Biotransformation of the 14C-hydrogenated analog of phenazepam in inbred mice

The elimination of 14C-hydrogenated analogue of phenazepam and its metabolites was studied in inbred C57B1/6 (B/6), BALB-c (C) and CBA mice. The kinetics of two-phase elimination of 14C-compounds with urine and feces was similar in all the above mouse strains. The excretion was realized mainly with feces, exceeding elimination with urine 5-6-fold in B6 mice and 10-11-fold in C and CBA mice. Some interstrain quantitative differences in metabolite composition were found. No metabolite was detected whose concentration would indicate a predominating direction of biotransformation of 14C-hydrogenated phenazepam analogue or distinguish it from other mouse strains. In the organism of mice, 14C-hydrogenated phenazepam analogue undergoes active aromatic hydroxylation and methoxylation via heterocycle (and possibly via chlorine-containing ring). At the same time dehydrogenation of 14C-hydrogenated phenazepam analogue molecule was not recorded.     

Experimental study on interactions between selenium and tin in mice

The organ distributions of tin and selenium, and their excretion into urine and feces, were determined in mice. There were four groups; (A) control, (B) Sn (5 μmol/kg/d) ip injection, (C) Se (5 μmol/kg/d) sc injection, and (D) Sn plus Se (5 μmol/kg/d, each). Animals received injections once a day for 12 consecutive days. The results were the following (1) Simultaneous injection of Sn and Se enhanced accumulation of both elements in the body, i.e., in group B, 14.1% of the total injected amount of Sn was excreted into urine and feces; in group C, 46.2% of total injected Se was excreted into urine and feces; in group D, 10.9% of total Sn and 37.5% of total Se were found in excreta. (2) Large amounts of Sn were found in bone, liver, spleen, and kidney in group B. When Se was administered jointly with Sn, the concentrations of Sn in bone and liver were suppressed, whereas those in spleen and pancreas were increased. (3) The effects of Se-injections at this dose on concentrations of Se in organs were small. (4) In plasma, chemical reduction of selenite by stannous chloride was not observed.     

Excretion of estrone,estradiol, and progesterone in the urine and feces of the female cotton-top tamarin (Saguinus oedipus oedipus)

The excretion of three gonadal steroids was studied in the urine and feces of female cotton-top tamarins (Saguinus oedipus oedipus). Each steroid, ¹⁴C-estrone, ¹⁴C-estradiol, and ¹⁴C-progesterone, was injected into a separate female cotton-top tamarin. Urine and feces were collected at 8 hr intervals for 5 days on the three tamarins. Samples were analyzed to determine the proportion of free and conjugated steroids. Steroid excretion patterns were determined by sequential ether extraction, enzyme hydrolysis, and chromatography. Labeled estrone was excreted in a slow and continuous manner into the urine (57%) and feces (43%) with 90% of the steroid conjugated. The nonconjugated form had an elution profile identical to ³H estrone, but the conjugated portion was not completely hydrolyzed by enzyme. Labeled estradiol was excreted primarily in the urine (87%) and was released rapidly. Over 90% of the injected ¹⁴C-estradiol was excreted in urine as a conjugate, of which 41% was converted to an estrone conjugate and the remaining 59% was excreted as a polar estradiol conjugate. Labeled progesterone was excreted primarily in the feces (95%), 61% of which was free steroid. Four to six individual peaks of radioactivity were found when using celite chromatography and high performance liquid chromatography (HPLC), indicating that progesterone is metabolized into several urinary and fecal metabolites. One of these peaks matched ³H-progesterone and others may be pregnanediols, pregnanetriols, and 17-hydroxyprogesterone. These steroidal excretion patterns help explain the atypical hormonal patterns seen during the tamarin ovarian cycle.     

A non-invasive technique for analyzing fecal cortisol metabolites in snowshoe hares (<Emphasis Type="Italic">Lepus americanus</Emphasis>)

