Optimization of Enzyme Assisted-ultrasonic Extraction of Sinigrin in Thlaspi arvense Seeds by Response Surface Methodology
In order to obtain active ingredient sinigrin with high efficiency and high yield from seeds of Thlaspi arvense,the enzymolysis assisted-ultrasonic extraction method was used.The content of sinigrin was determined by high performance liquid chromatography(HPLC).Based on the single factor experiment,enzymolysis time,enzymolysis temperature and enzyme addition amount were the key factors,and the yield of sinigrin was taken as the response value.The Design-Expert 11 software was applied and the Both-Behnken Design response surface methodology was used to optimize the enzymolysis assisted ultrasonic extraction process of sinigrin.The yield of sinigrin was compared with that of ultrasonic and Soxhlet extraction.The results showed that Agilent HC-C18 column(25 mm×4.6 mm×5 μm),mobile phase A methanol,mobile phase B 0.5%phosphoric acid aqueous solution,detection wavelength 215 nm,column temperature 25℃,flow rate 1 mL·min-1,sample size 10 μL,and the same degree elution for 10 min,the separation effect and peak shape were better.Using the screened alkaline protease as enzyme preparation,the optimal extraction conditions were that the enzymolysis temperature was 49℃,the enzyme dosage was 2.90%,the enzymolysis time was 18 min,the yield of sinigrin was 2.679 mg·g-1 under optimal conditions,which was no significant difference with the theoretical value(2.687 mg·g-1).The yield of sinigrin was significantly higher than that of ultrasonic extraction(2.036 mg·g-1)and Soxhlet extraction(1.702 mg·g-1)(P<0.05).The enzymolysis assisted-ultrasonic extraction process of sinigrin from T.arvense was simple,energy saving,eco-friendly,high efficiency and high yield,and it could be applied to the production and utilization of sinigrin from T.arvense,which laid a theoretical foundation for the in-depth development of T.arvense.
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