The dose-dependent effect of BHT (butylated hydroxytoluene) on vitamin K-dependent blood coagulation in rats.
Earlier studies have reported a reduction of vitamin K-dependent blood clotting factor activity and incidence of haemorrhagic death in rats fed butylated hydroxytoluene (BHT); however, the vitamin K status of the animals used in these studies was claimed to be inadequate. The aim of the study reported here was to determine the effect of BHT on vitamin K-dependent clotting factors in vitamin K-sufficient and vitamin K-supplemented rats. Rats given BHT (3000 mg/kg body weight) for up to 21 days, in a diet containing a minimum of 3 ppm vitamin K3 (six times the recommended requirement), showed decreased vitamin K-dependent blood clotting factor activities, demonstrated by increases in factor-specific clotting time assays. Clotting times were prolonged within 7 days, significantly increased within 14 days (P < 0.001) and maximally increased 5.5-fold at 21 days (P < 0.05). Supplementation with a further 250 ppm vitamin K3 reversed this effect. BHT did not increase prothrombin time (PT), the usual index of clotting. However, in a similar 14-day investigation, a small but significant increase in PT (up to 151%, P < 0.005) was seen within 7 days. Further vitamin K supplementation was incapable of reversing this effect completely. A similar trend was shown by activated partial thromboplastin time. The 1/51 dilution Thrombotest, a more sensitive indicator of vitamin K-dependent clotting factor activity in the rat, was significantly increased (more than four fold, P < 0.01) within 7 days. This increase was fully reversed by further vitamin K supplementation. Prolongation of Thrombotest time was significant at a BHT dose level of 600 mg/kg body weight per day and this could be reversed by further supplementation of only 3.0 ppm vitamin K. However, at dose levels of 125 mg BHT/kg body weight per day or less, no clotting defects were observed. These studies confirm that chronic administration of more than 600 mg BHT/kg/day to rats supplied with recommended amounts of vitamin K can depress clotting factors and precipitate haemorrhagic deaths. When further vitamin K is provided, these deaths could be prevented even though not all clotting abnormalities may be reversed. This study disapproves the proposal that BHT-related clotting factor defects are confined to rats of inadequate vitamin K status, but shows that such effects do not occur at dose levels lower than 600 mg/kg/day. The results further indicate that rats receiving a high dose of BHT have a higher vitamin K requirement than would otherwise be considered necessary. However, as BHT produces no clotting defects in these animals receiving an intake 1000 times the acceptable daily intake, such clotting effects are most unlikely to indicate a human safety problem for BHT.
Cottrell
S
S
DH Department of Toxicology, St Bartholomew's Hospital Medical College, St Bartholomew's Centre for Research, London, UK.
Andrews
C M
CM
Clayton
D
D
Powell
C J
CJ
D000818:Animals; D001777:Blood Coagulation / Q000187:drug effects*; D002084:Butylated Hydroxytoluene / Q000494:pharmacology*; D004305:Dose-Response Relationship, Drug; D008297:Male; D010314:Partial Thromboplastin Time; D011517:Prothrombin Time; D051381:Rats; D017208:Rats, Wistar; D014812:Vitamin K / Q000494:pharmacology* / Q000502:physiology
D014812:Vitamin K; D002084:Butylated Hydroxytoluene