Pig Urine Research Articles

In vitro dermal absorption of pesticides: V. In vivo and in vitro comparison of the herbicide 2,4-dichlorophenoxyacetic acid in rat, guinea pig, pig, human and tissue-cultured skin

In vitro dermal absorption tests were conducted with the 14C-ring-labelled herbicide 2,4-dichlorophenoxyacetic acid (2,4-D), dissolved in acetone and applied to dermatomed skin (0.5 mm) of a number of species at dose rates of 7–8 μg/cm 2. Skin absorption was determined for 48 hr after exposure using an in vitro flow-through system. Skin absorption was calculated from the sum of the percentage recovery of 14C activity in the receiver solution and the percentage recovery in the methanol washes of the skin at 48 hr and the skin digest samples. Two receiver solutions, Ringer's saline, and Hanks' HEPES buffered saline with 4% serum albumin were used. Listed in decreasing order, the total percentage in vitro dermal absorptions obtained by 48 hr after exposure for the five skin types were: 47 ± 4.3% [tissue cultured Testskin; Hanks' receiver (HR)], 40 ± 4.5% (rat; HR), 19 ± 1.8% (human; HR), 14 ± 2.3% (hairless guinea pig; HR), 14 ± 8.8% (pig; Ringer's receiver). The percentage recovery of the radiolabel in soapy water skin washes at 24 hr, methanol washes and skin digests at 48 hr, and of 14C-labelled volatiles collected in air traps at 48 hr after exposure are reported. Comparative in vivo studies were conducted for 14 days after exposure and demonstrated 32 ± 3.9 and 28 ± 7.8% recovery of 14C in the urine of rats (dose rate, 3 μg/cm 2) and guinea pigs (dose rate, 4 μg/cm 2), respectively. Total faecal recovery was 2 ± 0.3 and 9 ± 3.5% for rats and guinea pigs, respectively. Analysis of tissue taken at autopsy 14 day after dosing demonstrated a total tissue recovery of 14C activity of 1 ± 0.1 and 2 ± 0.5% in rats and guinea pigs, respectively. Including the 14C activity extracted from the skin removed from the dose site at 14 days after exposure, the total recovery of dermally absorbed residues was 49 ± 10.4 and 40 ± 9.9% in rats and guinea pigs, respectively. Recovery of 14C activity from soapy water skin washes conducted at 24 hr after exposure was 28 ± 8.1 and 43 ± 9.0% for rats and guinea pigs, respectively. Recovery in skin patches was 18% (guinea pigs) and 26% (rats). In summary, the in vitro/in vivo concordance for the rat dermal absorption data was good but the in vitro data for hairless guinea pigs underestimated the in vivo absorption, and therefore for 2,4-D, rat skin may provide a better model of percutaneous absorption.

In vitro dermal absorption of pesticides: IV. In vivo and in vitro comparison with the organophosphorus insecticide diazinon in rat, guinea pig, pig, human and tissue-cultured skin

