Citric Acid Levels Research Articles

Antioxidant Effect of a Plant-Derived Extracellular Vesicles' Mix on Human Skin Fibroblasts: Induction of a Reparative Process.

Plant-Derived Extracellular Vesicles extracellular vesicles (PDEVs) from organic agriculture (without the use of pesticides and microbicides) contain high levels of antioxidants. Organic PDEVs have shown an increased antioxidant power compared to PDEVs from single plants, suggesting a synergistic effect of the bioactives constitutively expressed in the PDEVs from single fruits. With this study, we wanted to investigate the beneficial effects of a mix of PDEVs on human skin cells. We found detectable levels of citric acid, ascorbic acid, glutathione, catalase, and SOD in a mix of PDEVs deriving from five different fruits (grape, red orange, papaya, pomegranate, and tangerine). We then treated H2O2-conditioned fibroblasts with the mix of PDEVs. The results showed that the PDEVs' mixture reverted the H2O2-induced redox imbalance, restoring mitochondrial homeostasis, with a strong reduction of mitochondrial anion superoxide and an increase in sirtuin levels. The antioxidant action was consistent with wound repair on a lesion produced in a fibroblast's monolayer. This result was consistent with an increased level of vimentin and matrix metalloproteinase-9, whose expression is directly related to the efficiency of the reparative processes. These data support a beneficial role of PDEVs in both preventing and treating skin injuries through their potent antioxidant and reparative activities.

Distinct Metabolites in Atherosclerosis Based on Metabolomics: A Systematic Review and Meta-analysis Primarily in Chinese Population

AimsAtherosclerosis is a life-threatening disease that develops when a plaque builds up inside an artery and progresses silently. Identifying the early pathological changes and the biomarkers of atherosclerosis deserves attention. We aimed to systematically study and integrate the various metabolites of atherosclerosis in the level of disease to provide more evidences to support early prevention and treatment of atherosclerosis. Data synthesisThe protocol was registered with PROPSERO (CRD42023441845). We searched 14985 records via EMBASE, PubMed, Web of Science, WanFang data, VIP data, and CNKI databases. The collected metabolites were for qualitative and quantitative meta-analysis. The I2 statistic estimated heterogeneity, with over 50% considered to adopt the random-effects model. A total of 49 articles were included in the meta-analysis. We finally integrated 83 and 16 metabolites presented more than two times in inclusion studies, respectively in blood (plasma and serum) and urine. Among them, the level of citric acid (SMD=-10.35 [95%CI -15.03, -5.67], p<0.001), lactic acid (SMD= 6.32 [95%CI 0.12, 12.52], p<0.001) and TMAO (SMD= 1.40 [95%CI 0.27, 2.53], p<0.001) had significant differences between atherosclerosis and controls. And we observed blood stasis syndrome of atherosclerosis patients present arterial ischemia and energy disorder obviously. ConclusionsThe study provides an in-depth understanding of the roles of metabolites on atherosclerosis progression and prediction primarily in Chinese population, which contributing to development of prevention and therapeutic potential in the future.

Nontargeted Metabolomics to Understand the Impact of Modified Atmospheric Packaging on Metabolite Profiles of Cooked Normal-pH and Atypical Dark-Cutting Beef.

Background: Limited knowledge is currently available on the effects of modified atmospheric packaging (MAP) on the metabolite profiles of cooked beef. The objective was to evaluate the impact of packaging on the cooked color and cooked metabolite profile of normal-pH (normal bright-red color) and atypical-dark-cutting beef (inherently slightly dark-colored) longissimus lumborum muscle. Methods: Normal-pH (pH 5.56) and atypical dark-cutting (pH 5.63) loins (n = 6) were procured from a commercial meat processor. Steaks were randomly assigned to one of three different packaging methods: vacuum packaging, carbon monoxide (CO-MAP), and high oxygen (HiOx-MAP). Following 5 d of retail display, steaks were cooked to 71 °C on a clamshell-style grill, and samples were collected for untargeted metabolites using gas-chromatography mass spectrometry. Results: Raw atypical dark-cutting steaks were less red (p < 0.05) than raw normal-pH steaks. However, there were no differences in internal cooked color between normal-pH and atypical dark-cutting steaks. Steaks packaged in HiOx-MAP steaks had a lower (p < 0.05) cooked redness than vacuum and CO-MAP steaks. A total of 129 metabolite features were identified in the study. Serine and tryptophan were over-abundant in cooked atypical dark-cutting beef compared to raw atypical samples. Citric acid levels were greater in HiOx-MAP packaged beef compared with VP both in normal and atypical dark-cutting beef after cooking, while no differentially abundant metabolites were shared between vacuum and CO-MAP steaks after cooking. Discussion: A slight increase in pH did not influence metabolite profiles in different packaging. However, there were packaging effects within normal and atypical dark-cutting beef. Conclusions: This study suggests that packaging conditions change metabolite profiles, which can influence cooked metabolites. Therefore, the metabolomics approach can be used to better understand cooked color defects such as premature browning.

