ICU-metabolism.se  2010 

 

Analytical lab

 

In connection to the research group we have a analytical laboratory that is located in the Clinical Research Centre (KFC) of Karolinska University Hospital in Huddinge. In the lab we analyse samples from our mainly clinical metabolic studies. Incorporated into the laboratory is Karolinska Stable Isotope Core.

 

The lab is located in the lab-hotel part of KFC (Klinisk Forsknings Centrum or Clinical Research Centre) in the Novum building adjunct to the Karolinska University Hospital in Huddinge. There we have one office, one preparation lab and one equipment room. The equipment roon contains 2 HPLC systems and 2 GC-MS systems.

 

Within the lab we perform biochemical analyses to measure amino acid concentrations, glutahtione concentrations (both reduced and oxidised), metabolite concentrations (e.g. glucose, lactate, ATP, creatine phosphate etc.), mitochondrial respiration, mitochondrial enzyme activities (citrate synthase and complex I and IV), protease activities (26S proteasome, cathepsin, calpain etc.) and protein content. We also analyse gene expression but do this in collaboration with others. Occasionally we run Western blot analyses. BUT our speciality is the analyses of stable isotope tracer in biological material using mass spectrometry. Stable isotope tracers are used to quantify metabolic pathways in vivo in patients. After that these tracers are given to patinets or volunteers plasma or tissue samples are taken and these are analysed for stable isotope contents in our lab using GC-MS. We use stable isotope tracers to measure glucose turnover rates, whole body protein turnover, synthesis rates of immune cells, muscle tissue, albumin and mitochondria and turnover rates of specific amino acids (glutamine, leucine, phenylalanine). More information on the use of these tracer can be found on our tracer site.

 

We have started a project to add metabolomics measurements to the lab. We already have a HPLC method for quantifying all amino acids and have now a student from Dublin (Garret Rochford)establishing a GC-MS method for free fatty acids profiling. We have now bought and received a new toy (an UHPLC-MSMS) that will be used to expand our metabolomics analyses. First we will play around and will try to establish analyses of as many acyl-carnitines and fatty acids as possible. We will make sure we can quantify all our metabolomics analyses by using as many internal stable isotope labblelled standards as possible to buy. We will keep you updated on the process. To the left a picture of the new toy named Jamie.

 

 

 

Currently we have 4 people working in the lab permanently (Christina Hebert, Eva Nejman, Maria Klaude and Towe Jakobsson) and several students that come and do analyses occasionally. We have risk assessments for all routine methods.