The plasma with the endogenous metabolites was analyzed with a targeted quality, and quantitative-controlled metabolomics approach using the AbsoluteIDQ® p180 Kit (Biocrates Life Sciences AG, Innsbruck), as published previously [28 (link), 29 (link)]. With this kit up to 188 endogenous metabolites can be quantified, including 21 biogenic amines, 21 amino acids, 90 glycerophospholipids, 40 acylcarnitines, 15 sphingolipids, and hexoses. Liquid chromatography tandem mass spectrometry (LC–MS/MS) was applied to detect amino acids and biogenic amines, whereas using flow injection analysis tandem mass spectrometry (FIA-MS/MS) we could quantify acylcarnitines, sphingolipids, glycerophospholipids, and hexoses. Sample preparation was performed according to the user manual of the kit. Samples were randomized, including multiple quality control samples in the measurement sequence. The complete analytical process (sample registration, work list generation, and data processing) was carried out using the Biocrates MetIDQ™ software, version Boron 2693 (Biocrates Life Sciences AG, Innsbruck), which is an integral part of the AbsoluteIDQ® p180 Kit. Metabolite concentrations were calculated using the MetIDQ™ software and reported in μmol/l.
Absoluteidq p180 kit
Manufactured by Biocrates
Sourced in Austria, United States, Japan, Germany
The AbsoluteIDQ p180 kit is a targeted metabolomics assay developed by Biocrates. The kit provides a quantitative analysis of up to 188 metabolites from various chemical classes, including acylcarnitines, amino acids, biogenic amines, and lipids. The kit utilizes flow injection analysis-tandem mass spectrometry (FIA-MS/MS) technology to enable the simultaneous measurement of these metabolites in a single analysis.
Automatically generated - may contain errors
Lab products found in correlation
Metidq software, by Biocrates (17 mentions)
Acquity uplc, by Waters Corporation (9 mentions)
Qtrap 5500 mass spectrometer, by AB Sciex (9 mentions)
Analyst 1, by AB Sciex (7 mentions)
Tq s triple quadrupole ms ms, by Waters Corporation (6 mentions)
Pre column securityguard, by Phenomenex (5 mentions)
Qtrap 4500, by AB Sciex (5 mentions)
Api 4000, by AB Sciex (5 mentions)
Qtrap 5500, by AB Sciex (5 mentions)
Agilent 1260 infinity binary lc, by Agilent Technologies (4 mentions)
1260 series hplc, by Agilent Technologies (4 mentions)
Formic acid, by Merck Group (4 mentions)
Pyridine, by Merck Group (4 mentions)
Xevo tq s mass spectrometer, by Waters Corporation (3 mentions)
Triple quad 4500, by AB Sciex (3 mentions)
Surveyor hplc system, by Thermo Fisher Scientific (3 mentions)
Metidq version 5 4 8 db100 boron 2607, by Biocrates (3 mentions)
Sil htc, by Shimadzu (2 mentions)
Zorbax eclipse xdb c18, by Agilent Technologies (2 mentions)
Absciex 5500 qtrap, by Waters Corporation (2 mentions)
216 protocols using absoluteidq p180 kit
1
Targeted Metabolomic Analysis of Plasma
2
Targeted Metabolic Profiling by LC-MS/MS
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
A targeted quantitative approach using a combined direct-flow injection and
liquid chromatography tandem MS/MS assay (AbsoluteIDQ® p180 kit, Biocrates,
Innsbruck, Austria) was applied as previously published published19 (link). The method of AbsoluteIDQ® p180 kit conforms with the US Food and
Drug Administration Guideline ‘Guidance for industry: bioanalytical method
validation,’ which implies proof of reproducibility within a given error
range. The method combines derivatization and extraction of analytes with
the selective mass-spectrometric detection using multiple
reaction-monitoring pairs. Isotope-labeled internal standards are integrated
into the platform for metabolite absolute quantification. This strategy
allows simultaneous quantification of 186 metabolites (40 amino acids and
biogenic amines, 40 acylcarnitines, 90 glycerophospholipids, 15
sphingomyelins, 1 monosaccharide). The list of measurable metabolites using
the Biocrates Absolute IDQ® p180 kit and their biological relevance is
provided inTable S1 . Significant metabolite changes were evaluated
using univariate pairwise comparison Mann–Whitney–Wilcoxon test (JMP pro12,
SAS).
