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甘油三酯（Triglyceride，TG）又称三酰甘油或三油酸甘油酯，是三分子长链脂肪酸和一分子甘油形成的脂肪分子，是人体内含量最多的脂类，大部分组织均可以利用甘油三酯分解产物供给能量，同时肝脏、脂肪等组织还可以进行甘油三酯的合成。目前，检测甘油三脂的常用方法有酶法和化学法。酶法测定具有简便、快捷、微量且试剂稳定等优点，适用于手工和自动化测定。化学法是使用异丙醇等不同的有机溶剂从血清中抽提出甘油三酯，分层或者吸附去除磷脂等影响因素，再经皂化、氧化，由显色反应进行测定。目前最常用的化学法是乙酰丙酮比色法，其中包括吸附抽提法和分溶抽提法。
检测原理：组织匀浆液或血清中的甘油三脂经TG抽提液处理后，甘油三脂被保留在上层，再经皂化生成甘油，后被氧化剂氧化成甲醛，甲醛与乙酰丙酮在铵根离子存在时，发生缩合反应（Hantgsch反应），生成带荧光的黄色物质即3,5-二乙酰-1,4-双氢二甲基吡啶，用分光光度计在420nm处进行比色测定，与相同处理的标准管对比计算其含量。本方法所用试剂室温下可保存半年以上，比较稳定，在0.25~11.44mg/ml（0.28~12.93mmol/L）之间有良好的线性关系，检测范围较宽。本产品仅用于科研领域，不宜用于临床诊断或其他用途。
适用样本：人或动物的血清、血浆、脑脊液等
自备仪器和试剂
操作步骤
1、样品准备
1.1 血清样品
取样后直接用于检测。
1.2 组织样品
准确称取适量组织样品（质量为m），按质量 (g)：生理盐水 (ml)=1：4的比例，加入生理盐水，冰浴条件下手动或机械匀浆，获取匀浆液（体积为VT），取0.1ml匀浆液备用。
2、TG加样
按照下表设置空白管、对照管、测定管，溶液应按照顺序依次加入，并注意避免产生气泡；如果样品中的TG含量过高，可减少样品用量或适当稀释后再进行测定。
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3、TG测定
取出离心管，流水冷却；空白管调零，比色杯光径1cm，用分光光度计测定420nm处标准管和测定管的吸光度（A标准、A测定）。
4、结果计算
100ml血清中含有的甘油三酯的量：
TG (mg/100ml)=(A测定/A标准)×2×100/0.1=(A测定/A标准)×2000
100g组织中含有的甘油三酯的量：
TG (mg/100g)=(A测定/A标准)×2×VT/0.1×100/m=(A测定/A标准)×2000×VT/m
样品中甘油三酯的浓度：
TG (mg/ml)=(A测定/A标准)×2
参数说明：
A测定：测定管的吸光度
A标准：标准管的吸光度
VT：一定质量组织的匀浆液总体积 (ml)
m：实际取用的组织质量 (g)
5、参考范围
血清TG正常值：0.55~1.70mmol/L；临界值：2.30mmol/L；危险值：4.50mmol/L。
注意事项
1、本法可直接用于检测脑脊液中的TG含量和尿液中的TG含量。
2、取血样后应及时分离，以免红细胞膜磷脂被磷脂酶水解产生游离甘油，或者抗凝剂存在时使红细胞内水溢出而稀释血浆TG值。
3、分离血浆前，标本应放于冰水中，并尽快分离。
4、该方法的线性范围是0.3~12mg/mL， 0.6mg/mL以上比较准确，超过11mg/mL会有稍微偏差。因此，样品浓度超出上述范围应做相应处理。
5、显色后吸光度会随时间变化，故应及时比色，当标本过多时，可置冰箱中逐一比色。
6、皂化、氧化、显色时间和温度对显色结果及最终的吸光度均会造成影响，所以每批次测定都应同时做标准对照。
7、做血浆标本时，通常选用1mg/mL EDTA.2K做抗凝剂。
8、试剂开封后请尽快使用，以防影响后续实验效果。
9、为了您的安全和健康，请穿实验服并戴一次性手套操作。
Triglycerides (TG), also known as triacylglycerol or triolein, are fat molecules formed from three long-chain fatty acids and one glycerol molecule. They are the most abundant lipids in the human body. Most tissues can utilize the breakdown products of triglycerides for energy, while organs such as the liver and adipose tissue can also synthesize triglycerides. Currently, common methods for detecting triglycerides include enzymatic and chemical methods. The enzymatic method is simple, rapid, requires minimal sample volume, and offers stable reagents, making it suitable for both manual and automated assays. Chemical methods involve extracting triglycerides from serum using organic solvents such as isopropanol, removing interfering factors like phospholipids through layering or adsorption, followed by saponification, oxidation, and colorimetric determination. The most commonly used chemical method is the acetylacetone colorimetric method, which includes adsorption extraction and solvent extraction techniques.
