Sample Digestion Research Articles

Biomarker profiling in plants to distinguish between exposure to chlorine gas and bleach using LC-HRMS/MS and chemometrics

Since its first employment in World War I, chlorine gas has often been used as chemical warfare agent. Unfortunately, after suspected release, it is difficult to prove the use of chlorine as a chemical weapon and unambiguous verification is still challenging. Furthermore, similar evidence can be found for exposure to chlorine gas and other, less harmful chlorinating agents. Therefore, the current study aims to use untargeted high resolution mass spectrometric analysis of chlorinated biomarkers together with machine learning techniques to be able to differentiate between exposure of plants to various chlorinating agents. Green spire (Euonymus japonicus), stinging nettle (Urtica dioica), and feathergrass (Stipa tenuifolia) were exposed to 1000 and 7500 ppm chlorine gas and household bleach, pool bleach, and concentrated sodium hypochlorite. After sample preparation and digestion, the samples were analyzed by liquid chromatography high resolution tandem mass spectrometry (LC-HRMS/MS) and liquid chromatography tandem mass spectrometry (LC-MS/MS). More than 150 chlorinated compounds including plant fatty acids, proteins, and DNA adducts were tentatively identified. Principal component analysis (PCA) and linear discriminant analysis (LDA) showed clear discrimination between chlorine gas and bleach exposure and grouping of the samples according to chlorine concentration and type of bleach. The identity of a set of novel biomarkers was confirmed using commercially available or synthetic reference standards. Chlorodopamine, dichlorodopamine, and trichlorodopamine were identified as specific markers for chlorine gas exposure. Fenclonine (Cl-Phe), 3-chlorotyrosine (Cl-Tyr), 3,5-dichlorotyrosine (di-Cl-Tyr), and 5-chlorocytosine (Cl-Cyt) were more abundantly present in plants after chlorine contact. In contrast, the DNA adduct 2-amino-6-chloropurine (Cl-Ade) was identified in both types of samples at a similar level. None of these chlorinated biomarkers were observed in untreated samples. The DNA adducts Cl-Cyt and Cl-Ade could clearly be identified even three months after the actual exposure. This study demonstrates the feasibility of forensic biomarker profiling in plants to distinguish between exposure to chlorine gas and bleach.

High-Throughput Silica Nanoparticle Detection for Quality Control of Complex Early Life Nutrition Food Matrices.

The addition of nanomaterials to improve product properties has become a matter of course for many commodities: e.g., detergents, cosmetics, and food products. While this practice improves product characteristics, the increasing exposure and potential impact of nanomaterials (<100 nm) raise concerns regarding both the human body and the environment. Special attention should be taken for vulnerable individuals such as those who are ill, elder, or newborns. But detecting and quantifying nanoparticles in complex food matrices like early life nutrition (ELN) poses a significant challenge due to the presence of additional particles, emulsion-droplets, or micelles. There is a pressing demand for standardized protocols for nanoparticle quantification and the specification of "nanoparticle-free" formulations. To address this, silica nanoparticles (SiNPs), commonly used as anticaking agents (AA) in processed food, were employed as a model system to establish characterization methods with different levels of accuracy and sensitivity versus speed, sample handling, and automatization. Different acid treatments were applied for sample digestion, followed by size exclusion chromatography. Morphology, size, and number of NPs were measured by transmission electron microscopy, and the amount of Si was determined by microwave plasma atomic emission spectrometry. This successfully enabled distinguishing SiNP content in ELN food formulations with 2-4% AA from AA-free formulations and sorting SiNPs with diameters of 20, 50, and 80 nm. Moreover, the study revealed the significant influence of the ELN matrix on sample preparation, separation, and characterization steps, necessitating method adaptations compared to the reference (SiNP in water). In the future, we expect these methods to be implemented in standard quality control of formulation processes, which demand high-throughput analysis and automated evaluation.

