information about the changes in ikat production that occurred during the 19th century in Central Asia.

Inter-Varietal Variation in Phenolic Profile, Antioxidant, Anti-Inflammatory and Analgesic Activities of Two Brassica rapa Varieties: Influence on Pro-Inflammatory Mediators.

The present research study aims to appraise the potential of polyphenol-rich extracts from two Brassica rapa varieties on antioxidant, anti-inflammatory and analgesic activities using carrageenan-induced paw edema model in rats. Methanol extracts of peels and pulps of Brassica rapa yellow root (BRYR) and Brassica rapa white root (BRWR) were prepared using the soxhlet extraction technique. All four extracts were analyzed by reversed-phase high-pressure liquid chromatography (RP-HPLC) for the polyphenols, and results showed that 10 phenolic acids and 4 flavonoids were detected. Gallic acid was the major phenolic acid (174.6-642.3 mg/100 g of dry plant material) while catechin was the major (34.45-358.5 mg/100 g of dry plant material) flavonoid detected in the extracts. The total phenolic contents (TPC) of BRYR peel, BRWR peel, BRYR pulp and BRWR pulp extracts were in the range of 1.21-5.01 mg/g of dry plant material, measured as GAE, whereas the total flavonoid contents (TFC) were found in the range of 0.90-3.95 mg/g of dry plant material, measured as QE. BRYR peel extract exhibited the best DPPH radical scavenging activity (IC50, 3.85 µg/mL) and reducing potential as compared with other extracts. The in vivo anti-inflammatory potential was assessed by carrageenan-induced rat paw edema, and the analgesic potential was investigated by a hot plate test. Suppression of biochemical inflammatory biomarkers including C-reactive protein (CRP), rheumatoid factor (RF) and tumor necrosis factor (TNF-α), and interleukin-6 (IL-6) concentration were also determined. Results showed that BRYR peel extracts reduced paw edema and suppressed the production of TNF-α, IL-6, CRP and RF most significantly, followed by BRWR peel, BRYR pulp and BRWR pulp extracts. In addition, histopathology observation also supports the anti-inflammatory effect of peel extracts as being greater than that of root pulp extracts. Moreover, it was observed that the analgesic effect of the root-peel extracts was also more pronounced as compared with root-pulp extracts. It can be concluded that BRYR peel extract has higher phenolic contents and showed higher suppression of TNF-α, IL-6, CRP and RF, with strong antioxidant, anti-inflammatory and analgesic effects.

Hybrid Polymer-Inorganic Materials with Hyaluronic Acid as Controlled Antibiotic Release Systems.

In recent years, significant developments have taken place in scientific fields such as tissue and materials engineering, which allow for the development of new, intelligent biomaterials. An example of such biomaterials is drug delivery systems that release the active substance directly at the site where the therapeutic effect is required. In this research, polymeric materials and ceramic-polymer composites were developed as carriers for the antibiotic clindamycin. The preparation and characterization of biomaterials based on hyaluronic acid, collagen, and nano brushite obtained using the photocrosslinking technique under UV (ultraviolet) light are described. Physical and chemical analyses of the materials obtained were carried out using Fourier transform infrared spectroscopy (FT-IR) and optical microscopy. The sorption capacities were determined and subjected to in vitro incubation in simulated biological environments such as Ringer's solution, simulated body fluid (SBF), phosphate-buffered saline (PBS), and distilled water. The antibiotic release rate was also measured. The study confirmed higher swelling capacity for materials with no addition of a ceramic phase, thus it can be concluded that brushite inhibits the penetration of the liquid medium into the interior of the samples, leading to faster absorption of the liquid medium. In addition, incubation tests confirmed preliminary biocompatibility. No drastic changes in pH values were observed, which suggests that the materials are stable under these conditions. The release rate of the antibiotic from the biomaterial into the incubation medium was determined using high-pressure liquid chromatography (HPLC). The concentration of the antibiotic in the incubation fluid increased steadily following a 14-day incubation in PBS, indicating continuous antibiotic release. Based on the results, it can be concluded that the developed polymeric material demonstrates potential for use as a carrier for the active substance.

