Urinary Delta-aminolevulinic Acid Research Articles

Interaction of Zinc, Methionine or Their Combination with Lead at Gastrointestinal or Post‐Absorptive Level in Rats

The ability of zinc, methionine or their combination given by gavage to prevent or treat experimental oral lead intoxication in rats was investigated. Simultaneous oral supplementation with zinc plus methionine was found to be most effective in reducing lead induced inhibition of delta-aminolevulinic acid dehydratase (ALAD) activity in blood, elevation of urinary delta-aminolevulinic acid (ALA) excretion and in enhancing the hepatic glutathione (GSH) contents. The combination was also most effective in reducing the accumulation of lead in blood, liver and kidney compared to zinc or methionine alone. Prevention was more effective than treatment after lead exposure which may be caused by a decrease in the absorption of lead in the gastrointestinal tract in the presence of zinc and/or methionine.

Optimized fluorometric determination of urinary delta-aminolevulinic acid by using pre-column derivatization, and identification of the derivative

We previously reported the usefulness of a fluorometric method to determine urinary delta-aminolevulinic acid (ALA) concentrations by using post-column derivatization to monitor the effect of lead exposure. We have further improved the method by introducing pre-column derivatization by using reaction of ALA with acetylacetone and formaldehyde. Response of the hematopoietic system to lead exposure can now be easily detected at blood lead concentrations as low as 162 micrograms/L. The fluorescent ALA derivative, a new aromatic product, 2-methylideneamino-3,5-diacetyl-4,6-dimethylphenylpropionic acid, is separated on octadecyl silica column by high-performance liquid chromatography and the fluorescence intensity is detected with a fluorophotometer. Sample recoveries for 12 urine samples from workers exposed to lead and unexposed controls were 91.9-110.2%. The results obtained by the pre-column derivatization method agreed with those by the post-column derivatization method. The new method increases the sensitivity to a detection limit to 10 micrograms of delta-aminolevulinic acid per milliliter of urine and is simple enough to be used for routine monitoring of the biological effect of exposure to low concentrations of lead.

Feline Precursor. porphyria, characterized by persistent delta. Aminolevulinic aciduria

AbstractA female domestic shorthaired cat had frequent seizures and vague gastrointestinal signs. Lead poisoning was suspected because of increased urinary delta aminolevulinic acid concentration. However, abnormalities continued despite repeated chelation therapy and blood lead concentration was normal. Delta aminolevulinic aciduria persisted, but concentrations of porphobilinogen in urine and of porphyrins in blood, urine and faeces were normal. An inherited porphyric disorder was suspected, involving deficiency of aminolevulinic acid dehydrase, an enzyme active in haem biosynthesis. Accumulation of aminolevulinic acid in vivo could explain the observed signs.

Studies on erythrocyte pyrimidine 5?-nucleotidase (P5N) test and its evaluation in workers occupationally exposed to lead

An erythrocyte pyrimidine 5'-nucleotidase (P5N) test was performed for 171 workers occupationally exposed to lead. Erythrocyte P5N activity was markedly inhibited by exposure to lead. Among several biological indicators (erythrocyte P5N, delta-aminolevulinic acid dehydratase (ALAD), protoporphyrin (PROTO), urinary delta-aminolevulinic acid (ALA), coproporphyrin (COPRO)), the P5N activity had the highest correlation with the concentration of lead in blood (r = -0.77). A significant inhibition of erythrocyte P5N was found in groups of lead workers with blood-lead levels of more than 10 to 19 micrograms/dl. This P5N inhibition started before any changes occurred in urinary ALA and COPRO. A 45 to 50% inhibition of P5N corresponded to the blood-lead value (50 micrograms/dl) of the BEI recommended by ACGIH. In some lead workers, erythrocyte nucleotides (mainly CTP and UTP) were determined. The data indicated that a marked accumulation of these nucleotides had occurred, and their levels correlated negatively with P5N activity and positively with blood lead.

