Targeted metabolite analysis

Introduction:

Targeted analysis is quantitative analysis of known metabolites, drugs and their metabolites in blood, body fluids, cell line, human and animals. The metabolites are usually extracted with organic solvents and enriched/cleaned-up by solid-phase extraction and liquid-liquid extraction. Liquid chromatography is used to further separate the metabolites before mass spectrometric detection to reduce matrix interference. Triple quadrupole mass spectrometer under MRM mode of operation is commonly used in targeted analysis. A standard calibration curve is established with known standard metabolites of various concentrations for absolute quantification of the target compound.


Triple quadrupole mass spectrometer has very high detection sensitivity and specificity. The ThermoFisher TSQ Altis triple quadrupole mass spectrometer at BMSC has detection sensitivity at 0.01 ppb level, it can be used for the determination of trace amounts of metabolites from minute amounts of samples. BMSC will host a SCIEX 7500 ultra-high-sensitivity triple quadrupole mass spectrometer in the near future, which will boost sensitivity by another 1~2 order of magnitudes.

Sample submission:

To ensure accuracy of experimental results, the samples submitted should meet the following requirements: at least 100~200 μL for body fluids (serum, plasma, cerebrospinal fluid, lymph, urine, saliva, etc.) , at least 5×106~1×107 for cells, 30~200 mg for animal tissue; 100 mg~1 g for plant tissue, and 1×107~1×108 for microbe.


Please provide detailed information about the target metabolites and related literature, including but not limited to the name of the target compound, CAS number, structural formula, literature or determination method from other sources.


    Please provide the standard target compound with high purity. If you need BMSC’s assistance in procurement of the standard compounds, please wait patiently, as some compounds may have a long time of delivery.

Sample processing:

The samples such as blood, tissue, cell, and body fluid are extracted with an organic extraction solution (80%-100% methanol or acetonitrile). The metabolites are extracted by homogenization, ultrasonication, or shaking. After centrifuging, the supernatant can be directly analyzed (after dilution when needed) by mass spectrometry. The supernatant can also be concentrated or purified by solid phase extraction (SPE) before mass spectrometry analysis.




 


 

Result output:

 The concentrations of target metabolites are quantified with internal standard or external standard methods with default quantitative data analysis software from the instrument vendor. The results will be sent by email. If you need the original raw data, please download from data storage server at HPC core facility.