To develop non-invasive techniques for monitoring steroid stress hormones in the feces of free-living animals, extensive knowledge of their metabolism and excretion is essential. Here, we conducted four studies to validate the use of an enzyme immunoassay for monitoring fecal cortisol metabolites in snowshoe hares (Lepus americanus). First, we injected 11 hares with radioactive cortisol and collected all voided urine and feces for 4 days. Radioactive metabolites were recovered predominantly in the urine (59%), with only 8% recovered in the feces. Peak radioactivity was detected an average of 3.5 and 5.7 h after injection in the urine and feces, respectively. Second, we investigated diurnal rhythms in fecal cortisol metabolites by measuring recovered radioactivity 2 days after the radioactive cortisol injection. The total amount of radioactivity recovered showed a strong diurnal rhythm, but the amount of radioactivity excreted per gram of feces did not, remaining constant. Third, we injected hares with dexamethasone to suppress fecal cortisol metabolites and 2 days later with adrenocorticotropic hormone to increase fecal cortisol metabolites. Dexamethasone decreased fecal cortisol metabolites concentrations by 61% and adrenocorticotropic hormone increased them by 1,000%, 8–12 h after injection. Fourth, we exposed hares to a simulated predator (dog). This increased the fecal cortisol metabolites concentrations by 175% compared with baseline concentrations 8–12 h after exposure. Thus, this enzyme immunoassay provides a robust foundation for non-invasive field studies of stress in hares.     

In vivo metabolism of leukotriene C4 in man: urinary excretion of leukotriene E4

Five - 20 nmoles of [5,6,8,9,11,12,14,15-3H8]leukotriene C4 was injected into three male volunteers. Forty-eight percent of the administered 3H was recovered from urine and 8% from feces, within a 72 hr period. Of the total urinary radioactivity 44% was excreted during the first hour after injection. This activity was mainly found in one compound, designated "I". The radioactivity excreted into urine later than one hour after injection, consisted partly of Compound I and two additional components, and partly of polar, non-volatile material. Compound I was identified as leukotriene E4 by UV-spectroscopy and cochromatographies in three high performance liquid chromatography systems with synthetic reference compounds. A total of 13% of administered radioactivity was excreted in urine as leukotriene E4.     

Dopamine β-hydroxylase: Determination of half-life and appearance in lymph fluid after IV injection of 125I-DBH into rats

A 4 day half-life of dopamine beta-hydroxylase (DBH) was determined for rats injected IV with ¹²⁵I-rat DBH from the slow exponential component of radioactivity appearing in plasma, urine, feces and combined urine and feces. Half-life estimates for ¹²⁵I-rat DBH injected IV into WKY and SHR animals did not differ from Sprague Dawley (Zivic Miller) rats. Radioactivity declined in parallel in plasma, urine and feces following IV ¹²⁵I-rat DBH administration and each radioactivity falloff curve could be resolved into two components. The slow phase of the decline of radioactivity excreted into urine and feces from which DBH half-life was calculated occurred between 5 and 25 days after ¹²⁵I-rat DBH injection. The early fast phase which is associated with distribution of the exogenous protein in body fluids and tissues continued for approximately the first 140 hr after DBH injection. The distribution characteristics of IV administered active bovine DBH and ¹²⁵I-rat DBH into the lymphatic system were examined. After active bovine DBH or ¹²⁵I-rat DBH was injected IV into rats, active DBH or radioactivity, respectively, appeared in lymph fluid (thoracic duct) within 20 min; reached peak concentrations within 90 min, and thereafter, declined in parallel with the plasma concentration. The concentration of radioactivity in plasma and lymph fluid were found to be unequal at 9 hr but were equivalent 68–75 hrs after IV injection of ¹²⁵I-rat DBH. Based on the amount of active DBH or radioactivity which accumulates in lymph fluid it is clear that'a substantial amount (> 50%) of the DBH in blood circulates through the lymphatic channels. Analysis of parallel experiments with labelled serum albumin indicate that use of these methods to study plasma proteins do provide sensitive measures of biological half-life and lymphatic distribution characteristics. Specifically for DBH, the results of our study suggest that DBH normally circulates in plasma and lymph fluid with a biological half-life of 4 days.     

Excretion of radiolabeled estradiol in the cat (Felis catus,L): A preliminary report

