To test the hypothesis that intra-hepatic availability of fatty acid could modify the rate of suppression of endogenous glucose production (EGP), acipimox or placebo was administered before and during a test meal. We used a modified isotopic methodology to measure EGP in 11 healthy subjects and ¹H MR spectroscopic measurement of hepatic triglyceride stores was also undertaken. Acipimox suppressed plasma FFA markedly before the meal (0.05 ± 0.01 mmol/l at - 10min, p=0) and throughout the post-prandial period (0.03 ± 0.01 mmol/l at 150 min). Mean peak plasma glucose was significantly lower after the meal on acipimox days (8.9 ± 0.4 vs. 10.1 ± 0.5 mmol/l p<0.01), as was mean peak serum insulin (653.1 ± 99.9 vs. 909 ± 118 pmol/l; p<0.01). Fasting EGP was similar (11.15 ± 0.58 µmol /kg/min placebo vs. 11.17 ± 0.89 mg/kg/min acipimox). The rate of suppression of EGP following the meal was almost identical on the two test days (4.36 ± 1.52 vs. 3.69 ± 1.21 µmol /kg/min at 40 min). There was a significant negative correlation between the acipimox-induced decrease in peak plasma glucose and liver triglyceride content (r = -0.827, p=0.002), suggesting that when levels of liver fat were low, inhibition of lipolysis was able to affect glucose homeostasis. Acute pharmacologic sequestration of fatty acids in triglyceride stores improves postprandial glucose homeostasis without effect on the immediate postprandial suppression of EGP.