The effect of ascorbic acid, storage period and packaging material on the formation of volatile N-nitrosamine in sausages.

This study aims to determine the amounts of seven volatile N-nitrosamine (VNA) derivatives which are in the risk group, in processed sausages. It also aimed to investigate the effects of the amount of added ascorbic acid, on the VNA level during the sausage manufacturing processes. For this purpose, meat doughs were prepared with two different levels of ascorbic acid. These meat doughs were put into the production process and packaged in two different packages. Thus, VNA derivatives and their amounts were determined according to production stages (heat treatments), packaging method (vacuum, MAP), storage process (Day 1, 7, 15, 30, 60, 90). As a result, it was found that the sausage product carries risk of VNA formation from the beginning of its production until the last day of storage before consumption for up to 90 days.

Recent advances in process engineering and upcoming applications of metal-organic frameworks.

Progress in metal-organic frameworks (MOFs) has advanced from fundamental chemistry to engineering processes and applications, resulting in new industrial opportunities. The unique features of MOFs, such as their permanent porosity, high surface area, and structural flexibility, continue to draw industrial interest outside the traditional MOF field, both to solve existing challenges and to create new businesses. In this context, diverse research has been directed toward commercializing MOFs, but such studies have been performed according to a variety of individual goals. Therefore, there have been limited opportunities to share the challenges, goals, and findings with most of the MOF field. In this review, we examine the issues and demands for MOF commercialization and investigate recent advances in MOF process engineering and applications. Specifically, we discuss the criteria for MOF commercialization from the views of stability, producibility, regulations, and production cost. This review covers progress in the mass production and formation of MOFs along with future applications that are not currently well known but have high potential for new areas of MOF commercialization.

Phytochemical constituents and anticancer activities of Tarchonanthus camphoratus essential oils grown in Saudi Arabia.

Tarchonanthus Camphoratus L. is traditionally known for its various medicinal purposes. In this study, the T. camphoratus essential oil (TCEO) was isolated via steam distillation, and its chemical constituents were determined using GC-MS. The in vitro antiproliferative effects of TCEO on A549, HepG2, MCF-7 cancer cells, and HUVEC non-tumor cells was investigated using an MTT assay. Flow cytometry analysis was conducted to evaluate cell cycle distribution using propidium iodide staining, and cell death mode using Annexin V-FITC/PI assays. The expression of some apoptosis related genes was investigated using qRT-PCR. Major constituents of TCEO included fenchol, borneol, 3-cyclohexene-1-methanol and 3-ethyl-3-methyl. Cell viability test showed that TCEO is highly effective against MCF-7 cells with IC50 12.5 µg/mL. Cell cycle arrest at the G1/S phase, and apoptosis mediation were evident in the presence of TCEO. Gene expression analysis of several pro-apoptotic and anti-apoptotic genes revealed the initiation of apoptosis in TCEO-MCF-7 cells. In conclusion, our study confirms the antiproliferative activity of the T. camphoratus essential oil.

Comparative metabolomics of Japanese Black cattle beef and other meats using gas chromatography-mass spectrometry.

Progress in metabolomic analysis now allows the evaluation of food quality. This study aims to identify the metabolites in meat from livestock using a metabolomic approach. Using gas chromatography-mass spectrometry (GC/MS), many metabolites were reproducibly detected in meats, and distinct differences between livestock species (cattle, pigs, and chickens) were indicated. A comparison of metabolites between tissues types (muscle, intramuscular fat, and intermuscular fat) in marbled beef of Japanese Black cattle revealed that most metabolites are abundant in the muscle tissue. Several metabolites (medium-chain fatty acids, etc.) involved in triacylglycerol synthesis were uniquely detected in fat tissue. Additionally, the results of multivariate analysis suggest that GC/MS analysis of metabolites can distinguish between cattle breeds. These results provide useful information for the analysis of meat quality using GC/MS-based metabolomic analysis.ABBREVIATIONS: GC/MS: gas chromatography-mass spectrometry; NMR: nuclear magnetic resonance; MS: mass spectrometry; IS: 2-isopropylmalic acid; MSTFA: N-Methyl-N-trimethylsilyltrifluoroacetamide; CV: coefficient of variation; TBS: Tris-buffered saline; MHC: myosin fast type; PCA: principal component analysis; OPLS-DA: orthogonal partial least-squares discriminant analysis; O2PLS: two-way orthogonal partial least-squares.

