Effect of dissociation energy on ion formation and sensitivity of an analytical method for determination of chlorpyrifos in human blood, using gas chromatography–mass spectrometer (GC–MS in MS/MS)

Abstract

A sensitive GC–MS method in MS/MS ion preparation was developed for quantitative estimation and qualitative determination of chlorpyrifos in human blood samples. This paper describes the combined effect of dissociation energy on ion formation of chlorpyrifos and sensitivity of this analytical method. Chlorpyrifos was extracted using methanol/hexane mixture from 0.2 ml human blood, deactivated with saturated acidic salt solution. The extract was then re-concentrated and analyzed by electron impact (EI) MS/MS gas chromatography–mass spectrometer. The MS/MS spectra of chlorpyrifos ion were recorded on different dissociation energy (30–100 V) to establish the structural confirmation and to demonstrate the effect of this energy on sensitivity, S/ N ratio and detection limit for quantification of chlorpyrifos, which is first time reported. At different exciting amplitude (30–100 V), different behaviors of base peak, sensitivity, S/ N ratio and detection limit of this method were observed for quantification of chlorpyrifos. The mass spectra recorded at dissociation energy <70 V, in between 70–80 V and >80 V correspond to the m/ z 314 (100%), m/ z 286 (100%) and m/ z 258 (100%), respectively. The sensitivity, signal and detection limit of this method increased on 95 V at m/ z 258. Therefore, m/ z 258 was used for the quantification of chlorpyrifos. The detection limit for quantification was 0.1 ng/ml with S/ N: 2 in human blood. The linear calibration curve with the correlation coefficient ( r > 0.99) was obtained. The percentage recoveries from 95.33% to 107.67% were observed for chlorpyrifos from human blood. The blood samples were collected at different time intervals. The concentration of chlorpyrifos in poisoning case was 3300, 3000, 2200, 1000, 600, 300 ng/ml on day 1, 3, 6, 8, 10 and 12, respectively. On the 12th day of exposure of chlorpyrifos, 90.9% reduction in concentration was observed. On day 14th the chlorpyrifos was not detected in the blood of the same subject. Thus the present method is useful for detection of chlorpyrifos in critical care practices and also provides tremendous selectivity advantages due to matrix elimination in the parent ion isolation step in blood sample analysis for chlorpyrifos.