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产品介绍
酮体是肝脏脂肪酸氧化分解的中间产物乙酰乙酸、β-羟基丁酸及丙酮三者统称。酮体中丙酮的生成量相当小，生成后即被吸收。乙酰乙酸和β-羟基丁酸则经血流进入肝外组织，在此被氧化，经柠檬酸循环提供更多能量给那些组织使用，例如骨、心肌和肾皮质。本试剂盒主要用于定性鉴定人、动物肝组织、肌肉组织中酮体含量。
本试剂盒检测原理如下：以辛酸钾为底物，在铵离子存在的碱性环境中酮体可与亚硝酸铁氰化钠作用生成紫色化合物。
产品组分
T1522042 | 组分名称 | 物理外观 | 50T | 储存 | 单次测试使用量 |
T1522042A | 组织匀浆液 | 液体 | 250 mL | RT. | 按需 |
T1522042B | 辛酸钾溶液 | 液体 | 5 mL | RT. 避光 | 按需 |
T1522042C | 酮体酸化液 | 液体 | 50 mL | RT. | 1 mL |
T1522042D | 酮粉 | 粉末 | 20 g | RT. 避光 | 0.3 g |
使用方法
1. 辛酸钾工作液配制
按辛酸钾溶液：蒸馏水=1：49的比例混合，即得辛酸钾工作液。
2. 样本处理
取鼠或兔子1只，用剪刀断头处死，使血液流尽，立即取出肝脏、肌肉组织，用预冷的组
织匀浆液冲洗数次，将其剪成碎末或用匀浆器匀浆备用。
3. 样本检测
| 测定管1 | 测定管2 | 测定管3 |
肝脏 | 0.3 g | - | - |
肌肉 | - | 0.3 g | - |
肝脏＋肌肉 | - | - | 0.3 g |
辛酸钾工作液 | 3 mL | 3 mL | 3 mL |
37℃水浴30 min。 | |||
酮体酸化液 | 1 mL | 1 mL | 1 mL |
混匀后静置10 min，取各管上清液 10 滴，分别置于 3 支新的EP管内；各加入 10 滴蒸馏水混匀。 | |||
酮粉 | 0.3 g | 0.3 g | 0.3 g |
静置，观察各管颜色变化。 |
4. 结果判定
肝脏管中呈紫色；肝脏+肌肉管呈淡紫色或粉色；肌肉管颜色未有变化。
注意事项
1. 酮体浓度高时，紫色明显；酮体浓度低时，出现淡紫色或淡粉色。
2. 处理肝脏的器皿不能与处理肌肉的器皿混用。
3. 如果肝脏管与肝脏+肌肉管颜色无法区分，可再各加少许等量的酮粉。
4. 为了您的安全和健康，请穿实验服并戴一次性手套操作。
Introduction
Ketone bodies are the collective term for three intermediate products of fatty acid oxidation in the liver: acetoacetic acid, β-hydroxybutyrate, and acetone. Acetone is produced in very small amounts and is rapidly absorbed upon formation. Acetoacetate and β-hydroxybutyrate enter extrahepatic tissues via the bloodstream, where they are oxidized through the citric acid cycle to provide additional energy to these tissues, such as bone, myocardium, and renal cortex. This kit is primarily used for the qualitative detection of ketone levels in human and animal liver and muscle tissues.
The detection principle of this kit is as follows: using potassium octanoate as the substrate, ketones react with sodium nitroprusside in an alkaline environment containing ammonium ions to form a purple compound.
Kit Contents
T1522042 | Components Name | Appearance | 50T | Storage | Quantity Per Test |
T1522042A | Tissue Homogenization Solution | Liquid | 250 mL | RT. | As needed |
T1522042B | Potassium Octanoate Solution | Liquid | 5 mL | RT. Store in the dark. | As needed |
T1522042C | Ketone Body Acidification Solution | Liquid | 50 mL | RT. | 1 mL |
T1522042D | Ketone Powder | Powder | 20 g | RT. Store in the dark. | 0.3 g |
Instructions for Use
1. Preparation of Potassium Octanoate Working Solution
Mix potassium octanoate solution with distilled water in a ratio of 1:49 to obtain the working solution.
2. Sample Preparation
Take one mouse or rabbit, decapitate it using scissors to ensure complete blood drainage. Immediately remove the liver and muscle tissues, rinse them several times with pre-chilled tissue homogenization solution, then cut into small pieces or homogenize using a homogenizer for later use.
3. Sample Measurement
| Assay tube 1 | Assay tube 2 | Assay tube 3 |
Liver | 0.3 g | - | - |
Muscle | - | 0.3 g | - |
Liver + Muscle | - | - | 0.3 g |
Potassium Octanoate Working Solution | 3 mL | 3 mL | 3 mL |
Incubate at 37℃ for 30 minutes. | |||
Ketone Body Acidification Solution | 1 mL | 1 mL | 1 mL |
Mix well and let stand for 10 minutes. Take 10 drops of the supernatant from each tube and place them separately in 3 new EP tubes; add 10 drops of distilled water and mix well. | |||
Ketone powder | 0.3 g | 0.3 g | 0.3 g |
Let stand and observe the color change of each tube. |
4. Result Judgment
The liver-containing tube shows purple color; the liver-plus-muscle tube appears light purple or pink; no color change is observed in the muscle-containing tube.
Precautions
1. A distinct purple color is observed at high ketone-body concentrations; light purple or pink appears at low ketone-body concentrations.
2. Glassware used for liver sample processing must not be shared with those for muscle sample processing.
3. If the color of the liver-containing tube cannot be distinguished from that of the liver-plus-muscle tube, add an equal small amount of ketone powder to each tube.
4. For your safety and health, please wear a lab coat and disposable gloves during the operation.
T1522042 | 组分名称 | 物理外观 | 50T | 储存 | 单次测试使用量 |
T1522042A | 组织匀浆液 | 液体 | 250 mL | RT. | 按需 |
T1522042B | 辛酸钾溶液 | 液体 | 5 mL | RT. 避光 | 按需 |
T1522042C | 酮体酸化液 | 液体 | 50 mL | RT. | 1 mL |
T1522042D | 酮粉 | 粉末 | 20 g | RT. 避光 | 0.3 g |
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