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测定血液中异烟肼的浓度,是检查异烟肼片剂在生物体内生物利用度的一个重要课题。通常测定血清异烟肼浓度的方法有萤光光度法,分光光度法,比色法及微生物法,其中以萤光光度法测测定的灵敏度为最高。本试验工作主要是参考Miceli介绍的萤光光度微量测定法,作了适当的修改,加大试样及试剂的用量以便于操作,测定健康人口服异烟肼后11小时内血液中异烟肼的浓度变化。本方法是将待测的血清加三氯醋酸沉淀蛋白质后取上清液,加入含缓冲剂的水杨醛溶液,在室温下生成异烟肼水杨醛的腙。过量的水杨醛在50℃与亚硫酸氢钠起加成反应而除去,然后用异丁醇提取所生的腙,用萤光分光光度计作测定,其萤光强度与异烟肼的浓度成正比。
Determining the concentration of isoniazid in the blood is an important issue for examining the bioavailability of isoniazid tablets in vivo. Fluorescence spectrophotometry, spectrophotometry, colorimetric and microbiological methods are commonly used to determine the concentration of isoniazid in serum, and the highest sensitivity is determined by fluorescence spectrophotometry. The main work of this experiment is to refer to Miceli fluorescent photometric microassay, made the appropriate changes, increase the amount of samples and reagents for easy operation, determination of healthy people after oral isoniazid within 11 hours of isoniazid in the blood The concentration changes. In the method, supernatant is precipitated after the serum to be tested is added with trichloroacetic acid, the salicylaldehyde solution containing buffer is added, and the hydrazone of isoniazid salicylaldehyde is generated at room temperature. Excess salicylaldehyde was removed by addition reaction with sodium bisulfite at 50 ° C, and the resulting hydrazone was extracted with isobutanol. Fluorescence spectrophotometer was used for the determination. The fluorescence intensity and isoniazid concentration Is proportional.