Absorption, distribution and excretion The detoxifying function of our body ...

As our body generates new cells to keep itself in good health, it also attempts to cleanse itself by the elimination of toxins through the skin, lungs, kidneys, liver, and colon. If one organ becomes congested, the others must work harder to keep the system balanced. The body doesn't just have to deal with the toxins we consume, but those from external sources, such as include environmental pollutants, polluted air and water. The liver is the main site of conjugation with glycine in both man and most experimental animals (rabbits, rats), with the exception of the dog. In sheep, where the kidney is the main site ofbiosyntheses (Snapper et al., 1924; Friedmann & Tachau, 1911), there is an apparent reduced capability of conjugating benzoic acid with glycine. Infusion of increasing amounts of benzoic acid into the rumen at levels up to 1.8 g/kg led to a progressive fall in conjugation and increasing excretion of free benzoic acid in the urine. Doses of 1.1 and 1.8 g/kg were toxic leading to death. Potassium deficiency also occurred as shown by the usual symptoms of severe muscular weaknessand tremors (Martin, 1966). For many years, a liver function test was used in man based on the urinary excretion of hippuric acid after a test dose of benzoic acid (6 g orally or 1.5-2.O g intravenously). Hence there exists a large amount of experience on the excretion of benzoic acid and hippuric acid in man. In the blood, benzoates existin the free state and are not bound to proteins (Knoefel & Huang,1956). In the dog, the kidney clearance was estimated to be 0.90-1.89% (Knoefel & Huang, 1956). Normal urinary excretion of hippuric acid in man was estimated to be 1.0-1.25 g/day, equivalent to 0.7-1.7 g of benzoic acid (Stein et al., 1954). Other determinations of the normal excretion in man and rat yield values lying between 1-3 mg/kg bw (Armstrong et al., 1955). The maximum rate of the hippuric acid excretion after ingestion of benzoic acid was observed to be 17 mg/minute and for benzoyl glucuronic acid, 0.67 mg/minute equivalent to 24 g/day calculated as benzoic acid (Schachter, 1957). Up to 10 g of benzoate is quantitatively excreted by man (Barnes, 1959). At high intake levels, up to 36% sodium benzoate is conjugated with glucuronic acid and all metabolites eliminated completely within 14 hours (Schachter, 1957). Seventy-five to 80% of administered benzoic acid is eliminated by man in six hours (Quick, 1931). Sodium benzoate also decreases uric acid (Quick, 1931), urea, and ammonia excretion in man (Lewis, 1914). Precursors of endogenous benzoate are phenylalanine and tyrosine. Experiments with labelled phenylalanine showed that about 1-2% is metabolized by this pathway. Rabbits given 50-400 mg/kg bw per day of deuterio-phenylalanine for six to 12 days, humans given 14-28 mg/kg bw per day for four to six days, and guinea-pigs given 300 mg/kg bw per day for 12 days, were examined (Bernard et al., 1955). 1-14C acetate, however, did not produce labelled benzoic acid in rabbits and guineapigs (Bernard et al., 1955). 3- 14C phenylalanine given intraperitoneally to rats, produced 0.6-1% of activity as urinary hippuric acid (Altman et al., 1954).

1,3,4,5-tetrahydroxycyclohexanoic acid (quinic acid) may also serve as a precursor of benzoic acid in intermediary metabolism (Dickens & Pearson, 1951). Several human subjects were given 6 g quinic acid orally or 250 g prunes and excreted hippuric acid in increased amounts during the following 24 hours (Quick, 1931). When deuterio-benzoic acid was administered to man and rats it was excreted with its deuterium content unchanged. Feeding guinea-pigs, with body fluids enriched in D20, with hydro-aromatic compounds led to urinary excretion of deuterio-benzoic acid with high D content. A similarly prepared rat, when fed 750 mg hydroxy benzoic acid over five days, excreted urinary benzoic acid enriched in D. When human subjects and guinea-pigs were given quinic acid over several days, 47-72% was converted to benzoic acid and excreted in the urine (Bernard et al., 1955). Four rats irradiated with 700 roentgens and four controls were given intraperitoneally carboxyl-14C-labelled sodium benzoate and fasted. Irradiation had no effect on the conjugating ability but the irradiated rats excreted less labelled hippuric acid due to dilution by endogenously produced benzoic acid (Schreier et al., 1954). Benzoic acid inhibits pepsin digestion and sodium benzoate inhibits trypsin digestion of fibrin but they have no effect on amylase or lipase. Trypsin digestion of casein is only initially depressed by sodium benzoate (Kluge, 1933). Benzoic acid is a specific powerful inhibitor of the D-amino-acid oxidase (50% inhibition by 10-4M) (Klein & Kamin, 1964). Concentrations in the range of 10-3M exert some unspecific inhibitory effects on the metabolism of fatty acids, e.g. on acetoacetate formation (Avigan et al., 1955). Benzoic acid is rapidly absorbed (Schanker et al., 1958) and rapidly and completely excreted in the urine (Schachter, 1957; Barnes, 1959). One healthy man given 6, 9, 13.9, 34.7 and 69.3 mmol of sodium benzoate showed a complete elimination of the drug within 10-14 hours (Schachter, 1957). Cumulation does not occur, as shown by experiments, on the distribution and elimination of sodium benzoate-1-C14 administered i.p., orally, or s.c. to the rat. Practically quantitative excretion occurs in the urine within one to two days, less than 1% of radioactivity appears in the faeces, and a few appear in organs. All radioactivity was identified as labelled benzoic acid (Lang & Lang, 1956). Orally or s.c. administered labelled benzoic acid appeared at 90% in the urine as hippuric acid, 0.1% of radioactivity occurred in the expired CO2, and 2% remained in the carcass (Bernard et al., 1955). Two urinary metabolites of benzoic acid are known, namely hippuric acid and benzoyl-glucuronic acid. Conjugation with glycine and glucuronic acid occurs in preference to oxidation because benzoic acid strongly inhibits fatty oxidation in the liver. In man, rabbit and rat, benzoic acid is almost entirely excreted as hippuric acid, whereas dogs excrete more conjugated glucuronic acid than hippuric acid (Williams, 1959). Sheep are less able to excrete free benzoic

acid in their urine (Martin, 1966). The urine of man, pig, rabbit and sheep contains up to 10% of benzoyl-glucuronic acid. The maximum urinary excretory rate achieved depends on the dose of benzoate given. Limiting values of hippuric acid excretion were approached in man at a dose of 13.9 mmol (Schachter, 1957). Limitations in availability of glycine account for this (Quick, 1933). In the rat the tolerance of large doses of benzoic acid depends on the addition of adequate amounts of glycine to the diet leaving sufficient glycine for protein synthesis. Normally preformed glycine is used though some is synthesized as well by the rat (Quick, 1931; Barnes, 1959). When rats were fed 1.5% benzoic acid (as the sodium salt) in the diet, they excreted 95% and more of the drug as hippuric acid in the urine. As the benzoate in the diet was increased to 3.75%, the ratio of hippuric acid to total benzoic acid in the urine decreased. Additional glycine raised elimination to 86-99%. The only other derivative, found in significant amounts in the urine, was benzoyl glucuronide (Griffith, 1929). Dogs and rabbits excrete hippuric acid independent of the route of administration of benzoic acid (Quick, 1931).