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Amplite® Colorimetric Phenolic Compounds Assay Kit

Product key features

  • High Sensitivity: Detects phenolic content with a limit of 0.02 mM catechin equivalents (CEs)
  • Enhanced Specificity: Minimizes interference from non-phenolic reducing agents, ensuring accurate measurements
  • Broad Applicability: Compatible with complex matrices such as plant extracts, foods, and beverages
  • High-Throughput Ready: Optimized for large-scale and automated workflows

Product description

The Amplite® Colorimetric Phenolic Compounds Assay Kit is a highly sensitive and specific assay for quantifying total phenolic content in diverse biological samples. The assay is based on the reaction between phenolic compounds and diazonium salts under alkaline conditions, forming a stable diazo chromophore that can be quantified by absorbance at 480 nm. Unlike the Folin-Ciocalteu method, this assay minimizes interference from non-phenolic reducing agents such as sulfites, reducing sugars, and ascorbic acid, ensuring greater specificity for phenolic compounds. With a detection limit of 0.02 mM catechin equivalents (CEs), the assay is compatible with high-throughput workflows and is validated for applications involving complex matrices such as fruits, vegetables, beverages (e.g., tea, wine, coffee), processed food products, plant extracts, and herbal or nutraceutical formulations.

Phenolic compounds are a diverse group of phytochemicals widely distributed in dietary and medicinal plants, including fruits, vegetables, cereals, and beverages such as tea, coffee, and wine. These compounds, encompassing phenolic acids, flavonoids, stilbenoids, lignans, and polyphenols, play essential roles in plants, such as UV protection, herbivore deterrence, and developmental signaling. In humans, their potent antioxidant properties have been linked to reduced risks of cardiovascular diseases, cancer, and neurodegenerative disorders. Additionally, their antimicrobial, anti-inflammatory, and immunomodulatory effects underscore their significance in both biomedical research and dietary science.

Example protocol

AT A GLANCE

Protocol Summary
  1. Prepare and add standards and samples (100 µL).

  2. Add 20 µL Phenolic Compound Probe to the wells of standards and samples.

  3. Add 80 µL Phenolic Compound Assay Buffer to the wells of standards and samples.

  4. Incubate the reaction at room temperature for 5 to 30 minutes.

  5. Monitor the absorbance with an absorbance plate reader at 480 nm.

Note: Bring kit components to room temperature before starting the experiment.

PREPARATION OF STANDARD SOLUTIONS

For convenience, use the Serial Dilution Planner:
https://www.aatbio.com/tools/serial-dilution/11332

Preparation of Standard Curve
Add 10 µL of 10 mM Catechin Standard solution (Component C) to 990 µL H2O to prepare 100 µM Catechin solution (STD7). Then take 300 µL of the STD7 solution and perform 1:2 serial dilutions in H2O to prepare the remaining serially diluted Catechin Standards (STD6-STD1), resulting in concentration ranges from 100 µM to 1.55 µM.

SAMPLE EXPERIMENTAL PROTOCOL

STD1
STD1
TS
TS
STD2
STD2
STD3
STD3
STD4
STD4
STD5
STD5
STD6
STD6
STD7
STD7
BL
BL

Table: 1 Layout of Catechin standards and test samples in a clear bottom 96- wells microplate. Catechin standards (STD7-STD1: 100 µM to 1.55 µM), TS= Test Samples, BL = 0 µM Catechin.

The following protocol can be used as a guideline and should be optimized accordingly.

  1. Prepare the standards and test samples according to the recommended protocol, and add 100 µL of each into the wells of a microplate.

  2. Add 20 µL Phenolic Compound Probe (Component A) to each well containing standards and samples.

  3. Add 80 µL Phenolic Compound Assay Buffer (Component B) to the wells of standards and samples.

  4. Incubate the reaction at room temperature for 5 to 30 minutes.

  5. Monitor the absorbance with an absorbance plate reader at 480 nm.

References

View all 50 references: Citation Explorer
Antioxidant Features of Humic Products by ABTS Assay.
Authors: Verrillo, Mariavittoria and Cozzolino, Vincenza and Spaccini, Riccardo
Journal: Methods in molecular biology (Clifton, N.J.) (2025): 223-227
A sensitive electrochemical biosensor based on Pd@PdPtCo mesoporous nanopolyhedras as signal amplifiers for assay of cardiac troponin I.
Authors: Wang, Miao and Sun, He-Nan and Liu, Xing-Yu and Liu, Mingjun and Li, Shan-Shan
Journal: Bioelectrochemistry (Amsterdam, Netherlands) (2025): 108838
A novel, portable, and cost-effective turbidimetric sensor for sensitive alkaline phosphatase activity assay.
Authors: Wang, Jikai and Xie, Zhulan and Xie, Haitao and Mo, Ziyi and Wang, Weiguo and Zhu, Yanli
Journal: Biosensors & bioelectronics (2025): 116857
Practical NIR Assay Derived from Cyanine to Evaluate Intracellular H2S in Living Cell Imaging.
Authors: Ye, Chenqian and Wang, Axue and Lu, Yuxin and Lin, Xinye and Huang, Luqiang and Li, Daliang
Journal: Sensors (Basel, Switzerland) (2024)
Headspace-SERS assay for early mildewing tobacco leaves.
Authors: Cao, Jiaying and Wang, Zhiguo and Jiang, Yuning and Zhou, Huimin and Liang, Qiuju and Guo, Xiaoyu and Wen, Ying and Yang, Haifeng
Journal: Talanta (2024): 126681
Page updated on June 9, 2025

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Catalog Number11332
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Storage, safety and handling

H-phraseH303, H313, H333
Hazard symbolXN
Intended useResearch Use Only (RUO)
R-phraseR20, R21, R22

Platform

Absorbance microplate reader

Absorbance480 nm
Recommended plateWhite plate, Clear bottom

Components

Catechin (µM) dose response was measured with Amplite® Colorimetric Phenolic Compounds (PC) Assay Kit in a 96-well white plate using a SpectraMax microplate reader (Molecular Devices). The signal was acquired at 480 nm.
Catechin (µM) dose response was measured with Amplite® Colorimetric Phenolic Compounds (PC) Assay Kit in a 96-well white plate using a SpectraMax microplate reader (Molecular Devices). The signal was acquired at 480 nm.
Catechin (µM) dose response was measured with Amplite® Colorimetric Phenolic Compounds (PC) Assay Kit in a 96-well white plate using a SpectraMax microplate reader (Molecular Devices). The signal was acquired at 480 nm.