Low soil pH enhances fruit acidity by inhibiting citric acid degradation in lemon (Citrus lemon L.)

Fruit acidity significantly influences fruit flavor, but the specific impact of soil pH on fruit acidity remains unclear. This study investigated the effects of various soil pH levels on fruit acidity and citric acid (CA) metabolism in lemon (Citrus limon L.). High soil pH (pH 8) decreased total soluble solids concentrations in lemon fruits, while low soil pH (pH 4) increased titratable acid and CA concentrations. Although low soil pH reduced the synthesis of CA due to the decreased citrate synthase and phosphoenolpyruvate carboxylase activities, the elevated fruit acidity under low soil pH conditions is not directly related to CA synthesis. Instead, low soil pH was found to suppress the activity of cytosolic aconitase (Cyt-ACO), an iron-dependent enzyme, indicating a potential role for CA degradation inhibition in low soil pH-induced CA accumulation. Furthermore, low soil pH significantly reduced cytosolic iron (Cyt-Fe) concentration, which was positively correlated with Cyt-ACO activity. In conclusion, low soil pH contributes to increasing fruit acidity in lemon, partially by inhibiting CA degradation due to the reduced Cyt-Fe concentrations. Our work unravels the influence of soil pH on CA accumulation and provides important clues for modulating CA levels through microelement fertilization in citrus.

Influence of Combined Supplemental Lighting and Nutrient Solution Concentration on Fruit Production and Quality of Cherry Tomato

We conducted an analysis on the combined effects of two light conditions (L1: greenhouse natural lighting; L2: greenhouse natural lighting plus supplemental lighting (SL)) and three nutrient solution concentrations (EC, NS1: 3.2 dS/m; NS2: 3.7 dS/m; NS3: 4.2 dS/m) on the growth, fruit production, and quality of two cherry tomato cultivars with different fruit coloring (‘Baiyu’ and ‘Qianxi’). The plants subjected to NS2 exhibited enhanced growth, photosynthetic parameters, and fruit production. The utilization of SL further enhanced stem diameter, leaf number, and single fruit weight, resulting in higher fruit weight per plant in ‘Baiyu’, which was not observed in ‘Qianxi’. The growth, fruit size, and fruit weight of both cultivars cultivated under NS3 conditions were suppressed, while these fruits exhibited elevated levels of total soluble solids (TSS), soluble sugars, vitamin C, polyphenols, fructose, glucose, sucrose, citric acid, and carotenoids. These levels were further enhanced by SL treatment. The improvement of fruit quality through the application of SL was found to be cultivar and EC dependent. In ‘Baiyu’, SL at NS1 significantly enhanced the accumulation of fruit water, minerals (N, P, K, Ca, and Mg), TSS, vitamin C, fructose, sucrose, and carotenoids. However, this effect was not observed in ‘Qianxi’. The combination of SL and EC 4.2 dS/m (NS3) generally contributes to the enhancement of fruit quality, while SL and EC 3.7 dS/m can ensure consistent fruit production. The yellowish-white fruit cultivar exhibited higher levels of soluble sugars, vitamin C, and polyphenols under L2NS3 conditions compared to the red fruit cultivar, whereas the carotenoid content showed an opposite trend. The findings are anticipated to establish a theoretical foundation for the consistent annual cultivation of cherry tomatoes in protected horticultural settings.

Engineering the Rhizosphere Microbiome with Plant Growth Promoting Bacteria for Modulation of the Plant Metabolome.