Isovaleronitrile co-induced with its precursor, l-leucine, by herbivory in the common evening primrose stimulates foraging behavior of the predatory blue shield bug.

Herbivore-induced plant volatiles play important roles in plant-insect and plant-plant interactions. The common evening primrose, Oenothera biennis, is often infested by the flea beetle, Altica oleracea, on which the predatory blue shield bug, Zicrona caerulea, is usually found. This observation suggests that the predatory bug can discriminate infested plants from intact ones to locate its prey. In this study, l-leucine-derived nitrogen-containing compounds [isovaleronitrile (3-methylbutanenitrile), (E/Z)-isovaleraldoxime and 3-methyl-1-nitrobutane] and some terpenes were identified as a characteristic volatile blend from herbivore-infested O. biennis leaves by gas chromatography/mass spectrometry, chemical synthesis, and incorporation assays using deuterium-labeled l-leucine. Volatile emission was also elicited by exogenous methyl jasmonate (MeJA), but not by mechanical damage. l-Leucine accumulated temporarily in O. biennis leaves after MeJA treatment prior to isovaleronitrile emission. Behavioral assays revealed that Z. caerulea showed a strong preference for herbivore-infested leaves, their volatiles, and isovaleronitrile in laboratory conditions.

Smelling the difference: separation of healthy and infected button mushrooms via microbial volatile organic compounds.

In the literature there is a lack of consensus regarding mushroom volatiles; most of the studies identify only a few volatiles. This study deals with button mushrooms, their emitted volatiles, and the main changes during infections (green mould and cobweb disease) in a time series experiment. Emitted volatile profiles were determined using HS-SPME-GC-MS coupled analytical technique. The separation of healthy and infected mushroom samples was done using different multivariate statistical methods (PCA, PLS-DA, HeatMap). The main volatile compounds were also determined. As a result, several compounds were found to successfully distinguish healthy (bisabolene, cymene, myrtenol, d-limonene, etc.) and infected (thujopsene, cedr-8-ene, chamigrene, patchulane, longifolene, etc.), mushroom samples, and an early disease detection was achieved. Results can be used for further investigation of infected mushroom identification in an early stage in packaged mushroom products. Furthermore, these results could help to identify infections in commercially available mushrooms, thus increasing shelf-life in super/hypermarkets.

Characterization and modelling of odor-active compounds release behavior from Fu-brick tea during boiling-water extraction by molecular sensory science approach.

The odor-active compounds in Fu-brick tea and their release behaviors during boiling-water were studied. Fifty-one odor-active compounds were identified, and their release behaviors were identified by continuously recovering 16 sections of condensed water combined with sensory, instrumental, and nonlinear curve fitting analysis. The odor intensities of condensed water and the concentrations of odor-active compounds could be significantly fitted (p < 0.01) to power-function type curves. Hydrocarbons showed the fastest release rate, while organic acids showed the slowest. The release rates had very little correlation with their concentrations, molecular weights, and boiling points. Most odor-active compounds (≥70 %) released need to evaporate more than 24 % of the added water during boiling-water extraction. Meanwhile, on the basis of odor activity value (OAV) calculation, the aroma recombination experiments were performed to explore the odor-active compounds that made major contributions to the formation of the aroma profile of each condensed water.

A web application to support the coordination of reflexive, interpretative toxicology testing.

