Chemical Composition of Commercial Cannabis.

Cannabis is widely used for medicinal and recreational purposes. As a result, there is increased interest in its chemical components and their physiological effects. However, current information on cannabis chemistry is often outdated or scattered across many books and journals. To address this issue, we used modern metabolomics techniques and modern bioinformatics techniques to compile a comprehensive list of >6000 chemical constituents in commercial cannabis. The metabolomics methods included a combination of high- and low-resolution liquid chromatography-mass spectrometry (MS), gas chromatography-MS, and inductively coupled plasma-MS. The bioinformatics methods included computer-aided text mining and computational genome-scale metabolic inference. This information, along with detailed compound descriptions, physicochemical data, known physiological effects, protein targets, and referential compound spectra, has been made available through a publicly accessible database called the Cannabis Compound Database (https://cannabisdatabase.ca). Such a centralized, open-access resource should prove to be quite useful for the cannabis community.

Evaluation of amino acids and other related metabolites levels in end-stage renal disease (ESRD) patients on hemodialysis by LC/MS/MS and GC/MS.

End-stage renal disease (ESRD) is a rapidly increasing health problem, and every year, about 2 million ESRD cases are reported worldwide. Hemodialysis (HD) is the vital renal reinstatement therapy for ESRD, and HD patterns play a crucial role in patients' health. Plasma metabolomics is the potential approach to understanding the HD process, effectiveness, and patterns. The lack of protein vitality is a primary problem for HD patients, and the quantities of amino acids intracellularly and in the blood are considered to be a symbolic index of protein metabolism and nutrition conditions. In the current study, LC/MS/MS and GC/MS methods were developed for 29 targeted plasma metabolites and validated as per ICH bioanalytical method validation M10 guidelines. The 29 metabolites included 20 proteinogenic amino acids and nine other related metabolites. The methods were employed to measure the absolute quantities (µM) of the targeted metabolites in HD patients (n=60) before and after dialysis (PRE-HD and POST-HD), and compared with the healthy control (HC) group (n=60). Phenylacetylglutamine was found to be higher in both PRE-HD (72.88±14.5 µM) and POST-HD (26.62±7.9 µM), when compared to HC (1.61±0.6 µM). On the other hand, glutamic acid was lower in PRE-HD (14.90±6.5 µM), and POST-HD (13.6±6.1 µM) than that of HC (245.4±37.8 µM). The dialytic loss was found to be 52-45% for arginine, lysine, and histidine, while it was 38-26% for glycine, cysteine, proline, alanine, threonine, glutamine, valine, and methionine. The dialytic loss was low (≤12%) for aspartic acid, glutamic acid, asparagine, leucine, tyrosine, tryptophan, and isoleucine. Graphical abstract adapted from mass spectrometry templates by Biorender.com retrieved from https://app.biorender.com/biorender-templates .

ESI-MS/MS analysis of protonated N-methyl amino acids and their immonium ions.

Methylation is one of the important posttranslational modifications of biological systems. At the metabolite level, the methylation process is expected to convert bioactive compounds such as amino acids, fatty acids, lipids, sugars, and other organic acids into their methylated forms. A few of the methylated amino acids are identified and have been proved as potential biomarkers for several metabolic disorders by using mass spectrometry-based metabolomics workstation. As it is possible to encounter all the N-methyl forms of the proteinogenic amino acids in plant/biological systems, it is essential to have analytical data of all N-methyl amino acids for their detection and identification. In earlier studies, we have reported the ESI-MS/MS data of all methylated proteinogenic amino acids, except that of mono-N-methyl amino acids. In this study, the N-methyl amino acids of all the amino acids (1-21; including one isomeric pair) were synthesized and characterized by ESI-MS/MS, LC/MS/MS, and HRMS. These data could be useful for detection and identification of N-methyl amino acids in biological systems for future metabolomics studies. The MS/MS spectra of [M + H]+ ions of most N-methyl amino acids showed respective immonium ions by the loss of (H2 O, CO). The other most common product ions detected were [MH-(NH2 CH3 ]+ , [MH-(RH)]+ (where R = side chain group) ions, and the selective structure indicative product ions due to side chain and N-methyl group. The isomeric/isobaric N-methyl amino acids could easily be differentiated by their distinct MS/MS spectra. Further, the MS/MS of immonium ions inferred side chain structure and methyl group on α-nitrogen of the N-methyl amino acids.

Transformation of polyphenols found in pigmented gluten-free flours during in vitro large intestinal fermentation.

In this work, 18 gluten-free flours (prepared from cereals, pseudocereals and legumes), differing in pigmentation, were screened for their phenolic profiles, cooked and, then, subjected to digestion and large intestinal fermentation in vitro. A combined targeted/untargeted metabolomic approach was used to elucidate the microbial biotransformation processes of polyphenols following digestion. This preliminary work demonstrated an increase in 3,5-dihydroxybenzoic acid (on average from 0.67 up to 1.30 µmol/g dry matter) throughout large intestinal fermentation of pseudocereals (esp. quinoa), due to their high alkylresorcinol contents. Isoflavones were converted into equol- or O-desmethylangolensin- derivatives, whereas anthocyanins were degraded into lower-molecular-weight phenolics (i.e., protocatechuic aldehyde and 4-hydroxybenzoic acid, with the latter exhibiting the highest increase over time). A decreasing trend was observed for antioxidant activities (i.e., FRAP and ORAC values) moving from digested to faecal fermented samples. These findings highlight that gluten-free flours are able to deliver bioaccessible polyphenols to the colon.

