1.Determination of Free 4-hydroxyproline with Dansylchloride by HPLC in Human Urine.
Keou Weon LEE ; Kyung Jong LEE ; Young Bong CHO
Korean Journal of Preventive Medicine 2002;35(4):282-286
OBJECTIVES: The level of 4-hydroxyproline (4-Hyp) in human urine was measured using high performance liquid chromatography (HPLC) with a fluorescence detector. This method is useful for medical examinations and investigating the radicals induced by physical, chemical, mental stresses. This method is superior to many published several methods in terms of its low cost and ability to analyze many samples. METHODS: The urine from workers in a tire manufacturing company (22 male pre- and post-shift workers) and 18 office-workers as controls were analyzed. Data concerning age, the cumulative drinking amount and the cumulative smoking amount was collected with a questionnaire. The optimum applied amount of dansyl-Cl, the optimum reaction temperature and time, the recoveries and the optimum pH of the eluent and buffer were determined. 4-Hyp from human urine was derivatized with dansyl-Cl (dimethylamino-naphthalene-1-sulfonyl chloride) after removing the alpha-amino acid by a treatment with phthalic dicarboxaldehyde (OPA) and cleaned with Bond Elut C18 column. The 4-Hyp derivatives were separated on a reversed phase column by gradient elution with a phosphate buffer (5 m mol, pH 8.0) and acetonitrile, and detected by fluorescence measurements at 340 nm (excitation) and 538 nm (emission). RESUJLTS: The detection limit for the urinary free 4-Hyp was 0.364 micro mol/l. The recovery rate of 4-Hyp was 99.7 %, and the effective pH of the phosphate buffer and borate buffer were 3.0 and 8.0, respectively. From statistical analysis, age, drinking and smoking did not affect the urinary free 4-Hyp in both the controls and workers. The range of urinary 4-Hyp in the controls, pre-shift, and post-shift workers were 0.33-16.44, N.D-49.06, and 0.32-56.27 micro mol/l. From the pared-sample t-test, the urinary 4-Hyp levels in post-shift workers (11.82+/-16.73 n mol/mg Cre) were 2-fold higher than in pre-shift workers (5.36+/-5.53 n mol/mg Cre) and controls (4.91+/-4.89 n mol/mg Cre). CONCLUSIONS: This method was developed with high sensitivity, accuracy, and precision. The present method was effectively applied to analyze the urinary free 4-Hyp in both controls and workers.
Chromatography, High Pressure Liquid*
;
Chromatography, Liquid
;
Drinking
;
Fluorescence
;
Humans*
;
Hydrogen-Ion Concentration
;
Hydroxyproline*
;
Limit of Detection
;
Male
;
Surveys and Questionnaires
;
Smoke
;
Smoking
2.Determination of Free 4-hydroxyproline with Dansylchloride by HPLC in Human Urine.
Keou Weon LEE ; Kyung Jong LEE ; Young Bong CHO
Korean Journal of Preventive Medicine 2002;35(4):282-286
OBJECTIVES: The level of 4-hydroxyproline (4-Hyp) in human urine was measured using high performance liquid chromatography (HPLC) with a fluorescence detector. This method is useful for medical examinations and investigating the radicals induced by physical, chemical, mental stresses. This method is superior to many published several methods in terms of its low cost and ability to analyze many samples. METHODS: The urine from workers in a tire manufacturing company (22 male pre- and post-shift workers) and 18 office-workers as controls were analyzed. Data concerning age, the cumulative drinking amount and the cumulative smoking amount was collected with a questionnaire. The optimum applied amount of dansyl-Cl, the optimum reaction temperature and time, the recoveries and the optimum pH of the eluent and buffer were determined. 4-Hyp from human urine was derivatized with dansyl-Cl (dimethylamino-naphthalene-1-sulfonyl chloride) after removing the alpha-amino acid by a treatment with phthalic dicarboxaldehyde (OPA) and cleaned with Bond Elut C18 column. The 4-Hyp derivatives were separated on a reversed phase column by gradient elution with a phosphate buffer (5 m mol, pH 8.0) and acetonitrile, and detected by fluorescence measurements at 340 nm (excitation) and 538 nm (emission). RESUJLTS: The detection limit for the urinary free 4-Hyp was 0.364 micro mol/l. The recovery rate of 4-Hyp was 99.7 %, and the effective pH of the phosphate buffer and borate buffer were 3.0 and 8.0, respectively. From statistical analysis, age, drinking and smoking did not affect the urinary free 4-Hyp in both the controls and workers. The range of urinary 4-Hyp in the controls, pre-shift, and post-shift workers were 0.33-16.44, N.D-49.06, and 0.32-56.27 micro mol/l. From the pared-sample t-test, the urinary 4-Hyp levels in post-shift workers (11.82+/-16.73 n mol/mg Cre) were 2-fold higher than in pre-shift workers (5.36+/-5.53 n mol/mg Cre) and controls (4.91+/-4.89 n mol/mg Cre). CONCLUSIONS: This method was developed with high sensitivity, accuracy, and precision. The present method was effectively applied to analyze the urinary free 4-Hyp in both controls and workers.
