• Journal of Electrochemical Science and Technology
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• v.2 no.1
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• pp.57-61
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• 2011

This paper describes an electrochemical immunoassay for simple, fast and quantitative detection of a urinary hippuric acid which is one of major biological indicator in toluene-exposed humans. The electrochemical system of immunoassay was based on the directly osmium complex conjugated with hippuric acid. With the competition between free hippuric acid (HA) and the osmium-hippuric acid conjugate (Os-HA) to bind with antibody hippuric acid (Anti-HA) coated onto gold nanoparticles, the electrical signals were proportional to urinary hippuric acid (HA) in the range of 0.01-5 mg/mL which is enough range to be used for in-field or point-of-care (POC) diagnosis. The proposed electrochemical method can be extended to the applications to detect a wide range of different small molecules in the field of health care.

• Korean Journal of Clinical Laboratory Science
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• v.44 no.1
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• pp.17-23
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• 2012

This study is assessed the urinary hippuric acid (HA) concentration in toluene-exposed workers. Toluene is widely used in the coating, printing, painting and petroleum industries. We analyzed the hippuric acid level of toluene-exposed workers (males: 198, females: 63) from 2008 to 2010 and examined the relationship between hippuric acid and the characteristics of their work. The measurements of the urinary hippuric acid were confirmed by high-performance liquid chromatography. Among the subjects, males had on average more work experience than females (males: 9.17 years, females: 4.45 years), but females showed a higher hippuric acid concentration levels than males (males: 0.26 g/L, females: 0.75 g/L). The mean of the HA concentration according to the age group was 0.19 g/L in 30's, 0.30 g/L in 40's, 0.54 g/L in 50's, 1.36 g/L in 60's. Those in their 60's had the highest concentration. The mean of HA concentration according to the type of work was 0.70 g/L for the coating, 0.52 g/L for the painting, and 0.16 g/L for the printing industries, revealing that the workers in the coating industry had the highest concentration. By the highest order, the mean of HA concentration by working period was as follows; less than 5 years, ${\leq}5\;years{\sim}<10\;years$. The factors that influence the HA concentration are not only how long one is exposed to toluene but also the type of work, worker's age, and their gender.

• Journal of the Korean Electrochemical Society
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• v.17 no.3
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• pp.172-178
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• 2014

In this work, we describe an electrochemical immunosensor for simple, fast and quantitative detection of a urinary hippuric acid (HA). Urinary HA, of molecular weight 180 DA, is one of the major metabolites and biological indicators in toluene-exposed humans. Simple and ubiquitous monitoring of exposure to toluene is very important in occupational health care. We propose the electrochemical immunoassay based on the dopamine-antigen conjugate for detecting hippuric acid. Our electrochemical immunoassay system employs a conjugate of dopamine (DA) as an electrochemical active molecule and hippuric acid (HA) as an antigen. As an electrochemical aspect, dopamine (DA) containing two hydroxyl group can show excellent redox signal. Also, dopamine-tethered hippuric acid (DA-HA) shows the reversible redox signal in the immunoassay. The competition between HA and DA-HA generated electric signals proportional to HA concentration. The electrochemical immunoassay was performed with DA-HA on the screen printed carbon electrodes (SPCEs), and then applies the mixture antigen (HA) and HA-antibody. The electrical signals were proportional to HA in the range of 0.010~2.500 mg/mL which is enough range to be used for the point-of-care.

• Journal of Environmental Science International
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• v.10 no.6
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• pp.445-450
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• 2001

This study was carried out In Investigate the correlation between toluene In air and hippuric acid In human urine, which 1.: based on the results of the health check-in and the measure of working environment, was investigated for two years. The concentration of toluene in air for the five working places in Yang-San area were ranged from 39.05±10.31 ppm to 4.04±3.38ppm and the mean value of those was 16.89±14.76ppm. The mean value was 16.89±14.76ppm which is below the permissible limit of toluene. The concentration of hippuric acid(HA) In the worker's urine were between 0.47g/ℓ and 0.76g/ℓ. There was mutual correlation (Υ=0.94) between the concentration of toluene in air and that of hippuric acid in urine. In case of male workers, the mean value of hippurlc acid In urine was 0.56g/ f and the female workers was 0.57g/ℓ that showed higher than the male's. However, there was no relationship between the workshop and the distinction of sex. From the mean value of toluene in air(T) and that of hippuric acid(H), we obtained the formula, Η=0.3logΤ+0.235.