Plant-growth-promoting bacteria (PGPB) have beneficial effects on plants. They can promote growth and enhance plant defense against abiotic stress and disease, and these effects are associated with changes in the plant metabolite profile. The research problem addressed in this study was the impact of inoculation with PGPB on the metabolite profile of Salicornia europaea L. across controlled and field conditions. Salicornia europaea seeds, inoculated with Brevibacterium casei EB3 and Pseudomonas oryzihabitans RL18, were grown in controlled laboratory experiments and in a natural field setting. The metabolite composition of the aboveground tissues was analyzed using GC-MS and UHPLC-MS. PGPB inoculation promoted a reconfiguration in plant metabolism in both environments. Under controlled laboratory conditions, inoculation contributed to increased biomass production and the reinforcement of immune responses by significantly increasing the levels of unsaturated fatty acids, sugars, citric acid, acetic acid, chlorogenic acids, and quercetin. In field conditions, the inoculated plants exhibited a distinct phytochemical profile, with increased glucose, fructose, and phenolic compounds, especially hydroxybenzoic acid, quercetin, and apigenin, alongside decreased unsaturated fatty acids, suggesting higher stress levels. The metabolic response shifted from growth enhancement to stress resistance in the latter context. As a common pattern to both laboratory and field conditions, biopriming induced metabolic reprogramming towards the expression of apigenin, quercetin, formononetin, caffeic acid, and caffeoylquinic acid, metabolites that enhance the plant's tolerance to abiotic and biotic stress. This study unveils the intricate metabolic adaptations of Salicornia europaea under controlled and field conditions, highlighting PGPB's potential to redesign the metabolite profile of the plant. Elevated-stress-related metabolites may fortify plant defense mechanisms, laying the groundwork for stress-resistant crop development through PGPB-based inoculants, especially in saline agriculture.

Isocitrate lyase promotes Puccinia striiformis f. sp. tritici susceptibility in wheat (Triticum aestivum) by suppressing accumulation of glyoxylate cycle intermediates.

Plant fungal parasites manipulate host metabolism to support their own survival. Among the many central metabolic pathways altered during infection, the glyoxylate cycle is frequently upregulated in both fungi and their host plants. Here, we examined the response of the glyoxylate cycle in bread wheat (Triticum aestivum) to infection by the obligate biotrophic fungal pathogen Puccinia striiformis f. sp. tritici (Pst). Gene expression analysis revealed that wheat genes encoding the two unique enzymes of the glyoxylate cycle, isocitrate lyase (TaICL) and malate synthase, diverged in their expression between susceptible and resistant Pst interactions. Focusing on TaICL, we determined that the TaICL B homoeolog is specifically upregulated during early stages of a successful Pst infection. Furthermore, disruption of the B homoeolog alone was sufficient to significantly perturb Pst disease progression. Indeed, Pst infection of the TaICL-B disruption mutant (TaICL-BY400*) was inhibited early during initial penetration, with the TaICL-BY400* line also accumulating high levels of malic acid, citric acid, and aconitic acid. Exogenous application of malic acid or aconitic acid also suppressed Pst infection, with trans-aconitic acid treatment having the most pronounced effect by decreasing fungal biomass 15-fold. Thus, enhanced TaICL-B expression during Pst infection may lower accumulation of malic acid and aconitic acid to promote Pst proliferation. As exogenous application of aconitic acid and malic acid has previously been shown to inhibit other critical pests and pathogens, we propose TaICL as a potential target for disruption in resistance breeding that could have wide-reaching protective benefits for wheat and beyond.

Exploring the impact of fermentation on bioactive compounds in two different types of carrot pomace

This study assessed the fermentation process of black and orange-colored carrot pomaces utilizing three varieties of lactic acid bacteria: Lactobacillus plantarum, Lactobacillus casei, and Lactobacillus acidophilus. The fermentation process influenced the titratable acidity and lactic acid production, with L. plantarum resulting in the highest increases, particularly noticeable in orange-colored carrot pomace (OCP). Initially, citric acid levels increased, while ascorbic acid levels decreased in all cultures. Black carrot pomace (BCP) fermented by L. casei exhibited the highest phenolic content (611.42 mg GAE/100 g dw) and antioxidant activity on day 3. The total phenolic content of OCP was the highest on day 3 with both L. casei and L. plantarum. The phenolic acid profiles varied by carrot type, with BCP being rich in neochlorogenic and chlorogenic acids, and OCP in gallic and p-coumaric acids. High anthocyanin content was found in OCP, especially cyanidin derivatives, peaking with L. casei. Similarly, OCP showed increased levels of both α-carotene and β-carotene on day 3 with L. casei. Carotenoid analysis revealed that BCP contained only α-carotene, with the highest level observed after 4 days of fermentation with L. plantarum. This study demonstrates that selective fermentation with lactic acid bacteria can alter the phytochemical profile of carrot pomaces, potentially enhancing the nutritional quality in various food applications.