Chromatography, High Pressure Liquid*
;
Chromatography, Liquid
;
Drinking
;
Fluorescence
;
Humans*
;
Hydrogen-Ion Concentration
;
Hydroxyproline*
;
Limit of Detection
;
Male
;
Surveys and Questionnaires
;
Smoke
;
Smoking
3.Effects of Intake of Alcohol on the Concentration of Urinary 4-hydroxyproline.
Keou Weon LEE ; Kyung Jong LEE ; Jae Beom PARK ; Kyu Yeub JANG ; Chul Woo BANG ; Young Bong CHO
Korean Journal of Aerospace and Environmental Medicine 2003;13(3):144-148
BACKGROUND: We studied whether alcohol intake influences in urinary 4-Hyp concentration and whether 4-Hyp concentration is change by time in healthy office workers for 2 weeks. METHODS: Three healthy male workers, who has a same job, recruited for this study, aged from 29 to 33 years, who had been any diagnosed or treated to the alcohol or bone related diseases. Urine samples were picked about 30 ml in screw capped vials for 2 weeks, and analysing method were used to previous published method for determination of urinary 4-Hyp with dansyl chloride. RESULTS: From the ANOVA-test results about objects' Hyp concentration that classify by urine picking time, When subjects did not ingest alcohol, urinary 4-Hyp concentration was difference according to subjects for-work and post-work. But, did not show difference when subjects ingested alcohol. While, T-test result that do alcohol intake existence and nonexistence and urine picking time by variable, urinary 4-Hyp concentration did not show difference for-work and post-work, but there was difference in post-work that is next day that ingest alcohol. When did not ingest alcohol, mean concentration of urinary 4-Hyp increased go by post-work (3.80+/-6.47 nmol/mg creatinine) in pre-work (1.46+/-1.61 nmol/mg creatinine). But when ingested alcohol, mean concentration of urinary 4-Hyp decreased by the next day morning (3.32+/-6.36 nmol/mg creatinine), order of for-work (1.54+/-1.59 nmol/mg creatinine) in alcohol intake future generations post-work (5.09+/-6.41 nmol/mg creatinine). CONCLUSION: We confirmed that alcohol is influencing in urinary 4-Hyp concentration of three healthy male workers, who has a same job, for 2 weeks, and mean concentration of 4-Hyp increasing go by the post-work in the pre-work, could confirm that influence in the next day morning if ingest alcohol in the evening.
Humans
;
Hydroxyproline*
;
Male
;
Social Responsibility
4.Relationship between Phenylglyoxylic Acid in Urine and Postural Body Sway in Styrene Exposed Workers.
Kyung Jong LEE ; Jae Bum PARK ; Keou Weon LEE ; Kwang Jin LIM ; Kyoo Yup JANG ; Cheol Woo BANG
Korean Journal of Occupational and Environmental Medicine 2002;14(4):459-467
OBJECTIVE: Until now,no effective screening tools have been available for evaluating the neurotoxicity of organic solvents and metals. The aim of this study was to evaluate the usefulness of posturography as a screening tool for the chronic neurotoxicity of organic solvents. METHOD: 36 workers in 4 septic tank manufacturers,who were exposed to styrene over a period of 1 year (exposed group),and 15 hospital volunteer manual workers were examined by posturography.The subjects' physical,medical,and occupational characteristics were obtained by means of a physical examination and a questionnaire. We excluded from both groups those persons who had psychiatric problems, diabetes, neurologic symptoms, gait disturbance,or a history of stroke.The sway area of the exposed group was compared to that of the non-exposed group using bivariate and multiple regression analysis. We controlled a number of variables including age, alcohol consumption,smoking, weight, height, and body mass index. RESULT: The sway area of the exposed group was found to be higher than that of the non-exposed group after taking into consideration the effects of other characteristics by means of multiple regression analysis. CONCLUSION: We concluded the posturography would be an effective tool for the screening of chronic neurotoxicity in workers exposed to styrene.
Body Mass Index
;
Gait
;
Hospital Volunteers
;
Humans
;
Mass Screening
;
Metals
;
Neurologic Manifestations
;
Physical Examination
;
Questionnaires
;
Solvents
;
Styrene*