Background: Reflexive laboratory testing workflows can improve the assessment of patients receiving pain medications chronically, but complex workflows requiring pathologist input and interpretation may not be well-supported by traditional laboratory information systems. In this work, we describe the development of a web application that improves the efficiency of pathologists and laboratory staff in delivering actionable toxicology results. Method: Before designing the application, we set out to understand the entire workflow including the laboratory workflow and pathologist review. Additionally, we gathered requirements and specifications from stakeholders. Finally, to assess the performance of the implementation of the application, we surveyed stakeholders and documented the approximate amount of time that is required in each step of the workflow. Results: A web-based application was chosen for the ease of access for users. Relevant clinical data was routinely received and displayed in the application. The workflows in the laboratory and during the interpretation process served as the basis of the user interface. With the addition of auto-filing software, the return on investment was significant. The laboratory saved the equivalent of one full-time employee in time by automating file management and result entry. Discussion: Implementation of a purpose-built application to support reflex and interpretation workflows in a clinical pathology practice has led to a significant improvement in laboratory efficiency. Custom- and purpose-built applications can help reduce staff burnout, reduce transcription errors, and allow staff to focus on more critical issues around quality.

Quantitative tandem mass spectrometry in the clinical laboratory: Regulation and opportunity for validation of laboratory developed tests.

Tandem mass spectrometry is an important analytical tool for clinical laboratories, but tests developed and validated in-house (laboratory developed tests, or LDTs) require special consideration. In late 2022, the forecast for United States (U.S.) federal regulation of LDTs changed unexpectedly when the VALID Act was not passed by the U.S. Congress. This Act would have modified the Food and Drug Administration's (FDA's) role to increase regulatory oversight for LDT providers. In this revised context, we review optimization of quantitative mass spectrometry LDT validation and suggest avenues other than an additional FDA mandate to achieve uniform best practice. Common challenges, logistical barriers, and recommendations for easing the burden of best-quality quantitative mass spectrometry LDT method validation are discussed.

Antifungal potential, chemical composition of Chlorella vulgaris and SEM analysis of morphological changes in Fusarium oxysporum.

In pursuit of an environmentally benign fungicide alternative, the current study explored the antifungal activity of Chlorella vulgaris extracts against six plant pathogenic fungi (in vitro). The well diffusion agar method was used to investigate the growth inhibition of Fusarium oxysporum, Fusarium sp., Fusarium solani, A. flavus, A. niger, and A. alternata using the three C. vulgaris extracts viz. methanol (CvME), acetone (CvAE), and diethyl ether (CvDE). Different concentrations of CvDE were also investigated against F. oxysporum. The morphological modifications in F. oxysporum treated with CvDE (5 mg/kg) were studied using SEM and the chemical composition of CvDE was also determined by GC-MS analysis. All extracts, with the exception of A. alternata, were found to be effective in inhibiting the growth of plant pathogenic fungi. The CvDE extract, followed by CvME and CvAE, was found to be efficient against tested fungi. The CvDE was most effective against F. oxysporum with a 73.3% growth inhibition. The effects of various CvDE concentrations on F. oxysporum were found to be dosage dependent. The SEM micrograph revealed that CvDE-treated F. oxysporum had substantially less conidia than the control. The CvDE treatment damaged the mycelial structure as well. Major chemical components detected in CvDE were Heptaldehyde (15.7%), Octadecenoic acid, methyl ester (12.6%), Hexadecanoic acid (12%), 3-Decyn-2-Ol (10.98%), (E)-3,7,11,15-tetramethylhexadec-2-ene (9.76%), heptadecane-1,2,3,4,5-pentol (8.7%), Docosane, 4-methyl (7.28%).

Prevention of post-surgical adhesion bands by local administration of frankincense n-hexane extract.