COMPARATIVE APPRAISAL OF THE ACACIA (ACACIA NILOTICA L.) AND MESQUITE (PROSOPIS JULIFLORA) ON THE BASIS OF MINERAL CONTENTS AND CONTAMINANTS FOR THE CAMEL FEEDS

This study evaluates the nutritional value and quality of feed and forage for camel husbandry and productivity. It focuses on two forage plant species, Acacia and Mesquite, based on mineral nutrients and contaminants for grazing and camel health benefits. The study was conducted in Jam Mahar and Tharo Mari locations of Hyderabad, contaminated lands of Oil and Gas Development Company Limited. Leaf tissue samples were collected from three trees of each plant species, oven-dried at 70°C for 48 hours, and analyzed for crude protein, fats, fiber, and ash. The plant material was digested using diacetic acid and filtered properly. The filtrate of digested samples was analyzed on Atomic Absorption Spectrometer (Nova 400) Analytik Jena, Germany. The findings of the study revealed that Acacia leaf, including pod samples, illustrated more crude protein (20.1%), fats (16%) and crude fiber (16.4) in comparison to mesquite (Devi). Findings from macro and the micronutrients of the study revealed that mesquite leaf tissues contained 36.7% and 3% more K. and Ca as compared to acacia leaf samples. Inversely to K and Ca, the Mg and Fe contents of Acacia leaf tissues were 77% and 59% higher than mesquite leaf tissue. The study found that acacia leaves have potential to meet the nutritional needs of camels and address feed shortage issues during the monsoon dry season. The findings suggest further research on the benefits of acacia leaves as fodder and their potential use in combination with other grasses to enhance various nutrients, including protein, lipids, fiber, macronutrients, and micronutrients

In vitro digestibility and solubility of phosphorus of three plant-based meat analogues.

Interest in plant-based meat analogues has increased and can be expected to be applied to pet foods, which necessitates the understanding of the nutrient supply in those foods. Our primary aim was to advance our understanding of the digestive properties of sterilized plant-based meat analogues. The impact of the preparatory processing steps on the solubility of meat analogues was studied. Meat analogues were made by mixing water, salt, and wheat gluten with soy protein isolate, pea protein isolate, or faba bean concentrate. Mixed materials were processed into model meat analogues using shear cell technology. Products were canned in water or gravy and sterilized. An animal-based canned pet food was made as a reference. Products sampled at the processing steps (mixing, shearing, sterilization) were digested in vitro. Samples of digestate were taken at the gastric phase (0 and 120 min) and small intestinal phase (120, 200, 280, and 360 min) for analysis of protein hydrolysis. The extent digestion of nitrogen and dry matter was determined at the end of incubation. Total phosphorus, soluble phosphorus after acid treatment, and after acid and enzymatic treatment were determined. The degree of hydrolysis after gastric digestion was low but increased immediately in the small intestinal phase; products based on pea had the highest values (56%). Nitrogen digestibility was above 90% for all materials at each processing step, indicating that bioactive compounds were absent or inactivated in the protein isolates and concentrate. Phytate seemed to play a minor role in meat analogues, but phosphorus solubility was influenced by processing. Shearing decreased soluble phosphorus, but this effect was partly reversed by sterilization. Nutrient digestibility as well as phosphorus solubility in plant-based products was higher than or comparable with the reference pet food. These findings show that the digestive properties of the tested plant-based meat analogues do not limit the supply of amino acids and phosphorus.

Microplastic contamination in rivers: a survey from the Nan River, Thailand

BackgroundSince the 1970s, microplastic contamination has been discovered in various environments. These emerging pollutants threaten water quality and freshwater ecosystems. We aimed to assess microplastic contamination in the Nan River by quantifying their presence, characterising their morphology, and identifying their polymer composition. MethodsThe survey was conducted along the Nan River in northern Thailand, which is divided into three distinct zones based on land use (ie, community areas, agricultural areas, and natural areas). A wide array of samples, including surface water, sediment, and specimens from five aquatic species—Barbonymus altus, Laides hexanema, Kryptopterus cryptopterus, Pomacea canaliculata, and Pseudodon cambodjensis cambodjensis—were collected. The presence of microplastics was analysed via the wet peroxide oxidation process for sample digestion, morphological characteristics were assessed through microscopic examination, and Fourier transform infrared spectrophotometry was used to identify the polymer composition of the microplastics. FindingsNatural areas had the highest mean concentration of microplastics in surface water (23·67 pcs/L, SD 14·22), whereas community areas had the highest mean concentration in soil sediment (24·67 pcs/g, 15·04). No statistically significant differences in the amounts of microplastics were observed among the various land-use zones. Microplastics were predominantly found in the gastrointestinal tracts of fish specimens, with varying quantities observed among different aquatic species; B altus had the highest concentration (11·80 pcs/g, 8·98). The primary microplastic shapes identified were filaments (53·99%), fragments (35·58%), and cylindrical forms (10·43%). We identified ten different polymer types, with polypropylene, nylons, and polymethyl methacrylate being the most prevalent among them. InterpretationAnalysing microplastic pollution in this area brings attention to the issues affecting the rivers of Thailand and emphasises ecological risks, particularly microplastic presence in aquatic species. Because of the Nan River's ecological importance, we call for increased research and collaboration to address the increasing threat of microplastic pollution. FundingNaresuan University.

