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HEPES

CAS No.
7365-45-9
Chemical Name:
HEPES
Synonyms
HEPES BUFFER;4-(2-HYDROXYETHYL)-1-PIPERAZINEETHANESULFONIC ACID;HYDROXYETHYLPIPERAZINE ETHANE SULFONIC ACID;4-(2-HYDROXYETHYL)PIPERAZINE-1-ETHANESULFONIC ACID;hepes solution;Biological Buffer;HEPES buffer Solution;EPES;4-(2-hydroxyethyl)piperazin-1-ylethanesulphonic acid;2-[4-(2-HYDROXYETHYL)-1-PIPERAZINYL]ETHANESULFONIC ACID
CBNumber:
CB5408557
Molecular Formula:
C8H18N2O4S
Molecular Weight:
238.3
MDL Number:
MFCD00006158
MOL File:
7365-45-9.mol
MSDS File:
SDS
TDS File:
TDS
Last updated:2026-08-10 16:40:05
Product description Number Pack Size Price
HEPES BioPerformance Certified, ≥99.5% (titration), suitable for cell culture H4034 25KG $6780
HEPES ≥99.5% (titration) H3375 25KG $6460
HEPES ≥99.5% (titration) H3375 50KG $12200
HEPES reagent grade, ≥99% (titration), crystalline H23830 100 g $72.6
HEPES reagent grade, ≥99% (titration), crystalline H23830 500 g $297
More product size

HEPES Properties

Melting point 234-238 °C
Boiling point 408℃[at 101 325 Pa]
bulk density 560kg/m3
Density 1.07 g/mL at 20 °C
vapor pressure 0Pa at 25℃
refractive index n20/D 1.339
storage temp. 2-8°C
solubility H2O: 1 M at 20 °C, clear, colorless
form powder
color White
PH 5.0-6.5 (1Maqueous solution)
pka 7.5(at 25℃)
PH Range 7.0 - 7.6
biological source synthetic
Water Solubility Soluble
λmax λ: 260 nm Amax: 0.06
λ: 280 nm Amax: 0.04
Merck 14,4654
BRN 883043
Stability Stable. Combustible. Incompatible with strong oxidizing agents. Protect from moisture.
Major Application diagnostic assay manufacturing
Cosmetics Ingredients Functions BUFFERING
InChI 1S/C8H18N2O4S/c11-7-5-9-1-3-10(4-2-9)6-8-15(12,13)14/h11H,1-8H2,(H,12,13,14)
InChIKey JKMHFZQWWAIEOD-UHFFFAOYSA-N
SMILES OCCN1CCN(CC1)CCS(O)(=O)=O
LogP -3.85 at 20℃
CAS DataBase Reference 7365-45-9(CAS DataBase Reference)
EWG's Food Scores 1
FDA UNII RWW266YE9I
EPA Substance Registry System 1-Piperazineethanesulfonic acid, 4-(2-hydroxyethyl)- (7365-45-9)
Absorption ≤0.05 at 290 in H2O at 33%
UNSPSC Code 41116107
NACRES NA.75

SAFETY

Risk and Safety Statements

Symbol(GHS)  Exclamation Mark (GHS07)
GHS07
Signal word  Warning
Hazard statements  H302-H315-H319-H335
Precautionary statements  P261-P271-P280
PPE Eyeshields, Gloves, type N95 (US)
Hazard Codes  Xi
Risk Statements  36/37/38
Safety Statements  24/25-22-36-26
WGK Germany  1
RTECS  TL6809000
TSCA  TSCA listed
HS Code  29335995
Storage Class 13 - Non Combustible Solids
REACH Registrations Active
NFPA 704
1
2 1

HEPES price More Price(216)

Manufacturer Product number Product description CAS number Packaging Price Updated Buy
Sigma-Aldrich H4034 HEPES BioPerformance Certified, ≥99.5% (titration), suitable for cell culture 7365-45-9 25KG $6780 2026-04-30 Buy
Sigma-Aldrich H3375 HEPES ≥99.5% (titration) 7365-45-9 25KG $6460 2026-04-30 Buy
Sigma-Aldrich H3375 HEPES ≥99.5% (titration) 7365-45-9 50KG $12200 2026-04-30 Buy
Sigma-Aldrich H23830 HEPES reagent grade, ≥99% (titration), crystalline 7365-45-9 100 g $72.6 2026-04-30 Buy
Sigma-Aldrich H23830 HEPES reagent grade, ≥99% (titration), crystalline 7365-45-9 500 g $297 2026-04-30 Buy
Product number Packaging Price Buy
H4034 25KG $6780 Buy
H3375 25KG $6460 Buy
H3375 50KG $12200 Buy
H23830 100 g $72.6 Buy
H23830 500 g $297 Buy

