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Hippuric acid and o-cresol in the urine of workers exposed to toluene.

Factory workers, 74 males and 56 females exposed predominantly to toluene up to 129 ppm, were examined for the urinary excretion of hippuric acid and o-cresol. The time-weighted averages (TWA) of toluene exposure were measured by personal sampling with carbon felt dosimeters. A preliminary study revealed that the concentrations of hippuric acid and o-cresol in urine increased during work and both reach their peaks at the end of the shift. Correlation coefficients between the TWA of toluene concentration in air and hippuric acid concentration in urine collected at the end of the shift were 0.803 for the 74 males, and 0.830 for the 56 females, while the counterpart correlation coefficients between toluene and o-cresol were 0.607 for the 74 males, and 0.627 for the 56 females, suggesting that hippuric acid is more reliable than o-cresol as an index of toluene exposure. In the urine samples (4 to 8 samples per subject) collected during 8-h worktime from 11 males and 13 females, the urinary levels of o-cresol increased as a function of exposure time in parallel with those of hippuric acid, and the correlation coefficients between o-cresol and hippuric acid were significant (r = 0.834 approximately 0.987; P less than 0.05) when the urine samples from the same subjects were examined. The comparison of the slopes of 24 regression lines between o-cresol and hippuric acid in urine revealed that the maximal slope was almost 8 times as large as the minimal one. From 8 female workers, five urine samples each were collected during 8-h worktime on two consecutive Mondays and analyzed for the two metabolites. The slopes of the regression lines between o-cresol and hippuric acid in the samples from the same subject were identical, regardless of variation in exposure intensity. The findings indicate that an individual difference exists in the pattern of toluene metabolism, and that the ratio between aliphatic and aromatic oxidation is presumably set congenitally. Possible toxicological significance is discussed.

Air Pollutants, Occupational↗

Potentiation by piridoxilate of the synthesis of hippurate from benzoate in isolated rat hepatocytes. An approach to the determination of new pathways of nitrogen excretion in inborn errors of urea synthesis.

The synthesis of hippurate from benzoate as compared to ureagenesis has been investigated in isolated rat hepatocytes. Half-maximal synthesis of hippurate was observed at 0.3 mmol/l benzoate. In the presence of 1 mmol/l benzoate, hippurate synthesis proceeded linearly with time at a rate of 40 +/- 10 mumol X h-1 X g-1 dry weight. This provided less than 10% of nitrogen epuration supported by concomitant urea synthesis (350 +/- 82 mumol X h-1 X g-1 dry weight). The incorporation of benzoate to hippurate was markedly limited by the availability of glycine. Half-maximal hippurate synthesis was observed at 2 mmol/l glycine. In the absence of glycine, piridoxilate, a glyoxylate derivative, markedly potentiated hippurate synthesis. Half-maximal stimulation was observed at 10 mmol/l piridoxilate. In the presence of 10 mmol/l piridoxilate, hippurate synthesis (420 +/- 35 mumol X h-1 X g-1 dry weight) provided more than 50% of nitrogen epuration supported by urea synthesis. It is concluded that supplementation with nitrogen-free analogues of glycine such as piridoxilate are required to potentiate hippurate synthesis in an attempt to replace ureagenesis as an alternative pathway of waste nitrogen excretion in inborn errors of urea synthesis.

Alanine↗

Toxicokinetics of toluene and urinary excretion of hippuric acid after human exposure to 2H8-toluene.

Nine male volunteers were exposed to 2H8-toluene (200 mg/m3 for two hours during a workload of 50 W) via inspiratory air with the aid of a breathing valve and mouthpiece. Labelled toluene was used to differentiate between hippuric acid originating from exposure to toluene and hippuric acid normally excreted in urine. The total uptake of toluene was 2.2 (standard deviation (SD) 0.2) mmol, or 50% of the amount inhaled. Four hours after the end of exposure 1.4 (SD 0.3) mmol or 65% of the total uptake had been excreted in urine as 2H-hippuric acid and 20 hours after the end of exposure the cumulative excretion of 2H-hippuric acid was 1.8 (SD 0.3) mmol, or 78% of the total uptake. By contrast the cumulative excretion of labelled plus unlabelled hippuric acid exceeded the total uptake of toluene already after four hours. The excretion rate of 2H-hippuric acid was highest, about 5 mumol/min, during exposure and the SD between the subjects was low. The background concentrations of unlabelled hippuric acid in urine were high, however, and there were large differences between subjects. These findings confirm earlier indications that for low exposure, urinary hippuric acid concentration cannot be used for biological monitoring of exposure to toluene.

