TREHALOSE

Trehalose / α-D-Glucopyranosyl-α-D-glucopyranoside / Mycose / Tremalose
CAS Number: 99-20-7 (Anhydrous); 6138-23-4 (Dihydrate)
E Number: E963

1. PRODUCT IDENTIFICATION AND CHEMICAL IDENTITY

Parameter	Detail
Chemical Name (IUPAC)	α-D-Glucopyranosyl-α-D-glucopyranoside
Common Name	Trehalose
Other Names	Mycose, Tremalose
CAS Number (Anhydrous)	99-20-7
CAS Number (Dihydrate)	6138-23-4
E Number	E963
Molecular Formula (Anhydrous)	C₁₂H₂₂O₁₁
Molecular Formula (Dihydrate)	C₁₂H₂₂O₁₁·2H₂O
Molecular Weight (Anhydrous)	342.30 g/mol
Molecular Weight (Dihydrate)	378.33 g/mol
Appearance	White crystalline powder
Taste	Mildly sweet (approximately 45% of sucrose sweetness)

Trehalose is a non-reducing disaccharide formed by the α,α-1,1-glycosidic linkage of two glucose molecules. It is naturally found in many plants, fungi, yeast, and invertebrates. It is known for its exceptional cell protection and stabilization properties, particularly its ability to protect biological structures against desiccation, freezing, and osmotic stress (anhydrobiosis). Due to these unique properties, it has wide applications in food, cosmetics, pharmaceuticals, and biotechnology.

2. SYNONYMS AND OTHER NAMES

Type	Name
Common Names	Trehalose, Mycose, Tremalose
Chemical Names	α-D-Glucopyranosyl-α-D-glucopyranoside, α,α-Trehalose
Abbreviations	Tre
E Number	E963
CAS Number (Anhydrous)	99-20-7
CAS Number (Dihydrate)	6138-23-4
HS Code	2940.00

3. CHEMICAL STRUCTURE

3.1. Molecular Structure

Trehalose is a disaccharide composed of two α-D-glucose units linked by an α,α-1,1-glycosidic bond. This unique linkage involves the anomeric carbons of both glucose molecules, making the molecule non-reducing.

Structural Formula:

        CH₂OH                CH₂OH
         |                     |
         O                     O
        / \                   / \
       /   \                 /   \
      /     \               /     \
     H       H             H       H
     |       |             |       |
     C       C             C       C
    / \     / \           / \     / \
   /   \   /   \         /   \   /   \
  H     OH OH    H       H     OH OH    H
   \   /   \   /         \   /   \   /
    \ /     \ /           \ /     \ /
     C - O - C             C - O - C
     |       |             |       |
     H       OH            H       OH

Simplified Representation (α,α-1,1-Glycosidic Bond):

    Glc(α) — O — Glc(α)

3.2. Molecular Formula (Anhydrous)

C₁₂H₂₂O₁₁

3.3. Molecular Weight (Anhydrous)

342.30 g/mol

3.4. Key Structural Features

Feature	Description
Sugar Units	Two α-D-glucose units
Glycosidic Linkage	α,α-1,1-glycosidic bond (anomeric carbons of both glucose units)
Reducing Property	Non-reducing (no free anomeric carbon)
Configuration	α,α
Hydrate Forms	Anhydrous and dihydrate forms available

4. PHYSICAL AND CHEMICAL PROPERTIES

Property	Value / Description
Physical Form	White crystalline powder
Melting Point	~97°C (dihydrate); ~203°C (anhydrous)
Boiling Point	Decomposes before boiling
Water Solubility (20°C)	~69 g/100 mL (high solubility)
Solubility in Ethanol	Practically insoluble
Sweetness Level	Approximately 45% of sucrose
pH (10% aqueous solution)	5.0 – 6.7
Optical Rotation ([α]D²⁰)	+178° (c=1, water)
Stability	Highly stable to heat and acidic pH; non-reducing structure prevents Maillard reaction (browning)
Hygroscopicity	Low; dihydrate form is not hygroscopic
Density	~1.5 g/cm³

5. SOLUBILITY PROFILE

Solvent	Solubility	Conditions
Water	~69 g/100 mL	20°C
Water	Highly soluble	50°C
Ethanol	Practically insoluble	25°C
Methanol	Slightly soluble	25°C
Acetone	Insoluble	25°C
Ether	Insoluble	25°C
Chloroform	Insoluble	25°C