A Univariate Optimization Strategy for Pre-concentration of Cobalt(II) in Various Matrixes by a DLLME before Analysis Using FAAS

A procedure based on dispersive liquid-liquid microextraction (DLLME) for cobalt (Co) quantification in an Iraqi environmental matrix by flame atomic absorption spectroscopy (FAAS) was applied in this work. A case-study approach was chosen to obtain further in-depth information on the Co levels and to evaluate the effectiveness of N-salicylideneaniline (SAN) as a complexing agent for pre-concentration and extraction of Co. An univariate strategy was utilized to achieve the optimum extraction conditions. The estimated limits of detection (LOD) and quantification (LOQ) under optimum conditions were 1.04 and 3.47 µg L−1, respectively. The results achieved by the proposed system were compared with those using the microwave digestion/graphite furnace atomic absorption spectrometer (MWD/GF-AAS) for digest samples and also for some water samples (Direct GF-AAS). The proposed procedure was applied for analyzing eleven environmental samples. The detectable Co levels for water samples ranged from 0.72 to 4.30 µg L−1 with a relative standard deviation of 3.7–8.8%, while the concentration for solid samples ranged from 0.17–4.51 µg g−1 (2.4–11.8 RSD %). DLLME/FAAS proposed procedure is effective, simple, and has the benefit of minimizing the organic solvent consumption by a few microliters, which results in little waste.

Abstract 7092: Cancer biomarker discovery with scalable biofluid proteomics

Abstract Biofluid proteomics opens opportunities for the discovery of early detection, diagnostic, and prognostic cancer biomarkers through non-invasive or minimally invasive screening. Plasma and serum are accessible sample types, however, the high dynamic range of proteins creates analytical challenges for the sensitive detection of potential biomarkers. In this work, we compare two proteomic approaches for large-scale cancer biomarker discovery: the Olink Proximity Extension Assay and the Seer Proteograph Product Suite. Olink Explore 3072 is an immunoassay that gains specificity by using dual antibody recognition to measure approx. 3,000 proteins in each sample. Antibody pairs are tagged with oligonucleotides, which hybridize and are amplified before measurement by next generation sequencing. Seer Proteograph is an enrichment strategy that uses a proprietary mix of functionalized magnetic nanoparticles with affinity for certain amino acids and functional groups in native state proteins. The resulting peptides are measured using untargeted mass spectrometry to identify proteins at the peptide level, paving the way for the identification of proteoforms and peptide variants. Aliquots of human plasma from 78 samples representing a range of cancer indications were obtained through the Discovery Life Sciences biobank and used for analysis by both the Olink and Proteograph workflows. Olink analysis achieved Pearson correlation scores of greater than or equal to 0.8 when correlating each of seven process controls to each other for all approx. 3000 proteins. On average, greater than or equal to 75% of the proteins were detected across all panels in all sample groups except for Normal and Benign groups (64% and 74% respectively). After stringent FDR filtering to ensure only high-quality data was analyzed, the Seer Proteograph workflow quantified 4,323 proteins across all 78 samples. Pearson correlation scores of greater than or equal to 0.96 were achieved when comparing digestion controls, process controls, and sample cleanup controls. Both platforms achieved detection of proteins across the dynamic range of plasma, including low abundance proteins. The Olink Explore 3072 and Seer Proteograph platforms were found to be complementary, with approx. 15% overlap in measured proteins. Cancer biomarker discovery at the proteomic level is bolstered by recent advances in analytical capabilities, as revealed by the Olink Explore and Seer Proteograph platforms. Importantly, complementary implementation of these two workflows will enable comprehensive biomarker analysis. Citation Format: Danielle B. Gutierrez, Jessica L. Moore, Jaison Arivalagan, Tiffany Louie, Leslie Wilkinson, Ray Clemens. Cancer biomarker discovery with scalable biofluid proteomics [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 7092.