Adolescent↗

Ammonia production from hippurate by the rat kidney in vitro.

Hippurate is known to be synthesized from benzoate and glycine in the liver and kidney. It takes part in renal ammoniagenesis by modulating the activity of gamma-glutamyl transpeptidase (gamma GT). Due to its chemical structure, however, hippurate might also serve as a substrate of renal ammoniagenesis. Hippurate may yield ammonia either having been cleaved by hippuricase or by Erlenmeyer's reaction after condensation with an aldehyde. In order to elucidate the possibility of hippurate being a substrate of renal ammoniagenesis, experiments were carried out on cortical kidney slices and on isolated tubular segments of the rat. The incubation medium (pH 7.1) was enriched with 10 mmol/l hippurate spiked with 15N-hippurate, some of the known competitive inhibitors of hippuricase, acivicin and different aldehydes. Factors known to affect hippuricase or gamma GT did not interfere with renal ammonia production. Glyceraldehyde (up to 1.0 mmol/l) but not glycerate had a stimulating effect, especially on the ammoniagenesis from hippurate. In normal rats fed a vegetarian diet, 1% of the added 15N moiety was found to be 15NH3. Renal 15NH3 production was significantly greater if, prior to the experiments, the animals were either acidotic or had a reduced renal mass or were fed animal proteins. These results indicate that hippurate may, to a certain extent, serve as substrate for ammoniagenesis.

Acidosis, Renal Tubular↗

DNA relatedness among strains of Campylobacter jejuni and Campylobacter coli with divergent serogroup and hippurate reactions.

Eleven strains of Campylobacter from earlier fluorescent-antibody studies were examined by DNA hybridization to determine their species. Three of the strains hydrolyzed sodium hippurate, and eight did not. Four of the hippurate-negative strains were in Campylobacter jejuni serogroups, and the remaining strains were in both C. jejuni and Campylobacter coli serogroups. DNA relatedness to type strains of C. jejuni and C. coli indicated that all three of the hippurate-positive strains and two of the hippurate-negative strains were C. jejuni. The six remaining hippurate-negative strains were C. coli. Two of the hippurate-negative strains in C. jejuni serogroups were C. jejuni, and two were C. coli. Three of the strains in serogroups of both species were C. jejuni, and four were C. coli. These studies confirm that a few strains of C. jejuni are hippurate negative and show that identical or highly related antigens are found in C. coli and C. jejuni.

Campylobacter↗

Renal 131I-hippurate clearance overestimates true renal blood flow in the instrumented conscious dog.

We evaluated renal 131I-hippurate clearance (ERPFhip) as a measure of renal blood flow (RBF) in chronically instrumented conscious dogs. When adjusted for renal hippurate extraction (Ehip, 0.77 +/- 0.01) and hematocrit (Hct, 39.7 +/- 1%), calculated RBFhip (656 +/- 37 ml/min) markedly exceeded renal blood flow measured with renal artery blood flow probes (RBFprobe, 433 +/- 27 ml/min). The discrepancy could not be explained by flow probe calibration, because in vivo comparison of flow probe values with renal venous outflow showed only a slight underestimation of renal blood flow (slope 0.93, 95% confidence interval 0.89-0.97). Redistribution of hippurate from erythrocytes into renal venous plasma during or shortly after blood sampling led to an underestimation of Ehip by 4 +/- 1% and thus could only explain a small part of the difference. Extrarenal hippurate clearance was excluded, because the amount of 131I-hippurate cleared from plasma equaled that appearing in the urine (303 +/- 17 and 307 +/- 17 ml/min). Applying these corrections, we found that RBFhip still exceeded RBFprobe by 37 +/- 3%. These data indicate that renal blood flow measured by the hippurate clearance technique markedly overestimates true renal blood flow. Because other errors were excluded, a combination of sampling of nonrenal blood and intrarenal hippurate extraction from erythrocytes might play a role.

