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肝细胞细胞色素P-450含量测定广泛用于药理学、毒理学和环境医学等研究领域。文献上的方法多须使用超速离心、光谱扫描等仪器,使其应用受到一定限制。本文介绍用国产751G型分光光度计测定小鼠肝匀浆细胞色素P-450含量的简便方法。一、样品制备:取小鼠肝脏,拈去血液。用0.25M蔗糖Tris-HCl缓冲液制成匀浆,浓度为200mg/ml。滤过后用含150mM KCl、10mM MgCl_2的50mmTris-HCl缓冲液稀释至20mg/ml。通以一氧化碳后,将样品等量移入样品杯和参照杯,静置3分钟,再向样品杯加入连二亚硫酸钠、混匀,静置2分钟后用751G型分光光度计进行测量,取波长分别为450nm和490nm时的吸收度,依公式计算细胞色素P-450含量。二、一氧化碳的制备和通气时间的选择:一氧化碳可用甲酸脱水制取。通气(100个气泡/分)1～8分钟,细胞色素P-450的测定值维挣在同一水平。在
Determination of hepatocyte cytochrome P-450 is widely used in pharmacology, toxicology and environmental medicine and other research areas. Many methods in the literature must use ultracentrifugation, spectral scanning and other equipment, its application is subject to certain restrictions. This article describes the use of domestic 751G spectrophotometer determination of mouse liver homogenate cytochrome P-450 content of a simple method. First, the sample preparation: take the mouse liver, pick the blood. Homogenized with 0.25 M sucrose Tris-HCl buffer at a concentration of 200 mg / ml. After filtration, it was diluted to 20 mg / ml with 50 mM Tris-HCl buffer containing 150 mM KCl, 10 mM MgCl 2. Pass through the carbon monoxide, the sample is equivalent to the sample cup and reference cup, let stand for 3 minutes, and then added sodium dithionite sample cup, mix, stand 2 minutes after the measured with a 751G spectrophotometer, take the wavelength respectively For the absorbance at 450 nm and 490 nm, the cytochrome P-450 content was calculated according to the formula. Second, the preparation of carbon monoxide and the choice of ventilation time: carbon monoxide available formic acid dehydration system. Ventilation (100 bubbles / min) 1 to 8 minutes, the measured value of cytochrome P-450 earned at the same level Victoria. in