I used the Lichtenthaler and Wellburn (1983) protocol for chlorophyll and carotenoid extraction, using 0.5 g of fresh leaves, 10 mL of 80% acetone, and the coefficients and equations from this protocol. However, the final carotenoid results were negative. Is there any explanation for this? Could I have made a mistake in the protocol? I have followed this protocol many times, but this is the first time this has occurred.