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花色素苷（Anthocyanin），亦称花青素，属类黄酮化合物，是广泛存在于植物细胞液泡中的水溶性色素，赋予花、果实、叶片红、紫、蓝等颜色。其在植物体内的含量与器官颜色、发育阶段及采后衰老过程密切相关，直接影响农产品的加工性能、贮藏品质和营养价值。
检测原理：花青素溶于有机溶剂，以有机溶剂粗提花青素，花青素在酸性溶液中呈红色，颜色深浅与花青素含量呈正比，于530nm有最大吸收波，可利用酶标仪测定其吸光度，进而计算出花青素的含量，方法简单易行。由于植物中花色苷的种类多样（如矢车菊素Cyanidin、天竺葵素Pelargonidin等），其摩尔消光系数不同，本试剂盒不提供单一标准品，因此结果以相对吸光值或相对含量表示。如需测定绝对含量，请用户自备与样品对应的花色苷纯品作为标准。本试剂盒主要用于植物组织或果实中花色苷的提取与相对定量，仅用于科学研究，不适用于临床诊断或其他用途。
适用样本：植物组织或果实
| 产品号 | Component | 50T | Storage |
| A1509392 | Anthocyanin Assay Buffer | 500 mL | RT. |
自备仪器和试剂
实验材料：桃子、李子、苹果、杏等果实或其他植物组织
操作步骤（仅供参考）
1、花青素提取
方法一：
(1) 4℃预冷研钵或匀浆器，以及Anthocyanin Assay Buffer。
(2) 取植物组织，洗净，擦干，称取新鲜样品0.25g，剪碎，置于研钵或匀浆器中。
(3) 加入2~3mL Anthocyanin Assay Buffer，充分研磨或匀浆后转入10mL离心管，用Anthocyanin Assay Buffer冲洗研钵或匀浆器并转移至离心管，补加Anthocyanin Assay Buffer至8mL。
(4) 4℃避光静置20min，期间摇动2~3次，然后过滤至离心管，也可用离心机8000r/min离心3min，滤液（上清液）即为花青素粗提液。
方法二：
将样品按上述比例加入提取液中，置于32℃温箱浸提4~8h（期间可摇动数次）或室温下避光浸提24h，离心取上清即为花青素粗提液。
2、加样及测定
取1cm比色皿，用Anthocyanin Assay Buffer做空白对照，取花青素提取液（1~2mL）用分光光度计在530nm波长处读取吸光度值；如果花青素浓度过高，可以适当稀释后再进行测定，样品的检测最好能设置2平行孔，求平均值。
3、结果计算
花青素单位定义：以OD值为0.1时的花青素浓度称为一个单位（U）；计算方法如下：
组织样品的花青素含量 (U/g)=(A测定-A空白)×N/(0.1xW)
参数说明：
A测定：花青素提取液在530nm的OD值
A空白：空白对照在530nm的OD值
W：样品鲜重 (g)
N：稀释倍数
注：亦可用自备的花青素标准品做梯度稀释（0.1、0.2、0.3、0.4、0.5、1mg/mL），用Anthocyanin Assay Buffer做空白对照，绘制标准曲线，进而计算出样品中花青素的含量。
组织样品的花青素 (mg/g)=C×VT×N/W
液体样品的花青素 (mg/mL)=C×N
参数说明：
C：根据标准曲线求得的测定管花青素含量 (mg/mL)
VT：花青素粗提液总体积 (ml) =8
W：样品鲜重 (g)
N：稀释倍数
注意事项
1、为了避免花青素见光发生变化，操作时应尽量避光，研磨或匀浆应尽量缩短时间。
2、取样量、试剂用量应根据花青素含量适当调整。
3、Anthocyanin Assay Buffer应密闭保存，避免有效成分挥发。
4、如果没有分光光度计，也可以使用酶标仪测定。
5、为了您的安全和健康，请穿实验服并戴一次性手套操作。
6、试剂开封后请尽快使用，以防影响后续实验效果。
Anthocyanin, also known as anthocyanin pigment, belongs to the flavonoid class of compounds. It is a water-soluble pigment widely present in plant cell vacuoles, imparting red, purple, blue, and other colors to flowers, fruits, and leaves. Its content in plants is closely related to organ color, developmental stage, and post-harvest senescence, directly affecting the processing properties, storage quality, and nutritional value of agricultural products.
