I have started doing 1H-NMR for metabolite analysis in biological samples like cell lysate, mice tissue lysate, and mice serum. I am extracting the metabolites using 0.4M Perchloric acid which is further neutralized to pH 7.0 using 5M KOH. I am using 1 million cells, extraction is done in 200 uL volume and 100uL of the extract is used for NMR. The intensity of peaks in the spectra that i got after 1 hour of NMR (700 MHz) run was very low.
So, regarding same I have few question if anyone can respond. Is the no. of cells enough for such purpose? Will lyophilization based concentration of samples affect the metabolite profile in NMR? Is PCA extraction an appropriate method for extraction of hydrophilic metabolites? What is the most appropriate and efficient method for identification and absolute quantification of metabolites using NMR in both targeted and untargeted metabolomics?
Kindly check here www.tandfonline.com/doi/full/10.1080/17460441.2016.1189899?src=recsys
QTrap and triple TOF mass spectrometers can do both targeted and targeted metabolomic anaysis for you. No. of cells required for the analysis depends on the detector response of your analytes.
With regard to identification and quantification of metabolites with NMR spectroscopy have a look at our review:
Article Current Experimental, Bioinformatic and Statistical Methods ...