In vitro skin absorption tests are currently being developed as an alternative to in vivo animal tests for predicting the degree of occupational exposure to pesticides. In the study reported here, in vitro percutaneous absorption tests were conducted with the 14C-ring-labelled pesticide, diazinon, dissolved in acetone and applied to the dermatomed skin (0.5 mm) of a number of species at a dose rate of 9.5–16.7 μg/cm 2. Skin permeation was determined for 48 hr after exposure using an in vitro flow-through system. Skin permeation was calculated from the sum of the percentage recovery of 14C activity in the receiver solution and the percentage recovery obtained in methanol washes of the skin at 48 hr and in skin digests. Listed in decreasing order, the total percentage in vitro dermal absorptions (mean ± SD) obtained by 48 hr after exposure for the five skin types were: 47 ± 3.4% (rat), 36 ± 0.9% (tissue cultured Testskin), 33 ± 2.8% (hairless guinea pig), 20 ± 3.1% (human) and 15 ± 13.1% (pig). The percentage recoveries in soapy water skin washes at 24 hr, in methanol washes and skin digests at 48 hr and of 14C-labelled volatiles collected in air traps at 48 hr after exposure are reported. Comparative in vivo studies demonstrated 37 ± 0.8 and 24 ± 5.7% recovery of 14C in the urine of rats (dose rate, 6 μg/cm 2) and hairless guinea pigs (dose rate, 5 μg/cm 2), respectively, by 14 days after exposure. Total faecal recovery 14 days after exposure was 18 ± 0.4 and 4 ± 0.9% for rats and guinea pigs, respectively. Analysis of tissue taken at autopsy 14 days after exposure demonstrated a total tissue recovery of 0.6 ± 0.1% [ 14C]diazinon in rats and 1 ± 0.2% in hairless guinea pigs. The total recovery in skin removed from the dose site at 14 days after exposure was 0.2 ± 0.02% and 0.1 ± 0.05% in rats and hairless guinea pigs, respectively. Recovery of radioactivity from soapy water skin washes conducted at 24 hr after exposure was 21 ± 3.8% for rats and 2 ± 0.1% for hairless guinea pigs. Recovery in skin patches was 23 ± 5.4% and 73 ± 2.9% in rats and hairless guinea pigs, respectively. The in vitro data for dermal absorption of [ 14C]diazinon for rats (47 ± 3.4%) and hairless guinea pigs (33 ± 2.8%) were in good agreement with the data observed for rats (56 ± 1.03%) and hairless guinea pigs (28 ± 6.0%) in vivo. This study supported the use of in vitro skin absorption tests as an alternative to in vivo animal testing.

近交系Weiser-Maples系モルモットの基礎研究

Urinalysis, hematological and blood chemical examinations and measurement of organ weights were carried out for the purpose of collecting background data on Weiser-Maples (WM) which was established as an inbred strain of guinea pigs. The results were compared with those for the commercial guinea pigs, Std: Hartley (H) . In addition, the measurement methods for urinary specific gravity and whether centrifugation of urine influences the results of urinalysis or not were examined. The specific gravity values calculated from the volume and weight of urine were highly correlated with those from the refraction rate. The results of urinalysis of the supernatant of centrifuged urine were similar to those of urine not centrifuged, but the results suggested that the use of the supernatant provided a more accurate measurement of urine in guinea pigs. The positive rate for urinary protein was higher in the WM strain (38.9%) than in the H strain (0%) . Although the WM strain showed higher or lower values than the H strain in some items of hematological and blood chemical examinations and organ weights, we could not regard them as characteristic of the WM strain.

Isolation of an endogenous Na-pump specific inhibitor from normal pig urine: characterization and comparison with the inhibitor purified from bovine adrenal glands

An endogenous Na-pump specific inhibitor has been purified to homogeneity from normal pir urine using Amberlite XAD-2 adsorption chromatography followed by five steps of reverse phase HPLC. Although most of the dose response curves for this purified Na-pump inhibitor, designated uroxin, in the various assay systems paralleled those of authentic ouabain and the specific Na-pump inhibitor previously purified from bovine adrenal glands (designated adrexin C), the cross-reactivity curve with anti-ouabain antibodies did not. The retention time of uroxin on various types of reverse phase HPLC columns were also difference from those of plant-derived cardiotonic steroids and adrexin C. The cross-reaction curve of adrexin C was superimposable with that of ouabain, and adrexin C coeluted with ouabain from all of the HPLC columns tested. The results from physical and chemical characterization of both purified inhibitors suggest that uroxin is a novel Na-pump inhibitor which is structurally different from any of the known cardiotonic steroids or other substances previously reported to exhibit Na-pump inhibitory activity. The results also indicate that adrexin C is indistinguishable from ouabain. These results suggest that there are at least two different types of endogenous Na +,K +-ATPase inhibitors in the mammalian body.

Isolation and characterization of an endogenous Na+,K(+)-ATPase-specific inhibitor from pig urine.

