Standardized targeted metabolomics using the biocrates MxP® Quant 500 kit on Agilent 6495B and 6495C triple quadrupole LC/MS systems

Metabolomics provides crucial insights into physiological mechanisms in health and disease, integrating a combined read-out of genetics, environment, and lifestyle. Mass spectrometry is the technology of choice, allowing the quantification of metabolites with high selectivity and sensitivity. The range of mass spectrometry platforms is growing steadily, and so is our kit application portfolio. For the first time, the MxP® Quant 500 kit for broad, quantitative metabolic profiling is now available for Agilent triple quadrupole (TQ) instruments.

Over the last year, the MxP® Quant 500 kit became the leading metabolomics tool for broad metabolic profiling with a focus on host-microbiome-diet interaction. This ready-to-use kit provides a simple automated workflow for the quality-controlled analysis of 630 small molecules and lipids from 26 compound classes. With more than 230 predefined metabolite sums and ratios of metabolic pathways, it facilitates advanced biological data interpretation and increased statistical power. The MxP® Quant 500 kit is the most comprehensive metabolomics kit on the market – and now it’s also accessible to Agilent users.

Optimizing for excellence

The Agilent 6495B and 6495C TQ LC/MS systems are widely used instrument bases. To adapt the MxP® Quant 500 kit to these platforms, we’ve optimized all instrumental parameters, from sample preparation setup to mass spectrometric features. A comprehensive set of defined human plasma samples was used to validate the Agilent 6495B and 6495C TQ mass spectrometers, coupled to an Agilent 1290 Infinity II UHPLC. All 13 small molecule classes were analyzed by UHPLC-ESI-MS/MS, while hexoses and all 12 lipid classes were analyzed by FIA-MS/MS.

The results were impressive:
– All detected analytes had excellent intra- and inter-batch accuracy (85-115% for LC analytes, 80-120% for FIA analytes) and precision (CV <15% for LC analytes, CV <20% for FIA analytes).
– Highly comparable results were generated using the NIST standard reference material (SRM) 1950 across multiple 6495B and 6495C TQ LC/MS systems. This also allows for pooling of data between laboratories.
– In a typical human plasma sample, 540 metabolites and lipids were found to be routinely detected (>LOD) from a small sample volume of only 10 µl. We also saw impressive results testing other sample matrices, including human feces and mouse liver homogenate.

This means that Agilent platform users can rely on the MxP® Quant 500 kit for benefits such as:
– A ready-to-use solution including all consumables and instrument-specific acquisition methods
– An automated, software-guided workflow from sample registration to data interpretation
– Fast turnaround times to guarantee high throughput analyses
– Reliable standardized quantification of the full range of metabolites and lipids.

The MxP® Quant 500 kit combined with the Agilent 6495B or 6495C TQ LC/MS system provides high-quality results with excellent reproducibility. This robust solution provides a powerful tool for broad, quantitative metabolic profiling.

All details and results of the adaptation on the 6495C TQ LC/MS system can be found in the application note 35044.
To find out more about the MxP® Quant 500 kit, please visit the product page.