Bioactive compounds of fresh tea shoots plucked in different seasons: optimization of extraction of polyphenols

AbstractIn this study, the effects of harvest year and period on the total polyphenol (TP), radical scavenging activity (RSA), total flavonoid (TF) and ascorbic acid (AA) contents of fresh tea leaves were investigated. Extraction of polyphenols from the tea leaf was optimized using the response surface methodology (RSM). The effect of the independent variables ((solvent concentration, extraction time and solid to solvent (S/S) ratio)) on TP and RSA was studied using Box-Behnken design (BBD). The results showed that extraction parameters selected were significant in the extraction procedure regarding TP and RSA (p &lt; 0.05). The optimum extraction conditions were obtained as 1/60.86 of S/S ratio, 84.65% of ethanol concentration and the extraction time of 50 min. As a result of the extraction of tea leaves under optimum conditions, the TP, RSA and TF contents of the leaves harvested in September in 2020 were found to be higher. In addition, the individual phenolic compounds of these leaves were analyzed by high performance liquid chromatography (HPLC) and epigallocatechin gallate (EGCG) was detected the most (57.52 mg/g DM). After in-vitro digestion of the same samples, TP and RSA values ​​were significantly decreased, mostly at intestinal stage (p &lt; 0.05), and the bioaccessibility of the polyphenols was determined as 48.98%. As a result of AA analysis by HPLC of the, the highest contents for 2019 and 2020 were observed in the leaves harvested in May (41.92 mg/100 g DM) and those in July (38.55 mg/100 g DM), respectively.

ANALYTICAL PROCEDURES USING SEM/EDX EQUIPMENT FOR DETERMINING HEAVY METALS CONTENTS IN FLY ASH GENEREATED FROM DOMESTIC SOLID WASTE INCINERATORS: A CASESTUDY OF SOCSON WASTE-TO-POWER PLANT IN HANOI

The researching and establishing of as process for analyzing fly ash generated from a domestic solid waste treatment plant by SEM/EDX method were investigated. Three sampling series with twelve (12) fly ash samples collected at SocSon waste-to-power plant were used for: (i) analytical procedure establishment; (ii) standardization; and (iii) test analysis. At each measurement, a cross-check with one random sample using XRF method was carried out to verify the accuracy of the SEM/EDX measurements. As a result, a 6-step measurement procedure has been proposed, including: experimental preparation, sample weighting, liquid/solid phase separation, sample drying, sample measurement and data processing. And for procedure adjustment, liquid/solid phase separation were repeated twice while drying time was doubly increased from 24h to 48h to collect ll heavy metal contaminated residues. The proposed procedure could provide results in about 3-5 seconds without any requirement of sample digestion while most of the heavy metalspresent in fly ash, including low-content ones such as Ag, Cd and Cr were simultaneously displayed. The standardized measurement procedure was applied to test the hazardous properties of fly ash by comparing with QCVN 07:2009/BTNMT – National Technical Regulations on Hazardous waste thresholds. The results identified that fly ash could be primarily considered as hazardous waste with 5/14 heavy metal parameters exceed the limit from 1,7 to 9,5 times and needs to be collected and treated properly. This result is also quite similar to the studies that have been done in China, the US or Vietnam. Thus, SEM/EDX method could be used for analyzing heavy metal components in fly ash samples of solid waste treatment plants.

Optimization of Selective Hydrometallurgical Tantalum Recovery from E-Waste Using Zeolites.

To protect future high-tech metal demand, a selective and efficient recovery method for tantalum from a tantalum-rich e-waste component sample was developed. Ultrasound-assisted digestion of the component sample was optimized, and the highest dissolution rate was achieved using a mixture of 8 mol/L H2SO4 and HF at a temperature of 60 °C. The determined amount of tantalum was as high as 11000 ± 1000 mg/kg, which results in a high potential for recyclable tantalum. The other major elements of this complex e-waste fraction were silicon, iron, aluminum, and tin. Efficient recovery of tantalum from the leachate was performed using the zeolite material ZSM-5. Extremely high selectivity and a recovery rate of over 98% were obtained. In terms of adsorption efficiency, selectivity, and durability of the material, optimal adsorption was obtained using the diluted sample at 0.5 mol/L of H2SO4. The adsorption capacity of ZSM-5 for tantalum was determined to be 10.5 ± 0.6 mg/g, and tantalum was selectively eluted with 1:4 diluted ethanolamine with a yield of 87.2 ± 1.5%.