Animals↗

Determination of renal plasma flow by endogenous hippuric acid clearance in the conscious rat.

(1) The endogenous hippuric acid can be fluorometrically determined down to a minimum of 0.6 micrograms/ml in 10 microliters of rat serum. This enables the determination of the effective renal plasma flow (ERPF) by measurement of the renal clearance of the endogenous hippuric acid in the conscious rat. (2) Endogenous hippuric acid clearance (128 ml/min . m2 or 2.48 ml/min/100 g body weight) corresponds in its extent with the clearance after a single injection of unlabelled hippuric acid and is higher (20%) than the clearance after a single injection of 131I-hippuran. (3) Exogenous hippuric acid has a short biological half-time of 12-20 min and a distribution space of about 50%; therefore, alteration of its intermediary supply can disturb the estimate of the renal clearance of endogenous hippuric acid. (4) The daily rhythm of the renal excretion of endogenous hippuric acid shows a constant low excretion between 0800 and 1200 hours; this period is the best suited for clearance measurements.

Animals↗

Hippurate hydrolysis and Christie, Atkins, Munch-Peterson tests as epidemiological diagnostic tools for Streptococcus agalactiae carriage in pregnancy.

OBJECTIVE: To evaluate the Christie, Atkins, Munch-Peterson (CAMP) and hippurate hydrolysis reactions as diagnostic tools for Streptococcus agalactiae carriage in pregnancy. DESIGN: Observational, analytical case-control study. SETTING: Hospital-based study in a primary and a tertiary health care institution. PATIENTS: One hundred and six pregnant and 56 non-pregnant (controls) women were included in the study. The participants were of different socio-economic status. A volunteer sample was used. About 800 subjects were contacted and 162 participated in the study. RESULTS: The sensitivity of the screening test varied from 25% for the CAMP test to 77.78% for the hippurate hydrolysis reaction. The specificity was the same for both tests at (50%). A significant difference in positivity between the CAMP and hippurate hydrolysis reactions (95% confidence limit, P < 0.05) was observed. The predictive values of the positive test were 66.6% (CAMP) and 87.55% (hippurate hydrolysis) while the negative test were 14.29% (CAMP) and 33.30% (hippurate hydrolysis). Pregnant women had 0.33 chances of being GBS carriers with the CAMP compared to 3.5 with the hippurate hydrolysis. CONCLUSION: The hippurate hydrolysis test is highly recommended since the reagents are easily available and the organism was easily isolated using this method. The presence of GBS in the anorectum and endocervix is likely to induce systemic and local immunity in the female genital tract. This can contribute to the development of a mucosal vaccine for GBS diseases.

Adult↗

The effect of cigarette smoking on urinary hippuric acid concentration in Thai workers with occupational exposure to toluene.

Urine hippuric acid determination is helpful for monitoring of group of workers at risk for exposure to toluene. However, some problems about the external source of variation are mentioned. Some studies have indicated that smoking is an important external source of variation for determination of urine hippuric acid level while the others stated the opposite findings. This study was conducted in an attempt to study the difference of urine hippuric acid between smoking and non smoking subjects in a press workers group. Urine samples were obtained from 46 workers (all male) who worked as press workers in the same press factory in Bangkok. The individuals were classified as control (non smoking, N = 26) and experimental (smoking, N = 20) according to their smoking. All samples were analyzed for hippuric acid level. The average urine hippuric acid level for the control were (0.35 +/- 0.31 mg/gCr) and experimental group (0.40 + 0.45 mg/gCr) were respectively. No significant difference was found between urine hippuric acid level between both groups. The data from the current study indicates that smoking does not influence the urinary hippuric acid levels in this study group.

Adult↗

Clinical evaluation of Tc-99m N,N'-bis(mercaptoacetyl)-2,3-diaminopropanoate as a replacement for I-131 hippurate: concise communication.

