计算溶液所需的质量、体积或浓度。
氨基酸 (AA) 含量检测试剂盒 (茚三酮, 比色法)
有货
为什么选择此级别
BioReagent,比色法,适用于分析 级 ，适用于对基线干扰要求严格的色谱和分析工作流程。
储存与运输
2-8°C储存。低温运输 。请查阅批次 COA 获取详细规格。
质量文档
SDS、COA、产品数据表及规格说明书均可下载。可通过批号查询获取批次 COA。
文献证明
在色谱分析、有机合成和交叉偶联反应领域已被 0 篇同行评审文献引用。
概述
氨基酸作为组成蛋白质的基本单元，在生物代谢过程中起着关键作用。动物肝脏、肾脏是氨基酸代谢的主要器官，尿液中氨基酸的变化可以反映体内氨基酸的代谢情况，以及肝脏、肾脏的生理状态；植物体内氨基酸含量对研究植物在不同时期氮代谢的变化、植物根系生理、植物对氮素的吸收、运输、同化及营养状况等具有重要意义。
氨基酸中的α-氨基在加热条件及弱酸环境下能够与水合茚三酮反应生成蓝紫色化合物，产物在570 nm处具有特征吸收峰，通过吸光值变化即可定量检测氨基酸的含量。
产品组分
A1515867 | 组份 | 物理状态 | 100T | 储存条件 | 单次测试使用量 |
A1515867A | 提取液 | 液体 | 120 mL | 2-8℃ | 300 μL |
A1515867B | 检测缓冲液 | 液体 | 60 mL | 2-8℃ | 500 μL |
A1515867C | 水合茚三酮 | 粉末 | 2 EA | 2-8℃, 避光. | 200 μL |
A1515867D | 维生素C | 粉末 | 2 EA | 2-8℃, 避光. | 500 μL |
A1515867E | 标准品 | 粉末 | 1 EA | 2-8℃, 避光. | 250 μL |
注:
组分A和组分B有腐蚀性和挥发性，应小心操作，同时应密闭保存，防止挥发；
组分C和组分D有毒且有刺激性，建议在使用过程中做好个人防护
自备仪器和试剂
分光光度计（能测570 nm处的吸光度）
离心管或试管、比色皿
离心机、水浴锅、研钵或匀浆器、磁力搅拌器或超声波
无氨蒸馏水或去离子水、95%乙醇、60%乙醇
使用说明
1. 氨基酸的提取
注意：推荐使用新鲜样本，如果不立即进行实验，样本可在-80℃保存1个月。
① 动物组织：称取约0.1 g动物组织样本，加入1 mL 提取液，室温充分匀浆，转移到1.5 mL EP管中（推荐使用螺旋盖EP管），盖紧后（防止水分散失）置于沸水浴15 min；自来水冷却后，10000 g，室温离心10 min，取上清液，待测。
② 植物组织：称取约0.1 g植物组织样本，加入1 mL 提取液捣碎，室温超声波破碎5 min（功率20%或200W，超声3 s，间隔7 s，重复30次），转移到1.5 mL EP管中（推荐使用螺旋盖EP管）， 盖紧后（防止水分散失）置于沸水浴15 min；自来水冷却后，10000 g，室温离心10 min，取上清液， 待测。
③ 细菌或细胞：收集5×10⁶个细菌或细胞，用冷PBS清洗细菌或细胞，800 g离心2 min，弃上清液。加入1 mL 提取液，室温超声波破碎细菌或细胞5 min（功率20%或200 W，超声3 s，间隔7 s，重复30次），转移到1.5 mL EP管中（推荐使用螺旋盖EP管），盖紧后（防止水分散失）置于沸水浴15 min；自来水冷却后，10000 g，室温离心10 min，取上清液，待测。
④ 血清（浆）、细胞上清、尿液等液体：吸取0.5 mL液体加入0.5 mL 提取液（推荐使用螺旋盖EP管），盖紧后（防止水分散失）置于沸水浴15 min；自来水冷却后，10000 g，室温离心10 min，取上清液，待测。
2. 试剂准备
2.1 配制茚三酮显色液
取一瓶水合茚三酮，加入 10mL 95%乙醇充分溶解，即为茚三酮显色液。未用完的试剂可4℃避光保存1周。
注：水合茚三酮有毒且有刺激性，建议在使用过程中做好个人防护。
2.2 配制维生素C工作液
取一瓶维生素C，准确溶解于30 mL检测缓冲液中，混匀。一次性用不完，可分装小管后-20℃保存1周有效。该试剂盒提供的维生素C及其配制的工作液为过量。
2.3 氨基酸标准品配制
称取10mg氨基酸标准品，加入1.332 mL去离子水，充分溶解后得到7.5 mg/mL的氨基酸标准溶液， 未用完的试剂可4℃避光保存1个月。
使用7.5 mg/mL 标准溶液，按照下表所示，进一步稀释标准品。每次实验都要做一次标准品检测，制作标曲；稀释后的标准品溶液不稳定，必须在4小时内使用。
序号 | 标准品 (μL) | 提取液（μL） | 浓度 (μg/mL) |
1 | 25 µL of 7.5 mg/mL | 975 | 187.5 |
2 | 200 µL of 187.5 μg/mL | 200 | 93.75 |
3 | 200 µL of 93.75 μg/mL | 200 | 46.875 |