The time course and end products of estradiol metabolism were studied in the domestic cat, which has been chosen as a model for steroid metabolism studies in nondomestic felidae. Radiolabeled estradiol was injected intravenously into three adult female cats; one had a spontaneous estrus, one was induced with follicle-stimulating hormone, and one had been ovariohysterectomized; feces, urine, and blood were collected daily, and the radioactivity content was determined. Feces and urine contained 47 and 1% of the injected dose (0.33 μCi), respectively. Metabolites appeared earlier in the urine than in feces (d 1 vs d 2 postinjection), and excretion was completed on d 5; no radioactivity was detected in plasma 24 h postinjection. Estradiol metabolites were excreted as unconjugated estrogens (22%) and as conjugates hydrolyzable with β-glucuronidase and acid solvolysis (7 and 50%, respectively); the remaining 14% were not recoverable with any of the above methods. The major portion of the conjugates was estradiol-17β (64–80%) while 11–16% appeared as estrone. Endogenous cycles related to the spontaneous and induced ovarian activity were monitored by observation of estrous behavior, vaginal epithelium cornification, and plasma estradiol determination. The reproductive state of each animal had no effect on the time course or type of metabolite excreted. We found low proportions of injected radioactivity excreted in the urine and high residual levels remaining after hydrolysis and extraction in the feces. These findings suggest that although feces are an abundant source of estradiol metabolite in the cat, and probably in the exotic felidae, development of noninvasive methods for monitoring ovarian cycles in these species will depend on more efficient methods for urine hydrolysis, on the resolution of problems encountered in fecal steroid analysis, or on the identification of metabolites which may be measured directly in the urine without hydrolysis or extraction.     

Urinary and fecal immunoglobulin A, cortisol and 11-17 dioxoandrostanes, and serum cortisol in metabolic cage housed female cynomolgus monkeys (Macaca fascicularis)

BACKGROUND AND METHODS: Quantitative enzyme-immunoassays of urinary and fecal immunoglobulin A (IgA), cortisol and 11-17-dioxoandrostanes (11,17-DOA), and serum cortisol in eight metabolic-cage-housed female cynomolgus monkeys were performed. The monkeys were divided into two groups, B and NB. Group B animals were blood sampled every 6 hours, whereas Group NB animals were not handled/blood sampled. RESULTS: No differences were recorded between the amounts of feces and urine excreted by the two groups. Group B animals excreted more urinary cortisol than did Group NB animals indicating that restraint-blood sampling resulted in a stress response. Excreted amounts of IgA and 11,17-DOA (urine and feces) did not differ between the groups. CONCLUSIONS: Urinary cortisol was a reliable marker of the stress associated with repeated blood sampling. Declining amounts of excreted urinary cortisol indicated that cynomolgus monkeys acclimated quickly to repeated blood sampling in metabolism cages. Within and between animal variation in amounts of feces voided demonstrated the importance of expressing fecal markers as 'amounts excreted per time unit per kg body weight' rather than just measuring the concentrations in fecal samples.     

Metabolic kinetics and the biological action of 238Pu in the body of rabbits after intratracheal administration

A study was made of the distribution and biological effect of 238Pu nitrate intratracheally administered to rabbits. The skeleton and liver were the main organs in which 238Pu was secondarily deposited to make 63.5 and 12.9%, respectively, of the total amount administered. For 60 days of observation 15.3% of the amount administered were excreted in feces and urine. With 238Pu dose of 520 kBq/kg acute radiation sickness developed while at a dose of 4 kBq/kg the life span of animals did not vary from the control.     

The fate of cyclamate in man and other species

1. (14)C-labelled cyclamate has been administered to guinea pigs, rabbits, rats and humans. When given orally to these species on a cyclamate-free diet, cyclamate is excreted unchanged. In guinea pigs some 65% of a single dose is excreted in the urine and 30% in the faeces, the corresponding values for rats being 40 and 50%, for man, 30-50% and 40-60%, and for rabbits, 90 and 5%, the excretion being over a period of 2-3 days. 2. Cyclamate appears to be readily absorbed by rabbits but less readily by guinea pigs, rats and humans. 3. If these animals, including man, are placed on a diet containing cyclamate they develop the ability to convert orally administered cyclamate into cyclohexylamine and consequently into the metabolites of the latter. The extent to which this ability develops is variable, the development occurring more readily in rats than in rabbits or guinea pigs. In three human subjects, one developed the ability quite markedly in 10 days whereas two others did not in 30 days. Removal of the cyclamate from the diet caused a diminution in the ability to convert cyclamate into the amine. 4. In rats that had developed the ability to metabolize orally administered cyclamate, intraperitoneally injected cyclamate was not metabolized and was excreted unchanged in the urine. The biliary excretion of injected cyclamate in rats was very small, i.e. about 0.3% of the dose. 5. The ability of animals to convert cyclamate into cyclohexylamine appears to depend upon a continuous intake of cyclamate and on some factor in the gastrointestinal tract, probably the gut flora.     

The metabolic fate of chlormadinone acetate in the baboon.