A Na+,K(+)-ATPase inhibitor was purified from 88.6 l pig urine with a yield of approximately 10 micrograms. It inhibits the ouabain-sensitive uptake of 86Rb into human erythrocytes and the specific binding of ouabain to Na+,K(+)-ATPase. It also exhibits cross-reactivity to anti-ouabain serum. The purification procedure consisted of adsorption chromatography on an Amberlite XAD-2 column, preparative scale C18 low-pressure liquid chromatography (LPLC), and five steps of HPLC with five different types of reverse-phase columns. The dose dependence of the purified substance for the inhibition of ouabain-sensitive Na+,K(+)-ATPase activity and 86Rb uptake into human erythrocytes, and for the ouabain-displacing activity, paralleled those of ouabain, spanning two orders of magnitude in concentration range. However, the curve obtained from the cross-immunoreactivity with anti-ouabain serum did not parallel that of ouabain. The inhibitory potencies of the purified substance against the Na(+)-pump and ouabain-binding were diminished with increasing K+ concentration, exhibiting characteristics typical of cardiac glycosides. This substance had no effect on Ca(2+)-ATPase activity in human erythrocyte plasma membrane and skeletal-muscle sarcoplasmic reticulum, nor on Mg(2+)-ATPase activity. Acid treatment with 6 M HCl at 115 degrees C for less than 1 min destroyed approximately 82% of the inhibitory activity of the purified substance against Na(+)-pump activity. Alkaline treatment with 0.2 M NaOH at 23 degrees C for 2 h and heat treatment at 150 degrees C for 30 min partially destroyed the inhibitory activity. Boiling for 10 min and digestion by various enzymes did not affect the activity. Molecular mass was estimated to be 620 Da by gel-filtration column chromatography. Preliminary MS analysis suggested that the purified substance has a molecular mass of 625 Da. An 1H-NMR study revealed that this substance does not contain a tertiary methyl group. The results suggest that the purified Na+,K(+)-ATPase inhibitor is not a peptide and is distinct from any of the known cardiotonic steroids or various substances previously reported to exhibit Na+,K(+)-ATPase inhibitory activity. Thus, the purified substance may be a novel endogenous regulator of Na+,K(+)-ATPase.

A Novel Endogenous Sodium-Pump Inhibitor in Pig Urine

To clarify the chemical nature and pathophysiological significance of an endogenous, specific Na-pump inhibitor, extracts of various tissues have been examined. The activities were extracted by an Amberlite XAD-2 adsorbent and fractionated by high-performance liquid chromatography (HPLC) on an ODS column. Among many tissues and fractions tested, the extract of pig urine has been found to contain a large amount of 86Rb uptake inhibitory activities. The strongest inhibitory activity for 86Rb influx into erythrocytes also exhibited a cross-reactivity with anti-ouabain antibodies. This activity has been purified to homogeneity using five steps of reverse-phase HPLC. Although most of the dose-response curves for this purified Na-pump inhibitor, designated as uroxin, in the various assay systems paralleled those of ouabain and the inhibitor purified from bovine adrenal glands (designated as adrexin C), the cross-reactivity curve with anti-ouabain antibodies did not. The retention time of uroxin on an ODS HPLC column was also different from that of digoxin, ouabain, or adrexin C. 1H nuclear magnetic resonance spectroscopic study suggests that uroxin is a novel Na-pump inhibitor that is structurally different from any of the known cardiotonic steroids or the substances previously reported to exhibit Na-pump inhibitory activity. Thus, uroxin may be a new type of endogenous regulator for the Na pump.

Epidermal growth factor concentrations in pig tissues and body fluids measured using a homologous radioimmunoassay.

A homologous radioimmunoassay for the measurement of epidermal growth factor (EGF) levels in pig tissues and body fluids has been developed using an antiserum to recombinant porcine EGF. The assay is highly specific, showing no cross-reactivity with a variety of other polypeptides including the structurally related protein, transforming growth factor-alpha. Furthermore, < 1% cross-reactivity was observed with mouse EGF emphasizing the necessity for homologous assays for EGF measurement. Immunoreactive EGF was present in extracts of pig kidney and pancreas (3.44 +/- 0.43 and 0.76 +/- 0.13 (S.E.M.) pmol/g wet weight respectively), but was not detected in extracts of submaxillary gland or liver. Although immunoreactive EGF was not detectable in uterine, allantoic or ovarian follicular fluids, colostrum contained EGF at biologically active concentrations (0.84 +/- 0.15 nmol/l). Immunoreactive EGF was also present in pig urine, with similar concentrations in samples from male or female animals. In addition, pig urine inhibited the binding of 125I-labelled EGF to 3T3 fibroblasts and stimulated DNA synthesis in quiescent monolayers of these cells, indicating that the immunoreactive material in urine is biologically active. Quantitative comparisons of the data presented here with that published previously indicate considerable species variation in the EGF levels of various tissues and body fluids.

