The use of p-chloromercuribenzoic acid in the determination of ascorbic acid with 2:6-dichlorophenolindophenol.
The use of p-chloromercuribenzoic acid in the determination of ascorbic acid with 2:6-dichlorophenolindophenol.
复制标题
对氯汞苯甲酸在2:6-二氯苯酚靛酚测定抗坏血酸中的应用
DOI：
10.1042/bj0620675
复制
发表时间：
1956
期刊：
影响因子：
--
通讯作者：
B. Iggo
中科院分区：
文献类型：
--
作者：
J. Owen;B. Iggo
材料与方法l -脱氢抗坏血酸是用醌氧化抗坏血酸制备的（Paterson, 1950）。2: 6-二氯苯酚-吲哚酚（吲哚酚）钠盐原液，含100 mg /100 ml，每周配制，保持在5℃。在不超过4天的时间间隔内，加入20%（w/v）的偏磷酸原液，并保持在5℃。根据Whitmore & Woodward（1946）的方法制备对氯苯甲酸（CMB），溶液中含有200mg ./ 100ml .的005N-NaOH。在20ml中加入50mg锌粉，摇匀30min，制得同型半胱氨酸溶液。同型半胱氨酸溶液（L. Light and Co.）， 68毫克/100毫升，在2%（w/v）偏磷酸中过滤掉多余的锌。用标准碘溶液滴定，还原率达90%。将Sb25加热产生的H2S鼓泡制得硫化氢溶液。在浓缩的。HCI，成2%（w/v）偏磷酸。用KMnO4滴定法测定了硫化氢的浓度。吸附试剂：用于色谱分析的氧化铝。佛罗里达土，富勒土，轻质高岭土和酸洗硅藻土（均来自英国药厂有限公司），佛罗里达（佛罗里达公司，沃伦，宾夕法尼亚州）。Celite （Johns Manville and Co Ltd, London）。根据Dalgliesh（1955）的方法制备去活性炭。Plauma。将2 vol血浆加入3 vol 3%（w/v）偏磷酸中制备人血浆无蛋白滤液（oxa- ated或肝素化）。红细胞。用一氧化碳初步处理后，制备了人红细胞的无蛋白滤过物（Butler & Cushman, 1940）。全血（20 ml）（草酸化或肝素化）离心，去除白细胞层。将红细胞重新悬浮在血浆中，然后将其置于血压计（500毫升）中，将甲酸钠和conc生成的一氧化碳稳定流注入血压计。硫酸。眼压计旋转15分钟。离心后立即加入2 vol红细胞至3vol。6%（w/v）偏磷酸。
MATERIALS AND METHODSReaent8. L-Dehydroascorbicacid was prepared by oxidation of ascorbic acid with quinone (Paterson, 1950). A stock solution of the sodium salt of 2: 6-dichlorophenol-indophenol (indophenol) containing 100 mg./100 ml. was made up weekly and kept at 5. A stock solution [20%(w/v)] of metaphosphoric acid was made up at intervals of not longer than 4 days and kept at 5. p-Chloromercuribenzoic acid (CMB) was prepared according to the method of Whitmore & Woodward (1946) and a solution containing 200 mg./100 ml. in 005N-NaOH was used. A solution of homocysteine was prepared by shaking for 30 min. 50 mg. of zinc dust in 20ml. of a solution of homocysteine (L. Light and Co.), 68 mg./100 ml., in 2%(w/v) metaphosphoric acid and filtering off the excess of zinc. Titration against standard iodine solution showed that reduction was 90% complete. A solution of hydrogen sulphide was prepared by bubbling H2S, generated by warming Sb25. in conc. HCI, into 2%(w/v) metaphosphoric acid. The concentration of hydrogen sulphide was determined by titration against KMnO4.Adsorption reagents: aluminium oxide for chromato-graphic analysis. Floridin earth, fuller's earth, light kaolin and acid-washed kieselguhr (all from British Drug Houses, Ltd.), Florisil (Floridin Co., Warren, Pa., USA), Celite (Johns Manville and Co. Ltd., London). Deactivated charcoal was prepared according to the method of Dalgliesh (1955). Plauma. Protein-free filtrates of human plasma (oxa-lated or heparinized) were prepared by adding 2 vol. of plasma to 3 vol. of 3%(w/v) metaphosphoric acid. Erythrocytes. Protein-freefiltrates of human erythro-cytes were prepared after preliminary treatment with carbon monoxide (Butler & Cushman, 1940). Whole blood (20 ml.)(oxalated or heparinized) was centrifuged and the leucocyte layer removed. The erythrocytes, resuspended in the plasma, were then placed in a tonometer (500 ml.), into whichwas led a steadystream ofcarbonmonoxidegenerated from sodium formate and conc. H2SO4. The tonometer was rotated for 15 min. After centrifuging, 2 vol. of erythro-cytes was added without delay to 3vol. of 6%(w/v) metaphosphoric acid.