• Bulletin of the Korean Chemical Society
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• v.33 no.5
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• pp.1485-1490
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• 2012

An electrochemical immunoassay based on osmium-hippuric acid (HA) conjugate films onto the electrode is presented for the detection of urinary HA. This is the first report on the use of the oxidative electropolymerization of 5-amino-1,10-phenanthroline (5-$NH_2$-phen) for immobilizing an antigen, osmium-conjugated HA. As a redox mediator, [Os(5-amino-1,10-phenanthroline)$_2$(4-aminomethylpyridine-HA)Cl]$^{+/2+}$ (Os-phen-HA) was successfully synthesized and electropolymerized onto the screen-printed carbon electrodes (SPCEs). The interaction between osmium-HA conjugate films and antibody-HA ($anti$-HA) was performed by cyclic voltammetry (CV) and differential pulse voltammetry (DPV). The electrical signals were linearly proportional to urinary HA in the range of 0.1-5.0 mg/mL, which is sufficient for use as an immunosensor using a cutoff concentration of 2.0 mg/mL in urine samples. The proposed electrochemical immunoassay method can be extended to various applications for detecting a wide range of different small antigens in the health care area.

• Bulletin of the Korean Chemical Society
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• v.34 no.2
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• pp.595-599
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• 2013

The coordination of pyridyl-N to pentacyanoferrate for the detection of small organic antigens in solution is presented. The unique contribution of this paper is the direct conjugation of pyridyl-N in small organic antigens to pentacyanoferrate. Pentacyanoferrate is promising as an electrochemical label owing to its good electro-chemical properties, which can be utilized to generate an electrical signal in homogeneous electrochemical immunoassays. The facilely synthesized pyridyl-N to pentacyanoferrate was characterized by the electrochemical and spectroscopic methods. Hippuric acid (HA) has been detected competitively on the interaction of free HA and pentacyanoferrate-(4-aminomethylpyridine-hippuric acid) (Fe-HA) to its antibody, with the detection limit of 0.50 ${\mu}g\;mL^{-1}$. While pentacyanoferrate-based immunoassay is in its simplicity and infancy, the proposed immunoassay offers attractive opportunities for developing pyridyl-N-based the electrochemical detection of small organic antigens in the health care area.

• Journal of Environmental Impact Assessment
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• v.24 no.2
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• pp.154-162
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• 2015

Correlation between occupational exposure levels to toluene and urinary hippuric acid (HA) concentrations were studied for 124 workers at 54 work places. The highest indoor toluene concentration measured at printing process was 81.116 ppm, and their geometric average concentration was as high as 12.304 ppm. The geometric average concentration of hippuric acid in urine samples from workers who are exposed to toluene was 0.714 g/g creatinine. This is almost five times higher than the average HA concentration from non-exposure workers. Printing workers showed the average HA concentration of 1.145 g/g creatinine from their urine samples. It is the highest concentration among the workers exposed to toluene. The correlation coefficient between HA concentrations in urine and indoor toluene concentration at work places was relatively high as r=0.624 (P<0.01). But the correlations of HA with sex, smoking, drinking, age and employment history was relatively low. We can express the regression equation for the urinary HA concentration which is Y = 0.037X + 0.562 as exposure toluene concentration is X. The urinary HA concentrations showed significantly increase depend on indoor toluene concentration at work place.

• Safety and Health at Work
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• v.2 no.3
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• pp.229-235
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• 2011

Objectives: In this study, we summarized the External Quality Assessment Scheme (EQAS) for the biological monitoring of occupational exposure to toxic chemicals which started in 1995 and continued until a $31^{st}$ round robin in the spring of 2010. The program was performed twice per year until 2009, and this was changed to once a year since 2010. The objective of the program is to ensure the reliability of the data related to biological monitoring from analytical laboratories. Methods: One hundred and eighteen laboratories participated in the $31^{st}$ round robin. The program offers 5 items for inorganic analysis: lead in blood, cadmium in blood, manganese in blood, cadmium in urine, and mercury in urine. It also offers 10 items for organic analysis, including hippuric acid, methylhippuric acid, mandelic acid, phenylglyoxylic acid, N-methylformamide, N-methylacetamide, trichloroacetic acid, total trichloro-compounds, trans,trans-muconic acid, and 2,5-hexanedione in urine. Target values were determined by statistical analysis using consensus values. All the data, such as chromatograms and calibration curves, were reviewed by the committee. Results: The proficiency rate was below 70% prior to the first round robin and improved to over 90% for common items, such as PbB and HA, while those for other items still remained in the range of 60-90% and need to be improved up to 90%. Conclusion: The EQAS has taken a primary role in improving the reliability of analytical data. A total quality assurance scheme is suggested, including the validation of technical documentation for the whole analytical procedure.