Environmental analysis of nitrobenzene using newly synthesized anisotropic gold nanostructures: Reaction kinetics and electrocatalytic activity

In this article, we established to produce anisotropic, colloidally stable gold nanostructures (AuNS) by rapidly mixing aqueous solutions of gold(III) chloride trihydrate (HAuCl4) and citric acid (CA) as dual reducing and capping agents at room temperature without the aid of a templating agent. Furthermore, the impact of various anions and shape-directing agents on the morphology and reaction kinetics of citric acid-capped gold nanostructures (CA-AuNS) was investigated. Additionally, the electrocatalytic activity of AuNS towards nitrobenzene (NB) was examined using a CA-AuNS modified glassy carbon (GC) electrode. The rate of reaction for the formation of CA-AuNS increased rapidly with higher concentrations of CA, indicating a strong dependence on the reductant’s concentration. This process followed first-order kinetics, suggesting that the reaction rate is directly proportional to the citric acid concentration. Consequently, optimizing CA levels can effectively control the synthesis rate and yield of CA-AuNS. The presence of nitrite ions led to rapid aggregation of gold nanostructures (AuNS) and a quick reaction time. Chloride ions increased the size of the AuNS. As bromide ion concentration increased, the reaction time slowed significantly, resulting in spherical gold nanoparticles. Both acetate ions and polyvinylpyrrolidone (PVP) inhibited the growth of AuNS. High-resolution transmission electron microscopy (HR-TEM) images showed the formation of various shapes, including spherical, hexagonal, pentagonal, and boat-like structures. In the presence of cetyltrimethylammonium bromide (CTAB), both complex formation between CTAB and HAuCl4 and the formation of spherical gold nanoparticles occurred. Upon addition of silver nitrate (AgNO3) to the colloidal solution, the color shifted to pale violet, and the morphology transitioned from anisotropic to irregular. However, when ascorbic acid (AA) was utilized, it proved ideal for maintaining the morphology of gold nanostructures (AuNS). The analysis of X-ray diffraction spectroscopy (XRD), cyclic voltammetry (CV), electrochemical impedance spectroscopy (EIS), and Fourier transform infrared spectroscopy (FT-IR) further confirmed the synthesized CA-AuNS. The FT-IR spectra of CA before and after the formation of AuNS indicated the participation of carboxylic acid (–COOH) and hydroxyl (–OH) groups in the reduction process of Au3+ metal ions. The GC/CA-AuNS electrode exhibits a larger electroactive surface area due to the coating of AuNS onto the electrode surface, enhancing the electrode’s electrical conductivity by providing additional active sites for electron transfer. In addition, the electrocatalytic activity of NB was investigated in this study. The GC/CA-AuNS electrode had better electrocatalytic activity than the unmodified GC electrode. As a result, with a LOD of 28.2 nM (S/N = 3), this electrode was chosen for sensitive NB determination.

The association of circulating bioenergetic metabolites with healthy human aging

Aging is an inevitable and gradual decline in several biological functions. Mitochondrial dysfunction is one of the most important hallmarks of aging. In this context, alterations in metabolites associated with mitochondrial dysfunction may serve as a significant biomarker. This study aimed to investigate the existence of a relationship between the key metabolites involved in bioenergetics metabolism and aging. 53 volunteers ranged 20–85 years participated in the study. We tested the association between different tricarboxylic acid (TCA) cycle metabolites, fatty acid metabolism, and amino acid metabolism with age, sex, body composition, and proxy markers of aging such as walking speed, grip strength and chair test. We found that lactic acid negatively correlated with age while several fatty acid metabolites, such as azelaic, sebacic, and linoleic acids, showed positive correlations with age (p < 0.05). Sex-specific trends, such as glycerol, and dodecanoic acid, were also observed for certain metabolites. Furthermore, citric acid levels were found to have a significant association with physical function and body composition measures. Participants with higher citric acid levels displayed improved performance in physical tests and favorable body composition indices. Additionally, fumaric acid and adipic acid showed positive correlations with fat-free body mass, while sebacic acid was negatively associated with measures of fat mass. These findings underscore the importance of understanding the role of circulating bioenergetics metabolites with age, sex variations, and their potential implications in body composition and physical performance.

