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


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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    ABSTRACT: In anaesthetized, mechanically ventilated rats with brain death induced by epidural balloon inflation, we assessed the levels of cerebral myo-inositol in three brain areas 4 h after brain death had been confirmed. The levels were measured using HPLC, along with the water content. Myo-inositol levels were significantly increased in the cerebellum (P<0.05) and decreased in the brainstem (P<0.001), compared to findings in controls. Such changes can serve as hallmarks of brain death.
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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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    ABSTRACT: A method for simultaneous determination of glycerol and mannitol in various human tissues was devised and for this we used high-performance liquid chromatography (HPLC). Specimens were homogenized in a mixture of chloroform and methanol, phosphate buffer (pH 7.0) and pentaerythritol (IS) solution. After centrifugation, an aliquot of the aqueous layer was evaporated to dryness and derivatized with p-nitrobenzoyl chloride at 50 degrees C for 1h, then applied to HPLC with analytical conditions of: column, CAPCELL PAK C18 MG (250 mm x 3.0 mm i.d., 5 microm, Shiseido Co. Ltd., Tokyo, Japan); column temperature, 1-2 degrees C; mobile phase, 75% acetonitrile-distilled water containing 0.05% trifluoroacetic acid, 0.05% heptafluoro-n-butyric acid and 0.1% triethylamine; flow rate, 0.5 ml/min; wavelength, 260 nm. Calibration curves for both substances were linear in concentration ranges from 1 to 500 microg/0.1g and correlation coefficients exceeded 0.99. The relative standard deviation (R.S.D.) of the method was evaluated at concentrations of 10 and 100 microg/0.1g, and ranged from 0.84 to 10.6%. Using this method, we determined the regional distribution levels of glycerol and mannitol in various tissues from an autopsied brain dead man.
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    ABSTRACT: s-Triazoles have been utilized to produce reagents that can react with drugs containing carbonyl group and drugs that are susceptible to oxidation with periodic acid to produce carbonyl function such as diols and amino alcohols. In this study, glucosamine and mannitol were analyzed through oxidation with periodic acid to give formaldehyde which was allowed to condense with 4-Amino-5-hydrazino-4H[1,2,4]-triazole-3-thiol (AHTT) (I). The condensation product was further oxidized to yield a purple colored compound (II) with maximum absorption at 550 nm. Beer's law was obeyed in the range of 12.5-125 μg ml-1 for glucosamine and 25-150 μg ml-1 for mannitol. Both drugs were also successfully determined in their pharmaceutical formulations with mean percentage recoveries ±RSD ranged between 100.69-101.51% ±1.57-0.44 for glucosamine and 100.06% ±1.08 for mannitol.
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