A simple method is using dimethyl sulfoxide (DMSO) for chlorophyll extraction according to Barnes et al. (1992) and Shinano et al. (1996).

1) Barnes JD, Balaguer L, Manrique E, Elvira S, Davison AW (1992). A reappraisal of the use of DMSO for the extraction and determination of chlorophylls a and b in lichens and higher plants. Environ Exp Bot 32: 83–100

2) Shinano, T., Lei, T.T., Kawamukai, T., Inoue, M.T., Koike, T., and Tadano, T. Dimethylsulfoxide Method for the Extraction of Chlorophylls a and b from the Leaves of Wheat, Field Bean, Dwarf Bamboo and Oak, Photosynthetica, 1996, vol. 32, pp. 409–415

Two or three similar discs (about 0.8 cm in diameter) from each leaf are weighed and then incubated with 3 ml DMSO in a glass tube at 65°C until the tissue become colorless. The absorbance at 664.9 and 648.2 nm of the DMSO extract is determined with a spectrophotometer and the chlorophyll a and b concentrations of the leaves are calculated according to the below equations of Barnes et al. method

Chl a = 14,85 x A664.9 – 5,14 x A648.2 (μgr Chl a/ml)

Chl b = 25,48 x A648.2 – 7,36 x A664.9 (μgr Chl b/ml)

Finally, after the appropriate calculations you can express the chlorophyll concentration as:

mgr chl/gr fresh weight or

mgr chl/cm2 of leaf surface