α-Tocopherol Depletion Exacerbates Lipopolysaccharide-Induced Reduction of Grip Strength

Backgroundα-Tocopherol (αT) deficiency causes several neurologic disorders, such as spinocerebellar ataxia, peripheral neuropathy, and myopathy. Furthermore, decreased antibody production, impaired ex vivo T cell function, and elevated cytokine production are observed in humans and mice with αT deficiency. Although modeling αT deficiency in animals is challenging, αT depletion can be more readily achieved in α-tocopherol transfer protein-null (Ttpa-/-) mice than wild-type (WT) mice. Thus, the Ttpa-/- mouse model is a useful tool for studying metabolic consequences of low αT status. Optimizing this mouse model and selecting the reliable indicators/markers of deficiency are still needed. ObjectiveOur objective was to assess whether αT depletion alters lipopolysaccharide (LPS)-induced inflammatory response in the brain and/or grip strength used as a proxy for fatigue. MethodsWT and Ttpa-/- weanling littermates (n = 37–40/genotype) were fed an αT deficient diet ad libitum for 9 wk. Mice were then injected with LPS (10 μg/mouse) or saline (control) intraperitoneally and killed 4 h later. Concentrations of αT in diet and tissues were measured via high-pressure liquid chromatography. Grip strength was evaluated via a grip strength meter apparatus 2 d before and 3.5 h after LPS injection. Cerebellar and serum interleukin-6 (IL-6) concentrations were measured via enzyme-linked immunosorbent assay. ResultsαT concentrations in the liver, heart, and adipose tissue of WT mice were higher than Ttpa-/- mice. Although αT was detected in the brain, muscle, and serum of WT mice, it was undetectable in these tissues of Ttpa-/- mice. Cerebellar and serum concentrations of IL-6 were increased in LPS-treated groups but were not significantly affected by genotype. Grip strength was reduced in LPS-treated groups, an effect that was more pronounced in Ttpa-/- mice. ConclusionsSystemic LPS administration caused an acute inflammatory response with a concomitant decline in grip strength, especially in Ttpa-/- mice. αT depletion appears to exacerbate reductions in grip strength brought on by systemic inflammation.

Evaluation of the dyeability of hemp fabric with Anatolian black pine (Pinus nigra subsp. Pallassiana) and Scots pine (Pinus sylvestris L.)

The development of more sustainable textile dyeing techniques includes a lot of potential in utilizing bio-based plant-derived colorants. Hemp fabric has been dyed with an aqueous extract of pine cones at various dye concentrations to produce a brownish to darker color. Agricultural waste specimens, Anatolian black pine ( Pinus nigra subsp. Pallassiana), and Scots pine ( Pinus sylvestris L.) cones were used as natural dye sources on a sustainable basis. The dyed hemp fabrics were analyzed by different analytical and technical methods such as CIE L*a*b*, high-pressure liquid chromatography–diode-array detection, and scanning electron microscopy with an energy dispersive X-ray spectrometer. The color characteristics of all the dyed fabrics were measured by a CIE L*a*b* spectrophotometer. The reversed-phase high-pressure liquid chromatography–diode-array detection method with the diode-array detection method was utilized for the identification of the components of dyes present in the dyed fabrics. Scanning electron microscopy with an energy dispersive X-ray spectrometer was used for imaging and elemental analysis of the surfaces of the dyed hemp fabrics. This work has created a novelty direct dyeing method with pine cones without using any mordants and to promote the use of agricultural waste. The fastness values of washing, rubbing, and light were assessed and contrasted. The fastness results were good in general. Washing fastness values were excellent and the same for all fabrics between 4 and 5. Rubbing fastness is also generally good and of the desired grade. The chemical compositions of hemp and pine cones, as well as the formation of dyestuff fiber, were investigated by using Fourier transform infrared analysis.

Unlocking the potential of glyphosate-resistant bacterial strains in biodegradation and maize growth.

