Pharmaceutical Secondary Standard; Certified Reference Material

N-(2-Hydroxyethyl)piperazine-N′-(2-ethanesulfonic acid), 4-(2-Hydroxyethyl)piperazine-1-ethanesulfonic acid, HEPES
Empirical Formula (Hill Notation):
CAS Number:
Molecular Weight:
Beilstein/REAXYS Number:
EC Number:
MDL number:
PubChem Substance ID:

Quality Level


certified reference material
pharmaceutical secondary standard


current certificate can be downloaded


HPLC: suitable
gas chromatography (GC): suitable


5.0-6.5 (25 °C, 238 g/L)

useful pH range

6.8 - 8.2

pKa (25 °C)


Featured Industry

Pharmaceutical (small molecule)



SMILES string




InChI key


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General 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.

Pharmaceutical secondary standards for application in quality control, provide pharma laboratories and manufacturers with a convenient and cost-effective alternative to the preparation of in-house working standards.


These Secondary Standards are qualified as Certified Reference Materials. These are suitable for use in several analytical applications including but not limited to pharma release testing, pharma method development for qualitative and quantitative analyses, food and beverage quality control testing, and other calibration requirements.
  • HEPES has been used in a wide variety of applications, including tissue culture.
  • It is used to buffer cell culture media in air.
  • Finds its usage in invitro experiments on Mg.
  • Recognized as one of Dr.Good′s recommended buffers, used in some cell culture media as a buffering agent and also in various biochemical reactions.
  • Recently, in the production of radiopharmaceuticals, it is the buffer of choice for scientific labeling.

HEPES may also be used as:
  • Component of homogenization buffer to homogenize adipose tissue for liquid chromatography/mass spectrometry and in HN buffer for dissolving viral stock titer after dot blot hybridization.
  • Pharmaceutical reference standard for the determination of the analyte in bulk drug and pharmaceutical formulations by high performance liquid chromatography.

Analysis Note

These secondary standards offer multi-traceability to the USP, EP and BP primary standards, where they are available.

Other Notes

This Certified Reference Material (CRM) is produced and certified in accordance with ISO 17034 and ISO/IEC 17025. All information regarding the use of this CRM can be found on the certificate of analysis.


To see an example of a Certificate of Analysis for this material enter LRAC3253 in the slot below. This is an example certificate only and may not be the lot that you receive.


13 - Non Combustible Solids

WGK Germany


Flash Point(F)

Not applicable

Flash Point(C)

Not applicable

Certificate of Analysis

Certificate of Origin

Simple and reliable chromatographic technique for the determination of HEPES in Ga-68-labeled pharmaceuticals
Maus S, et al.
Journal of Nuclear Medicine, 56(3), 1168- 1168 (2015)
FGF21 Prevents Angiotensin II-Induced Hypertension and Vascular Dysfunction by Activation of ACE2/Angiotensin-(1?7) Axis in Mice.
Pan X, et al.
Cell Metabolism (2018)
Determination of HEPES in 68 Ga-labeled peptide solutions
Sasson R, et al.
J. Radioanal. Nucl. Chem., 283(3), 753- 756 (2010)
Development and evaluation of a rapid analysis for HEPES determination in 68 Ga-radiotracers
Pfaff S, et al.
EJNMMI Research, 8(1), 95-95 (2018)
Hydrogen ion buffers for biological research.
Good NE, et al.
Biochemistry, 5(2), 467-477 (1966)

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