An exploratory metabolomic study reveals the Dipsacus asper-Achyranthes bidentate herb pair against osteoarthritis by modulating imbalance in polyunsaturated fatty acids and energy metabolism

Osteoarthritis (OA) is a degenerative joint disease primarily affecting the cartilage. The therapeutic potential of the Dipsacus asper-Achyranthes bidentate herb pair for OA has been acknowledged, yet its precise mechanism remains elusive. In this study, we conducted a comprehensive analysis of metabolomic changes and therapeutic outcomes in osteoarthritic rats, employing a gas chromatography-mass spectrometry-based metabolomics approach in conjunction with histopathological and biochemical assessments. The rats were divided into six groups: control, model, positive control, Dipsacus asper treated, Achyranthes bidentata treated, and herb pair treated groups. Compared to the model group, significant reductions in levels of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), interleukin-6 (IL-6), and iNOS were observed in the treated groups. Multivariate statistical analyses were employed to investigate metabolite profile changes in serum samples and identify potential biomarkers, revealing 45 differential biomarkers, with eighteen validated using standard substances. These analytes exhibited excellent linearity across a wide concentration range (R2>0.9990), with intra- and inter-day precision RSD values below 4.69% and 4.83%, respectively. Recoveries of the eighteen analytes ranged from 93.97% to 106.59%, with RSD values under 5.72%, underscoring the method's reliability. Treatment with the herbal pair effectively restored levels of unsaturated fatty acids such as linoleic acid and arachidonic acid, along with glucogenic amino acids. Additionally, levels of phosphoric acid and citric acid were reversed, indicating restoration of energy metabolism. Collectively, these findings highlight the utility of metabolomic analysis in evaluating therapeutic efficacy and elucidating the underlying molecular mechanisms of herb pairs in OA treatment.

Formulation of Effervescent Tablet from Kelulut Bee (Trigona sp.) Pollen from East Kalimantan

Since the beginning of human history, bee honey has been used by humans. However, there are also other bee products such as bee pollen (BP), royal jelly, propolis, and beeswax. Previous research has identified that pollen contains amino acids, lipids, flavonoids, micronutrients and many fat-soluble vitamins such as vitamins A, E and D, as well as water-soluble vitamins such as vitamins B1, B2, B6 and C. The objective of this study is to formulate bee pollen as effervescent tablet. The method in this study used alkaline granulation which requires an oven process for 24 hours and the use of a combination of sodium bicarbonate with citric acid. Three formulas were optimized in this study. The stability test results showed that formula number 3 has an average moisture content of 0.5696±0.6154%, and average flow rate of 0.67±0.02054 g/sec, the average compressibility in formula number 1 is 0.046±0.0452%. Overall, it is also known that formula number 2 has the smallest dissolution time compared to other formulations. The effect of the combination of citric acid and sodium bicarbonate used in the formulation of effervescent tablets from bee pollen on the physical quality of the tablets, namely friability, hardness, and dissolving time, the higher levels of citric acid and sodium bicarbonate used do not have a significant effect on friability, hardness, and time dissolving effervescent tablets from bee pollen from kelulut bees (Trigona sp).

Colonization of root endophytic fungus Serendipita indica improves drought tolerance of Pinus taeda seedlings by regulating metabolome and proteome.