liquid chromatography tandem MS/MS assay (AbsoluteIDQ® p180 kit, Biocrates,
Innsbruck, Austria) was applied as previously published published19 (link). The method of AbsoluteIDQ® p180 kit conforms with the US Food and
Drug Administration Guideline ‘Guidance for industry: bioanalytical method
validation,’ which implies proof of reproducibility within a given error
range. The method combines derivatization and extraction of analytes with
the selective mass-spectrometric detection using multiple
reaction-monitoring pairs. Isotope-labeled internal standards are integrated
into the platform for metabolite absolute quantification. This strategy
allows simultaneous quantification of 186 metabolites (40 amino acids and
biogenic amines, 40 acylcarnitines, 90 glycerophospholipids, 15
sphingomyelins, 1 monosaccharide). The list of measurable metabolites using
the Biocrates Absolute IDQ® p180 kit and their biological relevance is
provided in
using univariate pairwise comparison Mann–Whitney–Wilcoxon test (JMP pro12,
SAS).
3
Absolute IDQ Metabolite Quantification
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Serum samples from participants in the KORA FF4 study were measured with the AbsoluteIDQTM p180 Kit (BIOCRATES Life Sciences AG, Innsbruck, Austria). Metabolite concentrations were adjusted for plate normalization factors (NFs) to minimize the plate effect. For each metabolite, the plate NFs were calculated by dividing the mean of reference samples in each plate with the mean of all reference samples in all measured plates. Metabolite concentrations were natural-log transformed and scaled to a mean value of zero and standard deviation (SD) of one to ensure comparability between the metabolites.
In the mouse study, creatinine, SM C18:1 and PC aa C38:0 values in plasma, liver, lung, adrenal glands, adipose tissue, cerebellum, and testis samples were determined with the AbsoluteIDQTM p180 Kit (BIOCRATES Life Sciences AG, Innsbruck, Austria) and in urine with the AbsoluteIDQTM p150 Kit (BIOCRATES). Tissue homogenization, extraction solvents, assay preparation, and LC-MS/MS measurements have been described elsewhere [55 (link)]. Since each tissue sample from db/db and WT mice was measured on the same kit plate, we did not conduct plate correction. Metabolite concentrations were natural-log transformed and then scaled to a mean value of zero and SD of one for each tissue.
In the mouse study, creatinine, SM C18:1 and PC aa C38:0 values in plasma, liver, lung, adrenal glands, adipose tissue, cerebellum, and testis samples were determined with the AbsoluteIDQTM p180 Kit (BIOCRATES Life Sciences AG, Innsbruck, Austria) and in urine with the AbsoluteIDQTM p150 Kit (BIOCRATES). Tissue homogenization, extraction solvents, assay preparation, and LC-MS/MS measurements have been described elsewhere [55 (link)]. Since each tissue sample from db/db and WT mice was measured on the same kit plate, we did not conduct plate correction. Metabolite concentrations were natural-log transformed and then scaled to a mean value of zero and SD of one for each tissue.
4
Bile Acid Profiling from Liver Samples
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The Bile Acids Kit (Biocrates Life Sciences AG) was used as described in the manufacturer's instructions. Metabolites were extracted from liver samples by adding H 2 O/acetonitrile (1:1, v/v) per mg of sample, followed by homogenization with a tissue slicer for 10 min at 30 Hz with four steel beads. Samples were centrifuged at 1400g for 2 min, and the supernatant was analyzed. The targeted analysis was performed by adding 10 l of extracted serum samples to the AbsoluteIDQ p180 Kit (Biocrates Life Science AG), following the vendor's instructions (68) .