Background: and purpose: The formation of postoperative intra-abdominal adhesion band formation may lead to severe complications. This study aimed to evaluate the preventive effect of local administration of frankincense n-hexane extract (FHE) on the formation of postsurgical adhesion bands. Materials and methods: FHE was extracted from the resin of a Boswellia sacra tree and its components were identified by gas chromatography-mass spectrometry (GC-MS). In an animal model, the expression levels of TNF-α and TGF-ß1 cytokines after application of FHE were assessed to check the inflammatory and fibrotic cues, respectively. Results: Following FHE compound analysis, in vivo experiments demonstrated that intraoperative local administration of FHE resulted in the prevention of adhesion band formation. The adhesion grades in the FHE-treated group were significantly lower than those in the negative control (NC) and the positive control (Interceed). The infiltration of inflammatory cells observed by histopathology revealed a significant anti-inflammatory potential of FHE. Furthermore, the gene expression results proved that significant suppression of TNF-α and TGF-ß1 was responsible for its antiadhesion properties. Conclusions: The study reported the potential of FHE as an ointment for the prevention of adhesion bands.

Puerarin improves skeletal muscle strength by regulating gut microbiota in young adult rats.

Background: Sarcopenia is an age-related skeletal muscle dysfunction syndrome that is lacking validated treatments. Maximizing muscle strength in young adulthood may be a promising way to prevent sarcopenia in the elderly. The phytomolecule puerarin has been extensively used in clinical practice and reported to increase energy metabolism in skeletal muscle by directly targeting the skeletal muscle fiber. However, the bioavailability of puerarin is very poor, and almost 93% of puerarin stays in the intestine until excretion. Therefore, we hypothesize that puerarin may regulate gut microbiota to improve skeletal muscle strength and/or mass in adults. Methods: Twenty three-month old male Sprague Dawley rats were divided into two groups according to average weights, puerarin group (puerarin dissolved in 0.5% CMC-Na, 150 â€‹mg/kg/day, N â€‹= â€‹10), and control group (equal volume 0.5% CMC-Na, N â€‹= â€‹10). The treatment lasted for 8 weeks. Muscle weight, muscle fiber types and cross-sectional area (CSA), ex vivo muscle contraction test and grip strength were measured. 16S rDNA sequencing was employed to evaluate the gut microbiota composition in the sample of cecal content. Short-chain fatty acids (SCFAs) in cecal and serum were analyzed by gas chromatography-mass spectrometry. Adenosine triphosphate (ATP) concentration in skeletal muscle was also detected. Pearson's correlation was used to analyze the relations between SCFAs, ATP concentration and muscle function. Results: After puerarin treatment, grip strength, the specific twitch force, and the tetanic forces in the soleus (SOL) and extensor digitorum longus (EDL) muscle were significantly higher than those of the control group. The percentage and CSA of type II muscle fiber in EDL was higher in the puerarin group than those in the control group. Puerarin treatment significantly changed the gut microbial constitutes. Two SCFAs-productive microbiota, the families Peptococcaceae and Closteridiales, were significantly higher in the puerarin group than those in the control group, while the ratio of Prevotellaceae/Bacteroidaceae (P/B), a muscle atrophy indicator, was lower in the puerarin group. As expected, there were significant linear correlations between the concentrations of SCFAs, including cecal total SCFAs, serum n-butyric acid and total SCFAs, and skeletal muscle strength and function, including the twitch force and tetanic force of SOL and EDL, as well as the forelimb grip strength. Conclusion: In conclusion, puerarin improved the forelimb grip strength and muscle contraction function in young adult rats. The underlying mechanism may include that puerarin increased SCFAs production by regulating gut microbiota, augmented ATP synthesis and skeletal muscle strength. The translational potential of this article : Our study finds that a clinical used phytomolecule puerarin has the potential of improving skeletal muscle strength in young adult rats. As puerarin has long-term clinical experience and shows good safety, it might be a potential candidate for developing muscle strengthening agents.

Detection of SARS-CoV-2 infection by exhaled breath spectral analysis: Introducing a ready-to-use point-of-care mass screening method.