The metabolic fate of chlormadinone acetate (17alpha-acetoxy-6-chloro-4, 6-pregnadiene-3, 20-dione; CAP) was studied in intact and biliary fistula baboons. The steroid was labeled with 3H at position 1 and with 14C at the carboxyl moiety of the 17alpha-acetate, thus affording the opportunity to ascertain the loss of the 17alpha-acetoxy group and the fate of both labels. The averages of the radioactivity excreted, given as percentages of the amounts injected, and the standard deviations were as follows: In the urine of intact animals after 6 hours, 5.7 +/- 0.2% and 5.5 +/- 0.7% of the 3H and 14C were recovered, respectively. After 6 days, there was 17.5% of the 3H and 16.2% of the 14C in the urine plus 15.3% of the 3H and 16.4% of the 14C in the feces. In baboons with biliary fistulas, the total radioactivity excreted was 7.8 +/- 0.7% of the 3H and 11.6% of the 14C in the urine, and 30.9 +/- 4.4% of the 3H and 30.7% of the 14C in the bile after 6 hours. Glucosiduronates were the predominant conjugates in the urine and bile. The similarity in the urinary excretion of radioactivity in the first 6 hours in intact and biliary fistula animals, the relatively low excretion of radioactivity in the bile and after 6 days in the urine, and the low fecal excretion suggest that the metabolites of CAP are not involved in an extensive enterohepatic circulation in the baboon. Deacetylation of the 17alpha-acetate in CAP was detected in the early collection periods of the urine and bile and constituted a very small percentage of the injected compound. No significant oxygenation of CAP at position 1 was detected. The metabolism of CAP is discussed and compared to our previously reported data on the metabolism of progesterone, ethynodiol diacetate and medroxyprogesterone acetate and the data on other progestogens reported in the literature. It appears that the excretion of CAP is significantly slower in the baboon than that of the other progestogens. The amounts of glucosiduronates of CAP and/or its metabolites formed in vivo are less than those formed with the other progestogens. Also, the extent of deacetylation of the 17alpha-acetate of CAP is much less than that of the 3beta-acetate of ethynodiol diacetate.     

Ntau-methylhistidine excretion is a valid index of myofibrillar protein breakdown in the guineapig (Cavia porcellus).

The recovery of radioactivity in the urine of guineapigs following a bolus intravenous dose of chromatographically pure 14C-Ntau-methylhistidine was measured in order to test whether the excretion of Ntau-methylhistidine (Ntau-MH) is a valid index of myofibrillar protein breakdown in these animals. Four male and four female guineapigs were dosed and after 7 days, 91.65+/-2.82% and 3.58+/-0.91% of injected radioactivity was recovered in the excreta and tissues, respectively. The average total recovery of 95.2+/-3.0% was not significantly different from 100%. Male guineapigs excreted the radioactivity more slowly than females (70% of the dose excreted within 74 h vs 39 h, respectively) but cumulative excretion at 7 days was the same for each sex. Chromatographic analysis of the urine showed almost all of the radioactivity to be associated with a single peak corresponding to Ntau-MH, indicating a lack of significant metabolism. These data show that although the clearance of 14C-Ntau-MH is slower than in rats or humans the urinary excretion of Ntau-MH is a valid index for myofibrillar protein degradation in the guineapig.     

Assessment of urine and fecal testosterone metabolite excretion in Chinchilla lanigera males

Endemic chinchilla (Chinchilla spp.) populations are nearly extinct in the wild (South America). In captive animals (Chinchilla lanigera and C. brevicaudata), reproduction is characterized by poor fertility and limited by seasonal breeding patterns. Techniques applied for studying male reproductive physiology in these species are often invasive and stressful (i.e. repeated blood sampling for sexual steroids analysis). To evaluate endocrine testicular function, the present experiments were designed to (a) determine the main route of testosterone excretion (14C-testosterone infusion in four males); (b) validate urine and fecal testosterone metabolite measurements (HPLC was used to separate metabolites and immunoreactivity was assessed in all metabolites using a commercial testosterone radioimmunoassay, and parallelism, accuracy and precision tests were conducted to validate the immunoassay); and (c) investigate the biological relevance of the techniques applied (quantification of testosterone metabolite excretion into urine and feces from five males injected with hCG and comparison between 10 males and 10 females). Radiolabelled metabolites of 14C-testosterone were excreted, 84.7+/-4.2 % in urine and 15.2+/-3.9 % in feces. A total of 82.7+/-4.2% of urinary and 45.7+/-13.6% of fecal radioactivity was excreted over the first 24 h period post-infusion (metabolite concentration peaked at 8.2+/-2.5 h and 22.0+/-7.0 h, respectively). Several urinary and fecal androgen metabolites were separated by HPLC but only fecal metabolites were associated with native testosterone; however, there was immunoreactivity in more than one metabolite derived from 14C-testosterone. After hCG administration, an increase in androgen metabolite excretion was observed (p<0.05). Males excreted greater amounts daily of urinary androgen metabolites as compared with females (p<0.05); this difference was not evident in feces. Results of the present study indicate that the procedure used is a reliable and non-invasive method to repeatedly monitor variations in testicular endocrine activity in this species. It can be a useful tool that would help ensure the survival of the wild populations as well as to provide the basis for a more efficient use by the fur industry.     