Studies on endogenous formation of N-nitroso compounds in the guinea pig supplemented with proline or thioproline and sodium nitrate

The endogenous formation of N-nitrosoproline (NPRO) and N-nitrosothioproline (NTPRO, N-nitrosothiazolidine-4-carboxylic acid) was studied by monitoring their excretion in the urine of guinea pigs given oral doses of 10 mg proline or thioproline after supplementation with 34 mg (0.4 mmol) sodium nitrate. In order to estimate the conversion of nitrate to nitrate, the animals were also supplemented with 3.5 mg (0.05 mmol) sodium nitrate instead of sodium nitrate. In animals fed commercial diets, the excretion of NPRO and NTPRO under supplementation with sodium nitrate was 2.0 μg and 28.7 μg/animal/day, respectively, whereas the excretion under supplementation with sodium nitrate was 0.7 μg and 13.3 μg/animal/day, respectively. The higher excretion of NTPRO than NPRO in each case shows that thioproline is more effective for nitrite trapping than proline. The animals supplemented with nitrate excreted more than twice the amounts of NPRO or NTPRO than those supplemented with nitrite. It is assumed, therefore, that more than 0.1 mmol nitrate is reduced to nitrite and takes part in the endogenous nitrosation of the guinea pig. When various concentrations of l-ascorbic acid (AsA), known to inhibit the formation of N-nitroso compounds, were also administered orally to animals immediately after supplementation with sodium nitrate, the NPRO excretion decreased with increasing AsA concentration. These data indicate that the guinea pig, which is unable to synthesize AsA as well as humans, may be an appropriate animal model for evaluation of the endogenous nitrosation ability of humans ingesting nitrate.

Vitamin C activity of 2-O-alpha-D-glucopyranosyl-L-ascorbic acid in guinea pigs.

The vitamin C activity of 2-O-alpha-D-glucopyranosyl-L-ascorbic acid (AA-2G), which is one of chemically stable derivatives of L-ascorbic acid (AsA), in guinea pigs was investigated. Male guinea pigs were divided into 9 groups and fed AsA-deficient diet for 24 days with the following supplement: AA-2G- or AsA-supplemented groups were orally supplemented with 0.96, 1.92, 9.6 and 192 AA-2G mg/animal/day or equimolar amounts of AsA (0.5, 1, 5 and 100 mg/animal/day, respectively); AsA-deficient group received neither of them. The body weight gain, serum alkaline phosphatase activity, and the concentration of AsA and AA-2G in the liver, adrenals and urine of the guinea pigs were measured at the end of the experimental period. The AA-2G-supplemented guinea pigs showed similar body weight gain to the animals supplemented with equimolar amount of AsA. Serum alkaline phosphatase activity in both AA-2G- and AsA-supplemented groups was significantly higher than that of AsA-deficient group. But there was no significant difference between the groups supplemented with AA-2G and the equimolar amount of AsA. AA-2G-supplemented guinea pigs showed no apparent symptoms of scurvy. In AA-2G-supplemented groups, AA-2G was not detected in the liver, adrenals and urine, but AsA was found and the AsA concentration increased with increasing AA-2G dosage. The AsA concentration in the tissues of each AA-2G-supplemented groups was higher than that of AsA-deficient group, which was similar to that of the groups supplemented with equimolar amount of AsA. These results showed that AA-2G has the same vitamin C activity as AsA on a molar basis for the orally supplemented guinea pigs.

Analysis of lamotrigine and lamotrigine 2-N-glucuronide in guinea pig blood and urine by reserved-phase ion-pairing liquid chromatography

