Optimization and Comparative Study of Different Extraction Methods of Sixteen Fatty Acids of L. from Twelve Different Producing Areas of the Qinghai-Tibetan Plateau.

In this study, supercritical fluid extraction (SFE), ultrasonic-assisted extraction (UAE), and microwave-assisted extraction (MAE) were applied to explore the most suitable extraction method for fatty acids of L. from 12 different producing areas of the Qinghai-Tibetan Plateau. Meanwhile, the important experimental parameters that influence the extraction process were investigated and optimized via a Box-Behnken design (BBD) for response surface methodology (RSM). Under optimal extraction conditions, 16 fatty acids of L. were analyzed via high-performance liquid chromatography (HPLC) with fluorescence detection, using 2-(4-amino)-phenyl-1-hydrogen-phenanthrene [9,10-d] imidazole as the fluorescence reagent. The results showed that the amounts of total fatty acids in sample 6 by applying SFE, UAE, and MAE were, respectively, 16.58 ± 0.14 mg/g, 18.11 ± 0.13 mg/g, and 15.09 ± 0.11 mg/g. As an environmental protection technology, SFE removed higher amounts of fatty acids than did MAE, but lower amounts of fatty acids than did UAE. In addition, the contents of the 16 fatty acids of L. from the 12 different producing areas Qinghai-Tibetan Plateau were significantly different. The differences were closely related to local altitudes and to climatic factors that corresponded to different altitudes (e.g., annual mean temperature, annual mean precipitation, annual evaporation, annual sunshine duration, annual solar radiation.). The temperature indices, photosynthetic radiation, ultraviolet radiation, soil factors, and other factors were different due to the different altitudes in the growing areas of L., which resulted in different nutrient contents.