GAS-LIQUID CHROMATOGRAPHIC ISOLATION AND IDENTIFICATION OF SEVEN BARBITURATES AND GLUTETHIMIDE FROM LIVER AND URINE

GLC of barbitone, amytal, pentobarbital, secobarbital, thiobarbitone, heptabarbitone and glutethimide is reported by using glass column packed with 1.5% SE-52 on chromosorb W (60～80 mesh)and stainless steel column packed with 1.5% OV-17 on shimalite W (80～100 mesh). The gas chromatograph is equipped with FID using N₂ as the carrier gas, at a flow of 25 ml per minute (SE-52) and 50 ml per minute (OV-17). The column is maintained at 200℃ and the injection port and the detector at 250℃. The method is successfully applied to the isolation and identification of some barbiturates from urine and liver of human, dog or rabbit.Extraction procedure: Triturate 5 g of liver in a conical flask and add 0.3 ml of 6N HCl, 50 ml of ether. Shake vigorously and pour the organic phase into the refining column(containing 2 g of active charcoal, 20 g of aluminium oxide for chromatography and 20 g of anhydrous sodium sulfate). Gollect the eluate into a separator and extract with 0.45 N NaOH. Acidify the alkaline extract by 6 N HCl, then extract successively with 20 ml and 15 ml of ether. Deaquate the combined ether solution by anhydrous sodium sulfate. Evaporate the ether and dissolve the residue in 0.5 ml of methylene chloride. Inject 0.6～1.0 μl of this solution into gas chromatograph.