Comparison of extraction methods for chlorophyllide formation with three different species ( Glebionis coronaria , Pisum sativum L. and Prunus sargentii Rehd.). Leaves were homogenized in acetone at room temperatures (“room temperatures”) or sub-zero temperatures (“sub-zero temperatures”), respectively, by grinding leaves with steel stainless beads. Alternatively, leaves were immersed in pure acetone at 4°C (“acetone 4°C”) or in pure DMF at 4°C (“DMF 4°C”), respectively, without the grinding procedure. In the last pair of experiments, leaves were boiled for 5 (“boiling 5 sec”) or 10 sec (“boiling 10 sec”), and were subsequently immersed in acetone at 4°C for chlorophyll extraction. Details of the methods is described in the Method section. A, C and E. Levels of chlorophyll a and chlorophyllide a per gram FW of leaves. B, D and F. Chlorophyllide a levels in sample extracts expressed as the ratio of chlorophyllide a to the sum of chlorophyll a and chlorophyllide a. Chl a, chlorophyll a. Chlide a, chlorophyllide a. Error bars indicate standard deviations. Sample size, n = 3.