Decolorization Protocol
GUIDELINE
- Decolorization refers to the process of removing brightly colored organic impurities from the sample mixture. The procedure is usually carried out in the solution phase after the solid product and impurities are dissolved in a suitable solvent.
- Before beginning of decolorization, it is important to know whether the observed color is really an impurity or just the natural color of the desired product. Perform decolorization only after you have determined the color is due to an impurity.
METHODS
Decolorization Depends on the Solubility of Pigment in Different Solvents
- Water extraction alcohol precipitation. Removing a small part of water-soluble pigment.
- Alcohol lifting water. Removing most fat-soluble pigment.
- Acid-base precipitation method. When the impurity pigment is some flavonoids, anthraquinone and other phenolic acid components, which can precipitate by adjusting the solution below PH3.

Decolorization Depends on the Difference of Adsorb Ability
- Physical adsorption. Polar adsorbent, such as silica gel, alumina, which can remove hydrophilic pigment. Non-polar adsorbents, such as activated carbon, pulp, talc, diatomaceous earth, which can remove lipophilic pigment.
- Chemical adsorption. For example, alkaline alumina can be used to remove some phenolic acidic pigments such as flavonoids and anthraquinones. Ion exchange resin method, phenolic acid pigments such as flavone and anthraquinone can be removed by anion exchange resin.
- Semi-chemical adsorption. Polyamide and macro-porous resin. Adsorption principle for hydrogen bonding, macro-porous resin and part of the van der Waals force. Polyamide can form hydrogen bonds with phenolic and flavonoid hydroxyl groups through the amide carbonyl group in the molecule. The free amine group on the amide bond can also form hydrogen bonds with the carbonyl group on the quinones and fatty carboxylic acids.
Decolorization by Precipitation Method
- Calcium ions in lime milk can combine with some components to form calcium chelates and calcium salt precipitates.
- Under the action of sulfuric acid, the calcium salts formed by flavonoids, anthraquinones, phenols, saponins, some alkaloids and calcium ions can be decomposed and then dissolved into water.
- However, tannin, some proteins, organic acids, polar pigments, polysaccharides and so on cannot be decomposed.
Decolorization by Flocculant Method
- Polyacrylamide, often used in beverage process, sugar process, fermentation process.
- Disodium hydrogen phosphate, trisodium phosphate, often used in beverage process, fermentation process.
- Sulfuric acid, often used in beer process, fermentation process, starch process, dairy processing technology.
- Zinc sulfate is often used in century egg process, beer process, fermentation process.
- Ferrous sulfate, often used in beverage and beer processes.
NOTES
- Don't add activated charcoal to a solution that is near boiling. Violent foaming may result that can dangerously propel the solution out of the flask.
- Different decolorization methods can be used interchangeably.
RELATED PRODUCTS & SERVICES
For research use only. Not for any other purpose.
Resources
- FAQ
- Posters & Downloads
- Protocol
- Cell Culture Guide
- Technical Bulletins
-
Explore & Learn
-
Cell Biology
- Monocytes vs. Macrophages
- How to Detect and Remove Endotoxins in Biologics?
- Comparison of Different Methods to Measure Cell Viability
- CFU Assay for Hematopoietic Cell
- Techniques for Cell Separation
- Biomarkers and Signaling Pathways in Tumor Stem Cells
- Circulating Tumor Cells as Cancer Biomarkers in the Clinic
- What Are Myeloid Cell Markers?
- How to Start Your Culture: Thawing Frozen Cells
- Comparison of the MSCs from Different Sources
- How to Isolate and Analyze Tumor-Infiltrating Leukocytes?
- Contamination of Cell Cultures & Treatment
- Generation and Applications of Neural Stem Cells
- Stem Cell Markers
- Cell Cryopreservation Techniques and Practices
- Guidelines for Cell Banking to Ensure the Safety of Biologics
- What Is Cell Proliferation and How to Analyze It?
- Direct vs. Indirect Cell-Based ELISA
- Critical Quality Attributes and Assays for Induced Pluripotent Stem Cells
- STR Profiling—The ID Card of Cell Line
- Comparison of Several Techniques for the Detection of Apoptotic Cells
- How to Assess the Migratory and Invasive Capacity of Cells?
- Cryopreservation of Cells Step by Step
- What are PBMCs?