• Bulletin of the Korean Chemical Society
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• v.30 no.9
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• pp.2021-2026
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• 2009

Toluene, xylene and styrene are volatile organic solvents that are commonly used in mixtures in many industries. Because these solvents are metabolized and then excreted in urine, their urinary metabolites are thought to be biomarkers of occupational exposure to these solvents. Therefore, a simple, rapid, and yet reliable analytical method for determining the metabolites is required for accurate biological monitoring. In the present study, a simple and rapid HPLC-UV method was developed for the simultaneous determination of eight major metabolites of toluene, xylene and styrene: hippuric acid (HA), mandelic acid (MA), o-, m- and p-methylhippuric acids (o-, m- and p-MHAs), and o-, m- and p-cresols. A monolithic column was employed as the stationary phase and several conditions, including flow rate, composition of mobile phase and column temperature, were variables for the optimization of the chromatographic resolution. All eight metabolites were successfully resolved within 5 minutes in 10% aqueous ethanol containing 0.3% acetic acid and 1.6% $\beta$-cyclodextrin, using a flow rate gradient of 1.0 - 5.0 mL/min at 25 ${^{\circ}C}$. The performance of this method was validated by linearity, intra- and inter-day accuracy, and precision. The linearity was observed with correlation coefficients of 0.9998 for HA, 0.9999 for MA, 0.9989 for o-MHA, 0.9998 for m-MHA, 0.9991 for p-MHA, 0.9997 for o-cresol, 0.9998 for m-cresol, and 0.9986 for p-cresol. The intra- and inter-day precision of the method were less than 5.89% (CV) and the accuracy ranged from 92.95 to 106.62%. The validity was further confirmed by analysis of reference samples that were prepared by the inter-laboratory quality assurance program of the Korea Occupational Safety and Health Agency (KOSHA, Seoul, Korea). All measured concentrations of the analytes agreed with the certified values.

• Proceedings of the Korean Environmental Health Society Conference
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• 2003.06a
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• pp.187-191
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• 2003

In this study, biomonitoring methods were developed to measure BTEXs exposure level in the air, metabolites of benzene and toluene in human urine, individual susceptibility markers in human blood for evaluation of the health effects about environmental pollution. We have also performed a small-scaled molecular epidemiology study on residents in Chuncheon and workers in workplace for these method applications. The workers in workplace were surveyed as study areas, and the residents in Chuncheon which is in the suburban area were surveyed as comparative areas in this study. Actually, 31 workers in as target group and 33 residences in as control group this epidemiological study. The results obtained from this study were as follows: 1. Benzene is a well-known carcinogen, it's median concentrations were 0.00024∼0.02057ppm at suburban area and 0.002∼00.654ppm at work place, These benzene concentrations were not exceed the OSHA(Occupational Safety and Health Administration) threshold benzene level of 1ppm in the states. 2. Metabolites product of benzene(t,t-Muconic Acid) and toluene(Hippuric Acid) were not significant both in suburban and workplace area. The median concentration of t,t-MA and HA were 0.0122, 1.44277g/g creatinine, respectively. 3. In the case of individual susceptibility markers as CYPlAl, 41.8% of them has homozygous wild type(W) and who has heterozygous variant type(H) was 35.4% and 22.8% of homozygous variant type(M) genetic type. In the case of CYP2E1, 62.82% of them has homozygous wild type(D) type, 34.62% of each has heterozygous variant type (DC) and 2.56% of them has homozygous variant type (CC). Who doesn't have GSTM1 gene was 46.25% and who has GSTM1 gene was 53.75％. Who doesn't have GSTT1 gene was 40.0％ in study groups and who has GSTT1 gene was 60.0%. Who has W genetic type, which is homozygous wild type of GSTP1, was 69.18% and H genetic type, which is heterozygous variant type was 28.4%. M genetic type which is homozygous variant type was 2.4%. 4. Concentration differences of metabolites such as t,t-MA and HA in urine, which is generated by individual susceptibility marker of GSTM1, GSTT1, GSTP1 gene of Phase I and CYP1A1, CYP2E1 gene of Phase II, was examined. As a result, GSTP1 and GSTM1 indicate slight differences depend on the amount of metabolites in urine, it was not statistically significant.