基于Design-Expert软件设计优化超声波耦合负压真空法提取枇杷花黄酮工艺及其生物活性研究

Optimization of ultrasound coupled negative pressure vacuum extraction of Eriobotrya Lindl.flower flavonoids based on Design-Expert software and its biological activity study

目的优化超声波耦合负压真空法提取枇杷花黄酮的工艺,并对其生物活性进行研究。方法以枇杷花黄酮得率为指标,在单因素实验基础上,通过Design-Expert软件设计对枇杷花黄酮的提取工艺条件进行优化。同时以对1,1-二苯基-2-苦基肼(1,1-diphenyl-2-bitter hydrazine,DPPH)自由基和羟基自由基的清除率来评价抗氧化作用,以对大肠杆菌、枯草芽孢杆菌和金黄色葡萄球菌的抑菌圈直径和最低抑菌浓度来评价抑菌作用,以对小鼠耳朵肿胀度、小鼠耳组织中炎症因子肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、白细胞介素1β(interleukin-1β,IL-1β)和白细胞介素-6(interleukin-6,IL-6)含量来评价抗炎作用。结果最佳提取工艺条件为:超声功率400W、超声温度50℃、超声时间32min,此条件下,枇杷花黄酮得率为8.76%。枇杷花黄酮对DPPH自由基和羟基自由基都具有较强的清除能力,并且对DPPH自由基、羟基自由基的半数抑制浓度(halfmaximal inhibitory concentration,IC_(50))分别为0.223 mg/mL和0.274 mg/mL。枇杷花黄酮对大肠杆菌、枯草芽孢杆菌和金黄色葡萄球菌都具有不同的抑菌作用,且最低抑菌浓度分别为0.4、0.8和0.4 mg/mL。枇杷花黄酮能够减轻小鼠耳朵肿胀度,明显降低小鼠耳组织中炎症因子TNF-α、IL-6和IL-1β的含量,表明枇杷花黄酮具有显著的抗炎作用。结论本研究得到的枇杷花黄酮具有明显的抗氧化、抑菌和抗炎作用,为枇杷花的开发利用提供了理论依据。

Objective To optimize the extraction process of Eriobotrya Lindl.flower flavonoids by ultrasound coupled negative pressure vacuum method,and study its biological activity.Methods With the yield of flavonoids from Eriobotrya Lindl.flowers as an indicator,the extraction process conditions of flavonoids from Eriobotrya Lindl.flowers were optimized through Design Expert software design on the basis of single factor experiments.At the same time,the antioxidant effect was evaluated by the scavenging rate of 1,1-diphenyl-2-bitter hydrazine(DPPH)radical and hydroxyl radical,and the antibacterial effect was evaluated by the diameters of the inhibition zone and the minimum inhibitory concentrations against Escherichia coli,Bacillus subtilis and Staphylococcus aureus,and the degree of mouse ear swelling,the inflammatory factor in mouse ear tissue,tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β)and interleukin-6(IL-6)content were used to evaluate the anti-inflammatory effect.Results The optimal extraction process conditions are:Ultrasound power was 400 W,ultrasound temperature was 50℃,ultrasound time was 32 minutes.Under these conditions,the yield of Eriobotrya Lindl.flower flavonoids was 8.76%.Eriobotrya Lindl.flower flavonoids had strong scavenging ability on DPPH and hydroxyl radicals,and the half maximal inhibitory concentration(IC_(50))for DPPH and hydroxyl radicals were 0.223 mg/mL and 0.274 mg/mL,respectively.Eriobotrya Lindl.flower flavonoids had different antibacterial effects on Escherichia coli,Bacillus subtilis and Staphylococcus aureus,and the minimum inhibitory concentrations were 0.4,0.8 and 0.4 mg/mL,respectively.Eriobotrya Lindl.flower flavonoids could reduce mouse ear swelling and significantly reduce the inflammatory factor of TNF-α,IL-6 and IL-1βcontent in mouse ear tissue.This indicated that Eriobotrya Lindl.flower flavonoids had significant anti-inflammatory effects.Conclusion The obtained Eriobotrya Lindl.flower flavonoids have significant antioxidant,antibacterial,and anti-inflammatory effects,which provides a theoretical basis for the development and utilization of Eriobotrya Lindl.flower.