HEPES Chemical Properties,Uses,Production

Description

HEPES has been described as one of the best all-purpose buffers available for biological research. At biological pH, the molecule is zwitterionic, and is effective as a buffer at pH 6.8 to 8.2 (pKa 7.55). It is typically used in cell culture at concentration between 5mM to 30 mM. HEPES has been used in a wide variety of applications, including tissue culture. It is commonly used to buffer cell culture media in air. HEPES finds its usage in in vitro experiments on Mg.

Chemical Properties

White crystalline powder

Uses

Non-toxic to cell. It is used as a hydrogen ion buffer, which can control the constant pH range for long term. The concentration is 10-50mmol/L. Generally, 20mmol/LHEPES in nutrient solution can get buffer ability.

Uses

HEPES is common buffer for biological sciences, particularly used in cell culture to maintain physiological pH. It acts a buffering component, which is used in the preparation of buffers. It is described as one of the best all-purpose buffers available for use in biological research.
HEPES has been used as a component of:
Hank′s balanced salt solution, Dulbecco′s modified eagle′s medium and no-glucose DMEM, which are used for the preparation of tissue slices.
Buffers used in localized conductance measurements on the basolateral side of culture inserts to examine cation selectivity.

Uses

HEPES is a biological buffer. It may be used in isoelectric focusing applications and as a buffer in the quantitative and selective measurement of antigen-antibody reactions. HEPES is reportedly superior to NaHCO3 in controlling pH in tissue and organ culture. HEPES is not recommended for use with the Folin-Ciocalteu protein assay. 

Definition

ChEBI: A HEPES that is ethanesulfonate substituted by a 4-(2-hydroxyethyl)piperazin-4-ium-1-yl group at position 2.

General Description

HEPES buffered saline is a tissue culture buffer, which is used to maintain the pH of cell culture. It is water soluble and atmosphere independent. HEPES destroys magnesium in sodium chloride solutions.

Biological Activity

Multi purpose buffer used in biological research.

Biological Activity

HEPES is a multi purpose HEPES buffer used in cell culture and other biological research. Working pH range in aqueous solution: 6.8 - 8.2. Does not form complexes with metal ions. Used in cell culture media.

Biotechnological Applications

HEPES serves as a buffering agent and transfection reagent that mediates cytoplasmic delivery of proteins into cells, and mixing it with proteins such as antibodies, recombinant proteins, and small peptides yields protein–HEPES mixtures suitable for transfection into cultured cells[1]. As a zwitterionic buffering agent in cell culture media, HEPES activates MiT/TFE-dependent lysosomal-autophagic gene networks and induces lysosome biogenesis, while its addition to culture media also triggers a lysosomal stress-related inflammatory phenotype and affects host-immune gene programs in RAW macrophages[2]. HEPES serves as a building block for molecular engineering of biomedical polymers, functioning as a pendant group in the zwitterionic polymer polyHEPES-pz intended for biomedical applications[3].

Synthesis

1-(2-Hydroxyethyl)piperazine

103-76-4

Sodium isethionate

1562-00-1

HEPES

7365-45-9

The general procedure for the synthesis of 4-hydroxyethylpiperazineethanesulfonic acid from N-hydroxyethylpiperazine and sodium 2-hydroxyethylsulfonate is as follows: 0.5 mole of N-hydroxyethylpiperazine, 0.565 mole of sodium 2-hydroxyethylsulfonate, and 500 ml of methanol were added to a 1000 ml four-necked flask fitted with a refluxing condenser tube, and the condensation reaction was carried out under sufficient stirring. The reaction was first carried out at 50 °C for 2.5 minutes, then gradually heated to reflux and continued for 3.0 hours. The reaction was cooled after completion of the reaction to give a reaction mixture containing 4-hydroxyethylpiperazine ethanesulfonate. The yield of 4-hydroxyethylpiperazine ethanesulfonate was 93.6% as analyzed by high performance liquid chromatography. A 20 liter solution of 4-hydroxyethylpiperazine ethanesulfonate at a mass concentration of 15 wt% was diluted to 5 wt% with deionized water under stirring, followed by removal of sodium chloride by dialysis. Then the purified HEPES product was obtained by evaporation concentration, cooling crystallization and filtration. Finally, the product was washed once with anhydrous methanol and dried under vacuum to obtain purified HEPES.A sample of purified HEPES was dissolved and tested for purity, which showed a purity of 99.9%, a yield of 85.6%, a pH of 6.46, and a concentration of chloride ions of 6.8 μg/g.