Validation of a Method for Surveillance of Nanoparticles in Mussels Using Single-Particle Inductively Coupled Plasma-Mass Spectrometry.

Determining the concentration of nanoparticles (NPs) in marine organisms is important for evaluating their environmental impact and to assess potential food safety risks to human health. The current work aimed at developing an in-house method based on single-particle inductively coupled plasma-mass spectrometry (SP-ICP-MS) suitable for surveillance of NPs in mussels. A new low-cost and simple protease mixture was utilized for sample digestion, and novel open-source data processing was used, establishing detection limits on a statistical basis using false-positive and false-negative probabilities. The method was validated for 30 and 60 nm gold NPs spiked to mussels as a proxy for seafood. Recoveries were 76-77% for particle mass concentration and 94-101% for particle number concentration. Intermediate precision was 8-9% for particle mass concentration and 7-8% for particle number concentration. The detection limit for size was 18 nm, for concentration 1.7 ng/g, and 4.2 × 105 particles/g mussel tissue. The performance characteristics of the method were satisfactory compared with numeric Codex criteria. Further, the method was applied to titanium-, chromium- and copper-based particles in mussels. The method demonstrates a new practical and cost-effective sample treatment, and streamlined, transparent, and reproducible data treatment for the routine surveillance of NPs in mussels.

Uncovering microplastics contamination in canned seafood

There is limited research on the occurrence of microplastics (MPs) in canned seafood. All types of canned seafood investigated in the present study were contaminated. After sample digestion in 30 % hydrogen peroxide, a total of 40 MPs were recovered. Fibers were the most common type, blue was the dominant colour, and Fourier Transform Infrared Spectroscopy (FTIR) identified polyester as the most common polymer. Considering all samples, an average of 3.5 ± 5.2 MPs/can was obtained, with octopus in tomato sauce and tuna in olive oil presenting the highest contamination (5.2 ± 7.5 MPs/can and 5.2 ± 5.1 MPs/can, respectively). Also, significant differences between the number of MPs in the seafood tissues and immersion liquids were verified. The present study demonstrates MPs occurrence in canned seafood, a potential contamination pathway for humans. More research on the different stages of the canning processing is vital for understanding MPs contamination in cans.

Qualitative and Quantitative Analytical Techniques of Nucleic Acid Modification Based on Mass Spectrometry for Biomarker Discovery.

Nucleic acid modifications play important roles in biological activities and disease occurrences, and have been considered as cancer biomarkers. Due to the relatively low amount of nucleic acid modifications in biological samples, it is necessary to develop sensitive and reliable qualitative and quantitative methods to reveal the content of any modifications. In this review, the key processes affecting the qualitative and quantitative analyses are discussed, such as sample digestion, nucleoside extraction, chemical labeling, chromatographic separation, mass spectrometry detection, and data processing. The improvement of the detection sensitivity and specificity of analytical methods based on mass spectrometry makes it possible to study low-abundance modifications and their biological functions. Some typical nucleic acid modifications and their potential as biomarkers are displayed, and efforts to improve diagnostic accuracy are discussed. Future perspectives are raised for this research field.

Physical Pretreatments of Cassava Chips Influenced Chemical Composition, Physicochemical Properties, and In Vitro Digestibility in Animal Models.

Physical pretreatment procedures can significantly influence the quality of food and feed raw materials. To increase the ability to be digested in animals, cassava chips were pretreated by four alternative methods (extrusion, microwave irradiation, gamma irradiation, or NaOH hydrolysis), and then the chemical composition, physicochemical properties, and in vitro digestibility of the pretreated samples were assessed and compared with unprocessed cassava chips (control). The chemical compositions (crude protein, ether extract, neutral detergent fiber, acid detergent fiber, ash, non-fiber carbohydrate, and gross energy) were significantly altered due to the pretreatment methods (p < 0.05). The nutritive profile was qualitatively changed when assessed through Fourier-transform infrared spectroscopy. Some physicochemical properties in association with enzymatic hydrolysis, which include pH, water solubility, water absorption capacity, thermal properties (differential scanning calorimetry), diffraction pattern (X-ray diffractometry), and microstructure (scanning electron microscopy), were significantly changed. In vitro carbohydrate digestibility based on digestive enzyme extracts from Nile tilapia (Oreochromis niloticus) suggests the microwaving method for cassava chips preparation, while microwaving, followed by extrusion or gamma irradiation, was suggested for broiler (Gallus gallus domesticus). There were no differences in the pepsin-cellulase digestibility values tested for the ruminant model. These findings suggest the use of pretreated cassava chips in animal feeding.