5
Targeted Metabolomics of CSF and Serum
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
CSF metabolite concentrations were measured on a triple-quadrupole mass spectrometer (API4000, Sciex, Framingham, MA, USA) with an electrospray-ionization ion source coupled to a high-performance liquid chromatography system (SIL-HTc, Shimadzu, Japan) and the AbsoluteIDQ™ p180 kit and MetIDQ™ software (Biocrates Life Sciences, Innsbruck, Austria), as described in detail in [17 (link)]. Serum concentrations of phosphatidylcholines were measured on an AB SCIEX 5500 QTrap™ mass spectrometer (AB SCIEX, Darmstadt, Germany) using the MxP™ Quant 500 kit (Biocrates), following the manufacturer’s protocols (https://biocrates.com/mxp-quant-500-kit , accessed on 15 December 2020). Details about internal standards and quality controls (QC) are given in the manufacturer’s application note [22 ]. The Quant 500 kit measures the same glycerophospholipids as the p180 kit and is downward compatible with it, ensuring comparability of the data.
6
Quantitative Metabolite Analysis in Serum
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Quantitative analysis of other metabolites in serum samples, including amino acids, biogenic amines, acylcarnitines, phosphatidylcholines, lysophosphatidylcholines and sphingolipids was performed using the Biocrates AbsoluteIDQ® p180 kit, according to the manufacturer guidelines19 (link). Samples were analysed using flow injection analysis (FIA)-MS/MS and LC-MS/MS for different metabolite groups.
7
Metabolite Profiling of HCT116 Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
After compound treatment for 24 h at 37 °C and 21% O2 (n=3), HCT116 cells were washed twice with cold sodium chloride (0.9%) and incubated for 15 min with 1 ml 80% methanol at −80 °C. Subsequently, cells were harvested using a cell scraper and transferred together with the methanol into a new tube. Wells were washed once with 500 μl 80% methanol and added into the same tube. The resulting extracts were freeze-dried and resolved in 100 μl 100% methanol followed by a centrifugation step. Later, 10 μl of these extracts were used for target metabolite profiling by using the LC-MS based AbsoluteIDQ p180 Kit (Biocrates Life Sciences AG, Innsbruck, Austria). All samples were processed according to the manufacturer's instruction, and measurements were performed with a UHPLC-MS/MS System (Shimadzu UHPLC, Shimadzu, Kyoto, Japan and Sciex 5500 mass spectrometer, Sciex, Framingham, MA, USA). Multivariate data analysis was carried out by using the Umetrics SIMCA-P software (MKS Data Analytics Solutions, Malmö, Sweden).
8
Targeted Metabolomics for Acylcarnitine Quantification
9
Bile Acid, Amino Acid, and SCFA Quantification
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The AbsoluteIDQ Bile Acid Kit (Biocrates Life Sciences AG) was used for bile acid analyses. Liquid chromatography-mass spectrometry (LC-MS/MS) measurements were carried out by MRM acquisition on a Waters Acquity UPLC System and a QTRAP 5500 (AB Sciex). Data were processed with Analyst Software (1.6.2) and MetIDQ Software (Biocrates Life Sciences AG). For measurements of amino acids and amines, the AbsoluteIDQ p180 Kit (Biocrates Life Sciences AG) was used on a QTRAP mass spectrometer (MS) applying electrospray ionization (ESI) (ABI Sciex API5500Q-TRAP). After separation through a precolumn (Security Guard, Phenomenex, C18, 4 × 3 mm; Phenomenex) and hyphenated reverse phase column (Agilent, Zorbax Eclipse XDB C18, 3.0 × 100 mm, 3.5 µm), analytes were quantified by multi reaction monitoring (MRM) which was standardized by applying spiked-in isotopically labelled standards in positive and negative mode. For data processing, MetIQ software (Biocrates Life Sciences AG) was used. The isotope-labeled chemical derivatization method described by Han et al. [89 (link)] was modified for quantification of SCFA. SCFA were chromatographically separated on an Acquity UPLC BEH C18 column (1.7 μm) (Waters) using H2O (0.01% FA) and acetonitrile (0.01% FA). Analytes were quantified and identified by the scheduled MRM method.
10
Targeted Serum Metabolomics in Children
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!