The effect of a synthetic steroid (trienbolone) on the rate of release and excretion of subcutaneously administered estradiol in calves (18).

With the objective of obtaining values for the rate of release and excretion of subcutaneously implanted estradiol and to relate them to the metabolic effect of the hormone, a study was carried out with young Jersey bull calves. After subcutaneous administration of lactose tablets (implants) containing tritiated estradiol (4 mCi in 20 mg estradiol) the activity was followed in plasma, urine and feces for 107 days. Three calves received implants containing 140 mg trienbolone in addition to the 20 mg estradiol. In the first group maximum plasma concentration of estradiol-17 beta was 3 nmol/1. In the other group it was only 0.33 nmol/1. In calves receiving estradiol as the only steroid, 95% of the activity was excreted within 20 days after implantation. In the other group collection of urine and feces had to be carried out for 107 days in order to account for all the implanted activity. No 3H could be detected in urine and feces samples collected from the estradiol group more than 31 days ater implantation. The feces and urine samples collected from calves in the estradiol-trienbolone group 107 days after implantation contained from 1.4 - 3 nCi per gram. The remarkably decreasing effect of trienbolone on the release of estradiol and its possible importance for the effect of subcutaneously administered estradiol are discussed.     

Bacterially synthesized folate and supplemental folic acid are absorbed across the large intestine of piglets

A large pool of folate exists in the large intestine of humans. Preliminary evidence, primarily in vitro, suggests that this folate may be bioavailable. The purpose of this study was to test the hypothesis that supplemental folic acid and bacterially synthesized folate are absorbed across the large intestine of piglets. The pig was used as an animal model because it resembles the human in terms of folate absorption, at least in the small intestine. A tracer of [3H]-folic acid or [3H]-para-aminobenzoic acid ([3H]-PABA), a precursor of bacterially synthesized folate, was injected into the cecum of 11-day-old piglets. Feces and urine were collected for 3 days. Thereafter, piglets were killed, and livers and kidneys harvested. [3H]-Folate was isolated from biological samples by affinity chromatography using immobilized milk folate binding proteins and counted using a scintillation counter. In piglets injected with [3H]-folic acid, the feces, liver, urine and kidneys accounted for 82.1%, 12.3%, 3.9% and 1.7% of recovered [3H]-folate, respectively. In piglets injected with [3H]-PABA, the amount of recovered bacterially synthesized folate in the feces, liver and urine was 85.1%, 0.4% and 14.6%, respectively. Twenty-three percent and 13% of tritium were recovered in samples examined (liver, kidney, fecal and urine) from piglets injected with [3H]-folic acid and [3H]-PABA, respectively. Using our estimates of [3H]-folic acid absorption and the total and percent monoglutamyl folate content of piglet feces, we predict that at least 18% of the dietary folate requirement for the piglet could be met by folate absorption across the large intestine.     

Excretion, Metabolism and Tissue Distribution of A Spin Trapping Agent, α-Phenyl-N-Tert-Butyl-Nitrone (Pbn) in Rats

The objective of this study is using radiolabelled PBN to determine the tissue distribution, excretion, and metabolism of PBN in rats in order to evaluate the effective time to trap free radical in appropriate tissue(s). Our results demonstrated that PBN is rapidly absorbed when it is injected intraperitoneally in the animal. PBN can be used as an effective spin trapping agent for a variety of tissues since it is evenly distributed among a wide range of tissues measured. Since there is no difference in the tissue concentrations and distribution pattern of PBN at 15, 30 and 60min after injection of PBN. it is appropriate to choose any of these time intervals to terminate the experiment and extract the spin adduct. The excretion of PBN, however, is slow. The majority of the radioactivity (70%) was excreted by the first 3 days. Only 5.7% of radioactivity was collected from 3 to 14 days. The remaining 25% of the radioactivity may be in the form of expired ¹⁴CO₂. Trace amounts of radioactivity were recovered in the feces. PBN has probably only one major form of metabolite excreted in the urine. A small amount of the parent compound, however, was also excreted in the urine. The chemical structure of the metabolite(s) is still unknown.     