Lamotrigine is an investigational anticonsulvant drug undergoing clinical trials. A simultaneous assay was developed to quantitate lamotrigine and its major metabolite, lamotrigine 2-N-glucuronide, from guinea pig whole blood. The extraction procedure and reversed-phase high-performance liquid chromatographic (HPLC) assay employed sodium dodecylsulfate (SDS) as an ion-pairing reagent to selectively separate lamotrigine and lamotrigine 2-N-glucuronide from endogenous blood components, other anti-convulsant drugs, and their metabolites. The mobile phase was composed of acetonitrile—50 m M phosphoric acid (pH 2.2) containing 10 m M SDS (33:67, v/v), and components were detected at 277 nm. The total coefficients of variance (C.V.) for the blood assay were ≤ 9.4% for lamotrigine (0.25–20.0 μg/ml) and ≤ 13.4% for the glucuronide metabolite (0.25–10.0 μg/ml). Separate assays for lamotrigine and its glucuronide in urine were developed. In order to quantitate low levels of lamotrigine in guinea pig urine, lamotrigine was extracted with tert.-butyl methyl ether—ethyl acetate (1:1). The total C.V. for lamotrigine quantitation in urine was ≤ 7.5% (0.10–10.0 μg/ml). For the determination of lamotrigine 2-N-glucuronide, urine was diluted with an SDS—phosphoric acid buffer (1:4) and injected directly onto the HPLC system, total C.V. ≤ 4.2% (0.5–50 μg/ml).

Evidence for the in vivo formation of ascorbic acid 2- O- α-glucoside in guinea pigs and rats

In vivo formation of ascorbic acid 2- O- α-glucoside (AA-2G) in guinea pigs and rats given ascorbic acid (AA) orally in combination with maltose was examined. A metabolite of AA which has the same HPLC retention characteristics as authentic AA-2G was detected in the blood, urine and liver of guinea pigs 1–2 hr after their administration. The metabolite was isolated from the urine by Chromatographie procedures and identified as AA-2G by its UV spectrum, non-reducibility, susceptibility to α-glucosidase hydrolysis, HPLC profile and elementary analysis. The same glucoside was also synthesized by rats and found in the urine, although it could not be determined qualitatively in the blood. AA-2G-forming activities of tissue homogenates from both animals were apparently correlated with their α-glucosidase activities and, moreover, both activities were completely inhibited by a specific neutral α-glucosidase inhibitor. Thus, we conclude that AA-2G is a possible metabolite produced by enzymatic α-glucosidation after a combined administration of AA and maltose to guinea pigs and rats.

Identification and characterization of 19-nortestosterone in urine of meat-producing animals

To monitor the illegal use of 19-nortestosterone as an anabolizing agent in meat-production, the Belgian Institute of Veterinary Expertise applies a strategy of urine control by radioimmunoassay, positive samples being confirmed by thin-layer chromatography. We have evaluated this control strategy, using gas chromatography—mass spectrometry to confirm the presence of 19-nortestosterone, or its metabolite oestrane-diol, in positive samples from radioimmunoassay. Our results show that the effective way of proceeding remains reliable in cattle, for mature and immature males as well as non-pregnant females, and in pigs, for pregnant and non-pregnant sows. The possible presence of endogenous 19-nortestosterone in cattle, in pregnant cows urine, and in pigs, in boars and in cryptorchid pigs, impedes the control of the use of 19-nortestosterone on these samples. False-positive (not confirmed by gas chromatography—mass spectrometry) results were produced by radioimmunoassay in the urine of castrated pigs and sheep.

Metabolism of mephentermine in male guinea pigs and male mice

1. Urinary metabolites of mephentermine (MP), after i.p. administration of MP to male Hartley guinea pigs and mice, were identified by g.l.c.-electron impact (EI) mass spectrometry. Excretion of urinary radioactivity, and metabolites of 3H-MP, after i.p. administration, were determined by preparative t.l.c.-liquid scintillation counting. 2. About 27% of the radioactivity administered was excreted in the 24 h urine of guinea pigs, and 36% dose was excreted in 5 days. In mice, about 47% of the radioactivity was excreted in the 24 h urine, and 52% in 5 days. 3. Excretion rates of metabolites detected in the 24 h urine of guinea pigs were phentermine (Ph, 7.8%), a conjugate of N-hydroxyphentermine (N-hydroxy-Ph, 3.6%), p-hydroxyphentermine (p-hydroxy-Ph, 1.0%) and its conjugate (2.9%), and other metabolites (conjugates of MP and Ph, N-hydroxymephentermine (N-hydroxy-MP) and its conjugate, p-hydroxymephentermine (p-hydroxy-MP) and its conjugate, and N-hydroxy-Ph; less than 1.0%). The rates of excretion for mice were Ph (11.7%), conjugates of p-hydroxy-MP (3.1%), Ph (2.7%) and p-hydroxy-Ph (1.6%), and N-hydroxy-Ph (1.2%) and other metabolites (conjugates of MP and N-hydroxy-Ph, N-hydroxy-MP and its conjugate, p-hydroxy-Ph, and p-hydroxy-MP; less than 1.0%). 4. These results indicate that MP administered to mice is metabolized mainly to Ph and p-hydroxy-MP by N-demethylation and p-hydroxylation of the parent compound, and in guinea pigs the primary metabolic reaction of MP is N-demethylation producing Ph.(ABSTRACT TRUNCATED AT 250 WORDS)

