Details concerning the determination of the serum metabolome and the results related to feeding and parity are given in Pacífico et al. (4 (link)). Briefly, 10 μl of aliquots of serum samples were processed using a targeted metabolomics approach based on the Biocrates MxP® Quant 500 kit (Biocrates Life Sciences AG, Innsbruck, Austria). Analysis of serum metabolome as well as of reference standards and quality controls (provided by the manufacturer) was carried out by ultra-high-performance liquid chromatography (uHPLC) and flow injection analysis (FIA), both coupled to tandem mass spectrometry. An Agilent 1290 series UHPLC system coupled to a 6500+ QTrap mass spectrometer equipped with an Ion-Drive Turbo V® ESI source (both Sciex, Foster City, CA, USA) was used for the analysis. Chromatographic and mass spectrometric parameters were set according to manufacturer's instructions. Data analysis was carried out in Analyst 1.6.3 (Sciex) for LC-MS/MS data and in the Biocrates MetIDQ software for FIA-MS/MS data.
Mxp quant 500 kit
Manufactured by Biocrates
Sourced in Austria, Germany, United States
The MxP® Quant 500 kit is a targeted metabolomics assay developed by Biocrates. It is designed to quantify up to 500 metabolites simultaneously from a single sample. The kit utilizes mass spectrometry technology to provide reliable and accurate measurement of a wide range of metabolites, including amino acids, lipids, and other small molecules.
Automatically generated - may contain errors
Lab products found in correlation
Metidq software, by Biocrates (5 mentions)
5500 qtrap instrument triple quadrupole mass spectrometer, by AB Sciex (3 mentions)
Qtrap 5500, by AB Sciex (3 mentions)
Analyst 1, by AB Sciex (2 mentions)
Metidq, by Biocrates (2 mentions)
Qtrap 5500 mass spectrometer, by AB Sciex (2 mentions)
1290 infinity 2 lc, by AB Sciex (2 mentions)
Sciex 5500 qtrap mass spectrometer, by AB Sciex (2 mentions)
Omni bead ruptor 24, by Omni International (2 mentions)
Kit running solvent, by Biocrates (2 mentions)
Xevo tq xs mass spectrometer, by Waters Corporation (2 mentions)
Agilent 1290 series uhplc system, by Agilent Technologies (1 mentions)
Qtrap 6500 mass spectrometer, by AB Sciex (1 mentions)
6 500 qtrap instrument, by AB Sciex (1 mentions)
Sciex analyst, by AB Sciex (1 mentions)
5500 qtrap instrument, by AB Sciex (1 mentions)
Sil htc, by Shimadzu (1 mentions)
Api 4000, by AB Sciex (1 mentions)
Absoluteidq p180 kit, by Biocrates (1 mentions)
Agilent 1290 infinity 2, by Agilent Technologies (1 mentions)
43 protocols using mxp quant 500 kit
1
Serum Metabolomics Analysis of Feeding and Parity
2
Targeted Metabolomic Analysis of Plasma
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The targeted metabolomic analysis of plasma samples was carried out by tandem mass spectrometry with the Biocrates MxP® Quant 500 Kit (Biocrates, Innsbruck, Austria). Six hundred and thirty endogenous metabolites belonging to 26 biochemical classes, including amino acids, fatty acids, lysophosphatidylcholines, and phosphatidylcholines, have been quantified in the kit. A detailed description of experimental metabolomics measurement techniques can be found at https://patents.google.com/patent/ EP1875401B1 and https://patents.google.com/patent/ EP1897014B1.
In brief, sample preparation was conducted using a 96-well device to quantify metabolite profiles. In this device, inserts have been impregnated with internal standards. And, a predefined sample amount has been added. Next, some analytes were derivatized with a phenyl isothiocyanate (PITC) solution, and after derivatization, the analytes were extracted by organic solvents and then diluted. Finally, the extracts were examined by Liquid chromatography–mass spectrometry (LC–MS/MS) and Flow Injection Analysis Tandem Mass Spectrometry (FIA-MS/MS) methods via a 6,500 QTRAP® instrument (AB Sciex, Singapore).
