Determination of the main naphthoquinones in Onosma hookeri Clarke. var. longiforum Duthie and its optimization of the ultrasound-assisted extraction using response surface methodology.

In this study, besides isovaleryl shikonin, another shikonin derivative, tigloylshikonin, was also isolated from the roots of Onosma hookeri Clarke. var. longiforum Duthie as a main naphthoquinone constituent for the first time. Then optimization of the ultrasonic-assisted extraction was done by Box-Behnken design-response surface methodology on the basis of single-factor experiments. The optimized conditions were 72% (v/v) ethanol and the material to solution ratio was 1:37(g/mL) at 52 °C for 77 min. Under these conditions, the extraction yield of ethanol extract was 36.74 ± 0.32%, the contents of isovaleryl shikonin and tigloylshikonin reached 0.94 ± 0.03% and 2.23 ± 0.06%, respectively. Notably, in that optimized condition, the yield of isovaleryl shikonin increased by approximately 7.64-fold than the previous report. In the in vitro antioxidant activity assay, the optimal ethanol extract exhibited similar 2,2-diphenyl-1-picrylhydrazyl (DPPH) scavenging activity as butylated hydroxy toluene (BHT), but slightly weaker 2,2'-azino-bis-3-ethylbenzthiazoline-6-sulphonic acid (ABTS) scavenging activity and total antioxidant capacity than that of BHT. However, the active polar fraction, the ethyl acetate fraction, which is enriched with naphthoquinone constituents, performs as a better antioxidant agent than BHT. Therefore, both of them could be considered as a naturally sourced antioxidants compared to commercially available synthetic drugs. PRACTICAL APPLICATION: Onosma hookeri Clarke. var. longiforum Duthie, a traditional Chinese medicine and food item, has been in use since a long time. A systematic determination of the main naphthoquinones, and antioxidant capacity of the naphthoquinones-enriched ethanol extract and different polar fractions, was carried out in the present study. The results may provide theoretical basis for the claim that naphthoquinones-enriched ethanol extract and ethyl acetate fraction from the roots of Onosma hookeri Clarke. var. longiforum Duthie could be used as potential natural antioxidants in the pharmaceutical, food, and cosmetic industries.