- T Cell Activation and Expansion
- Quantification of Cytokines
- What Cell Lines Are Commonly Used in Biopharmaceutical Production?
- Isolation, Expansion, and Analysis of Natural Killer Cells
- Neural Differentiation from Induced Pluripotent Stem Cells
- IL-12 Family Cytokines and Their Immune Functions
- Tumor Stem Cells: Identification, Isolation and Therapeutic Interventions
- Multi-Differentiation of Peripheral Blood Mononuclear Cells
- How to Scale Up Single-Cell Clones?
- What are Mesothelial Cells?
- Major Problems Caused by the Use of Uncharacterized Cell Lines
- T Cell, NK Cell Differentiation from Induced Pluripotent Stem Cells
- Cell Culture Medium
- Human Primary Cells: Definition, Assay, Applications
- What are the Differences Between M1 and M2 Macrophages?
- Mesenchymal Stem Cells: A Comprehensive Exploration
- Enrichment, Isolation and Characterization of Circulating Tumor Cells (CTCs)
- Tips For Cell Cryopreservation
- How to Decide Between 2D and 3D Cell Cultures?
- Organoid Differentiation from Induced Pluripotent Stem Cells
- Unveiling the Molecular Secrets of Adipogenesis in MSCs
- Troubleshooting Cell Culture Contamination: A Comprehensive Guide
- CHO Cell Line Development
- How to Eliminate Mycoplasma From Cell Cultures?
- Strategies for Enrichment of Circulating Tumor Cells (CTCs)
- How to Isolate PBMCs from Whole Blood?
- How to Handle Mycoplasma in Cell Culture?
- How to Interpret ddPCR Results: Common Pitfalls and Troubleshooting
- ddPCR Assay Design for Gene Rearrangement Detection
- Cell Culture Troubleshooting: Molecular Diagnostics & QC for Preclinical R&D
- Cell Immortalization Step by Step
- Adherent and Suspension Cell Culture
- Optimization Strategies of Cell-Based Assays
- Live Cell Imaging: Unveiling the Dynamic World of Cellular Processes
- Overview of Cell Apoptosis Assays
- Cell-Based High-Throughput Screening Techniques
- From Collection to Cure: How ACT Works in Cancer Immunotherapy
- Role of Cell-Based Assays in Drug Discovery and Development
- What are White Blood Cells?
- Types of Cell Therapy for Cancer
- Immunogenicity Testing: ELISA and MSD Assays
- From Blur to Clarity: Solving Resolution Limits in Live Cell Imaging
- ddPCR vs qPCR vs NGS: Which Platform Fits Your Research?
- Key Techniques in Primary, Immortalized and Stable Cell Line Development
- From Primary to Immortalized: Navigating Key Cell Lines in Biomedical Research
- Cell Viability, Proliferation and Cytotoxicity Assays
- Eosinophils vs. Basophils vs. Neutrophils
- What Are CAR T Cells?
- Cultivated Meat: What to Know?
- What Are the Pros and Cons of Adoptive Cell Therapy?
- 3D-Cell Model in Cell-Based Assay
- How to Maximize Efficiency in Cell-Based High-Throughput Screening?
- Understanding Immunogenicity Assays: A Comprehensive Guide
- Exploring Cell Dynamics: Migration, Invasion, Adhesion, Angiogenesis, and EMT Assays
- A Complete Guide to Immortalized Cancer Cell Lines in Cancer Research
- Mastering Cell Culture and Cryopreservation: Key Strategies for Optimal Cell Viability and Stability
- Spheroid vs. Organoid: Choosing the Right 3D Model for Your Research
-
Histology
- Fluorescent Nuclear Staining Dyes
- Stains Used in Histology
- Troubleshooting in Fluorescent Staining
- Immunohistochemistry Controls
- Overview of the FFPE Cell Pellet Product Lines
- How to Apply NGS Technologies to FFPE Tissues?
- Overview of Common Tracking Labels for MSCs
- Comparison of Membrane Stains vs. Cell Surface Stains
- Microscope Platforms
- Cell Lysates: Composition, Properties, and Preparation
- Multiple Animal Tissue Arrays
- Immunohistochemistry Troubleshooting
- Cell and Tissue Fixation
- Tips for Choosing the Right Protease Inhibitor
- Mitochondrial Staining
- Guides for Live Cell Imaging Dyes
- Instructions for Tumour Tissue Collection, Storage and Dissociation
- Common Immunohistochemistry Stains and Their Role in Cancer Diagnosis
- How Immunohistochemistry Makes the Invisible Brain Visible?