Alterations in tissue Pb distribution and hematopoietic indices during advanced age

The inevitable degenerative processes accompanying advanced age suggest that senescent organisms might exhibit differences in toxicokinetics or an altered manifestation of toxic effects. This hypothesis was tested in the present experiment which compared tissue distribution and hematopoietic indices in groups of adult (8 months) and old male Fischer-344 rats (16 months) exposed to concentrations of 0, 250 or 500 ppm lead (Pb) acetate in drinking water for 7 months. A group of young (weanling) rats was included as a partial comparison. Blood lead (PbB), zinc protoporphyrin (ZPP) and urinary delta-aminolevulinic acid levels were determined after both 3 and 7 months of exposure. Organ weights, tissue Pb concentrations and urinary excretion of Pb, calcium, copper, and zinc were examined after 7 months of exposure. Tissue Pb distribution was markedly altered in old rats: bone and kidney Pb were reduced; liver Pb was substantially increased, even though PbB levels in adult and old rats were comparable at both measurement intervals, as was urinary Pb excretion at 7 months. Pb-induced elevation of ZPPs exhibited differential changes between 3 and 7 months: values in adults declined while levels in old rats increased or remained unchanged. These findings show that the responses of adult rats to Pb do not predict those of older rats and indicate the need for further characterization and elaboration of mechanisms of toxic effects with advanced age.

A new HPLC fluorimetric method to monitor urinary delta-aminolevulinic acid (ALA-U) levels in workers exposed to lead.

A new sensitive HPLC method for the determination of urinary delta-aminolevulinic acid (ALA-U) was used to evaluate the relationship between blood-lead (Pb-B) and ALA-U levels in male workers exposed to lead. The differences between the ALA-U levels determined by this method (ALA-U-HP) and by a colorimetric method (ALA-U-CL) are discussed. The HPLC method gave values similar to the ALA-U-CL values at high ALA-U level. However, at low blood-lead levels (58 +/- 22 micrograms/l, n = 23), the mean ALA-U-HP level corrected by urinary creatinine level was one-third of the corrected ALA-U-CL level (0.83 +/- 0.14 and 2.4 +/- 0.5 mg/g creatinine, respectively). A significant increase of the mean corrected ALA-U-HP level was observed at 162 +/- 22 micrograms/l Pb-B (P less than 0.05, n = 26), while that of ALA-U-CL was observed at 245 +/- 30 micrograms/l Pb-B (P less than 0.01, n = 37). The regression equation based on the logistic model fitted well to the relationship data between the Pb-B level and the percentage of the subjects with corrected ALA-U-HP above the cut-off point (1.12 mg/g creatinine) and the expected Pb-B level for 50% response was 270 micrograms/l Pb-B, while it did not fit well to the relationship data between Pb-B level and the percentage of the subjects with corrected ALA-U-CL above the cut-off point (3.5 mg/g creatinine).(ABSTRACT TRUNCATED AT 250 WORDS)

Excretion of porphyrin and porphyrin precursor during ethylene oxide inhalation

The effect of chronic inhalation of ethylene oxide on urinary coproporphyrin and delta-aminolevulinic acid were studied. When Wistar male rats were exposed to 500 ppm ethylene oxide three times a week, daily urine volume was increased by 200-300% from the first week to the fifth week of the experimental period. After exposure, daily coproporphyrin excretion and urinary coproporphyrin per mg of creatinine increased by 250% and 141%, respectively. On the other hand, daily excretion of delta-aminolevulinic acid in urine tended to increase but did not increase significantly by creatinine-correction. We think this is the first report of ethylene oxide induced experimental porphyria.

Relationship between inhibition of erythrocyte pyrimidine 5'-nucleotidase activity and biological response for porphyrin metabolism in workers occupationally exposed to lead.