Detection Principle
Triglycerides in tissue homogenates or serum are treated with TG extraction solution, retaining the triglycerides in the upper layer. They are then saponified to produce glycerol, which is oxidized by an oxidizing agent to form formaldehyde. In the presence of ammonium ions, formaldehyde condenses with acetylacetone (Hantzsch reaction) to form a yellow fluorescent compound, 3,5-diacetyl-1,4-dihydrodimethylpyridine. The absorbance is measured at 420 nm using a spectrophotometer, and the content is calculated by comparison with a similarly treated standard. The reagents used in this method can be stored at room temperature for over six months, demonstrating good stability. The assay shows a good linear relationship within the range of 0.25–11.44 mg/ml (0.28–12.93 mmol/L) and has a wide detection range. This product is intended for scientific research purposes only and is not suitable for clinical diagnosis or other uses.
Applicable Samples: Human or animal serum, plasma, cerebrospinal fluid, etc.
Reagents, consumables and Equipments not provided
Operating Steps
1. Sample Preparation
1.1 Serum Samples
Use directly for detection after collection.
1.2 Tissue Samples
Accurately weigh an appropriate amount of tissue sample (mass = m), add physiological saline at a mass (g) to volume (ml) ratio of 1:4, homogenize manually or mechanically under ice bath conditions to obtain a homogenate (total volume = V<sub>T</sub>). Take 0.1 ml of the homogenate for use.
2. TG Sample Addition
Set up blank, standard, and test tubes according to the table below. Add solutions in the specified order, taking care to avoid bubbles. If the TG content in the sample is too high, reduce the sample volume or dilute appropriately before measurement.
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3. TG Measurement
Remove the tubes and cool under running water. Zero the spectrophotometer with the blank tube, using a 1 cm light path cuvette, measure the absorbance of the standard and test tubes at 420 nm (A<sub>Standard</sub>, A<sub>Test</sub>).
4. Result Calculation
Triglyceride content in 100 ml of serum:
TG (mg/100 ml) = (ATest / AStandard) × 2 × 100 / 0.1 = (ATest / AStandard) × 2000
Triglyceride content in 100 g of tissue:
TG (mg/100 g) = (ATest / AStandard) × 2 × VT / 0.1 × 100 / m = (ATest / AStandard) × 2000 × VT / m
Triglyceride concentration in the sample:
TG (mg/ml) = (ATest / AStandard) × 2
Parameter Explanation:
ATest: Absorbance of the test tube
AStandard: Absorbance of the standard tube
V<sub>T</sub>: Total volume of the homogenate from a given mass of tissue (ml)
m: Actual mass of tissue used (g)
TG concentration conversion: 1 mg/mL = 1.13 mmol/L
5. Reference Range
Normal serum TG range: 0.55–1.70 mmol/L; Critical value: 2.30 mmol/L; Risk value: 4.50 mmol/L.
Precautions
This method can be directly used to detect TG levels in cerebrospinal fluid and urine.
Blood samples should be separated promptly to avoid hydrolysis of red blood cell membrane phospholipids by phospholipase, which produces free glycerol, or dilution of plasma TG values due to water release from red blood cells in the presence of anticoagulants.
Before plasma separation, samples should be placed in ice water and processed as soon as possible.
The linear range of this method is 0.3–12 mg/mL. Results are more accurate above 0.6 mg/mL, with slight deviations possible above 11 mg/mL. Samples exceeding this range should be processed accordingly.
Absorbance after color development changes over time, so measurements should be taken promptly. If there are many samples, they can be stored in the refrigerator and measured one by one.
Saponification, oxidation, color development time, and temperature can affect color development and final absorbance. Therefore, standard controls should be included in each batch of tests.
For plasma samples, 1 mg/mL EDTA·2K is typically used as the anticoagulant.
Please use the reagent as soon as possible after opening to avoid affecting subsequent experimental results.
For your safety and health, please wear a lab coat and disposable gloves during operation.
| T1509111 | Component | 50T | Storage |
| T1509111A | TG标准 (2mg/mL) | 1 mL | 2-8℃. Store in the dark. |
| T1509111B | TG抽提液 | 70 mL | RT. Store in the dark. |
| T1509111C | 酸性缓冲液 | 6 mL | RT. |
| T1509111D | 皂化剂 | 30 mL | RT. |
| T1509111E | TG氧化剂 | 30 mL | 2-8℃. Store in the dark. |
| T1509111F | TG显色剂 | 30 mL | 2-8℃. Store in the dark. |
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| 批号(Lot Number) | 证书类型 | 货号 |
|---|---|---|
 J2614475 |
分析证书 | T1509111 |
| F2602404 |
分析证书 | T1509111 |
| ZJ26F0332418 |
分析证书 | T1509111 |
| ZJ26F0332417 |
分析证书 | T1509111 |
| ZJ26F0332419 |
分析证书 | T1509111 |
| ZJ26F0332420 |
分析证书 | T1509111 |
| ZJ26F0332421 |
分析证书 | T1509111 |
| ZJ26F0332422 |
分析证书 | T1509111 |
危险品化学品经营许可证（带存储）