Glyphosate [N-(phosphonomethyl)-glycine] is a non-selective herbicide with a broad spectrum activity that is commonly used to control perennial vegetation in agricultural fields. The widespread utilization of glyphosate in agriculture leads to soil, water, and food crop contamination, resulting in human and environmental health consequences. Therefore, it is imperative to devise techniques for enhancing the degradation of glyphosate in soil. Rhizobacteria play a crucial role in degrading organic contaminants. Limited work has been done on exploring the capabilities of indigenously existing glyphosate-degrading rhizobacteria in Pakistani soils. This research attempts to discover whether native bacteria have the glyphosate-degrading ability for a sustainable solution to glyphosate contamination. Therefore, this study explored the potential of 11 native strains isolated from the soil with repeated glyphosate application history and showed resistance against glyphosate at higher concentrations (200 mg kg-1). Five out of eleven strains outperformed in glyphosate degradation and plant growth promotion. High-pressure liquid chromatography showed that, on average, these five strains degraded 98% glyphosate. In addition, these strains promote maize seed germination index and shoot and root fresh biomass up to 73 and 91%, respectively. Furthermore, inoculation gave an average increase of acid phosphatase (57.97%), alkaline phosphatase (1.76-fold), and dehydrogenase activity (1.75-fold) in glyphosate-contaminated soil. The findings indicated the importance of using indigenous rhizobacteria to degrade glyphosate. Therefore, by maintaining soil health, indigenous soil biodiversity can work effectively for the bioremediation of contaminated soils and sustainable crop production in a world facing food security.

Determination of tannin acyl hydrolase activity of Streptomyces mirabilis TBGS10 by measuring gallic acid using high-pressure liquid chromatography

Tannase is a hydrolytic enzyme known as tannin acyl hydrolase that acts on the ester bonds of hydrolyzable tannins and produces gallic acid. Tannase enzymes are obtained from various sources. Tannase from microbial sources, especially fungi like Aspergillus spp., has been used in the food, brewing, and pharmaceutical industries. Tannin acyl hydrolase activities of Actinomycetes are seldom reported, though they are known to produce a wide range of industrial enzymes. This communication deals with the tannase activity of a Streptomyces isolate from the shola forests of Munnar in the Western Ghats of Kerala. The isolate is identified as S. mirabilis TBGS10 based on morphological characters and 16s rDNA homology. The isolate showed promising tannase activity in plate assays, submerged, and solid-state fermentation conditions. High-pressure liquid chromatography determines the industrially important intermediate gallic acid produced through solid-state fermentation using cashew apple bagasse as substrate. The extract observed a gallic acid (GA) content of 142.624 g/mL with a retention time of 2.506 min. The tannase gene of the strain TBGS10 was PCR amplified using specific primers designed based on similar Streptomyces sequences available in the NCBI-GenBank

Chimgin from Ferula haussknechtii as AChE inhibitor and confirmation of the absolute configuration

Alzheimer’s disease (AD) is the most common cause of dementia worldwide and is classified as a neurodegenerative disorder. From a drug design perspective, natural products (NPs) are more drug-like and are highly compatible with biological systems compared to most synthetic libraries. NPs provide a more efficient and cost-effective approach to new drug discovery. However, the complexity of NPs makes their identification a challenging task. Chimgin, a bicyclic monoterpene with three chiral centers, exhibits a wide range of biological activity. Despite this, the exact structure of chimgin has remained unclear until now. In this study, we quantified the amount of chimgin in Ferula haussknechtii using analytical Reversed-phase high-pressure liquid chromatography equipped with photodiode array detector (RP-HPLC-PDA). Furthermore, we determined the absolute configuration of chimgin through electronic circular dichroism (ECD) spectroscopy and time-dependent density functional theory (TDDFT) calculations. Finally, we evaluated its inhibitory effect on AChE through in vitro and in silico studies. The extraction process yielded an output of 2.82 ± 0.10% with an exact amount of 0.62 ± 0.04 mg of chimgin per 100 g of plant. Based on the results of ECD and TDDFT calculation, the absolute configuration of chimgin was determined to be 1S, 2S, 4S. Chimgin exhibited an inhibitory effect on AChE with an IC50 of 37.43 µM and its mechanism of action was found to be competitive. Highlights Chimgin was isolated from the roots of Ferula haussknechtii. The amount of chimgin in the plant was determined by RP-HPLC-PDA. Its absolute configuration of chimgin was determined using ECD. In vitro acetylcholinesterase activity of the chimgin was evaluated. The docking and molecular dynamic simulation of chimgin was done. Communicated by Ramaswamy H. Sarma