On the enterohepatic cycle of triiodothyronine in rats; importance of the intestinal microflora

Until 70 h after a single iv injection of 10 uCi [125I]triiodothyronine (T3), normal rats excreted 15.8 +/- 2.8% of the radioactivity with the feces and 17.5 +/- 2.7% with the urine, while in intestine-decontaminated rats fecal and urinary excretion over this period amounted to 25.1 +/- 7.2% and 23.6 +/- 4.0% of administered radioactivity, respectively (mean +/- SD, n = 4). In fecal extracts of decontaminated rats 11.5 +/- 6.8% of the excreted radioactivity consisted of T3 glucuronide (T3G) and 10.9 +/- 2.8% of T3 sulfate (T3S), whereas no conjugates were detected in feces from normal rats. Until 26 h after ig administration of 10 uCi [125I]T3, integrated radioactivity in blood of decontaminated rats was 1.5 times higher than that in normal rats. However, after ig administration of 10 uCi [125I]T3G or [125I]T3S, radioactivity in blood of decontaminated rats was 4.9- and 2.8-fold lower, respectively, than in normal rats. The radioactivity in the serum of control animals was composed of T3 and iodide in proportions independent of the tracer injected, while T3 conjugates represented less than 10% of serum radioactivity. These results suggest an important role of the intestinal microflora in the enterohepatic circulation of T3 in rats.     

The metabolism of estriol in rabbits

Rabbits have been shown to excrete 6, 7-³H-estriol, its conjugates and metabolites preponderantly in the bile during the initial 4 hours following the I.V. injection of the labeled steroid. The amount of radioactivity excreted in the urine was $\text{1}\text{3}$ of that in the bile. Since in intact rabbits most of the injected radioactivity of ³H-estriol is excreted in the urine over a period of days (and very little in the feces), it appears that estriol and its conjugates and metabolites are involved in an efficient enterohepatic. circulation. In the bile, the preponderant metabolite of ³H-estriol was the 3-glucosiduronate. Even though the latter constituted a substantial part of the urinary metabolites, other conjugates and metabolites of estriol were present in considerable amounts. It is possible that the latter have resulted from gastro-intestinal and/or renal metabolism. Incubation of rabbit liver with estriol led to 75% conjugation with glucuronic acid in the 3-position.     

Passage of the intravenously administered 15N urea into the digestive tract and its excretion in the sheep.

The experiments performed on two wethers provided with simple rumen cannulas and reentrant cannulas, inserted into the proximal duodenum and ileum, showed a passage of 15N from labelled urea, injected intravenously, from the blood to the digestive tract. The amount of the 15N in the digesta was the highest in duodenum, slightly lower in the rumen and slightly lower in ileum. Approximately 50% of the injected 15N was excreted in urine. The amount of the 15N eliminated with feces was very small; 0.6 to 2.8% of the dose injected per day. About 73--84% of the 15N which passed the duodenum was absorbed in further parts of the digestive tract. It can be concluded that all parts of the digestive tract take part in utilization of the endogenous urea.     

Excretion,Metabolism and Tissue Distribution of A Spin Trapping Agent, α-Phenyl-N-Tert-Butyl-Nitrone (Pbn) in Rats

The objective of this study is using radiolabelled PBN to determine the tissue distribution, excretion, and metabolism of PBN in rats in order to evaluate the effective time to trap free radical in appropriate tissue(s). Our results demonstrated that PBN is rapidly absorbed when it is injected intraperitoneally in the animal. PBN can be used as an effective spin trapping agent for a variety of tissues since it is evenly distributed among a wide range of tissues measured. Since there is no difference in the tissue concentrations and distribution pattern of PBN at 15, 30 and 60min after injection of PBN. it is appropriate to choose any of these time intervals to terminate the experiment and extract the spin adduct. The excretion of PBN, however, is slow. The majority of the radioactivity (70%) was excreted by the first 3 days. Only 5.7% of radioactivity was collected from 3 to 14 days. The remaining 25% of the radioactivity may be in the form of expired ¹⁴CO₂. Trace amounts of radioactivity were recovered in the feces. PBN has probably only one major form of metabolite excreted in the urine. A small amount of the parent compound, however, was also excreted in the urine. The chemical structure of the metabolite(s) is still unknown.