Edible nuts deliver polyphenols and their transformation products to the large intestine: An in vitro fermentation model combining targeted/untargeted metabolomics.

The fate of polyphenols from edible tree nuts was investigated using a simulated in vitro intestinal fermentation system. The digested food matrix was fermented for 48 h and the changes in the phenolic profiles were evaluated by both untargeted UHPLC-QTOF and targeted UHPLC-Orbitrap mass spectrometry. The untargeted metabolomics approach allowed us to monitor the comprehensive changes in phenolic profiles from 0 up to 48 h of in vitro fermentation. Multivariate statistics (i.e., orthogonal projection to latent structures discriminant analysis) applied to this untargeted data allowed us to identify the most discriminating phenolic metabolites and to further understand the colonic transformation pathways involved. In particular, 13 putatively identified compounds derived from flavonoids, lignans and phenolic acids were found to have the highest discrimination potential. Six phenolic metabolites were then quantified by means of targeted metabolomics (using a UHPLC-Orbitrap). These metabolites included 3,4-dihydroxyphenylacetic acid, 4-hydroxybenzoic acid, hippuric acid, caffeic acid, protocatechuic acid and protocatechuic aldehyde. Using the targeted data, a clear matrix effect was observed over time, with an increase of some phenolic metabolites moving from 8 to 48 h of in vitro fermentation. Based on these data, catabolic pathways for colonic microbial degradation of flavonoids, hydroxycinnamic acids, tyrosols and lignans are proposed. Our findings show that edible tree nuts deliver polyphenols to the colon, where several microbial transformations occur that lead to smaller phenolic metabolites being observed. Furthermore, we found that the combined use of targeted and untargeted metabolomics can be particularly effective for investigating the fate of polyphenols in the large intestine.

Prostate Cancer Associated Lipid Signatures in Serum Studied by ESI-Tandem Mass Spectrometryas Potential New Biomarkers.

Prostate cancer (PCa) is one amongst the most common cancersin western men. Incidence rate ofPCa is on the rise worldwide. The present study deals with theserum lipidome profiling of patients diagnosed with PCa to identify potential new biomarkers. We employed ESI-MS/MS and GC-MS for identification of significantly altered lipids in cancer patient's serum compared to controls. Lipidomic data revealed 24 lipids are significantly altered in cancer patinet's serum (n = 18) compared to normal (n = 18) with no history of PCa. By using hierarchical clustering and principal component analysis (PCA) we could clearly separate cancer patients from control group. Correlation and partition analysis along with Formal Concept Analysis (FCA) have identified that PC (39:6) and FA (22:3) could classify samples with higher certainty. Both the lipids, PC (39:6) and FA (22:3) could influence the cataloging of patients with 100% sensitivity (all 18 control samples are classified correctly) and 77.7% specificity (of 18 tumor samples 4 samples are misclassified) with p-value of 1.612×10-6 in Fischer's exact test. Further, we performed GC-MS to denote fatty acids altered in PCa patients and found that alpha-linolenic acid (ALA) levels are altered in PCa. We also performed an in vitro proliferation assay to determine the effect of ALA in survival of classical human PCa cell lines LNCaP and PC3. We hereby report that the altered lipids PC (39:6) and FA (22:3) offer a new set of biomarkers in addition to the existing diagnostic tests that could significantly improve sensitivity and specificity in PCa diagnosis.

Chemical profiling and anti-psoriatic activity of methanolic extract of Andrographis nallamalayana J.L.Ellis.

Andrographis nallamalayana is being widely used as tribal medicine in the treatment of leucoderma and mouth ulcers. Chemical profiling of methanolic extract of the whole plant (PE), using GC-MS and LC-MS, revealed the presence of compounds viz. α-tocopherol, ß-sitosterol, tetradecanoic acid, monostearin, flavones/flavanones and their glycosides, chromones, etc. Topical application of imiquimod on the dorsal portion of male BALB/C mice resulted in the development of psoriatic symptoms (erythema, scaling, thickening and folding) with a mean disease activity index (DAI) of >7.0. Topical treatment with 100-µL PE (~6.4%/12.8%) formulations, for 12-days, resulted in the alleviation of disease symptoms. Compared to water-based formulations, emu oil-based formulation, PE400EO was found more effective in reducing the mean DAI (>84%), keratinocyte count (>65%) (p < 0.01) and interleukin-22 (~70%) (p < 0.05). We report, for the first time, anti-psoriatic activity of A. nallamalayana having great potential in developing a potent phytomedicine against psoriasis.