Optimization of a microwave-assisted extraction of secondary metabolites from crustose lichens with quantitative spectrophotodensitometry analysis.

A focused and rapid microwave-assisted extraction (MAE) process was carried out and optimized for secondary metabolites from crustose lichens using Taguchi experimental design and quantitative analysis on TLC by a Camag((R)) spectrophotodensitometer. The procedure was improved by quantitative determination of norstictic acid (NA), a common depsidone isolated from Pertusaria pseudocorallina (Sw.) Arn. Various experimental parameters that can potentially affect the NA extraction yields including extraction time, irradiation power, volume and the percentage of tetrahydrofuran (THF) were optimized. Results suggest that THF percentage and solvent volume were statistically the most significant factors. The optimal conditions were determined as follows: THF level of 100%, solvent volume of 15mL, microwave power of 100W and extraction time of 7min. Compared to the reflux method, MAE showed a drastic reduction of extraction time (7min vs. 3h) and solvent consumption (15mL vs. 30mL). The NA in total yield was 90% using the two methods. The optimal conditions were applied to other crustose lichens, Aspicilia radiosa, Diploicia canescens and Ochrolechia parella for the extraction of NA, diploicine (DP) and variolaric acid (VA), with 83%, 90% and 95% of recovery, respectively.