A rapid determination of erythrocyte pyrimidine 5'-nucleotidase (P5N) activity in lead workers was carried out using a high-performance liquid chromatograph (HPLC). The P5N activity had a good negative correlation with the concentration of lead in blood (PbB) ranging from 16 to 96 micrograms/dl (r = -0.82, n = 77). Further, the P5N was compared with other biological parameters: erythrocyte delta-aminolevulinic acid dehydratase (ALAD) activity, erythrocyte protoporphyrin (PROTO), urinary delta-aminolevulinic acid (ALA) and urinary coproporphyrin (COPRO). The correlation coefficients between P5N and ALAD, log PROTO, log ALA, and log COPRO were 0.59, -0.72, -0.65, and -0.61, respectively. On the other hand, the normal value of P5N obtained from 72 healthy subjects was 11.9 +/- 2.1 units; mumol uridine/h/g Hb (mean +/- SD), indicating that the lower limit of 95% confidence interval for normal P5N was about 8 units. When P5N was cut off at less than or equal to 8 units in 77 lead workers, the validity (sensitivity + specificity) for PbB greater than or equal to 40 micrograms/dl, PbB greater than or equal to 60 micrograms/dl, erythrocyte PROTO greater than or equal to 150 micrograms/dl RBC, urinary ALA greater than or equal to 6 mg/l, and urinary COPRO greater than or equal to 150 micrograms/l was 1.66, 1.76, 1.57, 1.68, and 1.60, respectively. From these results, it was confirmed that the erythrocyte P5N test is suitable for the biological monitoring of exposure to lead in a wide range, and its activity is useful in predicting the disturbance of porphyrin metabolism induced by lead.

Reevaluation of urinary excretion of coproporphyrins in lead-exposed workers.

Urinary concentrations of coproporphyrin I and III (CP I and III) were determined by high-performance liquid chromatography in 131 male workers exposed to lead, and the relationships between lead exposure and urinary coproporphyrins were reevaluated. CP I had a statistically significant correlation with lead in the blood (Pb-B), but it was not useful as an indicator of the effect of lead on heme metabolism. On the other hand, CP III had a good correlation with Pb-B and markedly increased when Pb-B levels exceeded 40 to 50 micrograms/100 ml. Both sensitivity and specificity were more than 80% when the health-based Pb-B limit and the screening level of CP III were fixed at 50 micrograms/100 ml and 50 micrograms/g creatinine, respectively. In conclusion, measurement of CP III is sufficiently sensitive and specific enough in practice for the early detection of health effects due to lead exposure in the same way as the measurement of delta-aminolevulinic acid in urine.

Optimized liquid-chromatographic method for fluorometric determination of urinary delta-aminolevulinic acid in workers exposed to lead.

We developed a fluorometric method for determining delta-aminolevulinic acid (ALA) in urine of lead workers. A high-performance liquid chromatograph (HPLC) equipped with a fluorescence HPLC monitor is used. The detection limit for aqueous ALA is 20 micrograms/L (15 pmol of ALA in the 100-microL sample). The working linear range of urinary ALA concentration was 0.1 to 100 mg/L. In 25 lead-exposed workers, ALA values by the present method for urine correlated well with those obtained by a conventional colorimetric method (r = 0.996). The advantage of the present method for micro-determination of urinary ALA is its high sensitivity.

Evaluation of a new specific analysis of urinary delta-aminolevulinic acid in man

A new specific method for urinary delta-aminolevulinic acid (ALA-U) measurement in man as a serial screening method for health control in occupational lead exposure was compared with a recommended, selective test (DAVIS method). The new test can be run fully automated, and the cost of material per sample is much lower than in the DAVIS method. ALA values obtained with this specific method were lower than those measured with the DAVIS method (= DAVIS values) in each case due to the elimination of interfering urinary compounds. In DAVIS values below the biological tolerance level (BAT-level) of 15 mg/l, the percentage of interfering compounds varied between 10 and 80%; above BAT level, these components represented about 20% of the total amount of ALA-U. A strong correlation can be demonstrated between specifically measured ALA values and the enzyme activity of ALA-dehydratase (ALA-D) in blood at lead levels (Pb-B) below 35 micrograms/dl (r = -0.768 for ALA-U/log ALA-D; n = 18).

