Chlorophyll Content Calculator

Chlorophyll is the green pigment that drives photosynthesis in plants, and measuring its concentration tells researchers about plant health, stress response, and productivity. The Chlorophyll Content Calculator uses spectrophotometric absorbance readings to quantify pigment levels in leaf tissue extracts. Enter your absorbance at 663 nm and 645 nm, cuvette path length, extract volume/tissue weight ratio, and extraction solvent to get Total Chlorophyll Content, along with Chlorophyll a, Chlorophyll b, and the Chl a/b ratio — all in mg/g fresh weight.

Absorbance reading at 663 nm wavelength

Absorbance reading at 645 nm wavelength

cm

Light path length through the sample

mL/g

Volume of extraction solvent per gram of tissue

Results

Total Chlorophyll Content

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Chlorophyll a

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Chlorophyll b

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Chl a/b Ratio

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Frequently Asked Questions

What wavelengths are used for chlorophyll measurement?

Chlorophyll content is typically measured using absorbance at 663 nm (chlorophyll a peak) and 645 nm (chlorophyll b peak). These wavelengths correspond to the maximum absorption of each chlorophyll type.

Which extraction solvent should I use?

80% acetone is the most commonly used solvent for chlorophyll extraction as it efficiently extracts both chlorophyll a and b. Methanol and ethanol are alternatives, but acetone generally provides better extraction efficiency.

What is a normal chlorophyll a/b ratio?

The chlorophyll a/b ratio typically ranges from 2.5 to 3.5 in healthy plants. Higher ratios may indicate stress conditions, while lower ratios can suggest shade adaptation or certain nutrient deficiencies.

How do I prepare samples for chlorophyll measurement?

Fresh leaf tissue should be ground in extraction solvent (typically 80% acetone) at a ratio of 1g tissue to 10mL solvent. Keep samples on ice and in darkness to prevent chlorophyll degradation during extraction.

What factors can affect chlorophyll measurements?

Light exposure, temperature, pH, and time can all affect chlorophyll stability. Always work in dim light, keep samples cold, and measure absorbance promptly after extraction to ensure accurate results.

Can I use this calculator for different plant species?

Yes, this calculator uses standard extinction coefficients that apply to most plant species. However, some species may have slight variations, so consider species-specific coefficients for critical research applications.

What should I do if my absorbance readings are too high?

If absorbance values exceed 2.0, dilute your extract with the same solvent used for extraction. High absorbance readings can lead to inaccurate measurements due to non-linear Beer-Lambert law behavior.