storage

Room temperature

Purification Methods

Crystallise the acid from hot EtOH and water. It is a useful buffer. [Beilstein 23 V 376.]

References

[1] Chen, S.-H., Chao, A., Tsai, C.-L., Sue, S.-C., Lin, C.-Y., Lee, Y.-Z., Hung, Y.-L., Chao, A.-S., Cheng, A.-J., Wang, H.-S., & Wang, T.-H. (2018). Utilization of HEPES for Enhancing Protein Transfection into Mammalian Cells. Molecular Therapy-Methods & Clinical Development, 13, 99–111. https://doi.org/10.1016/j.omtm.2018.12.005
[2] Tol, M. J., van der Lienden, M. J. C., Gabriel, T. L., Hagen, J. J., Scheij, S., Veenendaal, T., Klumperman, J., Donker-Koopman, W. E., Verhoeven, A. J., Overkleeft, H., Aerts, J. M., Argmann, C. A., & van Eijk, M. (2018). HEPES activates a MiT/TFE-dependent lysosomal-autophagic gene network in cultured cells: A call for caution. Autophagy, 14 3, 437–449. https://doi.org/10.1080/15548627.2017.1419118
[3] Tagad, H. D., Marin, A., Hlushko, R., & Andrianov, A. K. (2024). Hydrolytically Degradable Zwitterionic Polyphosphazene Containing HEPES Moieties as Side Groups. Biomacromolecules, 6791–6800. https://doi.org/10.1021/acs.biomac.4c01008

References

[1] Patent: CN106905262, 2017, A. Location in patent: Paragraph 0034; 0035; 0047

103-76-4
1562-00-1
7365-45-9
Synthesis of HEPES from 1-(2-Hydroxyethyl)piperazine and Sodium isethionate

HEPES Preparation Products And Raw materials

Raw materials

Preparation Products

Global( 1012)Suppliers
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Related Questions

Ask a question
Q:Why does HEPES buffer generate toxicity when exposed to light?STAR - Jul 6,2026
A:HEPES is a widely used amphoteric organic buffer. It is primarily employed in cell culture and molecular biology to maintain a stable physiological pH (typically between 6.8 and 8.2). This is because prolonged exposure to fluorescent lighting or sunlight causes HEPES to react with riboflavin, producing toxic hydrogen peroxide, which is subsequently toxic to sensitive cells.