A clinical comparison of Tc-99m N,N'-bis( mercaptoacetyl )-2,3- diaminopropanoate (Component A) (Tc-99m CO2-DADS-A) and I-131 hippurate was conducted in a series of five normal volunteers and 18 patients. Each subject was studied in one session with Tc-99m CO2-DADS-A and I-131-hippurate; digital and analog images were recorded for 30 min and after voiding. In the normal volunteers, digital images with Tc-99m CO2-DADS-A gave a kidney-to-background ratio at 3 min that was greater relative to I-131 hippurate, a leading-edge parenchymal transit time that was similar to I-131 hippurate, and a percent injected dose in the urine at 30 min that was slightly less than I-131 hippurate (p less than 0.05). In patients (serum creatinine 1.0 to 14.3 mg/dl), decreasing renal function impaired excretion of Tc-99m CO2-DADS-A more than that of I-131 hippurate (p less than 0.01). In analog images, Tc-99m CO2-DADS-A always gave superior spatial resolution. No evidence of hepatobiliary excretion was detected with either radiopharmaceutical. We conclude that Tc-99m CO2-DADS-A and similar compounds should be pursued as possible replacements for I-131 hippurate.

Alanine↗

[The iodine 131-hippurate clearance for the determination of the nephrotoxic effect of cis-dichlorodiamino-platin (II) (cis-platin, cis-DDP)].

In 25 patients, who received repeated doses of cis-DDP, we determined iodo-131-hippurate-clearance, creatinine clearance and serum creatinine to assess the nephrotoxic effect induced by cis-DDP. A highly significant decrease in the iodo-131-hippurate-clearance was found between the first and the following cycles. There was no significant change in serum creatinine and creatinine clearance however. The radio-hippurate-clearance showed a clinically not important reduction during several cytotoxic courses in 15/25 patients (60%). A clinically relevant impairment of the radio-hippurate-clearance occurred in 8/25 patients (32%), mainly before the third and fifth cytotoxic cycle. In two patients this clearance did not alter or was even rising. In 12 cases a diminuation of the iodo-131-hippurate-clearance was observed, which in only 5 cases correlated to a decreased creatinine clearance and in only two cases to an elevated serum creatinine. Therefore, we consider neither creatinine nor creatinine clearance nor the combination of both parameters to be appropriate to assess tubular nephropathy induced by cis-DDP. Radio-hippurate-clearance should play a superior role deciding on treatment stop or dose reduction of cis-DDP. The regression of cis-DDP-induced nephropathy was not evaluated.

Adult↗

Comparison of serine and hippurate as precursor equivalents during infusion of [15N]glycine for measurement of fractional synthetic rates of apolipoprotein B of very-low-density lipoprotein.

Enrichment in hippurate has been measured to indicate precursor enrichment during glycine tracer infusion studies to estimate fractional synthetic rates of individual hepatic export proteins. However, hippurate tends to overestimate precursor enrichment. Since glycine is rapidly converted to serine by liver cells, we compared tracer enrichment in hippurate and serine with that of glycine incorporated into apolipoprotein (apo) B-100. Ten healthy control subjects were studied in the postabsorptive state during an 8-hour primed-constant infusion of [15N]glycine (10 mumol.kg-1.h-1). Apo B of very-low-density lipoprotein (VLDL) was isolated by standard ultracentrifugation and isopropanol precipitation. Glycine and serine were isolated from plasma and hydrolyzed apo B, hippurate was isolated from plasma, and [15N]enrichment was determined by gas chromatography-mass spectrometry. Enrichment in serine and glycine isolated from apo B was identical at all time points, and their enrichment in apo B increased asymptotically, approaching an apparent plateau (mean +/- SD: 91% +/- 10% of calculated plateau at 8 hours) that was taken to represent hepatic protein precursor enrichment. Enrichment in both plasma serine and hippurate followed a biphasic pattern and continued to increase until the end of the study, raising the possibility that precursor enrichment had not reached a steady state during the study. The apo B plateau was lower (factor 0.76 +/- 0.27) than the final enrichment in hippurate and higher (factor 1.38 +/- 0.36) than that in plasma serine; however, predictions of protein precursor enrichment based on either metabolite were flawed by a large coefficient of variation (35% v 26%).(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

The synthesis of hippurate from benzoate and glycine by rat liver mitochondria. Submitochondrial localization and kinetics.

