Synthetic lipids can be deuterated in either the chains or head groups, or both. Specific deuteration has so far been restricted to:

• both (saturated) acyl chains;

• glycerol backbone only;

• sites in the head group

for practical details see:

Eibl, H. (1980) Chem. Phys. Lipids 26, 405-429

Harlos and Eibl, Biochemistry, 19, 895-899

Sixl and Watts (1982) Biochemistry, 24, 6446 - 12

Sixl and Watts (1983) PNAS, 80; 1613-1615

Synthetic phospholipids can have saturated acyl chains, or partially saturated acyl chains (C16:1; C18:1) are common) but their deuteration is a challenge due to oxidation during deuteration.

Some deuterated precursors including lipids and fatty acids, are available from various manufacturers, including Avanti (Birmingham, AL, US); Larandon, Sigma-Aldrich and Cambridge Isotopes.

Be aware that compounds bought in from the US carry a significant charge when shipped on ice, as well as possible import duty, (often more than the product costs) and so making large joint orders can save costs.

Bacterial growth in deuterated media produces a cell paste in which all bacterial phospholipids are uniformly labelled.

Such cell pastes can be subjected to extraction, partial purification to give a polar lipid fractions which can then, if so wished, be purified into individual lipid classes, using established protocols.

1. 15g of weight (pellets - Cells of E.Coli), suspended in 600ml of CH₃Cl : MeOH (2:1 v/v)

2. stirred for 4 hr at room temperature, and the supernatant was decanted after centrifugation (6° C, 10,000rpm for 15mins, with JA-10 rotor, Model J2-21 centrifuge, Beckman)

3. extraction was then repeated twice, first at room temperature (overnight), then at 37° C (overnight)

4. the extract was combined then passed through a sintered-glass filter, rotor evaporated, then dried under vacuum

5. Sample was then dissolved in a small amount of CH₃Cl, loaded onto a column (~ 15cm x 3cm) containing NaHCO₃ treated silicic acid.

6. Stepwise elution was carried out with CH₃Cl containing increasing proportions of methanol.

7. Two fraction are collected, one fraction (after ~ 198 ml) contained both PE + PG eluted at CH₃Cl : MeOH (8:1 and 6:1 v/v), and one fraction (after ~ 70ml) contained CL, eluted at CH₃Cl : MeOH (22:1 v/v)

8. All the fractions are monitored using TLC

9. Further purification on PE and PG mixture using silica gel only, the main eluted with solvent CH₃Cl : MeOH : (H₂O containing 10% of AcOH) (100 : 15 : 1 v/v) which contained PG (after ~ 210ml)

10. Then, the column was eluted with CH₃Cl : MeOH : (H₂O containing 10% of AcOH) (65 : 15 : 1 v/v) which contained PE (after ~ 126ml)

11. Final yield : PE, 26mg, PG, 38mg, CL 52mg of per-deuterated lipids

REF: Yasuhiro Kanemasa, Yuzuru Akamatsu and Shoshichi Nojima "Composition and Turnover of the Phospholipids in Escherichia Coli, Biochim. Biophys. Acta, 144 (1967), 382-390