In brief, sample preparation was conducted using a 96-well device to quantify metabolite profiles. In this device, inserts have been impregnated with internal standards. And, a predefined sample amount has been added. Next, some analytes were derivatized with a phenyl isothiocyanate (PITC) solution, and after derivatization, the analytes were extracted by organic solvents and then diluted. Finally, the extracts were examined by Liquid chromatography–mass spectrometry (LC–MS/MS) and Flow Injection Analysis Tandem Mass Spectrometry (FIA-MS/MS) methods via a 6,500 QTRAP® instrument (AB Sciex, Singapore).
3
Targeted Metabolomic Profiling of Serum
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Targeted metabolomics of serum samples collected from all participants between 24 and 48 h after diagnosis was conducted using Biocrates MxP® Quant 500 Kit (Biocrates, Innsbruck, Austria) measured by tandem mass spectrometry at Fraunhofer Institute for Toxicology and Experimental Medicine. Six hundred and thirty metabolites were assessed as part of the MetIDQ™ MetaboINDICATOR™ module designed specifically for MxP® Quant 500 kit data. Two hundred and two (202) metabolite indicators consisting of combinations of metabolite measurements that capture meaningful biological functions were also derived. Examples of such indicators include enzymatic activity based on substrate/product metabolite ratio and the sum of functionally/structurally similar metabolites. Flow injection Analysis Tandem Mass Spectrometry (FIA-MS/MS) was used to quantify lipids, and liquid chromatography-tandem mass spectrometry (LC-MS/MS) was used to quantify small molecules using 5500 QTRAP® instrument triple quadrupole mass spectrometer (AB Sciex, Darmstadt, Germany) as described previously (15 (link)). For the listed triacylglycerols, the “_” indicates that the positions (sn-1/sn-2/sn-3) of the fatty acid residues are unknown. Potential isomers are described by the manufacturer: https://biocrates.com/wp-content/uploads/2020/02/Biocrates_Q500_isomers_isobars.pdf .
4
Analyzing Gastric Metabolites After H. pylori Eradication
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We identified differential metabolites after H. pylori eradication utilizing the Biocrates MxP Quant 500 kit (Biocrates, Innsbruck, Austria), which is capable of quantifying 630 metabolites across the metabolic spectrum and based on a quality-controlled, highly reproducible analysis platform. The frozen gastric biopsy specimens were weighed and homogenized according to the guidelines of the manufacturer. The homogenates were centrifuged at 2 to 4°C. Supernatants were transferred to a 96-well-based Biocrates sample preparation plate which was impregnated with internal standards and dried under a nitrogen stream. Then 5% phenylisothiocyanate (PITC) solution was added for derivatization. After the derivatization, the target analytes were extracted with an organic solvent. The obtained extracts were analyzed for lipids by flow injection analysis tandem mass spectrometry (FIA-MS/MS) and small molecules by liquid chromatography-tandem mass spectrometry (LC-MS/MS).
5
Serum Metabolomics Using MxP Quant 500
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Targeted metabolomics of serum samples from participants was conducted using Biocrates MxP® Quant 500 Kit (Biocrates, Innsbruck, Austria) measured by tandem mass spectrometry at Fraunhofer Institute for Toxicology and Experimental Medicine. Six hundred and thirty metabolites were assessed as part of the MetIDQ™ MetaboINDICATOR™ module designed specifically for MxP® Quant 500 kit data. A total of 232 metabolite indicators, consisting of combinations of metabolite measurements that capture meaningful biological functions, were also derived. Examples of such indicators include enzymatic activity based on substrate/product metabolite ratio and the sum of functionally/structurally similar metabolites. A full list of metabolite indicators can be found in Supplementary File 1
6
Targeted Plasma Metabolomics by LC-MS/MS
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Tandem mass spectrometry was performed for targeted metabolomics of plasma samples using Biocrates MxP® Quant 500 Kit (Biocrates, Innsbruck, Austria) at the Fraunhofer Institute for Toxicology and Experimental Medicine (ITEM). FIA-MS/MS was utilized to measure lipids using a 5500 QTRAP® instrument with an ESI source (AB Sciex, Darmstadt, Germany). LC-MS/MS utilizing the same 5500 QTRAP® instrument was used for measuring small molecules as described previously (17 (link)). Appropriate MS software (Sciex Analyst®) was used to quantify data that was then imported into Biocrates MetIDQ™ software for calculation of analyte concentrations, assessment and compilation of data.