1. Rat liver mitochondria make hippurate at up to 4 nmol/min per mg of protein. The rate of synthesis supported by oxidation of glutamate with exogenous Pi present is identical with that supported by ATP plus oligomycin. Lower rates were obtained with other respiratory substrates, and when glutamate was used without Pi. 2. A matrix localization for hippurate synthesis is indicated by the latency of benzoyl-CoA synthetase and glycine N-acyltransferase to their extramitochondrial substrates, failure of exogenous benzoyl-CoA to inhibit incorporation of [14C]hippurate and inhibition of hippurate synthesis supported by ATP, but not glutamate, by carboxyatractyloside. 3. The relative activities of the individual enzymes and the mitochondrial content of benzoyl-CoA in the presence and absence of glycine suggest that hippurate synthesis is rate-limited by formation of benzoyl-CoA. 4. The increases in rates of ATP hydrolysis and of O2 consumption on the addition of benzoate and glycine were in good agreement with those required to support hippurate synthesis. The increase in respiration indicates that State-4 respiration [Chance & Williams (1957) Adv. Enzymol 17, 65-134] is not used, with these conditions, for ATP synthesis.

Adenosine Triphosphate↗

Correlation of a colorimetric and a HPLC method for the determination of serum hippuric acid concentrations in uremia.

Hippuric acid has been recognized as a potential marker of uremic toxicity in chronic renal failure. However, in most studies, serum hippuric acid concentrations have been determined by sophisticated methods, such as high-performance liquid chromatography. The present study was undertaken to evaluate whether the less complicated colorimetric determination method could replace such methods. Based on 21 different samples, the results obtained by both methods appeared to be correlated to each other in a highly significant way (total hippuric acid: r = 0.99, p less than 0.001; free hippuric acid; r = 1.00, p less than 0.001). Mean total and free hippuric acid concentrations and mean percent protein binding, obtained with both determination methods, were also identical. It is concluded that both the colorimetric method and high-performance liquid chromatography are equally reliable for the study of the concentration of hippuric acid in uremic serum and of its importance as a marker of the clinical and biochemical epiphenomena of uremic toxicity.

Chromatography, High Pressure Liquid↗

Hippurate participation in the inhibition of glucose utilization in renal failure.

Patients with renal insufficiency suffer from inhibited glucose utilization which can be tested in vitro by the incubation of tissues or isolated cells in diluted uraemic serum. 1. Systematic isolation from the serum of patients with renal insufficiency has revealed that hippurate participates in the inhibition of glucose utilization in the muscle and, to a lesser extent, in the kidney. It does not inhibit glucose utilization in the brain, red blood cells and platelets. 2. Maximal inhibition is 86.6% and AD50 is 0.432 mmol hippurate/l. These hippurate concentrations are achieved in the early stages of renal insufficiency. 3. While the inhibitory effect is evident at normal and elevated glucose concentrations, it does not manifest itself at hypoglycaemic concentrations. 4. A less pronounced inhibitory effect, compared with hippurate, is exerted also by O-OH hippurate, but it is absent altogether in P-amino-hippurate and O-OH hippuryl-beta-glucuronide. It is concluded that, through its inhibition of glucose utilization, hippurate may participate in the development of muscular weakness in patients with renal insufficiency.

Animals↗

[Renal clearance of hippurate in persons with chronic renal insufficiency].