BACKGROUND: The current SARS-CoV-2 pandemic created an urgent need for rapid, infection screening applied to large numbers of asymptomatic individuals. To date, nasal/throat swab polymerase chain reaction (PCR) is considered the "gold standard". However, this is inconducive to mass, point-of-care (POC) testing due to person discomfort during sampling and a prolonged result turnaround. Breath testing for disease specific organic compounds potentially offers a practical, rapid, non-invasive, POC solution. The study compares the Breath of Health, Ltd. (BOH) breath analysis system to PCR's ability to screen asymptomatic individuals for SARS-CoV-2 infection. The BOH system is mobile and combines Fourier-transform infrared (FTIR) spectroscopy with artificial intelligence (AI) to generate results within 2 min and 15 s. In contrast to prior SARS-CoV-2 breath analysis research, this study focuses on diagnosing SARS-CoV-2 via disease specific spectrometric profiles rather than through identifying the disease specific molecules. METHODS: Asymptomatic emergency room patients with suspected SARS-CoV-2 exposure in two leading Israeli hospitals were selected between February through April 2021. All were tested via nasal/throat-swab PCR and BOH breath analysis. In total, 297 patients were sampled (mean age 57·08 ± SD 18·86, 156 males, 139 females, 2 unknowns). Of these, 96 were PCR-positive (44 males, 50 females, 2 unknowns), 201 were PCR-negative (112 males, 89 females). One hundred samples were used for AI identification of SARS-CoV-2 distinguishing spectroscopic wave-number patterns and diagnostic algorithm creation. Algorithm validation was tested in 100 proof-of-concept samples (34 PCR-positive, 66 PCR-negative) by comparing PCR with AI algorithm-based breath-test results determined by a blinded medical expert. One hundred additional samples (12 true PCR-positive, 85 true PCR-negative, 3 confounder false PCR-positive [not included in the 297 total samples]) were evaluated by two blinded medical experts for further algorithm validation and inter-expert correlation. FINDINGS: The BOH system identified three distinguishing wave numbers for SARS-CoV-2 infection. In the first phase, the single expert identified the first 100 samples correctly, yielding a 1:1 FTIR/AI:PCR correlation. The two-expert second-phase also yielded 1:1 FTIR/AI:PCR correlation for 97 non-confounders and null correlation for the 3 confounders. Inter-expert correlation was 1:1 for all results. In total, the FTIR/AI algorithm demonstrated 100% sensitivity and specificity for SARS-CoV-2 detection when compared with PCR. INTERPRETATION: The SARS-CoV-2 method of breath analysis via FTIR with AI-based algorithm demonstrated high PCR correlation in screening for asymptomatic individuals. This is the first practical, rapid, POC breath analysis solution with such high PCR correlation in asymptomatic individuals. Further validation is required with a larger sample size. FUNDING: Breath of Health Ltd, Rehovot, Israel provided study funding.

Development and application of a High-Resolution mass spectrometry method for the detection of fentanyl analogs in urine and serum.

Introduction: The use of illicitly manufactured synthetic opioids, specifically fentanyl and its analogs, has escalated exponentially in the United States over the last decade. Due to the targeted nature of drug detection methods in clinical laboratories and the ever-evolving list of synthetic opioids of concern, alternative analytical approaches are needed. Methods: Using the fentanyl analog screening (FAS) kit produced by the Centers for Disease Control and Prevention (CDC), we developed a liquid chromatography-high resolution mass spectrometry (LC-HRMS) synthetic opioid spectral library and data acquisition method using information dependent acquisition of product ion spectra. Chromatographic retention times, limits of detection and matrix effects, in urine and serum, for the synthetic opioids in the FAS kit (n = 150) were established. All urine and serum specimens sent to a clinical toxicology laboratory for comprehensive drug testing in 2019 (n = 856) and 2021 (n = 878) were analyzed with the FAS LC-HRMS library to determine the prevalence of fentanyl analogs and other synthetic opioids, retrospectively (2019) and prospectively (2021). Results: The limit of detection (LOD) of each opioid ranged from 1 to 10 ng/mL (median, 2.5 ng/mL) in urine and 0.25-2.5 ng/mL (median, 0.5 ng/mL) in serum. Matrix effects ranged from -79 % to 86 % (median, -37 %) for urine, following dilution and direct analysis, and -80 % to 400 % (median, 0 %) for serum, following protein precipitation. The prevalence of fentanyl/fentanyl analogs in serum samples increased slightly from 2019 to 2021 while it remained the same in urine. There were only 2 samples identified that contained a fentanyl analog without the co-occurrence of fentanyl or fentanyl metabolites. Analysis of the established MS/MS spectral library revealed characteristic fragmentation patterns in most fentanyl analogs, which can be used for structure elucidation and drug identification of future analogs. Conclusions: The LC-HRMS method was capable of detecting fentanyl analogs in routine samples sent for comprehensive drug testing. The method can be adapted to accommodate testing needs for the evolving opioid epidemic.

