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Beer–Lambert Law Calculator

Solves the Beer–Lambert law A = ε·l·c for the selected quantity (absorbance, concentration, molar absorptivity or path length) from the other three, and reports the percent transmittance T = 100 × 10^(−A).

When to use

You have a spectrophotometer absorbance and need the concentration of the absorbing species from its molar absorptivity, the expected absorbance of a solution, or the molar absorptivity or cuvette path length from a calibration measurement.

Do not use when: Absorbance is above about 2, or the sample is concentrated, turbid or chemically interacting (the law is linear only for dilute, non-scattering solutions in monochromatic light); for concentration from a weighed mass use molarity.

Formula

A = molar_absorptivity_l_mol_cm × path_length_cm × concentration_mol_l; c = A / (ε × l); ε = A / (l × c); l = A / (ε × c); transmittance_percent = 100 × 10^(−A)

A is the decadic absorbance log10(I0/I) and ε the molar decadic absorption coefficient, so ε is in L·mol⁻¹·cm⁻¹ with l in cm and c in mol/L. Linearity assumes monochromatic light and a dilute (below about 0.01 mol/L), homogeneous, non-scattering sample; above A ≈ 1.5–2 stray light and detector limits make readings unreliable.

Inputs

ParameterTypeUnitRequiredDescription
modeenum: concentration | absorbance | molar_absorptivity | path_lengthdefault concentrationQuantity to solve for; the other three must be given (path_length_cm defaults to a 1 cm cuvette).
absorbancenumbernoBlank-corrected absorbance (optical density, base-10, dimensionless). Leave empty when solving for it. Range: ≥ 0, ≤ 10
molar_absorptivity_l_mol_cmnumberL/(mol·cm)noMolar (decadic) absorption coefficient at the measurement wavelength, e.g. 6220 for NADH at 340 nm. Leave empty when solving for it. Range: > 0, ≤ 10000000
path_length_cmnumbercmdefault 1Optical path length through the sample (inner cuvette width); standard cuvettes are 1 cm. Ignored when mode = path_length. Range: > 0, ≤ 1000
concentration_mol_lnumbermol/LnoMolar concentration of the absorbing species. Leave empty when solving for it. Range: > 0, ≤ 100

Outputs

OutputTypeUnitDescription
absorbancenumberBase-10 absorbance, given or solved.
molar_absorptivity_l_mol_cmnumberL/(mol·cm)Molar absorption coefficient, given or solved.
path_length_cmnumbercmOptical path length, given or solved.
concentration_mol_lnumbermol/LMolar concentration, given or solved.
concentration_scientificstringThe concentration in scientific notation, mol/L.
concentration_umol_lnumberµmol/LThe same concentration in micromoles per litre (µM).
transmittance_percentnumber%100 × 10^(−A): share of the incident light that passes through the sample.
solved_forstringName of the output that was computed: absorbance, molar_absorptivity_l_mol_cm, path_length_cm or concentration_mol_l.

Example

Absorbance of 1.0×10⁻⁴ M NADH at 340 nm (ε 6220, 1 cm cuvette): {"mode":"absorbance","molar_absorptivity_l_mol_cm":6220,"path_length_cm":1,"concentration_mol_l":0.0001}{"absorbance":0.622,"transmittance_percent":23.88,"concentration_umol_l":100,"solved_for":"absorbance"}

Concentration from A = 0.500 with ε 6220 in a 1 cm cuvette: {"mode":"concentration","absorbance":0.5,"molar_absorptivity_l_mol_cm":6220}{"concentration_mol_l":0.000080386,"concentration_scientific":"8.039e-5","concentration_umol_l":80.386,"transmittance_percent":31.62,"solved_for":"concentration_mol_l"}

GET https://tttkmbb.com/api/v1/calculate/beer-lambert?mode=absorbance&molar_absorptivity_l_mol_cm=6220&path_length_cm=1&concentration_mol_l=0.0001

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Sources

FAQ

What if my ε is in mM⁻¹cm⁻¹ or the concentration in mg/L?

Multiply an ε in mM⁻¹·cm⁻¹ by 1000 to get L·mol⁻¹·cm⁻¹ (NADH: 6.22 mM⁻¹cm⁻¹ = 6220). For mass concentrations divide by the molar mass to get mol/L, or use a specific absorbance in L·g⁻¹·cm⁻¹ and read the result as g/L.

Why is absorbance above 2 unreliable?

At A = 2 only 1 % of the light reaches the detector, so stray light and detector noise dominate and the response flattens; dilute the sample or use a shorter path length to bring A into the 0.1–1 range.

Is absorbance the same as optical density?

In practice yes for clear solutions: both are log10(I0/I). Optical density also includes light lost by scattering, which is why turbid samples deviate from the Beer–Lambert law.

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