In 8 subjects with chronic renal failure treated conservatively the renal hippurate clearance (CHip), polyfructosan (CPF) and creatinine (Ccr) clearance was assessed. The mean values of the investigated variables were as follows: CHip = 1.06 +/- 0.79 ml/s, CPF = 0.15 +/- 0.08 ml/s, Ccr = 0.27 +/- 0.17 ml/s. The ratios between the investigated clearance values were: CHip/CPF = 6.9 +/- 4.2 and CHip/Ccr = 4.2 +/- 1.4. The ratio of CHip/CPF was in some instances as high as 10 or more. Between values of CHip/CPF and the plasma hippurate concentration (PHip) a significant negative (non-linear correlation was found (p < 0.01). When the PHip values were very high, the value of the CHip/CPF ratio was close to 1. The recorded findings confirm that in subjects with chronic renal failure in the residual nephrons hippurate is synthetized and its urinary excretion in relation to glomerular filtration is increased. The findings suggest also that in these patients the tubular transport for hippurate is saturated. It will be necessary to take into account the possible saturation of tubular transport for hippurate when interpreting changes of drug excretion (i.e. those excreted by the same mechanism as hippurate) in subjects with chronic renal failure.

Adult↗

Participation of P-dependent and P-independent glutaminases in rat kidney ammoniagenesis and their modulation by metabolic acidosis, hippurate and insulin.

The key regulatory enzymes of kidney ammoniagenesis appear to be P-dependent (PDG) and P-independent (PIG) glutaminases. While the participation of PDG has been satisfactorily elucidated, the significance of PIG remains doubtful. Rat kidney cortex slices synthesized ammonia even under basal conditions. Metabolic acidosis, hippurate and insulin stimulated ammonia production. Under basal conditions, PDG activity in kidney homogenate, was twice as high as PIG activity. Metabolic acidosis stimulated ammonia production by the stimulation of both PDG (100%) and PIG (57%) activities. Hippurate stimulated only PIG activity both under basal conditions (90%) and in metabolic acidosis (52%), while it inhibited PDG activity only insignificantly under basal conditions and markedly (53%) in metabolic acidosis. Insulin stimulated both PIG and PDG activities under basal conditions as well as in metabolic acidosis and potentiated the PIG stimulation by hippurate while it potentiated the hippurate inhibition of PDG both under basal conditions and in acidotic rats. In conclusion, both PDG and PIG participate in ammoniagenesis and are stimulated by metabolic acidosis and insulin. Hippurate stimulates PIG, while it inhibits PDG in metabolic acidosis and even after insulin administration. The effect of hippurate appears to be of physiological interest.

Acidosis↗

The hippurate ratio as an indicator of functional hepatic reserve for resection of hepatocellular carcinoma in cirrhotic patients.

Predicting the ability of the cirrhotic liver to withstand resection remains a challenge for the surgeon. This study evaluates the use of the hippurate ratio, a novel assessment of glycine conjugation of para-aminobenzoic acid by the liver, as a preoperative indicator of functional hepatic reserve. Between 1998 and 2000, sixty-one cirrhotic patients were prospectively assessed for hepatic resection using the hippurate ratio, indocyanine green retention at 15 minutes (ICG R-15), and other standard measures of liver function. Twenty-six patients were excluded as candidates for resection on the basis of inadequate functional hepatic reserve. Patients excluded from resection had significantly higher ICG R-15 values (29% +/- 9% vs. 16% +/- 12%, P = 0.001), higher Child-Pugh scores (5.9 +/- 0.9 vs. 5.3 +/- 0.4, P = 0.01), and lower hippurate ratios (30% +/- 14% vs. 45% +/- 15%, P = 0.005). There was a significant correlation between the hippurate ratio and ICG R-15. Other indicators of liver function such as factor V, factor VII, albumin, bilirubin, prothrombin time, and transaminases were no different between patients who did and those who did not undergo resection. Of the 35 patients resected, there were seven (20%) who developed varying degrees of liver failure with three perioperative deaths (8.5%). Patients who had some degree of liver failure had significantly lower hippurate ratios than patients who had no liver failure (29% +/- 10% vs. 48% +/- 14%, P = 0.002). There was no difference in ICG R-15 values between patients who had liver failure and those who did not. The hippurate ratio offers information on hepatocellular reserve that is not provided by other measures of liver function and may allow better selection of cirrhotic patients for liver resection.

4-Aminobenzoic Acid↗