NIA Targeted Metabolomics Platform

About the Platform

The Biocrates MxP® Quant 500 kit, a targeted metabolomics assay, is run at the NIA site. The MxP® Quant 500 kit measures and reports quantitative values for specific small molecules and lipids – details, including the list of metabolites assayed, can be found here. Only those metabolites covered by the kit are measured and reported.

• Dozens of metabolites known to be synthesized and modulated by microbiota.
• Metabolites are reported with names and quantitative values.
• Up to 630 metabolites from 26 biochemical classes.

Other kits offered by Biocrates, including some that have more limited coverage and lower cost can also be run if preferred.

Kits come in different sizes (a 96-well plate kit has room for ~78 samples; a 56-well plate kit has room for ~40 samples), so please consult with the Core in selecting the appropriate kit for your project.

The Core needs payment from you to purchase kit(s) from Biocrates to run your samples. As of November 2nd, 2022, the cost for the 96-well MxP® Quant 500 kit is $7,841 (78 sample kit) and $5,175 for (40 sample kit). Alternative options include the Absolute IDQ® p180 kit which costs $5,500 for a 78 sample kit and $3,631 for a 40 samples kit.

We recognize that you may not wish to purchase a full kit as you may not have enough samples to warrant it. If your project does not fill a kit, the Core will attempt to merge your samples with those from other projects to limit cost to you (through cost-sharing with the other investigators). We will discuss logistics on a case-by-case basis, but, because of this, if you do not wish to purchase a full kit, the cost per sample and timing of your project run may vary somewhat depending on the availability of other samples.

Analytical Approach

Metabolites are measured using a Nexera HPLC system (Shimadzu) coupled to a 6500 QTRAP® mass spectrometer (AB Sciex) with an electrospray ionization source. Lipids, acyl carnitines, and hexoses are measured over two injections by flow injection analysis-tandem mass spectrometry (FIA-MS/MS) in positive ionization modes. Amino acids, amino acid related, carboxylic acids, fatty acids, indole derivatives, biogenic amines, bile acids, cresols, alkaloids, amine oxides, hormones, vitamins and cofactors, and nucleobases related metabolites are measured by liquid chromatography-tandem mass spectrometry (LC–MS/MS) from two injections in positive and negative ionization modes. Data from all injections are imported and quantified in Biocrates MetIDQ™ software. Analytes in the LC-MS/MS part are quantified using either external 7-point calibration curves with labeled standards or internally with labeled standards. Analytes in the FIA-MS/MS part are quantified using internal standards.

Accepted Samples

The types of samples accepted for analysis are listed below, along with the recommended (and minimum) material required. Additional sample types are welcome but will be accepted after consultation, on a case-by-case basis. Further, if minimum material amounts cannot be met, please contact us for guidance. Replicates and additional important technical issues relating to sample preparation can be discussed during initial project consultation.

Sample Types	Minimum Material	Recommended Material
Biofluids
Serum or Plasma	20 microliters	50 microliters
CSF	30 microliters	50 microliters
Urine	30 microliters	50 microliters
Cells mammalian cells (cell lysates)	2 – 3 million	5 million
Tissues
Brain, Muscle	~10 mg tissue	30 – 50 mg tissue

What is Included?

• All projects include consultation on experimental design. Pilot samples included at no cost at the discretion of the Core.
• Analytical measurement of samples by UHPLC-HRMS/MS.
• Sample preparation handled by the sample submitter (protocols will be provided) or by Core personnel on a case-by-case basis.
• Processing of raw data via formatting, quality control, and data cleaning.

Conditions

• Samples must be prepared according to Metabolomics Core specifications. Core reserves the right to refuse samples which do not adhere to guidance.
• We kindly request that you consider co-authorship or acknowledgement in any resulting work (manuscript, presentation, etc.)

Contact

All initial project consultations should be directed to the Trans-NIH Metabolomics Core Coordinator, Kirsten E. Overdahl, Ph.D.

Ruin Moaddel, Ph.D.
Staff Scientist

Tel 410-558-8294
moaddelru@mail.nih.gov

Kirsten E. Overdahl, Ph.D.
Chemist

Tel 984-287-4562
kirsten.overdahl@nih.gov