6. CHEMICAL PROPERTIES AND REACTIVITY

Property	Description
Chemical Character	Neutral (non-reducing sugar)
Glycosidic Bond	α,α-1,1 linkage; resistant to hydrolysis by α-glucosidases (trehalase cleaves it)
Hydrolysis	Hydrolyzed by trehalase enzyme to two glucose molecules
Acid Hydrolysis	Hydrolyzed by strong acids to glucose
Maillard Reaction	Does not undergo Maillard reaction (non-reducing)
Stability	Stable in acidic and neutral pH; high thermal stability
Incompatible Materials	Strong acids, strong oxidizing agents

7. REACTION EQUATIONS

7.1. Enzymatic Hydrolysis (Trehalase)

Trehalose + H₂O → 2 Glucose

7.2. Acid Hydrolysis

Trehalose + H₂O (H⁺) → 2 Glucose

7.3. Dehydration (Thermal Decomposition)

Trehalose → Decomposition Products (at high temperature)

8. FUNCTIONAL PROPERTIES

Property	Description
Bioprotection (Anhydrobiosis)	Protects biological membranes, proteins, and cells against desiccation, freezing, and high osmotic stress by replacing water molecules (water replacement hypothesis) and forming a vitreous (glassy) matrix inside cells.
Protein and Enzyme Stabilization	Protects proteins from thermal denaturation and aggregation; critical for vaccines, antibodies, and other biopharmaceuticals in liquid or lyophilized formulations.
Humectant	Increases skin moisture retention capacity in cosmetic formulations; helps reduce transepidermal water loss (TEWL).
Low Sweetness and Low Glycemic Index	Provides lower sweetness and glycemic index compared to sucrose; suitable for reduced-calorie and diabetic-friendly products.
Maillard Reaction Resistance	Non-reducing structure prevents browning reactions (Maillard) with amino acids during heat treatment; maintains color and flavor stability.
Odor and Taste Masking	Capable of masking undesirable bitter tastes and odors in some food and pharmaceutical formulations.

9. BIOLOGICAL MECHANISMS OF ACTION

Effect	Mechanism
Water Replacement	During dehydration, trehalose forms hydrogen bonds with polar head groups of membrane phospholipids and with proteins, replacing water molecules and maintaining native structure.
Vitrification (Glass Formation)	Trehalose forms a highly viscous, glassy (vitreous) matrix around biological structures, slowing down molecular motion and preventing crystallization and damage during freezing or drying.
Autophagy Induction	Trehalose induces autophagy, a cellular process that removes damaged components (misfolded proteins, damaged mitochondria), through activation of the mTOR-independent pathway.
Membrane Stabilization	Prevents membrane phase transitions and leakage by maintaining the liquid crystalline state of membranes under stress conditions.
Oxidative Stress Reduction	Reduces oxidative stress and protects cells from free radical damage.

10. SOURCES

Source	Description
Yeast	Saccharomyces cerevisiae and other yeasts produce trehalose as a stress protectant.
Fungi	Mushrooms and other fungi contain significant amounts of trehalose.
Plants	Selaginella (resurrection plant), wheat, corn, and other plants.
Invertebrates	Artemia (brine shrimp), tardigrades, and nematodes produce trehalose for anhydrobiosis.
Commercial Production	Enzymatic production from starch or fermentation with microorganisms.

11. PRODUCTION METHOD

Trehalose is commercially produced by enzymatic synthesis from starch or by fermentation.

11.1. Enzymatic Synthesis from Starch

Stage	Process	Description
1	Starch Liquefaction	Starch is liquefied with α-amylase to produce maltodextrins.
2	Saccharification	Maltodextrins are treated with amyloglucosidase to produce glucose.
3	Enzymatic Conversion	Glucose is converted to trehalose using trehalose synthase (from microorganisms).
4	Purification	Trehalose is purified by crystallization, filtration, and drying.
5	Packaging	The purified product is packaged as anhydrous or dihydrate form.

11.2. Fermentation

Stage	Process	Description
1	Fermentation	Microorganisms (e.g., Corynebacterium, Brevibacterium) are cultured in fermentation tanks with sugar and nutrient media.
2	Biosynthesis	Trehalose is biosynthesized by the microorganisms.
3	Separation	Trehalose is separated from the fermentation broth.
4	Purification	The product is purified to high purity.
5	Crystallization	Trehalose is crystallized in anhydrous or dihydrate form.
6	Drying	The purified product is dried to obtain final trehalose powder.