4 | 200 µL of 46.875 μg/mL | 200 | 23.4375 |
5 | 200 µL of 23.4375 μg/mL | 200 | 11.7188 |
6 | 200 µL of 11.71875 μg/mL | 200 | 5.8594 |
空白 | 0 | 200 | 0 |
3. 样本检测
3.1 分光光度计预热30 min以上，调节波长至570 nm。
3.2 在2 mL EP管（推荐使用螺旋盖EP管）中按照如下方式操作：
试剂 | 空白管 (μL) | 标准管 (μL) | 测定管 (μL) |
提取液 | 100 | 0 | 0 |
标准品 | 0 | 100 | 0 |
样品 | 0 | 0 | 100 |
茚三酮显色液 | 200 | 200 | 200 |
维生素C工作液 | 500 | 500 | 500 |
3.3 混匀后盖紧管盖（防止水分散失），置于沸水浴中保温5 min，冰浴冷却30 s后加入1.2 mL 60%乙醇，反复颠倒EP管数次。
3.4 空白管调零，于570 nm处测定吸光值，分别记为A空白、A标准和A测定，显色后务必在30 min内测完。
4. 结果计算
4.1 数据处理
计算ΔA测定=A测定-A空白，ΔA标准=A标准-A空白。
4.2 标准曲线绘制
以标准品浓度为x轴，ΔA标准为y轴，绘制标准曲线。
4.3 样本氨基酸含量的计算
(1) 按样本质量计算 氨基酸含量 (μg/g)=y÷(W÷V提取)×n=y÷N×n
(3) 按液体体积计算 氨基酸含量 (μg/mL)=y×2×n
(4) 按蛋白浓度计算 氨基酸含量 (μg/mg prot)=y÷Cpr×n
参数说明：
W: 样本质量，g；
V提取: 加入提取液体积，1 mL；
n: 样本的稀释倍数；
Cpr: 上清液蛋白质浓度，mg/mL；
N：细菌或细胞数量，以10⁴为单位（例如细菌数量为5×10⁶，N=500）；
2：提取液体时的稀释倍数，(0.5 mL+0.5 mL)/0.5 mL=2。
注意事项
1. 正式检测前，建议选择2-3个预期差异较大的样本进行预实验。
2. 对于组织样本及细胞样本等，可通过测定蛋白浓度来进行样本间结果均一化。推荐使用阿拉丁B665595 BCA蛋白定量试剂盒或R1491648 即用型BCA蛋白定量试剂盒。
3. 脯氨酸和羟脯氨酸与茚三酮反应在570 nm处无吸收峰，因此，570 nm处测定结果不含这两种氨基酸。建议使用P1515888 脯氨酸（PRO）含量检测试剂盒（茚三酮, 比色法）、P1515887 脯氨酸（PRO）含量检测试剂盒（茚三酮, 微量法）、H1515817 羟脯氨酸 (HYP) 含量检测试剂盒（Ehrlich, 微量法）。
4. 组分A和组分B有腐蚀性和挥发性，应小心操作，同时应密闭保存，防止挥发；组分C和组分D有毒且有刺激性，建议在使用过程中做好个人防护
5. 建议实验者自行建立标准曲线以提高结果准确度。如未自行建立标准曲线，可参考结果展示部分的典型标准曲线公式进行结果计算。
6. 生化检测试剂普遍具有刺激性、生物毒性等，为了您的安全和健康，请在实验全程做好生物安全防护，穿戴实验服、口罩、手套、头套等安全护具，在通风橱、生物安全柜中进行实验。
7. 该试剂盒仅用于科研领域，不宜用于临床诊断或其他用途。
As the fundamental building blocks of proteins, amino acids play pivotal roles in biological metabolism. The liver and kidneys serve as the primary organs for amino acid metabolism in animals. Variations in urinary amino acid levels reflect the body’s overall amino acid metabolic profile, as well as the physiological status of the liver and kidneys. In plants, measurements of endogenous amino acid concentrations are of great significance for investigating nitrogen metabolic shifts across developmental stages, root physiology, and the uptake, translocation, assimilation of nitrogen nutrients, alongside overall plant nutritional status.