- Histological Staining Techniques: From Traditional Chemical Staining to Immunohistochemistry
- How to Choose the Right Antibody for Immunohistochemistry (IHC)
- What You Must Know About Neuroscience IHC?
- Modern Histological Techniques
- From Specimen to Slide: Core Methods in Histological Practice
- Multiplexing Immunohistochemistry
- Comparing IHC, ICC, and IF: Which One Fits Your Research?
- Serum vs. Plasma
- How to Begin with Multiplex Immunohistochemistry (mIHC)
-
Exosome
- Collection of Exosome Samples and Precautions
- Current Research Status of Milk Exosomes
- How do PELN Deliver Drugs?
- Classification, Isolation Techniques and Characterization of Exosomes
- Emerging Technologies and Methodologies for Exosome Research
- How Important are Lipids in Exosome Composition and Biogenesis?
- Common Techniques for Exosome Nucleic Acid Extraction
- Exosome Size Measurement
- What are the Functions of Exosomal Proteins?
- How to Apply Exosomes in Clinical?
- Exosomes as Emerging Biomarker Tools for Diseases
- How to Perform Targeted Modification of Exosomes?
- Techniques for Exosome Quantification
- Production of Exosomes: Human Cell Lines and Cultivation Modes
- How to characterize exosomes?
- Summary of Approaches for Loading Cargo into Exosomes
- Exosome Transfection for Altering Biomolecular Delivery
- Exosome Antibodies
- Exosome Quality Control: How to Do It?
- Applications of MSC-EVs in Immune Regulation and Regeneration
- The Role of Exosomes in Cancer
- How to Enhancement Exosome Production?
- Unraveling Biogenesis and Composition of Exosomes
- What's the Potential of PELN in Disease Treatment?
- How to Label Exosomes?
- How to Efficiently Utilize MSC Exosomes for Disease Treatment?
-
ISH/FISH
- ISH probe labeling method
- Multiple Approaches to Karyotyping
- In Situ Hybridization Probes
- CARD-FISH: Illuminating Microbial Diversity
- Comprehensive Comparison of IHC, CISH, and FISH Techniques
- RNAscope ISH Technology
- Multiple Options for Proving Monoclonality
- FISH Techniques for Biofilm Detection
- Whole Chromosome Painting Probes for FISH
- Overview of Common FISH Techniques
- Guidelines for the Design of FISH Probes
- Small RNA Detection by ISH Methods
- Differences Between DNA and RNA Probes
- Overview of Oligo-FISH Technology
- FISH Tips and Troubleshooting
- How to Use FISH in Hematologic Neoplasms?
- What are the Differences between FISH, aCGH, and NGS?
- Comparative Genomic Hybridization and Its Applications
- Telomere Length Measurement Methods
- Different Types of FISH Probes for Oncology Research
- What Types of Multicolor FISH Probe Sets Are Available?
- What Is the Use of FISH in Solid Tumors?
- Reagents Used in FISH Experiments
- What are Single, Dual, and Multiplex ISH?
- Mapping of Transgenes by FISH
- ImmunoFISH: Integrates FISH and IL for Dual Detection
- 9 ISH Tips You Can't Ignore
-
Toxicokinetics & Pharmacokinetics
- Organoids in Drug Discovery: Revolutionizing Therapeutic Research
- How to Improve the Pharmacokinetic Properties of Peptides?
- What Are Metabolism-Mediated Drug-Drug Interactions?
- How to Improve Drug Plasma Stability?
- How Is the Cytotoxicity of Drugs Determined?
- Toxicokinetics vs. Pharmacokinetics
- Traditional vs. Novel Drug Delivery Methods
- Key Factors Influencing Brain Distribution of Drugs
- Overview of In Vitro Permeability Assays
- The Rise of In Vitro Testing in Drug Development
- Predictive Modeling of Metabolic Drug Toxicity
- Effects of Cytochrome P450 Metabolism on Drug Interactions
- How to Improve Drug Distribution in the Brain
- Key Considerations in Toxicokinetic
- Organ-on-a-Chip Systems for Drug Screening
- How to Design and Synthesize Antibody Drug Conjugates?