12. APPLICATIONS AND INDUSTRIAL USES

12.1. Food Industry (Most Important Application)

Application	Description	Typical Usage
Bakery Products	Humectant, texture improver, sweetness enhancer	0.5 – 10%
Confectionery	Prevents crystallization, improves mouthfeel	1 – 10%
Beverages	Sweetener, flavor stabilizer	0.5 – 5%
Frozen Foods	Cryoprotectant; prevents ice crystal formation	1 – 10%
Dried Foods	Prevents moisture loss and maintains texture	1 – 10%
Dairy Products	Texture improver, sweetener	1 – 5%

12.2. Pharmaceutical and Biotechnology

Application	Description	Typical Usage
Protein Stabilization	Stabilizes proteins, antibodies, and enzymes in liquid and lyophilized formulations	5 – 20% (lyophilized)
Vaccine Stabilization	Protects vaccines from thermal degradation	5 – 15%
Cell/Tissue Preservation	Cryoprotectant and lyoprotectant for cells, tissues, and organs	mM range in cell culture media
Drug Delivery Systems	Used in nanoparticle formulations and drug delivery systems	Variable
Excipient	Bulking agent, cryoprotectant, and stabilizer	Variable

12.3. Cosmetics and Personal Care

Application	Description	Typical Usage
Skin Care	Humectant, moisturizer, skin barrier support	1 – 5%
Anti-aging Products	Protects skin cells from oxidative stress	1 – 5%
Hair Care	Moisturizes hair, improves texture	0.5 – 2%
Sun Care	Protects skin cells from UV-induced stress	1 – 3%

12.4. Medical Research

Application	Description
Neuroprotection Research	Studied for potential neuroprotective effects in Alzheimer's, Parkinson's, and Huntington's disease models.
Ophthalmic Solutions	Used in dry eye treatments at 3-5% concentration.
Autophagy Induction	Research tool for studying autophagy and cellular clearance mechanisms.

13. TREHALOSE ANHYDROUS VS. DIHYDRATE

Property	Trehalose Anhydrous	Trehalose Dihydrate
CAS Number	99-20-7	6138-23-4
Molecular Formula	C₁₂H₂₂O₁₁	C₁₂H₂₂O₁₁·2H₂O
Molecular Weight	342.30 g/mol	378.33 g/mol
Water of Hydration	None	2 molecules
Melting Point	~203°C	~97°C
Hygroscopicity	Slightly hygroscopic	Not hygroscopic
Stability	More stable under dry conditions	Stable; does not absorb moisture
Solubility	Similar	Similar
Applications	Formulations requiring no additional water	General food and cosmetic applications

14. ALTERNATIVES AND COMPARISON

Alternative	Comparison
Sucrose	Higher sweetness and glycemic index; undergoes Maillard reaction; bioprotective properties are not as strong as trehalose.
Maltose	Reducing sugar; undergoes Maillard reaction; not as effective a bioprotectant as trehalose.
Mannitol	Lower solubility; has osmotic diuretic effect; different cell protection mechanism.
Sorbitol	High hygroscopicity; used as humectant but bioprotective properties are less pronounced than trehalose.
Glycerol	Common cryoprotectant; penetrates cells; does not form vitreous matrix like trehalose.
Summary	Trehalose's key advantages are its non-reducing structure (no Maillard reaction) and superior bioprotective properties (water replacement and vitrification).

15. HEALTH BENEFITS AND MECHANISMS

Benefit	Description
Cell Protection	Protects cells from damage caused by dehydration, freezing, and osmotic stress by maintaining membrane integrity and protein conformation.
Autophagy Induction	Induces autophagy (cellular self-cleaning) to remove damaged proteins and mitochondria; being researched for neurodegenerative diseases.
Antioxidant Properties	Reduces oxidative stress and protects cells from free radical damage.
Neuroprotective Effects	In animal models, trehalose promotes clearance of toxic protein aggregates in Alzheimer's, Parkinson's, and Huntington's disease.
Bone Health	Some studies suggest beneficial effects on bone metabolism.
Gastrointestinal Health	Can be hydrolyzed by trehalase enzyme; promotes healthy digestion.

16. CHEMICAL COMPATIBILITY TABLE

Substance / Environment	Compatibility	Description
Water	COMPATIBLE	Highly soluble; stable in aqueous solution.
Acids (HCl, H₂SO₄)	COMPATIBLE	Stable in acidic pH; hydrolyzes under strong acidic conditions.
Alkalis (NaOH, KOH)	COMPATIBLE	Stable.
Oxidizers	INCOMPATIBLE	May cause degradation.
Reducing Agents	COMPATIBLE	Generally compatible.
Proteins	COMPATIBLE	Stabilizes proteins; prevents aggregation.
Amino Acids	COMPATIBLE	Does not react (Maillard-resistant).
Air / Moisture	COMPATIBLE	Dihydrate form is not hygroscopic; anhydrous form may absorb moisture.
Light	COMPATIBLE	Stable; protect from UV for long-term storage.