Fate of [14C]arsanilic acid in pigs and chickens

Arsanilic acid uniformly labeled with {sup 14}C in the benzene ring was used to determine the metabolic fate of oral arsanilic acid in pigs and chickens. Arsanilic acid was well absorbed in both species, and urine was the predominant route of excretion. The bile was a minor (< 5% of the dose) route of excretion in pigs; however, biliary excretion was not measured in roosters. Arsanilic acid, N-acetylarsanilic acid, and (4-acetamidophenyl)dimethylarsine oxide were isolated from pig urine (17-39%, 15-29%, and < 5% of urinary {sup 14}C, respectively). Only 25% of the {sup 14}C in pig feces was extractable, and no metabolites could be isolated. Arsanilic acid was the only radioactive compound isolated from urine of colostomized roosters, and there was no suggestion of other metabolites from the isolation scheme employed. No attempt was made to isolate {sup 14}C compounds in feces from colostomized roosters or in excreta from normal roosters.

New metabolites of the drug 5-aminosalicylic acid. II. N-Formyl-5-aminosalicylic acid

1. A new metabolite of the drug 5-aminosalicylic acid (5-ASA) has been found in urine from pigs and in plasma of humans. The metabolite has been isolated from pig urine using an XAD-2 column and purified using preparative h.p.l.c. 2. The metabolite has been identified as N-formyl-5-ASA (5-formamidosalicylic acid) using 1H- and 13C-n.m.r. spectrometry and mass spectroscopy and the structure was confirmed by chemical synthesis. 3. N-Formyl-5-ASA is stable in human plasma and in potassium phosphate buffers between pH 3.0 and 9.0. It is hydrolysed below pH 3.0. 4. N-Formyl-5-ASA was readily formed in rat liver homogenate when 5-ASA and N-formyl-L-kynurenine were added. Thus N-formyl-5-ASA might be formed by the actions of formamidase in vivo. 5. N-Formyl-5-ASA has been found in human plasma from healthy volunteers dosed i.v. with 5-ASA (250 mg). N-beta-D-glucopyranosyl-5-ASA, N-acetyl-5-ASA and N-formyl-5-ASA were quantified in human plasma using a h.p.l.c. assay.

Metabolismin vivoof the tropane alkaloid, scopolamine, in several mammalian species

1. In vivo metabolism of scopolamine was studied in rats, mice, guinea pigs and rabbits. The structures of eight urinary metabolites including unchanged drug were elucidated by mass and nuclear magnetic resonance spectrometry. Determination of these metabolites was achieved by a g.l.c. method using a semi-capillary column. 2. The major metabolites in rats were the three phenolic metabolites, p-hydroxy-, m-hydroxy- and p-hydroxy-m-methoxy-scopolamine. 3. Significant intra-species difference of the metabolism was observed in rabbits. Tropic acid was the major metabolite in two rabbits out of three, while the other rabbit excreted mainly unchanged scopolamine, accompanied by five metabolites. Tropic acid was also the major metabolite in guinea pigs, but was of minor importance in mice. 4. The dehydrated metabolites, aposcopolamine and aponorscopolamine, were abundantly excreted in guinea pigs, moderately in mice, and least in rabbits and rats. 5. Excretion of glucuronide conjugates of scopolamine and norscopolamine were high in mice compared with other species. On the other hand, phenolic metabolites in rat urine; and tropic acid in rabbit and guinea pig urine, were excreted as the free forms. 6. These results indicate that scopolamine metabolism is highly species-specific.