Research on the targeted improvement of the yield of a new VB12-producing strain, Ensifer adhaerens S305, based on genomic and transcriptomic analysis

BackgroundVitamin B12 (VB12) has a wide range of applications and high economic value. In this study, a new strain with high VB12 production potential, Ensifer adhaerens S305, was identified in sewage. Because E. adhaerens strains have become the main strains for VB12 production via fermentation in recent years, the directional modification of the S305 strain to obtain a strain suitable for the industrial production of VB12 has great potential and commercial value.Results16S rRNA and genome-wide phylogenetic tree analysis combined with average nucleotide identity (ANI) analysis showed that the high-yielding VB12 strain was a E. adhaerens strain and that its VB12 synthesis pathway genes were highly similar to related genes of strains of this and other species, including E. adhaerens Casida A, Pseudomonas denitrificans SC 510, and E. adhaerens Corn53. High-pressure liquid chromatography (HPLC) results indicated that the VB12 yields of the S305 strain were more than double those of the Casida A strain under different medium components. Multiple genes with significantly upregulated and downregulated transcription were identified by comparing the transcription intensity of different genes through transcriptome sequencing. KEGG enrichment analysis of the porphyrin metabolism pathway identified 9 significantly upregulated and downregulated differentially expressed genes (DEGs) in the VB12 synthesis pathway, including 7 transcriptionally upregulated genes (cobA, cobT, hemA, cobJ, cobN, cobR, and cobP) that were episomally overexpressed in the Casida A strain. The results showed that the VB12 yield of the overexpressed strain was higher than that of the wild-type strain. Notably, the strains overexpressing the cobA and cobT genes exhibited the most significant increases in VB12 yield, i.e., 31.4% and 24.8%, respectively. The VB12 yield of the S305 strain in shake-flask culture was improved from 176.6 ± 8.21 mg/L to 245.6 ± 4.36 mg/L by integrating the cobA and cobT genes into the strain.ConclusionPhylogenetic tree and ANI analysis showed that the Ensifer and Sinorhizobium strains were quite different at the genome level; the overexpression and integrated expression of significantly upregulated genes in the VB12 synthesis pathway could increase the yield of VB12, further improving the VB12 yield of the E. adhaerens S305 strain.

Advances and challenges in conventional and modern techniques for halal food authentication: A review.

Food is one of the most necessary needs since human civilization. For Muslims, it is mandatory to consume halal food. From a halal authentication perspective, adulteration of food products is an emerging challenge worldwide. The demand for halal food consumption has resulted in an ever-increasing need for halal product validity. In the market, there are several food products in which actual ingredients and their source are not mentioned on the label and cannot be observed by the naked eye. Commonly nonhalal items include pig derivatives like lard, pork, and gelatin derivatives, dead meats, alcohol, blood, and prohibited animals. Purposely, various conventional and modern methods offer precise approaches to ensure the halalness and wholesomeness of food products. Conventional methods are physiochemical (dielectric) and electrophoresis. At the same time, modern techniques include high-pressure liquid chromatography (HPLC), gas chromatography (GC), electronic nose (E-Nose), polymerase chain reaction (PCR), enzyme-linked immunosorbent assay (ELISA), differential scanning calorimetry (DSC), nuclear magnetic resonance (NMR), near-infrared (NIR) spectroscopy, and Fourier transform infrared (FTIR) spectroscopy. This review intends to give an extensive and updated overview of conventional and modern analytical methods for ensuring food halal authenticity.

Simultaneous Lipid and Carotenoid Production via Rhodotorula paludigena CM33 Using Crude Glycerol as the Main Substrate: Pilot-Scale Experiments.