Analysis of key precursor peptides and flavor components of flaxseed derived Maillard reaction products based on iBAQ mass spectrometry and molecular sensory science.

Flaxseed derived Maillard reaction products (MRPs) have typical meaty flavor, but there is no report on comparison of their amino acids and peptides reactivity. The peptides and amino acids of flaxseed protein hydrolysates were separately collected by G-15 gel chromatography. Taste dilution analysis (TDA) showed that peptides-MRPs had high umami, mouthfulness, and continuity enhancement. Further, LC-MS/MS revealed that flaxseed protein hydrolysates consumed 41 peptides after Maillard reaction. Particularly, DLSFIP (Asp-Leu-Ser-Phe-Ile-Pro) and ELPGSP (Glu-Leu-Pro-Gly-Ser-Pro) accounted for 42.22% and 20.41% of total consumption, respectively. Aroma extract dilution analysis (AEDA) indicated that formation of sulfur-containing flavors was dependent on cysteine, while peptides were more reactive than amino acids for nitrogen-containing heterocycles. On the other hand, 11 flavor compounds with flavor dilution (FD) ≥ 64 were identified for flaxseed derived MRPs, such as 2-methylthiophene, 2-methyl-3-furanthiol, furfural, 2-furfurylthiol, 3-thiophenethiol, thieno[3,2-b] thiophene, 2,5-thiophenedicarboxaldehyde, 2-methylthieno[2,3-b] thiophene, 1-(2-methyl-3-furylthio)-ethanethiol, 2-methylthieno[3,2-b] thiophene, and bis(2-methyl-3-furyl)-disulfide. In addition, we further demonstrated the flavors formation mechanism of flaxseed derived MRPs.

Influence of seasonality on the physicochemical properties of Moringa oleifera Lam. Seed oil and their oleochemical potential.

The effect of abiotic factors, such as weather and climate can alter the properties of vegetable oils. In this work, the effects of the refining process and pre-drying of Moringa oleifera Lam. seeds collected in the dry and rainy seasons (seasonality) were evaluated on the characteristics of the extracted oils. The refined and crude dry season oils had lower acidity and moisture content than the dry and raw rainy season oils. Oleic acid (C18:1) showed the highest concentration in the different Moringa oils studied. The results suggest that dry season oils are more suitable for application as feedstocks in the cosmetic, food, medicinal and pharmaceutical industries. Furthermore, refining proved to be efficient in removing free fatty acids and in lightening the oil.

Chemical constituents of green teas processed from albino tea cultivars with white and yellow shoots.

Green tea processed from albino tea varieties often has umami taste and fresh aroma. This study identified green teas made from two types of albino tea cultivar, one having the white shoots (called Naibai, NB) and the other having the yellow shoots (called Huangjinya, HJY). Taste compounds analyses showed that galloylated catechins were highly concentrated in HJY green teas, whereas non-galloylated catechins and amino acids were more abundant in NB green teas. CsTA (involved in the catabolism of galloylated catechins) showed high expression in HJY tea shoots, resulting in gallic acid as a precursor for ß-glucogallin biosynthesis being abundant in HJY. CsPDX2.1 (responsible for theanine hydrolyzation) had a lower expression level in NB than HJY shoots. Fatty acid-derived volatiles (FADVs), glycosidically bound volatiles (GBVs) and carotenoid-derived volatiles (CDVs) were highly concentrated in HJY green teas, whereas amino acids-derived volatiles were highly concentrated in NB green teas.

