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H-50 buffer Preparation and Recipe

To prepare L  H-50 buffer

Table 1. Recipe

NameAmountConcentrationFormula/M.W.CAS #
HEPES(N-2-Hydroxyethylpiperazine-N'-2-ethanesulfonic acid)4.766 g0.02 MC8H18N2O4S/238.37365-45-9
Potassium Chloride3.727 g0.05 MKCl/74.557447-40-7
0.12 g1 mMMgSO4/120.377487-88-9
Sodium chloride0.584 g0.01 MNaCl/58.447647-14-5
Sodium bicarbonate0.42 g5 mMNaHCO3/84.01144-55-8
0.119 g1 mMNaH2PO4/119.987558-80-7
  1. Prepare 0.8 L of distilled water in a suitable container.
  2. Add 4.766 g of HEPES(N-2-Hydroxyethylpiperazine-N'-2-ethanesulfonic acid) to the solution.
  3. Add 3.727 g of Potassium Chloride to the solution.
  4. Add 0.12 g of Magnesium sulfate anhydrous to the solution.
  5. Add 0.584 g of Sodium chloride to the solution.
  6. Add 0.42 g of Sodium bicarbonate to the solution.
  7. Add 0.119 g of Sodium dihydrogen phosphate to the solution.
  8. Adjust the pH to 7.0.
  9. Add distilled water until the volume is 1 L.
  10. Autoclave and store cold or frozen.