Pinus taeda is an important forest tree species for plantations because of its rapid growth and high yield of oleoresins. Although P. taeda plantations distribute in warm and wet southern China, drought, sometime serious and long time, often occurs in the region. To explore drought tolerance of P. taeda and usage of beneficial microorganisms, P. taeda seedlings were planted in pots and were inoculated with root endophytic fungus Serendipita indica and finally were treated with drought stress for 53 d. Metabolome and proteome of their needles were analyzed. The results showed that S. indica inoculation of P. taeda seedlings under drought stress caused great changes in levels of some metabolites in their needles, especially some flavonoids and organic acids. Among them, the levels of eriocitrin, trans-aconitic acid, vitamin C, uric acid, alpha-ketoglutaric acid, vitamin A, stachydrine, coumalic acid, itaconic acid, calceolarioside B, 2-oxoglutaric acid, and citric acid were upregulated more than three times in inoculated seedlings under drought stress, compared to those of non-inoculated seedlings under drought stress. KEGG analysis showed that some pathways were enriched in inoculated seedlings under drought stress, such as flavonoid biosynthesis, ascorbate and aldarate metabolism, C5-branched dibasic acid metabolism. Proteome analysis revealed some specific differential proteins. Two proteins, namely, H9X056 and H9VDW5, only appeared in the needles of inoculated seedlings under drought stress. The protein H9VNE7 was upregulated more than 11.0 times as that of non-inoculated seedlings under drought stress. In addition, S. indica inoculation increased enrichment of water deficient-inducible proteins (such as LP3-1, LP3-2, LP3-3, and dehydrins) and those involved in ribosomal structures (such as A0A385JF23). Meanwhile, under drought stress, the inoculation caused great changes in biosynthesis and metabolism pathways, mainly including phenylpropanoid biosynthesis, cutin, suberine and wax biosynthesis, and 2-oxocarboxylic acid metabolism. In addition, there were positive relationships between accumulation of some metabolites and enrichment of proteins in P. taeda under drought stress. Altogether, our results showed great changes in metabolome and proteome in inoculated seedlings under drought stress and provided a guideline to further study functions of metabolites and proteins, especially those related to drought stress.

Correlation between seminal plasma biochemical markers and semen parameters in idiopathic oligoasthenoteratospermia: identification of biomarkers for L-carnitine therapy.

L-carnitine therapy for idiopathic sperm abnormalities exhibits variable effectiveness, and currently, there are no established criteria to predict patient response. This study investigated correlations between seminal plasma markers and semen parameters to identify biomarkers that can guide indications for L-carnitine therapy indications in patients with idiopathic sperm abnormalities. A retrospective review was conducted on 223 male patients with idiopathic oligoasthenoteratospermia, who sought medical attention at our clinic between January 2020 and October 2022. These patients underwent a pretreatment seminal plasma biochemical analysis, followed by a three-month continuous L-carnitine treatment. The correlation between seminal plasma biochemical parameters and pretreatment semen parameters was analyzed. Semen quality was compared between cases with normal and abnormal seminal plasma biochemical parameters, both pretreatment and posttreatment. The correlation between the changes in semen parameters after treatment and seminal plasma biochemical parameters were investigated. Correlation analyses revealed significant associations between all pretreatment semen parameters and seminal plasma biochemical markers, except for liquefying time and the ratio of normal morphology. Subgroup analysis, stratified by seminal fructose, zinc, citric acid, and neutral glycosidase levels, demonstrated that abnormal groups exhibited significantly different levels of semen parameters compared with the normal groups. The changing difference and changing ratio in the ratio of forward motile sperm showed a negative correlation with seminal fructose levels (r=-0.165 and -0.144). The changing difference in semen volume was negatively correlated with the level of seminal neutral glycosidase (r=-0.158). The changing ratio in semen volume, sperm concentration, total sperm count, and count of forward motile sperm all exhibited negative correlations with the levels of seminal neutral glycosidase (range from -0.178 to -0.224). Seminal plasma biochemical markers, particularly fructose and neutral glycosidase, may serve as valuable indicators for determining the eligibility of patients with idiopathic sperm abnormalities for L-carnitine therapy.

Effects of juicing methods on the bioactive compounds and flavor quality of 'Black-seed' pomegranate from three producing areas.

