[Determination of blood acetate by head space gas chromatography].
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Biomedical subjects
Publications and source records attributed to K Ameno.
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Positional nystagmus and body sway were measured for 8 hours following alcohol ingestion in 27 human volunteers. The intensity of positional alcohol nystagmus phase I (PAN-I) correlated well with blood alcohol concentration (BAC) along the time axis; the correlation coefficient between PAN-I and BAC 1 hour after alcohol ingestion was 0.62 (P less than .01). The minimum threshold BAC resulting in PAN-I was 0.23 mg/mL. Body sway was measured by stabilometry. The circumscribed area of posturography before and after alcohol ingestion was compared. Temporal changes in body sway and BAC were also closely correlated. The BAC threshold for increased body sway was estimated to be somewhere between 0.5 and 0.8 mg/mL, considerably higher than the threshold for PAN-I. Measurements of blood acetaldehyde, the dehydrated product of ethanol metabolism, showed no correlation with either the intensity of PAN or changes in body sway. The intensity of PAN and body sway measured at the time of peak BAC 1 hour after alcohol ingestion, however, showed significant positive correlation (r = .50, n = 25; P less than .01).
An experiment was carried out on rats to determine whether or not a skeletal muscle sample was suitable for the determination of ethanol concentration in a carcass. Gas chromatography was used to estimate the ethanol and n-propanol concentrations in the femoral muscle and intracardial blood. The ethanol concentration of each sample was corrected according to the moisture ratio of circulating blood, viz., 78.5%. The ethanol concentration ratio of blood to muscle was 1.03 two hours after ethanol administration. When the carcasses of rats pre-treated with ethanol were stored at 15 degrees C and 25 degrees C, respectively, the ethanol concentrations in muscle and blood increased with time. At all times the concentration was higher in blood than in muscle, and also higher in samples collected from the carcass stored at 25 degrees C than at 15 degrees C. When the control carcass was stored in the same manner, the postmortem production of ethanol was noticed in both blood and muscle. As in the experimental rats, the control rats exhibited a higher blood ethanol than muscle ethanol level. Again, the ethanol concentration was higher in samples collected from the carcass stored at 25 degrees C than at 15 degrees C. The ratio of ethanol to n-propanol was less than 20:1 in blood and less than 10:1 in muscle. These results suggest that skeletal muscle may be a suitable tissue for the postmortem detection of ethanol.
Cyanamide is a potent inhibitor of aldehyde dehydrogenase (ALDH: EC 1.2.1.3) used in the treatment of alcoholics. In the presence of ethanol, cyanamide causes an accumulation of acetaldehyde, a highly toxic metabolite of ethanol, with unpleasant side-effects. A similar accumulation is seen in some Oriental people with low ALDH activity. We have investigated the effects of ethanol and cyanamide administration on the activation of the hypothalamic-pituitary-adrenal (HPA) axis using in situ hybridization histochemistry and radioimmunoassay. Ethanol plus cyanamide resulted in a significant increase in corticotrophin-releasing factor and arginine vasopressin mRNA in the paraventricular nucleus, and pro-opiomelanocortin mRNA in the anterior pituitary. Plasma corticosterone concentrations were also significantly elevated following ethanol plus cyanamide administration. The blood concentration of acetaldehyde in the ethanol plus cyanamide group increased significantly. These results suggest that acetaldehyde, induced by blocking ethanol metabolism, is able to activate the HPA axis operating through a central mechanism.
The regulation mechanism of inhibition of intestinal ethanol absorption induced by high acetaldehyde (AcH) concentration in blood was investigated. We used atropine (AT), atropine methylbromide (ATMB), pirenzepine (PI), bethanechol (BE) and pilocarpine (PL) with or without cyanamide (CY; a potent inhibitor of aldehyde dehydrogenase, which induces high AcH concentration in blood). The K(a) (absorption rate constant) value after the CY-alone pretreatment was significantly lower than that in controls. In the high AcH-induced cases, the values of K(a) in AT and ATMB pretreatments were similar to controls, but the value of K(a) in PI pretreatment was lower than that in controls. The values of K(a) in the case of BE pretreatment with and without high AcH levels were lower than in controls. The K(a) value in the PL with CY was significantly lower than that with CY alone. However, its action was blocked by ATMB pretreatment. These results suggest that high blood AcH concentrations inhibit intestinal ethanol absorption through the peripheral cholinergic nerves via muscarinic receptors, except for the muscarinic M(1) receptor, compared to other subtypes of muscarinic receptors.
A simple, sensitive, and rapid quantitative method for Dipterex in serum is described. A SepPak C18 cartridge for the extraction and gas chromatography with flame thermionic detection for determination are used. The detection limit is 2.5 ng/mL, and linearity is obtained in the range 5-500 ng/mL.
We report a simple, sensitive, and rapid quantitation of Amitraz in plasma after Extrelut-3 column extraction by gas chromatography with nitrogen-phosphorus detection (GC-NPD). The plasma sample was diluted four-fold with borate buffer (0.01M, pH 11), put into an Extrelut-3 column, left for 15 min, and then eluted with 15 mL of n-hexane. The n-hexane eluate was evaporated under nitrogen gas flow at room temperature. The residue was reconstituted with 0.1 mL of acetone containing nitrazepam as an internal standard. A 2-microL aliquot was injected into a wide-bore capillary column GC-NPD. The detection limit was 0.5 ng/mL and linearity was obtained in the range of 1-200 ng/mL. Amitraz in the buffer at pH 11 remained stable in a freezer for one week at -20 degrees C. The GC-NPD method was found useful in studying the pharmacokinetics of a single dose intravenous administration of Amitraz to a dog.
A method for rapid detection and identification of bromvalerylurea (BVU), bromodiethylacetylurea (BDU), and allylisopropylacetylurea (AIU) in serum and urine by high-performance liquid chromatography (HPLC) with a multiwavelength UV detector after Sep-Pak C18 cartridge extraction is reported. A Jasco Finepak C18 reversed-phase column was used for the separation. Acetonitrile-distilled water (1:1, v/v) was used as a mobile phase. There was no significant absorption of the three hypnotics in the UV spectra (210-350 nm). However, the absorption of each was higher at the shorter wavelengths. The quantifications for the three hypnotics detected at 210 nm by the chromatogram were linear over the range 0.2-4 micrograms/mL and the detection limits of BVU, BDU, and AIU were 5, 10, and 10 ng as absolute amounts, respectively. The mean recovery yields of BVU, BDU, and AIU by Sep-Pak C18 cartridge extraction were 85.7 +/- 4.1, 98.6 +/- 2.2, and 95.1 +/- 3.5% (n = 5) in serum and 79.5 +/- 3.8, 95.7 +/- 1.8, and 93.0 +/- 4.2% (n = 5) in urine, respectively. An optimal system of thin-layer chromatography for the identification of the hypnotics is also discussed.
A rapid, simple method based on second-derivative spectroscopy of the simultaneous analysis of paraquat and diquat in serum and urine is described. Paraquat and diquat in serum were deproteinized with sulfosalicylic acid, and those in urine were reduced with NaOH-dithionite solution. A qualitative and quantitative analysis of reduced paraquat and diquat was made at the amplitude peaks of 396-403 nm and 454-464 nm in the second-derivative spectra, respectively. The entire procedure was completed within about 10 minutes for a serum sample and within about 5 minutes for a urine sample. Application of the proposed method on a poisoned patient is also reported.