计算溶液所需的质量、体积或浓度。
维生素C(VC)含量检测试剂盒（磷钼酸, 微量法）
有货
为什么选择此级别
BioReagent,比色法,生化试剂 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
避光,室温。常规运输 。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
维生素 C(Vitamin C)又称 L-抗坏血酸，是高等灵长类动物与其他少数生物的必需营养素，在生物体内维生素 C 是一种抗氧化剂，为酸性己糖衍生物，是烯醇式己糖酸内酯，保护身体免于自由基的威胁，同时也是一种辅酶，其广泛的食物来源为各类新鲜蔬果。Vc 有 L-型和D-型两种异构体，只有 L-型的才具有生理功能，还原型和氧化型都有生理活性。
维生素 C 检测试剂盒(磷钼酸，微量法)检测原理是在强酸和偏磷酸根离子存在条件下，钼酸铵能与还原型维生素 C 反应生成蓝色化合物，在一定浓度范围吸光度值与浓度呈线性关系， 以酶标仪 760nm 处检测吸光度，获得Vitamin C 含量.
产品组分
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使用方法
1. 配制组织匀浆液
按组织匀浆液 A:组织匀浆液 B:蒸馏水=8:1:91 的比例混匀即得。
2. 准备样品
取待测材料如青菜、水果、松针等,清洗擦干，准确称量1g,加入研磨器内，再加入少量组织匀浆液,研磨碎，留取上清，再次用组织匀浆液研磨, 最后一并倒入10 mL离心管，4000 r/min 离心5 min,取上清并补充组织匀浆液至10 mL, 充分混匀，即为待测液。如果样品维生素C浓度较低，可适当提高研磨时组织的质量。
3. 配制 MPA 工作液
称取 0.1 g MPA 粉末，充分溶解于 5 mL 酸性缓冲液 A 中，也可根据使用情况称取一定量的 MPA 按比例溶解即可，4℃保存 3~4 天有效。
4. 配制钼酸铵工作液
取钼酸铵粉末 0.25 g，充分溶解于 10 mL 蒸馏水中，也可根据使用情况称取一定量的钼酸铵加水按比例溶解即可，4℃避光保存。注意：钼酸铵溶于水会逐渐变化成乳白色浊液，则不能使用。
5. 配制系列 Vitamin C 标准
将 5 mg Vitamin C 标准用 1 mL 组织匀浆液溶解即得 Vitamin C 标准(5 mg/mL)，再用组织匀浆液稀释成 Vitamin C 标准(400 μg/mL) ，取干净离心管或试管，按下表进行操作，依次稀释。Vitamin C 标准(5 mg/ml)放置-20℃, 短期保存。
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6. 加样
按照下表设置空白孔、标准孔、测定孔，溶液应按顺序依次加入，并注意避免产生气泡。如果样品中的 Vitamin C 含量过高，可以减少样品用量或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。
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7. Vitamin C 测定
立即混匀，30℃水浴 20 min，以酶标仪测定 760 nm处系列标准孔、测定孔和空白孔的吸光度。
8. 计算
减去空白孔吸光度，以系列标准 Vitamin C 为横坐标，以对应的吸光度为纵坐标，绘制标准曲线，求得回归方程，以测定孔的吸光度代入回归方程求得样品维生素 C 含量。
100 g 样品中维生素 C 含量(mg) =(C×V)×100/(W×1000)×D=(C×V) /(W×10)×D
C:待测样品的吸光度值在标准曲线上查出的维生素C的浓度(μg/mL)
V:待测液的总体积(mL)=10
W:样品质量(g)=1
1000: μg换算成mg
D:待测样品的稀释倍数
注意事项：
1. Vitamin C 标准避免反复冻融，以免失效或效率下降，最好现配现用。
2. 组织匀浆液 A 保存温度过低时易产生结晶，可用温水浴溶解后再使用。
3. 钼酸铵工作液室温保存时间过长或者过期会出现白色粘稠物，不可使用，应换新的产品使用。不使用时建议 4℃保存。
4. 待测样品如不能及时测定，应置于 2~8℃保存，3 天内稳定。
5. 如果样品浓度过高，应用组织匀浆液稀释后重测，结果乘以稀释倍数。
结果展示
典型标准曲线
Vitamin C (VC), also known as L-ascorbic acid, is an essential nutrient for higher primates and a few other organisms. As an endogenous antioxidant, it is an acidic hexose derivative and an enol-form hexonic acid lactone that protects the body from free radical damage. It also functions as a coenzyme, and is widely sourced from various fresh fruits and vegetables. VC exists in two isomeric forms: L-form and D-form. Only the L-form possesses physiological activity, and both its reduced and oxidized states are biologically active.
The Vitamin C Content Assay Kit (Phosphomolybdic Acid, Micro-volume Method) works on the following principle: in the presence of strong acid and metaphosphate ions, ammonium molybdate reacts with reduced vitamin C to form a blue compound. Within a certain concentration range, absorbance shows a linear correlation with vitamin C concentration. The vitamin C content is quantified by measuring absorbance at 760 nm using a microplate reader.
