Refer to p 41
The amount of urinary δ-aminolevulinic acid was measured by using Wakopak® Wakosil-II 5C18 HG.
As one of special health checkup methods for workers engaged in work with lead, measurement of δ-aminolevulinic acid excreted in the urine is performed.
The reference standard was added to test urine, the amount of derivatized δ-aminolevulinic acid was measured, and the reference standard could be recovered satisfactorily.

Column : Wakopak® Wakosil-Ⅱ 5C18 HG
( 4.6 mm×150 mm)
Eluent : CH3OH/2.5 % CH3COOH=40/60(v/v)
Flow Rate : 0.8 mL/min. at 40℃
Detections :Em.473 nm, Ex.363 nm, RANGE=1, ATTEN=2
Sample : Std. solution( 0.5 mg/L), Urine
Inj.Vol. : 50 μL

Recovery of added reference standard
Amount of added
reference standard
2.5 mg/L 50 mg/L
Recovery (%) 96.20% 93.70%

[References]
Okayama: Clin. Chem. 36/8, 1494 (1990)

Refer to p 44
The amounts of urinary hippuric acids were measured by using Wakopak® Wakosil-II 5C18-100.
As one of special health checkup methods for health maintenance of workers engaged in work with organic solvents, measurement of hippuric acid, methylhippuric acid and manderic acid excreted in the urine is performed. These acids are metabolites of toluene,xylene and styrene, respectively, absorbed into the body. They are excreted in the urine, and their amounts are used as exposure indexes.
The quantities of these metabolites and creatinine can be determined by injecting 100-fold diluted urine directly into HPLC.
The peak of manderic acid having metallic ions and chelating performance can be detected more sharply.

Column : Wakopak® Wakosil-Ⅱ 5C18-100 (4.6 mm x 100 mm)
Eluent : CH3CN/20 mM NaH2PO4(pH 3.0) 10 mM β-Cyclodextrin=10/80(v/v) add. 3 mM SDS
Flow Rate : 1.0 mL/min. at 40°C
Detection : UV 210 nm
Sample : 1)Manderic Acid, 2)Hippulic Acid, 3)o -MHA, 4)p -MHA,
5)m -MHA, 6)Creatinine
Injection Volume: 10 μg/mL each, 10 μL
Reproducibility of analysis of hippuric acids
(area value)
  MA HA o -MHA p -MHA m -MHA Creatinine
SD 703 683 1068 1124 1252 1533
CV% 0.63 0.72 0.74 0.76 0.71 0.98




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