计算溶液所需的质量、体积或浓度。
花青素(总花色苷)含量检测试剂盒(pH示差，微量法)
有货
为什么选择此级别
BioReagent,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
花青素是广泛存在于自然界的植物水溶性天然色素，天然状态下极少游离，大多与各类单糖结合生成糖苷，即为花色苷。花色苷常见于陆生植物的根、茎、叶、花和果实，具备抗氧化活性，可清除自由基。
本试剂盒检测原理如下：花色苷的分子结构与颜色随 pH 发生改变，而样品中干扰杂质的光谱特征不随 pH 变化。pH 1.0 条件下，花色苷主要以红色的黄烊盐阳离子形式存在；pH 4.5 条件下，花色苷转变为无色的甲醇假碱。选取 pH 1.0 与 pH 4.5 这一组对花色苷吸光度差异大、且结构相对稳定的 pH 条件，测定校正浊度后的吸光度差值，该差值与花色苷含量成正比，即可计算样品中花色苷的总量（以矢车菊素-3-葡萄糖苷计）。
产品组分
A1524828 | 组分名称 | 物理外观 | 100T | 储存 | 单次测试使用量 |
A1524828A | 提取液 | 液体 | 50 mL | 2-8℃ | 按需 |
A1524828B | 试剂一 | 液体 | 20 mL | 2-8℃ | 180 μL |
A1524828C | 试剂二 | 液体 | 20 mL | 2-8℃ | 180 μL |
使用方法
1. 样本处理
称取约0.1g 样品（可根据样本类型适当调整待测样本量，具体比例可以参考文献），加入1mL 提取液，充分研磨匀浆后转移至EP管中，提取液定容至1mL，盖紧后60℃ 浸提30min，期间可振荡数次，室温12000rpm, 离心10 min, 取上清液待测。
2. 样本测定
按照下表进行样本测定。
| 测定孔1 | 测定孔2 |
待测样本（μL） | 20 | 20 |
试剂一（μL） | 180 | - |
试剂二（μL） | - | 180 |
混匀，静置5min，使用酶标仪分别测定530nm和700nm处OD值，其中测定孔1在530nm和700nm处OD值记为A1、A1’，测定孔2在530nm和700nm处OD值记为A2、A2’，计算ΔA=(A1-A1’) - (A2-A2’)。 |
3. 结果计算
（1） 使用96 孔板测定的计算公式如下
花色苷含量(μg/g 质量)=[ΔA×V÷(ε×d)×M×F×10⁶]÷W=33.4×ΔA×F÷W
V：提取液体积，0.001L；
ε：花色苷的摩尔消光系数，26900 L/mol/cm；
d：96 孔板光径，0.5 cm；
M：花色苷的相对分子质量：449.2 g/mol；
F：稀释倍数，此反应体系下为10；
10⁶ ：单位换算系数，1g= 10⁶ μg
W：样本质量：g
（2）使用微量玻璃比色皿测定的计算公式
将上述计算公式中光径d: 0.5 cm 调整为d: 1 cm 进行计算即可。
注意事项
1. 试剂二有轻微刺激性气味，建议在使用过程中做好个人防护。
2. 如果A1大于1，可适当加大稀释倍数，保证总体积200 μL 不变，如10 μL 上清液和190 μL 试剂一（相当于稀释20 倍）；如果A1小于0.1，可以适当缩小稀释倍数，保证总体积200 μL 不变，如100 μL 上清液和100 μL 试剂一（相当于稀释2 倍），使A1保持在0.1-1 范围内，可提高检测灵敏度；注意应同样调整上清液和试剂二体积比例，计算时以实际稀释倍数代入计算公式中。
3. 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。
4. 为了您的安全和健康，请穿实验服并戴一次性手套操作。
Anthocyanidins are water-soluble natural pigments widely distributed in plants. In nature, they rarely exist in free form; most are glycosides conjugated with various monosaccharides, namely anthocyanins. Anthocyanins are commonly found in the roots, stems, leaves, flowers and fruits of terrestrial plants and exhibit antioxidant activity to scavenge free radicals.