Rhodotorula paludigena CM33 is an oleaginous yeast that has been demonstrated to accumulate substantial quantities of intracellular lipids and carotenoids. In this study, crude glycerol, a by-product of biodiesel production, was used as a carbon source to enhance the accumulation of lipids and carotenoids in the cells. The culture conditions were first optimized using response surface methodology, which revealed that the carotenoid concentration and lipid content improved when the concentration of crude glycerol was 40 g/L. Different fermentation conditions were also investigated: batch, repeated-batch, and fed-batch conditions in a 500 L fermenter. For fed-batch fermentation, the maximum concentrations of biomass, lipids, and carotenoids obtained were 46.32 g/L, 37.65%, and 713.80 mg/L, respectively. A chemical-free carotenoid extraction method was also optimized using high-pressure homogenization and a microfluidizer device. The carotenoids were found to be mostly beta-carotene, which was confirmed by HPLC (high pressure liquid chromatography), LC-MS (liquid chromatography-mass spectrometry), and NMR (nuclear magnetic resonance). The results of this study indicate that crude glycerol can be used as a substrate to produce carotenoids, resulting in enhanced value of this biodiesel by-product.

Analytical and chemometric strategies for elucidation of yerba mate composition.

Yerba mate, a popular plant consumed mainly as an infusion, possesses nutritional and medicinal properties attributed to its secondary metabolites. This study aimed to develop strategies to elucidate the phenolic composition of yerba mate samples from Brazil, Argentina, Uruguay, and Paraguay. Optimization of ultrasonic-assisted extraction (UAE) was performed, and the extracted compounds were characterized using ultra-high-pressure liquid chromatography coupled with quadrupole/time-of-flight mass spectrometry (UHPLC-QTOF-MS), molecular fluorescence and high-pressure liquid chromatography with diode-array detection (HPLC-DAD). Chemometric analysis, including parallel factor analysis (PARAFAC) and principal component analysis (PCA) explored metabolite profiles and identify patterns. PARAFAC modelling of the molecular fluorescence results revealed higher pigment content in Brazilian samples, while other countries' samples exhibited higher phenolic content. PCA modeling of HPLC-DAD results indicated that cultivated yerba mate contained higher chlorogenic acids levels, and samples from Argentina, Paraguay, and Uruguay exhibited higher concentrations of chlorogenic acids and flavonoids.

Determination of isohydrafural and methyluracil in ear drops by high performance liquid chromatography

Abstract Objective The aim of current study was the development and validation of a reversed-phase high-pressure liquid chromatography (RPHPLC) method for the quantitative determination of two active ingredients, isohydrafural (IHF) and methyluracil (MU) in fixed dose combination ear drops. Methods: An efficient separation of the two compounds was achieved on a Teknokroma C1 5μm (150*4,6mm) column, with a methanol: water 60:40 (V: V) mobile phase, at 300C temperature and 0.6 mL/min flow-rate. The total analysis time was 5.5 min. Result: The verified validation parameters were: linearity, selectivity, specificity, precision (repeatability and reproducibility), robustness, limit of detection and quantification. Conclusions: Good separation and lack of interference from other chromatographic peaks, rapidanalysis times were obtained. The newly developed chromatographic method can be applied for the analysis of fixed dose combination ear drops with isohydrafural and methyluracil.

Supercritical fluid chromatography is not (only) normal-phase chromatography

Supercritical fluid chromatography (SFC), now using carbon dioxide as a major component of the mobile phase, has been known for over 60 years but still some misunderstandings remain about its capabilities. Amongst them, SFC is often described as a normal-phase chromatographic technique, based on different considerations: polarity of the stationary phase, elution order of the analytes, relative non-polarity of the mobile phase, non-linear retention behaviour, or adsorption retention mechanisms. All of these assumptions are true to a certain extent, and in certain circumstances. But also, all of these assumptions are wrong in different circumstances. In this paper, the criteria to categorize SFC as a normal-phase chromatographic method will be examined individually, considering all knowledge acquired from the early years of its development. Finally, it will appear that the “normal-phase” glass lens is greatly reducing the true extent of SFC's possibilities.

Identification of bioactive compounds and inhibitory effects of TNF-α and COX-2 in the extract from cultured three-spot seahorse (H. trimaculatus).