17. TOXICOLOGICAL PROFILE

Parameter	Value
Acute Oral Toxicity (LD50, Rat)	> 5000 mg/kg (very low toxicity)
Acute Dermal Toxicity	Low
Acute Inhalation Toxicity	Low risk; may cause mild respiratory irritation.
Skin Irritation	Mild irritant in powder form.
Eye Irritation	May cause mild eye irritation.
Skin Sensitization	Not classified as a sensitizer.
Mutagenicity	Negative.
Carcinogenicity	Not classified as a carcinogen.
Reproductive Toxicity	Not classified as a reproductive toxin.
Target Organs	No specific target organs identified.
High Dose Effects	High doses (>50 g/day) may cause gastrointestinal discomfort (osmotic diarrhea).
Trehalose Intolerance	Extremely rare condition due to trehalase enzyme deficiency.

18. GHS CLASSIFICATION AND LABELING

GHS Classification (Generally Not Classified as Hazardous):

Hazard Class	Category	H-Statement
Not classified as hazardous	-	-

Signal Word: None (not classified as hazardous)

Hazard Pictograms: None

Hazard Statements (H-Codes): None

Safety Phrases (Historically):

Code	Statement
S24/25	Avoid contact with skin and eyes.
S26	In case of contact with eyes, rinse immediately with plenty of water and seek medical advice.

19. PRECAUTIONARY STATEMENTS (P-CODES)

Code	Statement
P264	Wash hands thoroughly after handling.
P280	Wear protective gloves/protective clothing/eye protection/face protection.
P305+P351+P338	IF IN EYES: Rinse cautiously with water for several minutes. Remove contact lenses if present.
P337+P313	If eye irritation persists: Get medical advice/attention.
P501	Dispose of contents/container in accordance with local regulations.

20. FIRST AID MEASURES

Route of Exposure	Action to Be Taken
Inhalation	Move to fresh air. If respiratory irritation persists, seek medical attention.
Skin Contact	Wash with plenty of soap and water. Remove contaminated clothing. If irritation persists, consult a physician.
Eye Contact	Rinse thoroughly with plenty of water for at least 15 minutes. Remove contact lenses. If irritation persists, seek medical attention.
Ingestion	Rinse mouth with water. Drink plenty of water. Do not induce vomiting. If large amount is swallowed, seek medical attention.
General Note	If symptoms persist, always consult a healthcare facility.

21. FIRE-FIGHTING MEASURES

Parameter	Information
Fire Hazard	Not flammable; may decompose at high temperatures.
Hazards During Fire	Thermal decomposition produces carbon monoxide and carbon dioxide.
Suitable Extinguishing Media	Water spray, CO₂, dry chemical powder, alcohol-resistant foam.
Special Protective Equipment	Self-contained breathing apparatus (SCBA), full protective clothing.
Special Precautions	Prevent fire-fighting water from entering the environment.

22. ACCIDENTAL RELEASE MEASURES

Parameter	Information
Personal Protection	Dust mask, protective goggles, gloves, coverall.
Ventilation	Increase ventilation; use local exhaust in confined spaces.
Containment	Cover spilled area to prevent dust dispersion.
Cleaning Methods	Collect dry using vacuum or careful sweeping. Avoid washing with water (product is soluble and may spread).
Environmental Precautions	Prevent entry into drains, sewers, and water bodies.
Waste Disposal	Dispose of in accordance with local regulations.

23. STORAGE AND SHELF LIFE

Parameter	Information
Storage Conditions	Store in a cool (15-25°C), dry, and well-ventilated area, in tightly closed original packaging.
Container Requirements	Tightly closed, PE-lined bags or drums.
Materials to Avoid	Strong acids, strong oxidizers, moisture (for anhydrous form).
Shelf Life	24 months (under proper storage conditions).
Stability Note	Dihydrate form is not hygroscopic; anhydrous form may absorb moisture.
Packaging Options	1 kg, 5 kg, 10 kg, 25 kg PE-lined bags, fiber drums.

24. PACKAGING OPTIONS

Packaging Type	Quantity	Material
PE-lined kraft bag	1 – 5 kg	Paper / Polyethylene
PE-lined fiber drum	10 – 25 kg	Cardboard / Plastic
Plastic drum	25 kg	HDPE
Big-Bag	500 – 1000 kg	Polypropylene

25. TRANSPORT INFORMATION

Parameter	Information
UN Number	Not applicable (not classified as dangerous goods).
Hazard Class	Not classified as hazardous for transport.
Packing Group	Not applicable.
ADR/RID	Not classified as dangerous goods.
IMDG Code	Not classified as dangerous goods.
IATA (Air)	Not classified as dangerous goods.
Marine Pollutant	No.
Transport Temperature	Ambient temperature; protect from moisture.