The detection principle of this assay kit is described below: the molecular structure and color of anthocyanins change with pH, whereas the spectral characteristics of interfering impurities in samples remain pH-independent. At pH 1.0, anthocyanins predominantly exist as red flavylium cations. At pH 4.5, anthocyanins convert into colorless carbinol pseudobases. A pair of pH conditions (pH 1.0 and pH 4.5) are selected, at which anthocyanins show a large absorbance difference and relatively stable structure. The absorbance difference, corrected for turbidity, is measured. This difference is proportional to anthocyanin concentration, enabling quantification of total anthocyanin content in samples (expressed as cyanidin-3-glucoside equivalents).
Kit Contents
A1524828 | Component Name | Appearance | 100T | Storage | Quantity Per Test |
A1524828A | Extraction Solution | Liquid | 50 mL | 2-8℃ | As needed |
A1524828B | Reagent 1 | Liquid | 20 mL | 2-8℃ | 180 μL |
A1524828C | Reagent 2 | Liquid | 20 mL | 2-8℃ | 180 μL |
Instructions for Use
1. Sample Preparation
Weigh approximately 0.1 g of sample (sample mass may be adjusted according to sample type; refer to literature for appropriate ratios). Add 1 mL of Extraction Solution, grind and homogenize thoroughly, then transfer the mixture to an Eppendorf tube. Make up to 1 mL with Extraction Solution, cap tightly, and incubate at 60 °C for 30 min with occasional shaking. Centrifuge at 12000 rpm for 10 min at room temperature and retain the supernatant for assay.
2. Sample Measurement
Perform sample measurement according to the table below.
Reagent name (μL) | Assay Well 1 | Assay Well 2 |
Sample | 20 | 20 |
Reagent 1 | 180 | — |
Reagent 2 | — | 180 |
Mix thoroughly and let stand for 5 minutes. Read absorbance at 530 nm and 700 nm using a microplate reader. Record absorbance values of Assay Well 1 at 530 nm and 700 nm as A1 and A1’, and absorbance values of Assay Well 2 at 530 nm and 700 nm as A2 and A2’. Calculate ΔA = (A1 - A1’) - (A2 − A2’). |
3. Calculation of Results
(1) Formula for 96-well microplate assay
Total anthocyanin content (μg/g sample) = [ΔA × V ÷ (ε × d) × M × F × 10⁶] ÷ W = 33.4 × ΔA × F ÷ W
V: Volume of extraction solution, 0.001 L
ε: Molar extinction coefficient of anthocyanin, 26900 L/mol/cm
d: Optical path length of 96-well plate, 0.5 cm
M: Molecular weight of anthocyanin, 449.2 g/mol
F: Dilution factor; equals 10 in this reaction system
10⁶: Unit conversion factor (1 g = 10⁶ μg)
W: Weight of sample, g
(2) Formula for micro glass cuvette assay
Use the same formula above, with optical path length d changed from 0.5 cm to 1 cm.
Precautions
1. Reagent 2 has a slight pungent odor. Wear appropriate personal protective equipment during operation.
2. If A1 > 1, increase the dilution factor while maintaining a total volume of 200 μL. For example, mix 10 μL supernatant with 190 μL Reagent 1 (20-fold dilution). If A1 < 0.1, reduce the dilution factor while keeping the total volume at 200 μL. For example, mix 100 μL supernatant with 100 μL Reagent 1 (2-fold dilution). Keep A1 in the range of 0.1–1 to optimize detection sensitivity. Adjust the volume ratio of supernatant and Reagent 2 accordingly, and substitute the actual dilution factor into the calculation formula.
3. Before the formal test, it is recommended to select 2-3 samples with significant expected differences for a preliminary experiment.
4. For your safety and health, please wear a lab coat and disposable gloves during the operation.
规格
规格或纯度
BioReagent, 适用于分析
英文名称
Anthocyanin (Total Anthocyanins) Content Assay Kit (pH Differential, Micro Method)
应用
细胞代谢
检测方法
分光光度法
检测仪器
酶标仪
检测波长
530nm,700nm
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12个月。
组分与储存
A1524828 | 组分名称 | 物理外观 | 100T | 储存 | 单次测试使用量 |
A1524828A | 提取液 | 液体 | 50 mL | 2-8℃ | 按需 |
A1524828B | 试剂一 | 液体 | 20 mL | 2-8℃ | 180 μL |
A1524828C | 试剂一二 | 液体 | 20 mL | 2-8℃ | 180 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):
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技术文档和文章
溶液计算器
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