Effects of lead, zinc and copper absorption on peripheral nerve conduction in metal workers

To examine the antagonistic effects of lead, zinc and copper on the nervous system in man, we measured maximal motor and sensory conduction velocities (MCV and SCV, respectively) in the distal radial and median nerves in 20 gun metal founders with asymptomatically increased absorption of these metals [blood lead concentrations ranged from 16 to 64 micrograms/dl (mean 42)]; and analyzed metal effects on nerve conduction by multiple regression analysis. The MCVs in the radial and median nerves and the SCV in the forearm segment of the median nerve were significantly slowed; indicators of lead absorption (urinary delta-aminolevulinic acid and coproporphyrin) were inversely correlated with the SCV in the radial nerve. The SCVs in the radial nerve and in the hand segment of the median nerve, which were not significantly slowed, were positively related to indicators of copper absorption (plasma or erythrocyte copper). The SCV in the radial nerve was also positively related to the indicator of zinc absorption (plasma zinc); similarly, the MCVs in the radial and median nerves were positively related to the indicators of zinc absorption (erythrocyte or urinary zinc). These findings suggest that copper and zinc antagonize the subclinical effects of lead on peripheral nerve conduction velocities; the conduction velocity in the distal radial nerve is sensitive not only to asymptomatic lead absorption but also to zinc and copper absorption.

A chronobiological study of delta-amino levulinic acid urinary excretion.

Determination of urinary delta-amino levulinic acid (ALA) is now systematically used in occupational health to detect an excessive exposure to lead in professionally exposed workers. However, to determine whether circadian changes of the urinary excretion of ALA alter the significance of the test, we quantified the ALA levels in the urine of 19 healthy young adults. Urine samples were taken every 3 hr between 0700 and 2300 hr and ALA levels were determined by a spectrocolorimetric method. The data indicated that the 24-hr mean ALA level was: 1.81 mg/g creatinine. The peak values (2.24 +/- 0.24) were obtained between 1400 and 1700 hr whereas the lowest ALA levels were found between 2200 and 0300 hr. Cosinor analysis revealed a significant circadian rhythm while no significant difference could be found according to sex. Possible explanations of our findings are discussed.

Clinical Laboratory Manifestations of Exposure to Dioxin in Children

Following a major environmental accident near Seveso, Italy, on July 10, 1976, we attempted to determine if the 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) released into the atmosphere had any effect on the liver function and lipid metabolism of exposed children. From July 1976 to June 1982, we analyzed the results of more than 4500 laboratory tests (alanine aminotransferase, aspartate aminotransferase, gamma-glutamyltransferase, alkaline phosphatase, cholesterol, and triglycerides in plasma and delta-aminolevulinic acid in urine) in a population of about 1500 children aged 6 to 10 years at the moment of the accident. The children exposed to the highest concentration of TCDD showed alterations in serum gamma-glutamyltransferase and alanine aminotransferase activity compared with the control group. These differences were restricted to values inside limits set from the lower end of the normal range to slightly above it. The observed abnormalities were slight and disappeared with time.

Quantitative determination of urinary delta-aminolevulinic acid as an index of lead exposure by high-performance liquid chromatography.

Quantitative determination of urinary delta-aminolevulinic acid as an index of lead exposure by high-performance liquid chromatography.

Improved method for the determination of urinary delta-aminolevulinic acid in lead workers

An improved method for the determination of urinary delta-aminolevulinic acid (ALA) as an indicator of lead absorption is described. It is a modification of the analytical procedure described by Tomokuni and Ogata in 1972. In the present method, the spot urine from lead-exposed workers which was diluted 3-fold with distilled water beforehand was used as the sample to eliminate as much as possible the urinary matrix interference. In addition, methyl acetoacetate was used as the condensation reagent for ALA-pyrrole, because the color intensity of ALA-pyrrole was higher with the use of methyl acetoacetate than that with ethyl acetoacetate which was used in the original method. The urinary ALA value determined by the present method was similar to that measured using an internal-standard addition method. This method is suitable as a rapid and inexpensive screening procedure for the routine quantitative analysis of urinary ALA.

An outbreak of lead poisoning in dogs.

