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Destarched AIR (3 mg) was saponified using 500 μl of 2 M of NaOH with continuous mixing at 300 rpm in a thermomixer at 25°C for 24 h. The solution was then neutralized with 100 μl of concentrated hydrochloric acid (HCl) and extracted with 300 μl of ethyl acetate. Samples were dried and dissolved in 50% (v/v) methanol for HCA analysis using a Dionex Ultimate 3,000 high-performance liquid chromatography (HPLC) system. Details for the HPLC protocol and run parameters were as previously described, except that detection was at 305 nm (Bartley et al., 2013a).
Copyright and License information: ©2021 Saha, Lin, Thibivilliers, Xiong, Pan and Bartley.
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