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Hanks' salts

Hanks' salts is a collective group of salts rich in bicarbonate ions, formulated in 1940 by the microbiologist John H. Hanks.[1] Typically, they are used as a buffer system in cell culture media and aid in maintaining the optimum physiological pH (roughly 7.0–7.4) for cellular growth. Due to their poorly reactive nature and small concentration in solution, Hanks' salts are mainly used in media that are exposed to atmospheric conditions as opposed to CO2 incubation. Performing the latter drastically exceeds the buffer capacity of Hanks' salts and may result in cell death.


The recipe according to AATBIO[2]

Table 1. Required components

Component Amount Concentration
NaCl (mw: 58.44 g/mol) 8 g 0.14 M
KCl (mw: 74.55 g/mol) 400 mg 0.005 M
CaCl2 (mw: 110.98 g/mol) 140 mg 0.001 M
MgSO4-7H2O (mw: 246.47 g/mol) 100 mg 0.0004 M
MgCl2-6H2O (mw: 203.303 g/mol) 100 mg 0.0005 M
Na2HPO4-2H2O (mw: 177.99 g/mol) 60 mg 0.0003 M
KH2PO4 (mw: 136.086 g/mol) 60 mg 0.0004 M
D-Glucose (Dextrose) (mw: 180.156 g/mol) 1 g 0.006 M
NaHCO3 (mw: 84.01 g/mol) 350 mg 0.004 M
  1. Prepare 800 mL of distilled water in a suitable container.
  2. Add 8 g of NaCl to the solution.
  3. Add 400 mg of KCl to the solution.
  4. Add 140 mg of CaCl2 to the solution.
  5. Add 100 mg of MgSO4-7H2O to the solution.
  6. Add 100 mg of MgCl2-6H2O to the solution.
  7. Add 60 mg of Na2HPO4-2H2O to the solution.
  8. Add 60 mg of KH2PO4 to the solution.
  9. Add 1 g of D-Glucose (Dextrose) to the solution.
  10. Add 350 mg of NaHCO3 to the solution.
  11. Add distilled water until volume is 1 L.
  1. ^ Hanks JH, Wallace RE (1 June 1949). "Relation of oxygen and temperature in the preservation of tissues by refrigeration" (PDF). Proceedings of the Society for Experimental Biology and Medicine. 71 (2). New York, N.Y.: Society for Experimental Biology and Medicine: 196–200. PMID 18134009.
  2. ^ "HBSS (Hank's Balanced Salt Solution) Preparation and Recipe | AAT Bioquest". www.aatbio.com. Retrieved 2021-07-06.