Application of gas chromatography–tandem mass spectrometry (GC/MS/MS) for the analysis of deuterium enrichment of water

Abstract

Incorporation of deuterium from deuterium oxide (2H2O) into biological components is a commonly used approach in metabolic studies. Determining the dilution of deuterium in the body water pool (BW) can be used to estimate body composition. We describe three sensitive GC-MS/MS methods to measure water enrichment in BW . Samples were reacted with NaOH and U-13C3-acetone in an autosampler vial to promote deuterium exchange with U-13C3-acetone hydrogens. Headspace injections were made of U-13C3-acetone-saturated air onto a 30m DB-1MS column in EI-mode. Subjects ingested 30ml 2H2O and plasma samples were collected. BW was determined by standard equation. DXA scans were performed to calculate body mass, body volume and bone mineral content. A 4 compartmental model was used to estimate body composition (fat and fat free mass). Full scan experiments generated a m/z 45 peak and to a lesser extent a m/z 61 peak. Product fragment ions further monitored included 45 and 46 using selected ion monitoring (SIM;Method1), the 61>45 and 62>46 transition using multiple reaction monitoring (MRM;Method2) and the Neutral Loss, 62>45, transition (Method3). MRM methods were optimized for collision energy (CE) and collision-induced dissociation (CID) argon gas pressure with 6eV CE and 1.5 mTorr CID gas being optimal. Method2 was used for finally determination of 2H2O enrichment of subjects due to lower natural background. We have developed a sensitive method to determine 2H2O enrichment in body water to enable measurement of FM and FFM.

In Vitro Study of Deuterium Effect on Biological Properties of Human Cultured Adipose-Derived Stem Cells

“…And, conversely, in deuterated water, all processes are slowed down not only from the biological point of view within the cell, but also from the point of view of increasing the nonuniformity of water density, which slows down metabolic processes in the environment outside the cell [ 33 ]. Our data correlate with the data of other authors, where similar effects of water of different D/H isotope composition are observed in different systems in vitro and in vivo [ 34 – 36 ].…”

“…First of all, that concerns the medicine. It is well known that one of the pharmaceutical trends is the development of deuterium-containing drugs, which are deuterated analogues of already known proton-containing biologically active compounds [ 10 , 11 , 29 , 34 , 42 ]. The second direction refers to the role of deuterium-depleted water as an adjuvant in the treatment of cancer [ 12 , 13 ].…”

Comparing DNA enrichment of proliferating cells following administration of different stable isotopes of heavy water

“…in vitro cell media, mouse drinking water, mouse, and human total body water (urine)), as it can affect isotopic enrichments into DNA as well as potentially alter cellular metabolism 29 . Other investigators using stable heavy water for their research have determined the level in total body water (TBW) 27 , 28 , 30 – 39 using infrared absorption or by utilizing test methods for plasma and urine, which involve using a metal catalyst (e.g. uranium), high temperatures (e.g.…”