The gastrointestinal handling and metabolism of [1-13C]palmitic acid given as the free fatty acid was examined in six healthy women by measuring the excretion of 13C-label in stool and in breath as 13CO2. The gastrointestinal handling of [1-13C]palmitic acid was compared with the apparent absorption of dietary lipid by measuring lipid losses in stool. The variation both within and between subjects was determined by repeating the study in the same individuals on separate occasions. The time course for excretion of label in stool over the five-day study period followed a common pattern, with most of the label excreted over the first two days of the stool collection. 13C-Label excreted in stool over the five-day study period was 14.3 +/- 9.8% of that administered and on repeating the trial was 31.6 +/- 24.7% (not significantly different due to variability); there was poor agreement within subjects. Lipid excreted in stool expressed as a percentage of ingested lipid was 5.2 +/- 4.4% in Trial 1 and 5.9 +/- 4.0% in Trial 2, and was the same in each individual on repeating the trial. There was no clear relationship between the excretion of 13C-label and lipid in stool (Trial 1: R = -0.43, P > 0.40; Trial 2: R = -0.02, P > 0.97). On the first occasion, 22.0 +/- 4.5% of the administered label was excreted on breath over the 15-h study period and on repeating the trial was 15.8 +/- 9.5% (not significantly different) with poor repeatability in a given individual.(ABSTRACT TRUNCATED AT 250 WORDS)

A rapid automated method for isotopic analysis of 13C and 18O in CO2 has been developed. A variety of gas samples containing CO2 can be swept from serological tubes into a helium carrier flow; impurities are separated on a GC column so that a pure pulse of CO2 in He flows into the mass spectrometer. Isotopic ratio determinations are carried out as the pulse passes through the mass spectrometer, allowing a sample to be measured approximately every 4 min. A double, concentric needle-probe is used to flush the sample from the tube so that 100% sample recovery is achieved, maximizing sensitivity and preventing the possibility of fractionation. The precision of the technique, sigma(m-1), is better than 0.2% (0.0002 atom per cent excess) for 13C and 0.4% (0.83 p.p.m.) for 18O for 10 micromol of CO2 at natural abundance. Samples containing only atmospheric concentrations of CO2 can also be analyzed.

The objective was to determine whether rats could synthesize longer chain polyunsaturates from hexadecadienoate (16:2n-6) and hexadecatrienoate (16:3n-3). Rats were gavaged with uniformly 13C-labelled hexadecadienoate or hexadecatrienoate, euthanized 24 h later, and total lipids were extracted from liver and carcass. Gas chromatography/combustion/isotope ratio mass spectrometry was used to measure 13C levels in individual liver, carcass, and whole body fatty acids. 13C Enrichment was present in desaturated and chain-elongated polyunsaturates, including linoleate, arachidonate, alpha-linolenate, and docosahexaenoate at 12-13% of the dose of tracer given. 13C Enrichment from hexadecatrienoate was highest in carcass and liver alpha-linolenate, representing 3.5 and 17.9% of the total alpha-linolenate pool, respectively. For linoleate, arachidonate, or docosahexaenoate, the contribution of 13C did not exceed 0.2% of the total body pool. Green leafy vegetables common in the human diet were shown to contain up to 1.2% of total fatty acids as hexadecadienoate and 11.6% as hexadecatrienoate. Hence, humans consuming green vegetables probably synthesize a small proportion of their total body content of linoleate and alpha-linolenate.

SummaryThe relative advantages of LC-UVand LC-MS-MS methodologies for taxol as applied to toxicokinetic and pharmacokinetic studies is reviewed and some specific applications of the two methods used in our laboratories compared. The LC-MS-MS method was found to be ~enty-five times more sensitive than the LC-UV method. In essence where plasma concentrations are high and rapid turn round of data is not required the use of the LC-UV is more than adequate. However if sensitivity is an issue, limited amounts of plasma are available and/orwhere tight deadlines are pivotal LC-MS-MS is the method of choice.

Polyunsaturated fatty acids are needed for normal neonatal brain development, but the degree of conversion of the 18‐carbon polyunsaturated fatty acid precursors consumed in the diet to their respective 20‐and 22‐carbon polyunsaturates accumulating in the brain is not well known. In the present study, in vivo 13C nuclear magnetic resonance spectroscopy was used to monitor noninvasively the brain uptake and metabolism of a mixture of uniformly 13C‐enriched 16‐and 18‐carbon polyunsaturated fatty acid methyl esters injected intragastrically into neonatal rats. In vivo NMR spectra of the rat brain at postnatal days 10 and 17 had larger fatty acid signals than in uninjected controls, but changes in levels of individual fatty acids could not be distinguished. One day after injection of the U‐13C‐polyunsaturated fatty acid mixture, 13C enrichment (measured by isotope ratio mass spectrometry) was similar in brain phospholipids, free fatty acids, free cholesterol, and brain aqueous extract; 13C enrichment remained high in the phospholipids and cholesterol for 15 days. 13C enrichment was similar in the main fatty acids of the brain within 1 day of injection but 15 days later had declined in all except arachidonic acid while continuing to increase in docosahexaenoic acid. These changes in 13C enrichment in brain fatty acids paralleled the developmental changes in brain fatty acid composition. We conclude that, in the neonatal rat brain, dietary 16‐and 18‐carbon polyunsaturates are not only elongated and desaturated but are also utilized for de novo synthesis of long‐chain saturated and monounsaturated fatty acids and cholesterol.