Phenolic Characterization of a Purple Maize (Zea mays cv. "Moragro") by HPLC-QTOF-MS and Study of Its Bioaccessibility Using a Simulated In Vitro Digestion/Caco-2 Culture Model.

The present work aimed to characterize the phenolic and antioxidant content of the Argentinian purple maize "Moragro" cultivar. Additionally, the INFOGEST simulated in vitro digestion model was used to establish the effect of digestion on bioactive compounds. Finally, digestion samples were used to treat Caco-2 cells in the transwell model to better understand their bioavailability. Twenty-six phenolic compounds were found in purple maize cv. "Moragro", 15 nonanthocyanins and 11 anthocyanins. Several compounds were identified in maize for the first time, such as pyrogallol, citric acid, gallic acid, kaempferol 3-(6″-ferulylglucoside), and kaempferol 3-glucuronide. Anthocyanins accounted for 24.9% of total polyphenols, with the predominant anthocyanin being cyanidin-3-(6″ malonylglucoside). Catechin-(4,8)-cyanidin-3,5-diglucoside and catechin-(4,8)-cyanidin-3-malonylglucoside-5-glucoside were detected as characteristics of this American maize variety. Total polyphenol content (TPC; by the Folin-Ciocalteu method), HPLC-DAD/MSMS, and antioxidant activity [by DPPH and ferric-reducing antioxidant power (FRAP)] were evaluated throughout in vitro digestion. TPC, DPPH, and FRAP results were 2.71 mg gallic acid equivalents (GAE)/g, 24 μmol Trolox equiv/g, and 22 μmol Trolox eq/g, respectively. The in vitro digestion process did not cause significant differences in TPC. However, the antioxidant activity was significantly decreased. Moreover, the bioavailability of anthocyanins was studied, showing that a small fraction of polyphenols in their intact form was conserved at the end of digestion. Finally, a protective effect of digested maize polyphenols was observed in the Caco-2 cell viability. The results suggest that "Moragro" purple maize is a good source of bioavailable anthocyanins in the diet and an interesting source of this group of compounds for the food industry.

Advanced review of the contributing factors for the Microwave Digestion of food matrices for trace elemental analysis

Microwaves are electromagnetic waves which move at the speed of light. Samples directly exposed to this phenomenon are heated faster than conventional conduction heating methods. This is exploited in acid assisted digestion where a food sample in a suitable acid matrix can be completely destroyed leaving a clear solution of measurable elements. In pressurised systems the temperature can be rapidly increased far above the conventional boiling point of the solvent and the closed microwave vessel environment is ideal for volatile elements.There is a constant aspiration to achieve lower levels of detection and advances in instrument capabilities with specialised plasma based techniques for the identification and quantification of individual elements at ultra-trace levels have evolved. To realise this; sample preparation, digestion efficiency and sample introduction need to be optimised. Exogenous inputs such as preparation environment, reagent grade, the analyst and apparatus cleaning steps all have the potential to leave an elemental footprint on the sample and therefore contribute to the uncertainty of the analytical result. Knowledge of sample composition and subsequent interaction with microwaves and plasma are an important consideration for complete digestion without which the result is not accurate.

Comparisons of Different Digestion Methods for Heavy Metal Analysis from Fruits

Fruit juices are produced in home or industrially from horticultural crops by pressing the liquid part. They are rich in sugar, vitamins, and minerals like iron, copper, potassium, folate minerals, and vitamins A, B, and C which are essential for giving the body the nutrients it needs to stay healthy since fruits contain vital mineral components like copper (Cu), iron (Fe), and manganese (Mn), which is necessary for human growth and respiration. However, they may have heavy metals which may poison health risk and toxic even the presence is in little amount. Since fruit juices doesn’t pass through different processes, except extracting the liquid from the fruits of vegetables contamination and heavy metals affect human health. Before determination of heavy metals different procedures are applied for analysis. Digestion is the key component for determination of heavy metals from different samples. In this paper we are concerned on wet digestion methods for analysis. Closed system wet digestion is preferred since it lower the risk of contamination. There are different wet digestion types. Some of them are conventional wet digestion, ultraviolet digestion, ultrasound-assisted acid decomposition, conventional heating, microwave-assisted wet digestion etc. From thus, microwave digestion procedure was preferred for the digestion of samples for determination of heavy metals due to its ability to oxidize almost all of the organic samples.