Color, nutrients and flavor are the key characteristics of pomegranate juice, but they are susceptible to processing methods and raw materials. In this study, the effects of aril juicing and whole fruit juicing methods on the composition of 'Black-seed' pomegranate juice from three producing areas were studied, including physicochemical parameters, color attributes, organic acids, sugars, phenolic compounds, and volatile compounds. The whole fruit juicing method resulted in higher juice yields of pomegranate fruit with 69.01-72.59%, hue angle values were 5.95-6.45°, and the juice remained red. The highest level of citric acid (21.21 g L-1 ), total acids (24.78 g L-1 ), and total anthocyanin content (435.59 mg L-1 ) were found in whole fruit juice, and seven tannins were detected. The most abundant volatile compounds were (Z)-3-hexen-1-ol and 1-hexanol in all juice samples, with alcohol content increased and aldehydes content decreased by whole fruit juicing. Principal component analysis revealed that the 24 indexes (variable important in projection >1) clearly distinguished juice samples obtained by two juicing methods, with ellagic acid hexoside, (E)-2-heptenal, (+)-catechin, and octanoic acid having the best discriminatory potential. Overall, the effects of juicing method on 'Black-seed' pomegranate juice were greater than those of raw-material-producing areas. These results confirmed the potential for using the whole 'Black-seed' pomegranate for processing, and also provided a theoretical basis for the healthy product development and utilization of dark-color pomegranate varieties. © 2023 Society of Chemical Industry.

Determination of sugars and organic acids in diverse carob genotypes using HPLC techniques

Carob fruit is widely known for their abundance of health-boosting compounds like polyphenols, ascorbic acid, organic acids, and micronutrients. These compounds offer various benefits, including antioxidative, antimicrobial, antidiabetic, liver-protective, anti-inflammatory, anticancer, and heart-protective effects. In this research, High-Performance Liquid Chromatography (HPLC) methods were employed to assess sugar and organic acid levels in mature fruit pods of three distinct carob genotypes from the Kahramanmaras region in Turkey. The findings revealed that genotype-2 had the highest concentrations of glucose (1301 mg/100g) and fructose (875 mg/100g), genotype-3 exhibited the highest level of xylose (1548 mg/100g), while genotype-1 displayed the highest levels of sucrose (9183 mg/100g) and total sugars (12457 mg/100g). Regarding organic acids, genotype-1 exhibited the highest levels of oxalic acid (17.62 mg/100g), citric acid (612.50 mg/100g), and fumaric acid (8.0 mg/100g), while genotype-3 showed the highest levels of malic acid (234.92 mg/100g) and succinic acid (1089.76 mg/100g); however, genotype-2 had the highest amount of ascorbic acid (8.17 mg/100g). In conclusion, genotype-1 demonstrated the most favorable performance in terms of having the highest levels of total sugar and organic acids compared to the other two genotypes.

Rapid Detection of Adulteration in Boswellia Extracts with Citric Acid by UPLC–HRMS and 1H NMR

Boswellia serrata ole-gum-resin extracts (BSEs) are commonly used as food supplements, especially in osteoarthritis management. The quality standard is established by determining 11-keto-β-boswellic acid (KBA) and acetyl-11-keto-boswellic acid (AKBA) content using high-performance liquid chromatography (HPLC) or assessing the total boswellic acid (TBA) content by titrimetry. The limited geographical distribution of Boswellia species and increasing industrial demand could increase the risk of adulteration in Boswellia-containing products. In this study, 14 BSEs from commercial sources, used in food supplements, were analyzed in comparison with a USP Reference Standard extract. The KBA and AKBA content was determined by HPLC, whereas the TBA content was determined by titration. Targeted UHPLC-high-resolution mass spectrometry (HRMS) was applied to identify the carboxylic acid content in the samples. The 1H NMR spectra of extracts were also analyzed. Only two products met the criteria for KBA and AKBA content. Although, the TBA content complied with the expected amount, 10 extracts contained citric acid levels of 6–11% even though citric acid is not a cha­racteristic component of BSEs. Our results suggest undeclared addition of citric acid to comply with declared contents of TBA when using titration methods. Incorporation of citric acid to industrial samples – in order to alter the outcomes of the titration analysis – was demonstrated for the first time.

Metabolomic analysis of seminal fluids in infertile individuals.

