Article

Sensitive and simple determination of mannitol in human brain tissues by gas chromatography-mass spectrometry

Department of Forensic Pathology and Sciences, Graduate School of Medical Sciences, Kyushu University, Fukuoka, Japan.
Journal of chromatography. B, Biomedical sciences and applications 08/2001; 758(1):103-8. DOI: 10.1016/S0378-4347(01)00145-1
Source: PubMed

ABSTRACT A simple, reliable and sensitive gas chromatographic-mass spectrometric method was devised to determine the level of mannitol in various human brain tissues obtained at autopsy. Mannitol was extracted with 10% trichloroacetic acid solution which effectively precipitated brain tissues. The supernatant was washed with tert.-butyl methyl ether to remove other organic compounds and to neutralize the aqueous solution. Mannitol was then derivatized with 1-butaneboronic acid and subjected to GC-MS. Erythritol was used as an internal standard. For quantitation, selected ion monitoring with m/z 127 and 253 for mannitol and m/z 127 for internal standard were used. Calibration curves were linear in concentration range from 0.2 to 20 microg/0.1 g and correlation coefficients exceeded 0.99. The lower detection limit of mannitol in distilled water was 1 ng/0.1 g. Mannitol was detected in control brain tissues, as a biological compound, at a level of 50 ng/0.1 g. The precision of this method was examined with use of two different concentrations, 2 and 20 microg/0.1 g, and the relative standard deviation ranged from 0.8 to 8.3%. We used this method to determine mannitol in brain tissues from an autopsied individual who had been clinically diagnosed as being brain dead. Cardiac arrest occurred 4 days later.

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    • "Biochemical diagnoses of brain death greatly aid forensic specialists and clinical staffs [1]. We evaluated brain levels of various therapeutic drugs in patients with brain death [2] [3] [4] [5] [6]. For example, cerebral concentrations of mannitol, an anti-edema reagent, alters as the time of administration in a rodent model [7]. "
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    • "Tomiya et al. [9] reported that some sugar alcohols such as sorbitol and myo-inositol were detected as endogenous compounds in rat tissues. We also found sorbitol in human tissues [10]. Since phenylisocyanate derivatives of mannitol were not completely separated from that of sorbitol [11], we selected p-nitrobenzoyl chloride as derivatizing reagent. "
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Akiko Shimamoto