Accelerating Proteomics Using Broad Specificity Proteases.

Trypsin is the gold-standard protease in bottom-up proteomics, but many sequence stretches of the proteome are inaccessible to trypsin and standard LC-MS approaches. Thus, multienzyme strategies are used to maximize sequence coverage in post-translational modification profiling. We present fast and robust SP3- and STRAP-based protocols for the broad-specificity proteases subtilisin, proteinase K, and thermolysin. All three enzymes are remarkably fast, producing near-complete digests in 1-5 min, and cost 200-1000× less than proteomics-grade trypsin. Using FragPipe resolved a major challenge by drastically reducing the duration of the required "unspecific" searches. In-depth analyses of proteinase K, subtilisin, and thermolysin Jurkat digests identified 7374, 8178, and 8753 unique proteins with average sequence coverages of 21, 29, and 37%, including 10,000s of amino acids not reported in PeptideAtlas' >2400 experiments. While we could not identify distinct cleavage patterns, machine learning could distinguish true protease products from random cleavages, potentially enabling the prediction of cleavage products. Finally, proteinase K, subtilisin, and thermolysin enabled label-free quantitation of 3111, 3659, and 4196 unique Jurkat proteins, which in our hands is comparable to trypsin. Our data demonstrate that broad-specificity proteases enable quantitative proteomics of uncharted areas of the proteome. Their fast kinetics may allow "on-the-fly" digestion of samples in the future.

Identification of lentils (Lens culinaris Medik) from Eglouvi (Lefkada, Greece) based on rare earth elements profile combined with chemometrics

An analytical approach has been developed to verify the authenticity of premium lentils originating from Eglouvi, Lefkada, Greece. The method relies on the digestion of samples followed by the analysis of their rare earth elements (REEs) content. Lentils originating from Eglouvi exhibit higher content in most REEs compared to lentils from other regions as well as distinct Sc/Y and Sc/Yb concentration ratios. Principal component analysis effectively segregates “Eglouvi” lentils into a distinct cluster. Soft Independent Modelling of Class Analogy (SIMCA) successfully models “Eglouvi” lentils. Significant enhancement in model specificity was achieved upon inclusion of Sc/Y and Sc/Yb concentration ratios as additional variables. The model is capable of detecting adulteration in blends of Eglouvi lentils, with a minimum rejection threshold of 4.6% w/w for Greek lentil adulterants and 6.0% w/w for imported lentil adulterants.

Characterisation and agronomic evaluation of acidified food waste anaerobic digestate products

Raw liquid anaerobic digestate was synthesised into nutrient-dense solid digestates via acidification and evaporation. Acidification retained ammonium in the digestate whilst also donating the anion to free ammonium to form an ammonium salt. Digestate was treated with the addition of sulphuric, nitric, and phosphoric acid resulting in the formation of ammonium sulphate, ammonium nitrate and ammonium phosphate, respectively then evaporated into a solid fertiliser product. FTIR, XRD and SEM-EDS collectively confirm that the addition of acids completely converted the free ammonium in the raw digestate into their respective ammonium salt counterparts. Compounds of potassium chloride, silicon dioxide, calcium carbonate, magnesium ammonium phosphate, sodium nitrate, and sodium chloride were identified in all solid digestate samples. Plant growth and grain yield was higher in urea ammonium nitrate, raw liquid digestate and acidified digestate products compared to control and unacidified solid digestate. Urea ammonium nitrate and ammonium nitrate solid digestate had the highest dry shoot, likely due to the high available nitrogen found in both fertilisers. Overall, acidification and evaporation of liquid digestate can efficiently transform it into a valuable solid fertiliser with a high nutrient density. This process not only has the potential to mitigate handling and storage constraints of low nutrient density digestate in anaerobic digestion facilities but also offers a sustainable alternative to conventional fertilisers.