Under heating and weakly acidic conditions, the α-amino group of amino acids reacts with ninhydrin hydrate to form a blue-violet chromogenic complex, which exhibits a characteristic absorption peak at 570 nm. Amino acid concentrations can therefore be quantified by measuring changes in absorbance.
Components
A1515867 | Components | Appearance | 100T | Storage | Quantity Per Test |
A1515867A | Extraction Buffer | Liquid | 120 mL | 2-8℃ | 300 μL |
A1515867B | Assay Buffer | Liquid | 60 mL | 2-8℃. | 500 μL |
A1515867C | Ninhydrin | Solid | 2 EA | 2-8℃. Store in the dark. | 200 μL |
A1515867D | Ascorbic acid | Solid | 2 EA | 2-8℃. Store in the dark. | 500 μL |
A1515867E | Standard | Solid | 1 EA | 2-8℃. Store in the dark. | 250 μL |
Note:
Components A and B are corrosive and volatile. Handle with caution and store in tightly sealed containers to prevent volatilization.
Components C and D are toxic and irritant. Appropriate personal protective equipment shall be worn during handling.
Reagents, consumables and Equipments not provided
Spectrophotometer (able to measure absorbance at 570 nm)
Centrifuge tubes or test tubes, cuvettes
Centrifuge, water bath, mortar or homogenizer, magnetic stirrer or ultrasonic device
Ammonia-free distilled water or deionized water, 95% ethanol, 60% ethanol
Instructions for Use
1. Extraction of Amino Acids
Note: Fresh samples are recommended. Samples can be stored at -80 °C for up to 1 month if the assay cannot be performed immediately.
① Animal tissue
Weigh approximately 0.1 g animal tissue sample and add 1 mL extraction solution. Homogenize thoroughly at room temperature, then transfer the mixture into a 1.5 mL EP tube (screw-cap EP tubes are recommended). Tighten the cap to avoid water loss, and incubate the tube in a boiling water bath for 15 min. Cool the tube with tap water, centrifuge at 10,000 g for 10 min at room temperature, and collect the supernatant for subsequent measurement.
② Plant tissue
Weigh approximately 0.1 g plant tissue sample, add 1 mL extraction solution and grind the tissue. Perform ultrasonic lysis at room temperature for 5 min (power: 20% or 200 W; pulse setting: 3 s sonication, 7 s interval, 30 cycles total). Transfer the homogenate into a 1.5 mL screw-cap EP tube. Seal the tube tightly to prevent water evaporation, then heat in a boiling water bath for 15 min. Cool with tap water, centrifuge at 10,000 g for 10 min at room temperature, and retain the supernatant for testing.
③ Bacteria or cultured cells
Collect 5×10⁶ bacterial or mammalian cells. Wash the pellet with pre-cooled PBS, centrifuge at 800 g for 2 min, and discard the supernatant. Add 1 mL extraction solution and lyse the pellet by sonication at room temperature for 5 min (power: 20% or 200 W; pulse setting: 3 s sonication, 7 s interval, 30 cycles total). Transfer the lysate to a 1.5 mL screw-cap EP tube, seal tightly to avoid water loss, and heat in boiling water for 15 min. After cooling under tap water, centrifuge at 10,000 g for 10 min at room temperature, and collect the supernatant to be assayed.