Three-spot seahorse (Hippocampus trimaculatus) has been consumed as traditional Chinese medicine in Asian society. This study was designed to analyze the bioactive compounds of the solvent extracts from cultured three-spot seahorse by high pressure liquid chromatography coupled with electrospray ionization tandem mass spectrometry (HPLC-ESI/MS/MS). Subsequently, their biological activities were evaluated and confirmed by cell modes and Western blot analysis. Experimental results indicated that taurine and arginine were the primary bioactive compounds identified and quantified without pre- or post-column derivatization within 20 min retention time. The analytical method was established and validated with intraday/interday RSD from 0.25% to 3.34% and with recovery from 87.8% to 91.2%. As compared to other extracts, water layer extract (WLE) contained the most taurine and arginine contents of 6.807 and 0.437 mg/g (dry basis), respectively. In the meanwhile, WLE also showed anti-inflammatory activity on LPS-induced NO production and inhibited the protein expression of TNF-α and COX-2 by Western blot analysis with better cell viability.

Asymmetric Cu(I)-Catalyzed Conjugate Borylation of α,β-Unsaturated Acyl Silanes

AbstractWe report the development of a Cu(I)-catalyzed conjugate borylation of α,β-unsaturated acyl silanes using bis(pinacoloto)diboron. Racemic borylations of β-aryl- and β-alkyl-substituted silyl enones were achieved using ligand-free conditions to access β-borylated acyl silanes in up to 97% yield. Josiphos enabled the synthesis of enantioenriched boronic esters in up to 58% yield and 94% ee. The racemic reaction was demonstrated on 5.0 mmol scale and isolation of the boronic esters was achieved using simple filtration and normal-phase chromatography. This work supplements known methods to access boronic esters from electron-deficient olefins using Cu(I) catalysis.

The Impact of Different Withering Process Conditions on the Bioactivity and Quality of Black Tea from Azorean Camellia sinensis

The objective of this study was to investigate the variability of natural bioactive compounds, such as catechin, theaflavin, total phenolic content (TPC), and total flavonoid content (TFC), of Azorean black tea (Camellia sinensis L., O. Kuntze) as well as its antioxidant activities according to different withering times. The TPC, TFC, free radical scavenging activity (FRSA), ferric reducing antioxidant power (FRAP), and ferrous-ion-chelating (FIC) activities were determined by colorimetric methods, and catechin and theaflavin contents were analyzed by high-pressure liquid chromatography (HPLC). The FRSA shows similar results for the withering range of 6 to 16 h (hours). For FRAP, the best results were observed at 16 h, and for FIC, the highest value was at 20 h. The TPC and TFC showed the highest value at 9 h and the lowest at 20 h. For the total theaflavins, the highest results were obtained after 12 h of withering, and the lowest values were obtained at 16 and 20 h. According to the different withering times, the highest value of total catechin levels was at 12 h, while the lowest value was observed at 20 h. Regarding caffeine content, all samples presented similar results, with the exception of the 12 h time point. In conclusion, the best withering times were observed in the range of 9 to 16 h, showing decreased values at 20 h, with the exception of FIC.

Emotional states predict cellular immune system activity under conditions of life as it is lived: A multivariate time-series analysis approach.

The relationship between emotional states and immune system activity is characterized by bidirectional influences; however, limited information is available regarding the temporal dynamics of these effects. The goal of this investigation was to examine how these psychoimmunological interdependencies unfold over time under conditions of "life as it is lived". For this purpose, three healthy women collected their entire urine over a period of approximately two months at 12-h intervals (8 am-8 pm, 8 pm-8 am), resulting in a total of 112 to 126 consecutive measurements per subject. In addition, among other regular psychological assessments, the subjects completed the EWL-60-S, an emotional state questionnaire, each morning and evening. To assess the extent of T-helper type 1 immune activation, the neopterin per creatinine concentration was measured in the urine samples using high-pressure liquid chromatography. The dynamic relationships between the time series of the six emotional states (performance-related activity, general inactivity, extraversion/introversion, general feeling of comfort, emotional irritation, anxiety/depressiveness) and urinary neopterin levels were estimated in vector-autoregressive models and evaluated using Granger-causality tests, impulse-response functions and forecast error variance decompositions. The findings showed that emotional states explained up to 20% of the variance of urinary neopterin per creatinine levels, whereby most of the effects occurred within a period of approximately three days. Across all subjects, increases in anxiety/depressiveness and extraversion led to increases in neopterin levels, while a general feeling of comfort led to decreases in neopterin. These results emphasize the importance of the interdependencies between emotional states and immune system activity and showcase the potential that intensive longitudinal study designs offer for psychoneuroimmunology.