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Science

Beer-Lambert Law Absorbance Calculator

The Beer–Lambert law states that for a dilute solution and monochromatic light, absorbance is proportional to both concentration and path length. Pick the variable you want, supply the other three, and the calculator also reports the transmittance T = 10⁻ᴬ.

Result

Transmittance T T = 10⁻ᴬ. A = 1 → 10 % transmitted, A = 2 → 1 % transmitted.

Formula

A = ε × c × ℓ

A is dimensionless; ε is in M⁻¹·cm⁻¹ (L·mol⁻¹·cm⁻¹); c is in M (mol/L); ℓ is in cm (a standard cuvette is 1 cm).

Formula

A = ε × c × ℓ ; T = 10⁻ᴬ

Frequently asked

What is the difference between absorbance and transmittance?

Transmittance T = I / I₀ is the fraction of light passing through the sample (0 to 1, or 0–100 %). Absorbance A = −log₁₀ T linearises that ratio so it scales with concentration. T = 10 % ⇔ A = 1; T = 1 % ⇔ A = 2.

Where do I get the molar absorptivity ε?

ε depends on wavelength, solvent and pH and is typically published per-wavelength (e.g. "ε₂₈₀" or "ε at λmax"). Common values: NADH at 340 nm ≈ 6 220 M⁻¹·cm⁻¹; tryptophan at 280 nm ≈ 5 500 M⁻¹·cm⁻¹. If no literature value exists, build a standard curve (known c → measured A) and read ε from the slope.

My reading is A > 2 — why is the answer unreliable?

Above A ≈ 2, less than 1 % of the light reaches the detector, so stray light becomes a sizeable relative error. Molecules can also aggregate or shift in colour at high c, breaking linearity. The fix is to dilute (e.g. 10×) until A falls into the ideal 0.1–1 window and then multiply back by the dilution factor.

Can I enter the concentration in mM or µM directly?

The c input must be in M (mol/L): 1 mM = 1 × 10⁻³ M, 1 µM = 1 × 10⁻⁶ M, 1 nM = 1 × 10⁻⁹ M. So "50 µM" goes in as 0.00005 (or in scientific form 5e-5). Likewise ε must be in M⁻¹·cm⁻¹ (i.e. L·mol⁻¹·cm⁻¹).

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