④ Liquid samples: serum (plasma), cell culture supernatant, urine, etc.
Pipette 0.5 mL liquid sample and mix with 0.5 mL extraction solution in a 1.5 mL screw-cap EP tube. Tighten the cap to prevent water evaporation, then incubate in a boiling water bath for 15 min. Cool the tube with tap water, perform centrifugation at 10,000 g for 10 min at room temperature, and collect the supernatant for measurement.
2. Reagent Preparation
2.1 Preparation of Ninhydrin Chromogenic Solution
Dissolve one vial of ninhydrin hydrate completely in 10 mL of 95% ethanol to prepare the ninhydrin chromogenic solution. Unused reagent can be stored at 4℃ away from light for up to one week.
Note: Ninhydrin hydrate is toxic and irritating. Appropriate personal protective equipment must be worn during handling.
2.2 Preparation of Vitamin C Working Solution
Accurately dissolve one vial of vitamin C in 30 mL assay buffer and mix thoroughly. If the solution cannot be used up in one experiment, aliquot it into small tubes and store at -20℃; it remains valid for one week after preparation. The kit contains excess vitamin C powder and corresponding working solution volume.
2.3 Preparation of Amino Acid Standard Stock Solution
Weigh out 10 mg amino acid standard powder and dissolve fully in 1.332 mL deionized water to obtain a 7.5 mg/mL amino acid standard stock solution. Unused stock solution can be stored at 4℃ protected from light for up to one month.
Dilute the 7.5 mg/mL standard stock solution stepwise according to the table below. A standard curve shall be generated with serial diluted standards in every single assay. The diluted standard solutions are unstable and must be used within 4 hours after dilution.
No. | Standard (μL) | Extraction Buffer（μL） | Concentration (μg/mL) |
1 | 25 µL of 7.5 mg/mL | 975 | 187.5 |
2 | 200 µL of 187.5 μg/mL | 200 | 93.75 |
3 | 200 µL of 93.75 μg/mL | 200 | 46.875 |
4 | 200 µL of 46.875 μg/mL | 200 | 23.4375 |
5 | 200 µL of 23.4375 μg/mL | 200 | 11.7188 |
6 | 200 µL of 11.71875 μg/mL | 200 | 5.8594 |
Blank | 0 | 200 | 0 |
3. Sample Assay
3.1 Preheat the spectrophotometer for more than 30 min and set the wavelength to 570nm.
3.2 Carry out the operations in 2 mL EP tubes (screw-cap EP tubes are recommended) according to the table below:
Reagent | Blank Tube (μL) | Standard Tube (μL) | Test Tube (μL) |
Extraction Buffer | 100 | 0 | 0 |
Standard | 0 | 100 | 0 |
Sample | 0 | 0 | 100 |
Ninhydrin Chromogenic Solution | 200 | 200 | 200 |
Vitamin C Working Solution | 500 | 500 | 500 |
3.3 Mix thoroughly and tighten the tube cap to prevent water loss. Incubate in a boiling water bath for 5 min, cool in an ice bath for 30 s, add 1.2 mL of 60% ethanol, and invert the EP tube repeatedly several times.
3.4 Zero the instrument with the blank tube, then measure the absorbance at 570 nm, recorded as Ablank,Astandard and Asample respectively. The absorbance must be measured within 30 min after color development.
4.1 Data Processing
Calculate ΔAsample = Asample − Ablank
ΔAstandard = Astandard − Ablank
4.2 Standard Curve Plotting
Plot the standard curve with standard concentration as the X-axis and ΔAstandard as the Y-axis.
4.3 Calculation of Amino Acid Content in Samples
(1) Calculation based on sample weight
Amino acid content (μg/g) = Y ÷ (W ÷ V₍extraction₎) × n = Y ÷ W × n
(2) Calculation based on bacterial or cell count
Amino acid content (μg/10⁴ cells) = Y ÷ (N ÷ V₍extraction₎) × n = Y ÷ N × n
(3) Calculation based on liquid volume
Amino acid content (μg/mL) = Y × 2 × n
(4) Calculation based on protein concentration
Amino acid content (μg/mg prot) = Y ÷ C₍prot₎ × n
Parameter Definition
W: Sample weight, g
V₍extraction₎: Volume of extraction solution added, fixed at 1 mL
n: Dilution factor of the sample
C₍prot₎: Protein concentration of supernatant, mg/mL
N: Number of bacteria or cells, in units of 10⁴ (e.g., if the cell count is 5×10⁶, N = 500)
2: Dilution factor for liquid samples: (0.5 mL sample + 0.5 mL extraction solution) ÷ 0.5 mL sample = 2
Precautions
1. Before formal testing, it is recommended to conduct a pilot test with 2–3 samples with expected large differences in amino acid levels.