7
Serum Metabolomics Using Biocrates MxP Quant 500
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Using the Biocrates MxP® Quant 500 Kit (Biocrates, Innsbruck, Austria), assessed by tandem mass spectrometry at the Fraunhofer Institute for Toxicology and Experimental Medicine, targeted metabolomics was performed on serum samples obtained from all subjects between 24 and 48 h after diagnosis. As part of the MetIDQ™ MetaboINDICATOR™ module created especially for MxP® Quant 500 kit data, 630 metabolites were evaluated. Additionally, 202 metabolite-indicators were created from combinations of metabolite measurements that encapsulate significant biological processes, for example indicators of enzymatic activity (substrate/product metabolite ratio) and sum of functionally or structurally similar metabolites. Quantification of lipids and small molecules were performed using Flow injection Analysis Tandem Mass Spectrometry (FIA-MS/MS) and liquid chromatography-tandem mass spectrometry (LC-MS/MS) respectively, using 5500 QTRAP® instrument triple quadrupole mass spectrometer (AB Sciex, Darmstadt, Germany). The “_” signifies that the fatty acid residue locations (sn-1/sn-2/sn-3) for the listed triacylglycerols are unknown. Potential isomers are described by the manufacturer: https://biocrates.com/wp-content/uploads/2020/02/Biocrates_Q500_isomers_isobars.pdf .
8
Metabolomic Analysis of EV and Plasma
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Metabolomic analysis of 0.01 mL EV- or PFP sample was performed using a Biocrates MxP Quant 500 kit (Biocrates AG, Innsbruck, Austria) according to the manufacturer’s instructions, as detailed in the Online Supplementary Material. The analytical procedure was guided by the laboratory information management software Biocrates MetIDQ (version 7.13.11, DB109-Nitrogen-2850 Revision 31.995, Base Version 109, Runtime version 1.8.0_181, Runtime architecture amd64; Biocrates A.G., Innsbruck, Austria). Analytical system control and data acquisition were accomplished using Sciex Analyst version 1.5.3. The analytical results were evaluated by MetIDQ and results were analyzed in R74 . Metabolites were excluded from the analysis if less than half of the measurements (< 7 for CTRL and < 4 for VEH) were below the limit of detection (LOD) in both groups independently for EV and plasma samples. For the analytes included, values below the LOD were imputed with a normal distribution around half of the LOD. For correlation analysis, metabolite types for which at least twelve plasma-EV measurement pairs were available were analyzed with a linear regression model. For both linear regression and statistical analysis, data were log10 transformed.
9
Targeted Metabolomic Profiling in Biofluids
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A targeted metabolomic assessment was carried out following the manufacturer’s protocol using Biocrates’ MxP® Quant 500 kit (Biocrates, Innsbruck, Austria), which quantifies 630 distinct metabolites in human and rodent plasma (10 µl), human CSF (10 µl), and mouse brain (10 µl). For the mouse hypothalamus and hippocampus, tissue was homogenized following a previously published protocol [41 ] with slight modifications. Additional details can be found in the Supplementary Materials .
10
Targeted Metabolomics of Serum Samples
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The targeted metabolomics of serum samples collected from all participants within 24 to 48 hours after diagnosis was performed using Biocrates MxP® Quant 500 Kit (Biocrates, Innsbruck, Austria). Tandem mass spectrometry was performed at the Fraunhofer Institute for Toxicology and Experimental Medicine. We analyzed 630 metabolites as part of the MetIDQ™ MetaboINDICATOR™ module designed for MxP® Quant 500 kit. Lipid quantification was performed using Flow Injection Analysis Tandem Mass Spectrometry (FIA-MS/MS), and small molecule quantification was done using liquid chromatography-tandem mass spectrometry (LC-MS/MS) with the 5500 QTRAP® instrument triple quadrupole mass spectrometer (AB Sciex, Darmstadt, Germany) as previously described (32 (link), 33 (link)).
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