Determination of the propylene oxide–hemoglobin adduct by gas chromatography‐electron impact ionization mass spectrometry

Abstract

A gas chromatography-mass spectrometry method was developed for the determination of the propylene oxide (PO)-hemoglobin adduct. The adduct was identified as hydroxy propyl valine (HPV), and released from globin by the modified Edman degradation and extracted with ethyl ether. HPV and deuterated HPV (d6-HPV) were synthesized for identification and quality control. d6-HPV was used as an internal reference standard. The dried extract was completely derivatized with N-methyl-N-(trimethylsilyl) trifluoroacetamide (MSTFA). The method detection limit (MDL) of the assay was 10 pmole/g for HPV, based on assayed hemoglobin of 0.1 g. The method was applied to the determination of the PO-hemoglobin adduct formed in young female Sprague-Dawley rats after treatment for 4 and 5 weeks with 5 and 10 mM PO via drinking water. HPV was detected by the proposed procedure. After 4 weeks, the concentration of HPV was 6.75 nmole/g hemoglobin during treatment with 5 mM, and 80.26 nmole/g hemoglobin during treatment with 10 mM. The adduct level in 5 weeks increased up to about 51.47 nmole/g during treatment with 5 mM PO in the drinking water and up to about 120.27 nmole/g during treatment with 10 mM PO. This method was also applied to determine the concentrations of HPV in the blood of 20 persons living near the Ulsan petroleum industrial complex in Korea. As a result, HPV-hemoglobin adduct was detected in the concentration range 0-1100 pmol/g in the human blood samples.