Enter your Absorbance at 663 nm and Absorbance at 645 nm readings, along with your cuvette path length, extract volume/tissue weight ratio, and extraction solvent to calculate total chlorophyll content, broken down into Chlorophyll a, Chlorophyll b, and the Chl a/b ratio — all using the appropriate solvent-specific equations automatically.
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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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.
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.
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.
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.
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.
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.
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.