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Measurement of glutathione kinetics in neonates by liquid chromatography coupled to isotope ratio mass spectrometry (CAT#: STEM-ST-0073-LJX)

Introduction

A novel analytical method using liquid chromatography coupled to isotope ratio mass spectrometry (LC/IRMS) was developed for measuring the fractional synthesis rate (FSR) of glutathione (GSH) in neonates after infusion of [1-13C]-glycine as a tracer. After transformation of GSH into GSSG, its dimeric form, the intra-erythrocytic concentration and 13C-isotopic enrichment of GSH were determined using 200 µL of blood. The results showed that, using LC/IRMS, the concentration (range of µmol/mL) was reliably measured using norvaline as internal standard with precision better than 0.1 µmol/mL.




Principle

Isotope ratio mass spectrometry (IRMS) leverages magnetic sector mass spectrometry to enable high-precision measurement of the stable isotope content of a sample. Typical measurements target hydrogen, carbon, nitrogen, and oxygen analyses—although elements with masses up to and including sulfur can be measured. Solid, liquid, or gas phase samples are converted to simple gases then introduced to the IRMS. During analysis, an electron impact source ionizes sample-derived gas which is then accelerated down a flight tube, separated by mass, and quantified using a series of Faraday cups. The high precision of IRMS enables enumeration of even very small isotopic fractionation associated with physical, chemical, and biological transformations or natural abundance measurements.

Applications

For explaining the detailed molecular mechanisms behind biological processes
For understanding and quantifying nutrient and material exchanges between ecosystems
For providing ultra-precise stable isotope analyses
For understanding the geological history of the Earth
For food authenticity, forensic science, medical research and anti-doping testing

Procedure

1. Fill the reaction tube and install it, connect the gas path
2. Check for helium leaks
3. Heat up the reactor, wait for the reaction tube to burn stable, adjust the state of the equipment
4. Wrap the sample in a tin cup and test the sample
5. Store and process data

Materials

• Sample Type:
Glutathione in neonates

Notes

1.The approach is also valuable for quantifying the reactivity and progression of an applied stable isotope tracer to help determine reaction rates and final disposition of applied substrates.
2.IRMS offers a way of measuring isotopic variations with extremely high levels of accuracy. It can be used to detect isotope values of lighter elements with no issues, making it instrumental in the analysis of organic and natural samples.
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