- What factors influence drug distribution?
- Parameters of Pharmacokinetics: Absorption, Distribution, Metabolism, Excretion
- What Is the Role of the Blood-Brain Barrier in Drug Delivery?
- Physical and Chemical Properties of Drugs and Calculations
- Experimental Methods for Identifying Drug-Drug Interactions
- How to Conduct a Bioavailability Assessment?
- What are the Pharmacokinetic Properties of the Antisense Oligonucleotides?
- What Are Compartment Models in Pharmacokinetics?
- Methods of Parallel Artificial Membrane Permeability Assays
- Pharmacokinetics Considerations for Antibody Drug Conjugates
- Comparison of MDCK-MDR1 and Caco-2 Cell-Based Permeability Assays
- Unraveling the Role of hERG Channels in Drug Safety
- Pharmacokinetics of Therapeutic Peptides
- How Genotoxicity Testing Guides Safer Drug Development
- Top 5 Pitfalls in In Vitro ADME Assays and How to Avoid Them
- 2D vs 3D Cell Culture Models: Which Is Best for Drug Toxicity Testing?
- The Bioanalysis Masterclass: Labile Metabolites
- What Are Biomarkers in Drug Discovery?
- What Is Genotoxicity in Pharmacology? Mechanisms and Sources
- Preclinical Workflow for Drug Toxicity Testing
- Bioanalysis Errors: How to Spot and Fix Them Early
- Troubleshooting Common Issues in Drug Toxicity Testing
- A Complete Guide to CYP Reaction Phenotyping in 2026
- In Vitro ADME vs In Vivo ADME
- How to Interpret CYP Phenotyping Data
- Mastering the Noise: A Practical Guide to Minimizing Variability in Preclinical Studies
- Reaction Phenotyping vs. Metabolic Stability
- Comparing Plasma Protein Binding Methods
- The 8 Costliest Mistakes in Preclinical CYP Phenotyping
- The Essentials of Quantitative Bioanalysis
- Why iPSC-derived Cells are Useful in Toxicology?
- How Are Biomarkers Validated in Drug Development?
- When Should You Introduce ADME Tox Testing in Drug Development?
- Biomarkers vs. Functional Assays: Closing the Preclinical Gap
- Why Cardiotoxicity Matters in R&D?
- 6 Easy Steps to Get Your In Vitro ADME Done
- From Cells to Systems: Modern Approaches to Disease Modeling
- How Can You Optimize Drug Toxicity Assessment?
- How to Choose the Right In Vitro ADME Assays for Small-Molecule Drugs
- The 8 Types of Drug Toxicity Every Researcher Must Know
- What Are the Best Methods to Test Cardiotoxicity?
-
Disease Models
- Animal Models of Neurodegenerative Diseases
- Disease Models of Diabetes Mellitus
- Summary of Advantages and Limitations of Different Oncology Animal Models
- What Human Disease Models Are Available for Drug Development?
- Overview of Cardiovascular Disease Models in Drug Discovery
- Why Use PDX Models for Cancer Research?
- Preclinical Models of Acute Liver Failure
- Bridging the Translational Gap: A Systematic Framework for Model Selection
- Advanced Organoid Culture: A Practical Optimization Guide
- Are You Choosing the Right Preclinical Model? 10 Key Questions
- How to Match Models to Drug Modalities: Small Molecules vs. Biologics
- How to Select the Right Preclinical Model for Drug Development
- Organoids vs Organ-on-Chip: Which is More Predictive?
- Humanized Mouse Models: Core Technical Considerations
- Animal Models vs. NAMs: Understanding Modern Preclinical Research
- How to Select the Right Humanized Mouse Model for Immuno-Oncology
- Static vs. Dynamic In Vitro Models
- Implementing NAMs in Drug Development
- Oncology Model Strategy: From Screening to Validation
- Organ-on-a-Chip: Is Your Microfluidic Setup Ready for Preclinical Trials?
- Why Oncology Organoids Fail? How to Build Models That Work
-
Cell Biology
- Life Science Articles
- Download Center
- Trending Newsletter

