Hank's Buffered Salt Solution (HBSS) Recipe

Hank's buffered salt solution (HBSS) is a fairly complicated solution to make which is why a lot of people just buy it. First you make each solution, the premix, and then the final solution. All solutions should be stored at 4^oC.

Hank's Stock Solutions (HBSS)

Stock #1

1. Dissolve the following in 90ml of distilled H₂O
   • 8.0 g NaCl
   • 0.4 g KCl
   • 1.0 g glucose
2. qs to 100 ml with distilled H₂O

Stock #2

1. Dissolve the following in 90ml of distilled H₂O
   • 0.358 g Na₂HPO₄ (anhydrous)
   • 0.60 g KH₂PO₄
2. qs to 100 ml with distilled H₂O

Stock #3

1. Add 0.72 g CaCl₂ to 50ml of distilled H₂O

Stock #4

1. Add 1.23 g MgSO₄x7H₂O to 50ml of distilled H₂O

Stock #5

1. Add 0.35 g NaHCO₃ to 10ml of distilled H₂O

Hank's Buffered Salt Solution (HBSS) Premix

Combine the solutions in following in order:
10.0 ml Solution #1
1.0 ml Solution #2
1.0 ml Solution #3
86.0 ml distilled H₂O
1.0 ml Solution #4

Hank's Buffered Salt Solution (HBSS) full strength (mix prior to use)

9.9 ml Hank's Premix
0.1 ml Stock #5

Hank's Buffered Salt Solution (HBSS) (Full Strength with carbonate) composition

0.137 M NaCl
5.4 mM KCl
0.25 mM Na₂HPO₄
0.1g glucose
0.44 mM KH₂PO₄
1.3 mM CaCl₂
1.0 mM MgSO₄
4.2 mM NaHCO₃