2. For tissue and cell samples, protein concentration measurement can be used to normalize results across samples. Recommended kits: Aladdin BCA Protein Quantitation Kit (Cat. No. B665595) or Ready-to-Use BCA Protein Quantitation Kit (Cat. No. R1491648).
3. Proline and hydroxyproline produce no characteristic absorption peak at 570 nm after reaction with ninhydrin, so the measurement at 570 nm does not include these two amino acids. Specialized kits are recommended for their separate detection:
Cat. No. P1515888: Proline (PRO) Content Assay Kit (Ninhydrin Colorimetric Method)
Cat. No. P1515887: Proline (PRO) Content Assay Kit (Ninhydrin Microplate Method)
Cat. No. H1515817: Hydroxyproline (HYP) Content Assay Kit (Ehrlich Microplate Method)
4. Components A and B are corrosive and volatile. Handle with care and store hermetically sealed to avoid volatilization. Components C and D are toxic and irritant; adequate personal protective equipment shall be worn during operation.
5. Operators are advised to generate their own standard curve for higher test accuracy. If no self-generated curve is available, calculate results using the typical standard curve equation provided in the Result Demonstration section.
6. Biochemical assay reagents are generally irritant and biologically toxic. For your safety and health, full biosafety protection is required throughout the experiment, including lab coat, respirator mask, gloves and hair cap. All operations shall be performed inside a fume hood or biosafety cabinet.
7. This kit is intended for research use only and shall not be applied for clinical diagnosis or other medical purposes.
规格
别名
氨基酸检测试剂盒（茚三酮比色法） | 氨基酸 (AA) 含量检测试剂盒 比色法 | 氨基酸 (AA) 检测试剂盒 (茚三酮比色法) | 氨基酸 (AA) 含量检测试剂盒 | 氨基酸含量检测试剂盒
英文别名
Amino Acid (AA) Content Assay Kit | Micro Amino Acid (AA) Assay Kit
规格或纯度
BioReagent, 比色法, 适用于分析
英文名称
Amino Acid (AA) Content Assay Kit (Ninhydrin, Colorimetric Method)
应用
细胞代谢
样品类型
尿液, 植物组织, 药物, 血清, 食品
检测范围
5.86-187.5 μg/mL
检测方法
比色法
定量&半定量
定量
检测仪器
可见光分光光度计
检测波长
570nm
敏感性
Light-sensitive
检测时长
10 min
样本体积
1 mL
储存与运输
储存条件
2-8°C储存
运输条件
低温运输
稳定性与储存
各组分在相应的储存条件下保质期1年。
组分与储存
A1515867 | 组份 | 物理状态 | 100T | 储存条件 | 单次测试使用量 |
A1515867A | 提取液 | 液体 | 120 mL | 2-8℃ | 300 μL |
A1515867B | 检测缓冲液 | 液体 | 60 mL | 2-8℃. | 500 μL |
A1515867C | 水合茚三酮 | 粉末 | 2 EA | 2-8℃, 避光 | 200 μL |
A1515867D | 维生素C | 粉末 | 2 EA | 2-8℃, 避光 | 500 μL |
A1515867E | 标准品 | 粉末 | 1 EA | 2-8℃, 避光 | 250 μL |
技术文档
高级数据
质检证书（CoA,COO,BSE/TSE 和分析图谱)
C of A & Other Certificates(BSE/TSE, COO):
输入批号以搜索分析图谱:
通过匹配包装上的批号来查找并下载产品的 COA，每批产品都进行了严格的验证，您可放心使用！
| 批号(Lot Number) | 证书类型 | 货号 |
|---|---|---|
 ZJ26F0939006 |
分析证书 | A1515867 |
| ZJ26F0737307 |
分析证书 | A1515867 |
技术文档和文章
溶液计算器
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