26. REGULATORY STATUS

Region / Authority	Status
European Union	Approved as food additive E963.
USA (FDA)	GRAS (Generally Recognized as Safe).
Turkey	Approved as food additive; regulated by Turkish Food Codex.
Japan	Approved as food ingredient.
China	Approved as food additive.
Australia/New Zealand	Approved as food additive.
Kosher/Halal	Can be certified upon request.

27. SECTORAL SUITABILITY TABLE

Sector	Suitability	Description
Food Industry	HIGH	Humectant, texturizer, sweetener, stabilizer in bakery, confectionery, beverages, frozen foods.
Pharmaceuticals	HIGH	Protein and vaccine stabilizer, excipient, cryoprotectant.
Cosmetics	HIGH	Humectant, moisturizer, anti-aging, skin barrier support.
Biotechnology	HIGH	Cell culture, enzyme stabilization, research tools.
Medical Research	MEDIUM	Neuroprotection, autophagy induction research.

28. QUICK REFERENCE TABLE

Property	Value
CAS Number (Anhydrous)	99-20-7
CAS Number (Dihydrate)	6138-23-4
Molecular Formula (Anhydrous)	C₁₂H₂₂O₁₁
Molecular Weight (Anhydrous)	342.30 g/mol
Molecular Weight (Dihydrate)	378.33 g/mol
E Number	E963
Appearance	White crystalline powder
Taste	Mildly sweet (~45% of sucrose)
Melting Point (Dihydrate)	~97°C
Melting Point (Anhydrous)	~203°C
Water Solubility (20°C)	~69 g/100 mL
pH (10% solution)	5.0 – 6.7
Optical Rotation	+178°
Sweetness	~45% of sucrose
Main Uses	Food additive, protein stabilizer, humectant, excipient
Shelf Life	24 months
UN Number	Not applicable

29. CRITICAL NOTICES AND BEST PRACTICES

CRITICAL NOTICES:

Non-Reducing Structure: Trehalose does not undergo Maillard reaction (browning) due to its non-reducing nature. This is a key advantage in formulations where color and flavor stability are important during heat treatment.
Exceptional Bioprotective Properties: Trehalose is unique among disaccharides for its ability to protect cells, proteins, and membranes from desiccation, freezing, and osmotic stress. This property is utilized in food (freeze-dried products), pharmaceuticals (vaccine stabilization), and cosmetics (cell protection).
Form Selection: Anhydrous and dihydrate forms are available. Dihydrate is not hygroscopic and is preferred for general applications; anhydrous is used in moisture-sensitive formulations.
Trehalase Enzyme: Trehalose is hydrolyzed by trehalase enzyme in the small intestine. Trehalase deficiency is a rare condition that may cause intolerance to trehalose.
Gastrointestinal Effects: High doses (>50 g/day) may cause osmotic diarrhea in sensitive individuals. For most applications, typical consumption levels are well below this threshold.
Low Glycemic Index: Trehalose has a lower glycemic index than sucrose and glucose, making it suitable for diabetic-friendly products.
Regulatory Status: Approved as a food additive in many countries; always check local regulations before use in food applications.

BEST PRACTICE RECOMMENDATIONS:

Storage: Store in a cool (15-25°C), dry, and well-ventilated area. Keep containers tightly closed. Protect from moisture (for anhydrous form).
Personnel Protection: Use protective gloves, goggles, and dust mask. Ensure adequate ventilation.
Dosage Control: In food applications, recommended usage is typically 0.5-10%. For pharmaceutical formulations, usage varies based on specific application.
Formulation: For lyophilized formulations, use 5-20% trehalose as a stabilizer. For cell culture media, use mM concentrations.
Quality Assurance: Verify product purity and source (anhydrous vs. dihydrate). Test for heavy metals and microbial contamination.
Waste Management: Dispose of waste in accordance with local regulations.

LEGAL DISCLAIMER:

The information provided in this document is based on our current knowledge and experience and is presented in good faith; however, as all conditions of use are beyond our control, it does not constitute a binding specification or warranty. This Technical Data Sheet is for informational purposes only. Users are obligated to test the suitability of the product for their own applications. For complete safety, storage, handling, transport, waste, and regulatory compliance information, the official Safety Data Sheet (SDS/MSDS) provided by the manufacturer/supplier must be consulted.

Trehalose, Mycose, Tremalose, E963, 99-20-7