An outbreak of lead toxicosis in working dogs on a sheep farm is described. The affected dogs exhibited neurological signs which appeared to be initiated by strenuous physical activity. Radiological examination and estimation of urinary delta aminolevulinic acid (U-ALA) was not found to be useful for clinical diagnosis but blood lead estimation was helpful in detecting and confirming dogs that were exposed to lead. Gross pathological lesions were absent in a lead poisoned dog. However, histopathological lesions were seen in brain and bone. Acid-fast lead inclusion were not detected in liver and kidney epithelial cells but were present in osteoclasts.

Blood lead, urinary delta aminolevulinic acid and the diagnosis of lead poisoning in dogs.

Blood lead, urinary delta aminolevulinic acid and the diagnosis of lead poisoning in dogs.

Simple screening test for qualitative detection of increased delta-aminolevulinic acid in urine.

A simple qualitative procedure for demonstrating increased delta-aminolevulinic acid (ALA) in urine is needed to identify individuals with an inherited deficiency of the enzyme ALA dehydratase, a newly described porphyria-like syndrome. The present procedure includes two steps, the last of which is carried out only on samples that are positive in the first step. Urine is tested for pyrroles, preformed or formed by heating with acetylacetone, by looking for a pink color after p-dimethylaminobenzaldehyde is added. Samples showing the color are next tested for ALA by adsorbing natural pyrroles onto Dowex-1 in sodium acetate, heating the supernate with acetylacetone, adsorbing the resulting aminoketone pyrroles onto Dowex-1 in water, separating the ALA from other aminoketones by differential extraction, and again looking for a pink color with p-dimethylaminobenzaldehyde. Positive urines, those with concentrations of ALA greater than or equal to 10-15 mg/L, yield a pink color in each step. Samples with greater than 30 mg/L ALA show a magenta color. Aminoacetone in excess of 3 mg/L ALA also yields a pink color, but such a concentration in urine is unlikely. Many samples can easily be tested concurrently.

Investigations on neurotoxicity of chemical substances at the workplace. V. Determination of the motor and sensory nerve conduction velocity in persons occupationally exposed to lead.

A cross-sectional study was performed in order to investigate the influence of chronic lead-exposure on the peripheral nervous system. We examined 148 male workers of a storage battery manufacturing plant, who had been exposed to lead metal and inorganic lead compounds for 1 to 28 years (mean 11 years). Fifteen workers with non-occupational risks of peripheral neuropathy (former diseases, alcohol abuse, medication) were excluded from the study. The investigation program comprised: case history, physical examination, analyses of blood- and urine-samples and determination of maximal motor, mixed and sensory conduction velocity (NCV) of the ulnar and median nerve of the right forearm. Objectively no worker showed any signs of health effects related to lead exposure. The "Biological Monitoring" included the determination of (1) Blood-lead level (Pb-B), (2) Free erythrocyte porphyrins (FEP), (3) delta-Aminolevulinic acid dehydratase (ALA-D) and (4) delta-Aminolevulinic acid in urine (ALA-U). Further "time-weighted-average (TWA)"-values of Pb-B were calculated on the basis of several determinations over the period 1975-1981. The following "actual" ("TWA") median values resulted: Pb-B 53 micrograms/dl (54 micrograms/dl), ALA-U 5.6 mg/l (8.4 mg/l), FEP 2.0 mg/l (2.0 mg/l). The "Biologischer Arbeitsstoff Toleranz Wert (BAT)" of 70 micrograms/dl for Pb-B was exceeded in 15 workers (11%), and of 15 mg/l for ALA-U in 30 cases (23%). In comparison with age-matched controls, the lead workers showed a mild slowing of NCV with mean values between 0.8 and 2.0 m/s. Multiple stepwise regression analyses revealed statistically significant correlations between the four NCV and age as well as Pb-B. There were better correlations by using "TWA" than "actual" data of Pb-B. Consideration of the results of the regression analyses, together with an evaluation of the individual neurophysiological status as a function of internal lead exposure, a "dose-effect-relationship" was found only in the case of Pb-B exceeding 70 micrograms/dl. From our study it is concluded that chronic lead exposure resulting in blood-lead levels of below 70 micrograms/dl is no occupational risk causing a functionally significant slowing of nerve conduction velocities.