Buffer Recipe Calculator

Calculate the required concentrations and molar amounts of weak acid and conjugate base components using the Henderson-Hasselbalch equation.

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Buffer System Properties

Specify acid pKa, target pH, concentration, and volume.

Required Base / Acid Ratio

1.74 ratio

* Ratio of [conjugate base] to [weak acid].

Required Base (A⁻) 63.5 mM (31.8 mmol)
Required Acid (HA) 36.5 mM (18.2 mmol)
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Effective Range

Buffers work best when target pH is within ±1 unit of pKa. Outside this range, buffer capacity is rapidly exhausted.

Common Biological Buffers & pKa Values Reference Table

Buffer System pKₐ (at 25°C) Effective pH Range Primary Application
Acetate Buffer 4.76 3.8 – 5.8 Enzyme kinetics, protein precipitation
Phosphate (PBS) pKa2 7.20 6.2 – 8.2 Physiological cell culture, immunology
Tris Buffer 8.06 7.0 – 9.0 DNA/RNA biochemistry, PAGE electrophoresis
HEPES Buffer 7.48 6.8 – 8.2 Good's buffer for cell culture media

Methodology & Equations

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Henderson-Hasselbalch

pH = pKₐ + log₁₀([Base] / [Acid])

Calculates the logarithmic ratio of conjugate base ($[A^-]$) to weak acid ($[HA]$).

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Stoichiometric Split

[Acid] = C / (R + 1), [Base] = C × R / (R + 1)

Splits total concentration C based on ratio R = 10^(pH - pKa).

Frequently Asked Questions

What is the Henderson-Hasselbalch equation?

The Henderson-Hasselbalch equation is pH = pKa + log10([Base] / [Acid]). It calculates the required ratio of conjugate base ([A⁻]) to weak acid ([HA]) for a buffer solution at a given pH.

What is optimal buffer capacity?

Buffer capacity is highest when the target pH equals the acid's pKa (where [Base] = [Acid]). In ptice, effective buffer capacity spans pKa ± 1 pH unit.

Why should temperature be considered when preparing biological buffers?

The pKa of amine buffers like Tris decreases as temperature rises (-0.031 pH units/°C). A Tris buffer prepared at pH 8.0 at 25°C will shift to ~pH 8.6 at 4°C.

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