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Glutathione in Whole Blood LC-MS/MS Analysis Kit

Glutathione is a natural tripeptide (γ-L-glutamyl-L-cysteinylglycine) which participates in several physiological processes such as DNA synthesis, protein synthesis, cell membrane stabilization, amino acid transportation and xenobiotic detoxification. Glutathione exists in reduced form (GSH) and in oxidized form as disulfide (GSSG). The GSH/GSSG ratio is often used as a marker of the cellular redox state. GSH is a critical indicator protecting organisms against diseases and toxicities. Thus, measurements of GSH concentrations in biological samples are of importance, owing to its potential for disease diagnosis and providing valuable information for the understanding of GSH homeostasis.

Highlights

For the determination of total glutathionine level, a simple reduction step is apllied prior to protein precipitation. No need of chemical derivatisation

Total run time is 9.5 min.

Safeguarded by its own stable labelled isotopic internal standard

Small volume of patient’s sample is required

Long life span of HPLC column

Parameters

Total Glutathione (GSH+GSSG), Glutathione (GSH), Glutathionedisulfide (GSSG), GSH/GSSG

Matrix

Whole Blood

Sample Preparation

  • Step 1
    (Total Glutathione)
    Pipette 20 μL of whole blood or quality control sample into a glass centrifuge tube then, add 40 μL of Reagent-1 and vortex for 15 sec. Next, incubate in a water bath at 60oC for 15 min.
  • Step 2
    Let the tube cool down to the room temp and dilute by adding 1940 μl of Reagent-2, vortex 3 sec.
  • Step 3
    Transfer 50 μL of diluted sample into a new tube. Then, add 25 μl of internal standard and 225 μl of Reagent-3 respectively, vortex 3 sec. Subsequently, centrifuge at 4000 rpm for 3 min.
  • Step 4
    Decant the supernatant into HPLC vial prior to injection

Sample Preparation

  • Step 1
    (Free Glutathione)
    Pipette 20 μL of whole blood or quality control sample into a glass centrifuge tube and add 1980 μl of Reagent-2, vortex 3 sec.
  • Step 2
    Transfer 50 μL of diluted sample into a new tube. Then, add 25 μl of internal standard and 225 μl of Reagent-3 respectively, vortex 3 sec. Subsequently, centrifuge at 4000 rpm for 3 min.
  • Step 3
    Decant the supernatant into HPLC vial prior to injection

Chromatogram

ADAPTING DIAGNOSTICS IN CHROMATOGRAPHY COUPLED MASS SPECTROMETRY..
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