Product Components
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Assay Procedure
1. Preparation of Working Tissue Homogenate
Mix Tissue Homogenate Solution A, Tissue Homogenate Solution B and distilled water in a volume ratio of 8:1:91.
2. Sample Preparation
Take test samples (e.g., green vegetables, fruits, pine needles), rinse and blot dry. Accurately weigh 1 g of sample, transfer to a grinder, add a small amount of tissue homogenate and grind thoroughly. Collect the supernatant, re-grind the residue with additional tissue homogenate, then transfer all contents into a 10 mL centrifuge tube. Centrifuge at 4000 r/min for 5 min. Collect the supernatant, top up to 10 mL with tissue homogenate, and mix thoroughly to obtain the test solution.
If the vitamin C concentration in the sample is low, the sample weight for homogenization may be increased appropriately.
3. Preparation of MPA Working Solution
Weigh 0.1 g of MPA powder and dissolve completely in 5 mL of Acidic Buffer A. Alternatively, weigh an appropriate amount of MPA and dissolve proportionally according to actual usage. The solution is stable for 3–4 days at 4℃.
4. Preparation of Ammonium Molybdate Working Solution
Weigh 0.25 g of ammonium molybdate powder and dissolve completely in 10 mL of distilled water. Alternatively, weigh an appropriate amount and dissolve proportionally according to actual usage. Store at 4℃ protected from light.
Note: If the solution gradually turns milky white, it is invalid and must not be used.
5. Preparation of Vitamin C Standard Series
Dissolve 5 mg of Vitamin C standard in 1 mL of tissue homogenate to prepare a 5 mg/mL Vitamin C standard solution, then dilute with tissue homogenate to a 400 μg/mL working standard. Take clean centrifuge tubes and perform serial dilutions according to the table below. The 5 mg/mL Vitamin C standard solution can be stored at -20℃ for short-term preservation
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6. Sample Loading
Set up blank wells, standard wells and sample wells according to the table below. Add reagents sequentially in the indicated order and avoid generating bubbles. If the vitamin C content in the sample is too high, reduce the sample volume or dilute appropriately before assay. Duplicate wells are recommended for each sample, and the average value shall be used for calculation.
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7. Detection
Mix immediately, incubate in a water‑bath at 30 °C for 20 min, then measure the absorbance of serial standard wells, sample wells and blank wells at 760 nm using a microplate reader.
8. Calculation
Subtract the absorbance of blank wells. Take serial concentrations of standard Vitamin C as the abscissa (X‑axis) and the corresponding absorbance as the ordinate (Y‑axis) to plot the standard curve and obtain the regression equation. Substitute the absorbance of sample wells into the regression equation to calculate the Vitamin C content of samples.
Vitamin C content per 100 g sample (mg)= (C × V) × 100 / (W × 1000) × D = (C × V) / (W × 10) × D
- C: Vitamin C concentration of the test solution read from the standard curve (μg/mL)
- V: Total volume of the test solution (mL) = 10
- W: Sample weight (g) = 1
- 1000: Conversion factor from μg to mg
- D: Dilution factor of the test sample
Notes
1. Avoid repeated freeze-thaw cycles of the Vitamin C standard to prevent inactivation or activity loss. Fresh preparation before use is recommended.
2. Crystallization may occur in Tissue Homogenate Solution A if stored at excessively low temperature. Dissolve the crystals in a warm water bath before use.
3. White viscous precipitates may form in the ammonium molybdate working solution if stored at room temperature for too long or past expiration. Do not use such solution and replace with fresh reagent. Storage at 4℃ is recommended when not in use.
4. If the test sample cannot be assayed immediately, store at 2–8℃; it remains stable for 3 days.
5. If the sample concentration is too high, dilute the sample with tissue homogenate and retest. Multiply the final result by the dilution factor.
Results Presentation
Typical standard curve:
规格
规格或纯度
BioReagent, 比色法, 生化试剂
英文名称
Vitamin C (VC) Content Assay Kit (PMA, Micro Method)
应用
细胞代谢
样品类型
植物组织
检测范围
10-400 μg/mL
检测方法
比色法
检测仪器
酶标仪
检测波长
A760 nm
敏感性
Light-sensitive
检测时长
1 h
样本体积
40 μL
储存与运输
储存条件
避光,室温
运输条件
常规运输
稳定性与储存
各组分在相应的储存条件下保质期1年。
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:
技术文档和文章
溶液计算器
常见问题
本产品如何运输？
本产品按常温标准条件运输，无需温控包装。
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「BioReagent」表示该产品经过加工和检测，适用于生物和生化应用。规格可能包括内毒素限值、微生物控制、无菌性、低 DNase/RNase/蛋白酶活性，以及在相关情况下的生物活性验证。
本产品应如何储存？
请于室温、避光条件下保存。本品对光敏感，请保存在原始的避光或棕色容器中。
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每个订单都会随附分析证书（COA），您也可以从账户订单历史中下载。如需订购前的COA，请联系您的客户代表，或使用PDP上的"申请COA"按钮——我们将提供代表性批次的COA。
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