Aroma classification and characterization of Lactobacillus delbrueckii subsp. bulgaricus fermented milk.

The aroma of the fermented milk produced by twenty-eight Lactobacillus delbrueckii subsp. bulgaricus strains was evaluated via quantitative descriptive analysis. According to the sensory analysis results, the fermented milks were grouped into milky-type, cheesy-type, fermented-type and miscellaneous-type. The representative samples of cheese-type and fermented-type were analyzed by headspace-gas chromatography-ion mobility spectrometry (HS-GC-IMS) and flavoromics. A total of 95 volatile compounds were identified and particularly, 12 aroma-active compounds were detected by using gas chromatography-olfactometry-mass spectrometry (GC-O-MS). Among the different aroma types, 2,3-butanedione, δ-decalactone, acetaldehyde, butanoic acid, acetic acid and hexanoic acid were finally screened out as the key aroma-active compounds by quantitative and odor activity value (OAV) analysis combined with aroma recombination, omission and addition experiments. These findings were valuable in developing specific fermented milk products with different aroma profiles.

Quantitation of non-derivatized free amino acids for detecting inborn errors of metabolism by incorporating mixed-mode chromatography with tandem mass spectrometry.

Introduction: Amino acids are critical biomarkers for many inborn errors of metabolism, but amino acid analysis is challenging due to the range of chemical properties inherent in these small molecules. Techniques are available for amino acid analysis, but they can suffer from long run times, laborious derivatization, and/or poor resolution of isobaric compounds. Objective: To develop and validate a method for the quantitation of a non-derivatized free amino acid profile in both plasma and urine samples using mixed-mode chromatography and tandem mass spectrometry. Methods: Chromatographic conditions were optimized to separate leucine, isoleucine, and allo-isoleucine and maintain analytical runtime at less than 15 min. Sample preparation included a quick protein precipitation followed by LC-MS/MS analysis. Matrix effects, interferences, linearity, carryover, acceptable dilution limits, precision, accuracy, and stability were evaluated in both plasma and urine specimen types. Results: A total of 38 amino acids and related compounds were successfully quantitated with this method. In addition, argininosuccinic acid was qualitatively analyzed. A full clinical validation was performed that included method comparison to a reference laboratory for plasma and urine with Deming regression slopes ranging from 0.38 to 1.26. Conclusion: This method represents an alternative to derivatization-based methods, especially in urine samples where interference from metabolites and medications is prevalent.

Antioxidant and antidiabetic compounds identification in several Indonesian underutilized Zingiberaceae spices using SPME-GC/MS-based volatilomics and in silico methods.

This study aimed to identify compounds in 12 minor Zingiberaceae spices grown in Indonesia linked with in vitro α-glucosidase inhibitor and antioxidant (DPPH, FRAP, CUPRAC) activities using SPME-GC/MS volatilomics. The results illustrated that Zingiber aromaticum Val., Alpinia malaccensis (Burm.f.) Roscoe, Amomum compactum Sol. ex Maton, and Zingiber purpureum Roscoe had the highest α-glucosidase inhibitor and DPPH, FRAP, CUPRAC antioxidant activities, respectively. Also, the total phenolic content positively influenced DPPH, FRAP, and CUPRAC antioxidant activities. The strongest positive correlation with α-glucosidase inhibitor and DPPH antioxidant activities was found in eucalyptol; whereas o-cymene and terpinen-4-ol had the strongest correlations with FRAP and CUPRAC antioxidants, respectively. Furthermore, the molecular docking analysis revealed that all compounds with a strong correlation with α-glucosidase inhibitor activity (based on their OPLS VIP score) had binding energies (-5.06 - -6.26 kcal/mol) close to Acarbose (-10.11 kcal/mol). Thus, this study provided vital information on the volatile compounds in underutilized spices associated with their health beneficial properties.