Infertility impacts a substantial number of couples worldwide, and about 50% of cases are linked to male factors. The analysis of seminal fluid composition can improve diagnostic accuracy and offer deeper insights into the pathophysiology of male factor infertility. This study seeks to identify novel markers for diagnosing and treating male infertility by comparing organic acid profiles in the seminal fluid of individuals with normospermia, oligospermia, and azoospermia. Semen samples were collected from men with normospermia, oligospermia, and azoospermia. The organic acid profile in the seminal fluid was analyzed using liquid chromatography-mass spectrometry/mass spectrometry (LC/MS-MS). Data analysis was performed using SPSS and MetaboAnalyst. The study revealed significant differences in metabolite levels among normospermic, oligospermic, and azoospermic individuals. In groups with oligospermia, there were significant decreases in the levels of 2-OH-Isovaleric Acid, 3-Methyl-2-Oxovaleric Acid, Ethyl-Malonic Acid, Citric Acid, Oxoproline, Malic Acid, N-Acetyl-Aspartic Acid, Suberic Acid, Glutaconic Acid, and Succinic Acid. Similarly, individuals with azoospermia exhibited a notable reduction in the levels of Citric Acid, Malic Acid, and Suberic Acid. Furthermore, according to the Variable Importance in the Projection (VIP) score analysis, Ethyl-Malonic Acid, Glycolic Acid, and 3-Methyl-2-Oxovaleric Acid were identified as crucial factors for diagnosis and potential treatment strategies. The data obtained from the study highlights the significant potential of metabolites in assessing infertility and gaining a more in-depth understanding of the underlying pathological mechanisms.

Enhancing Vegetative Growth by Adding Phosphorus, Silicon, and Citric Acid to Pepper Plants Cultivated in Plastic Greenhouses

The research was conducted in a plastic greenhouse at the College of Agricultural Engineering Sciences, University of Baghdad - Jadiriyah Campus, during the 2021-2022 season, to study the effect of phosphorus, silicon, and citric acid on pepper plants using a factorial experiment design with three replicates. The first factor had three levels of phosphorus (0, 160, and 320 kg P2O5 per hectare), the second factor had three levels of potassium silicate (0, 75, and 100 kg per hectare), and the third factor had four levels of citric acid (0, 2, 4, and 6 kg per hectare). The statistical analysis showed that treatment P2S2C1 resulted in an increase in the main stem diameter (25.10 mm), treatment P3S3C1 led to an increase in the main branch diameter (16.10 mm), and treatment P3S3C2 showed an increase in the diameter of secondary and tertiary branches (13.50 mm and 8.90 mm, respectively). Treatment P2S3C1 resulted in an increased number of leaves and the dry weight of the total vegetative mass (1286.7 leaves and 415.0 g plant-1 respectively). Treatment P3S2C4 led to an increase in the dry weight of roots (25.47 g plant-1), treatment P2S3C4 showed an increased number of fruits (48.34 fruits plant-1), and treatment P3S3C4 resulted in an increased total yield (4.87 tons greenhouse-1).

Dietary citric acid decreased the theront number of Cryptocaryon irritans (Ciliata) in seawater-adapted tilapia (Oreochromis niloticus).

Cryptocaryoniasis remains a major parasitic disease and economic challenge for marine aquaculture. Cryptocaryoniasis in marine fish is caused by Cryptocaryon irritans (Ciliata). A theront is a motile, free-swimming stage in the life cycle of C. irritans, which is typically the infective stage that actively seeks out a host to initiate infection. Population density and growth rate of theronts were investigated in Nile tilapia, Oreochromis niloticus fed with citric acid-supplemented feed. The experiment involved feeding three diets with graded levels of citric acid (0, control diet, 0.5, 1 and 1.5 g kg-1 diet), to seawater-adapted Nile tilapia (O. niloticus) juveniles for 21 days. The results showed that citric acid in the fish feed had an impact on the theront number of C. irritans in a manner of dose-dependent. In the experimental cohort administered a diet supplemented with 1.5 g kg-1 citric acid, the population density of theronts was observed to be significantly reduced, measured at 29 ± 3.34, as opposed to 473.34 ± 16.48 in the control group at the culmination of the experiment. The observed population growth rate of theronts was significantly higher in the control group than in the group administered the citric acid feed (p < .005). The growth rate (r d-1 ) was 0.12 in control, 0.05 in 0.5 g kg-1 , 0.031 in 1 g kg-1 , and - 0.031 in 1.5 g kg-1 citric acid-supplemented groups. Fish growth and feed conversion ratio were not affected by the citric acid in the feed. In conclusion, the findings of this investigation provide a valuable addition to our understanding of the potential protective effects of citric acid supplementation for fish against the C. irritans parasite. This is evidenced by the observed reduction in theronts present in the water.