View Lastest Price from HEPES manufacturers

Image Update time Product Price Min. Order Purity Supply Ability Manufacturer
4-(2-Hydroxyethyl)-1-piperazineethanesulfonic acid pictures 2026-08-26 4-(2-Hydroxyethyl)-1-piperazineethanesulfonic acid
7365-45-9
1KG 99% 200mt Jinan Finer Chemical Co., Ltd
HEPES pictures 2026-08-26 HEPES
7365-45-9
$52.00 500g >=99.5% (titration) 100kg Shanghai Acmec Biochemical Technology Co., Ltd.
HEPES pictures 2026-08-26 HEPES
7365-45-9
$52.00 500g >=99.5% (titration) 100kg Shanghai Acmec Biochemical Technology Co., Ltd.
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  • HEPES
    7365-45-9
  • $52.00
  • >=99.5% (titration)
  • Shanghai Acmec Biochemical Technology Co., Ltd.
  • HEPES pictures
  • HEPES
    7365-45-9
  • $52.00
  • >=99.5% (titration)
  • Shanghai Acmec Biochemical Technology Co., Ltd.
HEPES 2-[4-(2-HYDROXYETHYL)-1-PIPERAZINYL]ETHANESULFONIC ACID MOLECULAR BIOLOGY GRADE HEPES 2-[4-(2-HYDROXYETHYL)-1-PIPERAZINYL]ETHANESULFONIC ACID 99+% N-(2-HYDROXYETHYL)-PIPERAZINE-N'-2-ETHANESULFONIC ACID(4-(2-HYDROXYETHYL)PIPERAZINE-1-ETHANESULFONIC ACID(HEPES-ACID)) Hepes,FreeAcid(N-[2-Hydroxyethyl]Piperaz 1-HeptaneSulfonicAcidSod.SaltGr N-(2-Hydroxyethyl)Piperazine-N'-(3-EthanesulfonicAcid) N-(2-Hydroxyethyl)Piperazine-N'-(2-EthanesulfonicAcid)(Hepes) Hepes,LowSodium,FreeAcid HEPES free acid, DNase, Rnase and Protease free, for molecular biology, 99+% Hydroxyethyl)-1-piperazineethyanesulfonic acid, 4-(2- 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid(HEPES) 2-(4-(2-Hydroxyethyl)piperazin-1-yl)ethanesulfonic acid 4-(2-hydroxyerhyl)piperazine-1-erhaesulfonic acid HEPES, for biochemistry, 99% 4-(2-Hydroxyethyl)-1-Piperazin Hydroxyethyl)piperazine-N'-2-ethane sulfonic acid 1-Piperazineethanesulfonic acid, 4-(2-hydroxyethyl)- N-(2-hydroxymethyl)piperazine-N`-(2-ethanesulfonic acid)(HEPES) N-2-HYDROXYETHYLPIPERAZINE-N-2-ETHANSULFONICACID(HEPES) 4-(2-Hydroxyethyl)piperazin-1-ylethansulfonsure 2-[4-(2-Hydroxyethyl)-1-piperazine]ethanesulfonic acid HEPES HEPES / 4-(2-HYDROXYETHYL)-1-PIPERAZINEETHANESULPHONIC ACID HEPES (4-(2-HYDROXYETHYL)PIPERAZINE-1-ETHANESULFONIC ACID) HEPES ULTRAPURE N-2-HYDROXYETHYLPIPERAZINE-N''-2-ETHANE SULPHONIC ACID >99.5% N-[2-HYDROXYERHYL]PIPERAZINE-N''-ETHANESULFONICACID(HEPES) HEPES free acid ultrapure N-2-Hydroxyethylpiperazine-N'-2-ethanesulphonic acid, low sodium >99.5% HEPES(N-[2-Hydroxyethyl]piperazine-N'-[Ethanesulfonic acid]),Free Acid N'-2-N-2-hydroxyethyl piperazineethanesulfonic acid HEPES, >=99.5% (titration) HEPES SynonyMs 4-(2-Hydroxyethyl)-1-piperazineethanesulfonic acid HEPES,4-(2-hydroxyerhyl)piperazine-1-erhanesulfonic acid HEPES Vetec(TM) reagent grade, 99.5% 4-CHLORO-1-NAPHTHOL ULTRA PURE GRADE POTASSIUM HYDROXIDE 0,05 MOL DUTY PAID POTASSIUM HYDROXIDE 0,5MOL/L DUTY PAID POTASSIUM HYDROXIDE 1 MOL/L DUTY PAID SELENIUM STAND MALATE DEHYDROGENASE ULTRA PURE GRADE PHOSPHOTUNGSTIC ACID HIGH PURITY GRADE 4-(2-hydroxyethyl)-1-piperazineethanesulfonicaci hepesfreeacid 2-[4-(2-HYDROXYETHYL)-1-PIPERAZINE]ETHANESULFONIC ACID LABOTEST-BB LT00138109 LABOTEST-BB LT00233156 HEPES HEPES-ACID 4-(2-Hydroxyethyl)-1-piperazineethanesulphonic acid hepes free acid molecular biology*reagent HEPES N-(2-Hydroxyethyl)piperazine-N-(2-ethanesulfonic acid) HEPES, 1M solution HEPES, Free Acid, MB Grade (1.12041) HEPES, Free Acid, ULTROL Grade HEPES, Molecular Biology Grade N-(2-Hydroxyethyl)piperazine-N-(2-ethanesulfonic acid), Molecular Biology Grade HEPES, Tissue Culture Tested N-(2-Hydroxyethyl)piperazine-N-(2-ethanesulfonic acid), Tissue Culture Tested N-(2-HYDROXYETHYL)PIPERAZINE N-2-ETHANESULFONIC ACID N-2-HYDROXYETHYLPIPERAZINE-N'-2-ETHANE SULPHONIC ACID