Hypoxanthine, Xanthine, and Uric Acid Concentrations in the Cerebrospinal Fluid, Plasma, and Urine of Hypoxemic Pigs

The concentrations of hypoxanthine, xanthine, and uric acid in plasma and cerebrospinal fluid (CSF), as well as the urinary output of hypoxanthine and xanthine, were measured in four groups of pigs (three groups with different degrees of hypoxemia and one control group). During hypoxemia with arterial O2 tension between 2.1 and 3.0 kPa [group 1, fractional inspired oxygen (FiO2) = 0.08], hypoxanthine increased in CSF from a mean basal value of 18.1 to 39.3 mumol/L at death (p less than 0.02), in plasma from 25.4 to 103.6 mumol/L (p less than 0.05), and in urine from 21.3 to 87.1 nmol/kg/min (p less than 0.02). Xanthine changed in a similar way: in CSF from 4.0 to 10.6 mumol/L (p less than 0.02), in plasma from 0.7 to 48.1 mumol/L (p less than 0.02), and in urine from 4.0 to 12.6 nmol/kg/min (p less than 0.05). Uric acid increased in CSF from 2.7 to 11.6 mumol/L (p less than 0.05), and in plasma from 15.4 to 125.0 mumol/L (p less than 0.02). During hypoxemia with arterial O2 tension between 3.0 and 4.0 kPa (group 2, FiO2 = 0.11), hypoxanthine increased in the CSF from 14.7 to 42.9 mumol/L (p less than 0.02). Plasma hypoxanthine increased from 20.3 to a maximum of 44.1 mumol/L (p less than 0.02), but decreased to initial values by the time of death. The urinary excretion of hypoxanthine increased from 13 to 54 nmol/kg/min (p less than 0.02).(ABSTRACT TRUNCATED AT 250 WORDS)

Thermospray high performance liquid chromatography/mass spectrometric identification of a bladder carcinogen metabolite isolated from guinea pig urine

An in vivo urinary metabolite of the bladder carcinogen 2-amino-4-(5-nitro-2-furyl) thiazole was isolated from guinea pig urine and was identified by direct analysis using thermospray mass spectrometry/high-performance liquid chromatography as 1-(2-amino-4-(5-nitro-2-furyl)-2-thiazolyl)-1-deoxy-beta-D-glucopyran uronic acid. The structure of this metabolite was also established by chemical synthesis. Both positive and negative ion thermospray mass spectrometry of the conjugate showed fragment ions resulting from cleavage across the pyran ring of the glucuronic acid comprising of aglycone moiety. These characteristic fragment ions may be diagnostic for identification of N-glucuronides from O-glucuronides.

Sulphadimidine in the urine of experimentally fed pigs.

An enzyme-linked immunoassay for the detection of sulphadimidine in unextracted pig urine is described. Twenty-four hour urine samples from six individually caged pigs, four treated and two controls, were examined during a 10-day treatment and a 12-day withdrawal period. The concentration of sulphadimidine in the urine of the treated pigs increased rapidly after feeding started and decreased on withdrawal. The maximum concentration in a control pig was 308 ng/ml, and this concentration was probably due to contamination of the environment. By the seventh day after withdrawal of the drug its concentration in the urine of the treated pigs was less than 500 ng/ml.

Excretion of Phenazopyridine and its Metabolites in the Urine of Humans, Rats, Mice, and Guinea Pigs

The metabolism of the urinary tract analgesic phenazopyridine [2, 6-diamino-3-(phenylazo)pyridine; PAP] was studied in the urine of humans, rats, mice, and guinea pigs. Urinary excretion was rapid in human and guinea pig, but in the rat and mouse it was slower and there was significant fecal excretion. Metabolism of PAP was extensive in all four species, and there were marked quantitative differences in the routes of metabolism. The extent of azo bond cleavage was high in the mouse and guinea pig, moderate in the rat, and low in humans. Hydroxylation of both the phenyl and pyridyl rings of PAP was observed in all species. In the human, 5-hydroxyl PAP was the major metabolite (48.3% of the dose). It was concluded that there are marked species differences in the metabolism of PAP, and that none of the species studied resembles the human; the rat comes closest, but cannot be considered a particularly good model.