Detection Principle
Anthocyanins are soluble in organic solvents. Crude extraction of anthocyanins is performed using an organic solvent. Anthocyanins appear red in an acidic solution, and the color intensity is proportional to the anthocyanin content. They exhibit a maximum absorption peak at 530 nm. The absorbance can be measured using a spectrophotometer, and the anthocyanin content can be calculated accordingly. This method is simple and easy to perform. Due to the diversity of anthocyanin types in plants (e.g., Cyanidin, Pelargonidin, etc.) and their different molar extinction coefficients, this kit does not provide a single standard. Therefore, results are expressed as relative absorbance or relative content. To determine the absolute content, users are requested to prepare their own pure anthocyanin standard corresponding to the sample. This kit is primarily used for the extraction and relative quantification of anthocyanins in plant tissues or fruits. It is for research use only and is not suitable for clinical diagnosis or other purposes.
Applicable Samples: Plant tissues or fruits
| Product Code | Component | 50T | Storage |
| A1509392 | Anthocyanin Assay Buffer | 500 mL | RT. |
Reagents, consumables and Equipments not provided
Operating Steps (For Reference Only)
1. Anthocyanin Extraction
Method One:
(1) Pre-cool a mortar or homogenizer, as well as the Anthocyanin Assay Buffer, to 4°C.
(2) Take plant tissue, wash and dry it. Weigh 0.25 g of fresh sample, cut into pieces, and place it in the mortar or homogenizer.
(3) Add 2–3 mL of Anthocyanin Assay Buffer. Grind or homogenize thoroughly, then transfer to a 10 mL centrifuge tube. Rinse the mortar/homogenizer with Anthocyanin Assay Buffer and transfer the rinses to the tube. Adjust the final volume to 8 mL with Anthocyanin Assay Buffer.
(4) Incubate at 4°C in the dark for 20 minutes, shaking 2–3 times during incubation. Then filter into a centrifuge tube. Alternatively, centrifuge at 8000 rpm for 3 minutes. The filtrate (supernatant) is the crude anthocyanin extract.
Method Two:
Add the sample to the extraction solution according to the ratio described above. Place in a 32°C incubator for 4–8 hours (shaking several times during incubation) or extract in the dark at room temperature for 24 hours. Centrifuge and collect the supernatant as the crude anthocyanin extract.
2. Sample Addition and Measurement
Using a 1 cm pathlength cuvette, use Anthocyanin Assay Buffer as the blank control. Take the anthocyanin extract (1–2 mL) and read the absorbance at 530 nm using a spectrophotometer. If the anthocyanin concentration is too high, dilute appropriately before measurement. It is recommended to set up duplicate measurements for each sample and take the average.
3. Result Calculation
Definition of Anthocyanin Unit: One unit (U) of anthocyanin is defined as the concentration that yields an OD value of 0.1.
Calculation method:
Anthocyanin content in tissue sample (U/g) = (ATest - ABlank) × N / (0.1 × W)
Parameter Explanation:
ATest: OD value of the anthocyanin extract at 530 nm
ABlank: OD value of the blank control at 530 nm
W: Fresh weight of the sample (g)
N: Dilution factor
Note: A standard curve can also be prepared using a user-provided anthocyanin standard with gradient dilutions (e.g., 0.1, 0.2, 0.3, 0.4, 0.5, 1.0 mg/mL), using Anthocyanin Assay Buffer as the blank control. The anthocyanin content in the sample can then be calculated from the curve.
Anthocyanin in tissue sample (mg/g) = C × VT × N / W
Anthocyanin in liquid sample (mg/mL) = C × N
Parameter Explanation:
C: Anthocyanin concentration of the test sample obtained from the standard curve (mg/mL)
VT: Total volume of the crude anthocyanin extract (mL) = 8
W: Fresh weight of the sample (g)
N: Dilution factor
Precautions
To prevent photodegradation of anthocyanins, perform operations in low light as much as possible, and keep grinding/homogenization time short.
Sample amount and reagent volumes should be adjusted appropriately based on the anthocyanin content.
Anthocyanin Assay Buffer should be stored tightly sealed to prevent evaporation of active components.
If a spectrophotometer is unavailable, a microplate reader can also be used for measurement.
For your safety and health, please wear a lab coat and disposable gloves during operation.
Please use the reagent as soon as possible after opening to avoid affecting subsequent experimental results.
| 产品号 | Component | 50T | Storage |
| A1509392 | Anthocyanin Assay Buffer | 500 mL | RT. |
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| 批号(Lot Number) | 证书类型 | 货号 |
|---|---|---|
 ZJ26F0838312 |
分析证书 | A1509392 |
| ZJ26F0333091 |
分析证书 | A1509392 |
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