Bioactive Components Isolation Based On The Integrated Ultrasonic/Microwave-assisted Preceded By Enzymatic Treatment From Two Species Of Lavandula

The integrated ultrasonic/microwave-assisted(US/MIC)were used to optimize the isolation processes of bioactive compounds from two species of Lavandula: L.pubescens(Lp)and L.angustifolia(La).The bio-nanocapsules of essential oils which obtained from La(La.EO)incorporated with whey protein isolate were prepared.To prepare the bioactive extractions,modern techniques including ultrasonic,microwave irradiations and integrated US/MIC were used.For polyphenolic crude extracts from Lp(Lp.PCE),three techniques of extraction were used.US/MIC at extraction conditions of 40 kHz(ultrasonic frequency),50 W(ultrasonic power),480 W(microwave power),and 5 min(treatment time).Ultrasonichomogenizer at extraction conditions of 20 kHz(ultrasonic frequency),150 W(ultrasonic power),and 5 min(treatment time).The extraction using conventional maceration was used as a control.Folin-Ciocalteu method was used to determine the total phenolic contents(TPC)(mg gallic acid equivalent/g dry matter).DPPH and ?-carotene/linoleic acid bleaching were used to investigate the anti-free radical activity of Lp.PCE.Rancimat assay was used to investigate the oxidative stability of bleached and deodorized palm olein oil(RBDPOo)before and after addition of Lp.PCE with and without the synergistic effect of citric acid(CA).For Lp.EO and La.EO isolation,the integrated techniques represented by US/MIC assisted preceded by enzymolysis were used to enhance the efficiency of isolation processes.Cellulase(CEase),hemicellulase(HEase),and combined of both enzymes(CEase/HEase)were used at 10 mg/ 100 m L of the liquid phase.Response Surface Methodology(RSM)depending on Box-Behnken Design(BBD)with 17 runs was employed.The Clevenger-type apparatus was employed to extract the essential oils,thus,determine the optimal enzymatic pre-treatment.GC-MS was used to analyze the chemical composition of Lp.EO and Lp.EO.SEM was used to observe the changes on the surface structure of the Lp and La plant tissues after enzymatic pre-treatments and after US/MIC treatment.To determine the free radical-scavenging activity,DPPH·-SA and ABTS·+SA assays were used.The anti-pathogenic bacterial activity of Lp.PCE,Lp.EO,and La.EO was investigated against negative and positive bacteria.La.EO was subjected to the encapsulation experiments.For nanocapsules preparation,sunflower oil(SFo)was used as a carrier material and whey protein isolate(WPI)was used as a wall material.Zetasizer Nano(Malvern Instruments,Westborough,MA)capable of electrophoresis measurements was used to determine the size distributions,polydispersity index,and electrical charge(Zeta potential).Morphological observation of nanoemulsions droplets and nanocapsules were carried out using CLSM,TEM and SEM.To investigate the incorporated between La.EO and WPI,XRD and FTIR analysis were employed.The TPCs of Lp.PCE(mg gallic acid equivalent/g dry matter)were found to be 253.87,216.96 and 203.41 for US/MIC extract,ultrasonic-homogenizer extract and maceration extract,respectively.The US/MIC extract achieved the higher scavenger effect of DPPH·(90.53%)with EC50(19.54 ?g/m L),and higher inhibition of ?-carotene/linoleate emulsion deterioration(94.44%)with IC50(30.62 ?g/m L).The activity of the US/MIC treatment could prolong the induction period(18.82 h)and oxidative stability index(1.67)of fresh refined,bleached and deodorized palm olein oil(RBDPOo)using Rancimat assay.There was an important synergist effect between citric acid and Lp extracts in improving the oxidative stability of fresh RBDPOo.Carvacrol was the major component in all treatments.CEase treatment achieved the highest carvacrol(29.22%).The changes in the morphological structure of Lp tissues were tracked using SEM.CEase pre-treatment achieved the highest yield% in comparison to the control(Ctrl),CEase/HEase and HEase treatments.The optimal isolation conditions were consisted of the concentration of enzymes(14.82mg/100 m L),sonication time(38.18min)and microwave power(333W).At optimal isolation conditions,the maximum yield%,DPPH·-SA% and ABTS·+SA(mM Trolox)were 0.67%,65.40% and 0.025,respectively.The enzymolysis pre-treatments could be used to enhance the release of secondary metabolites from plant matrix.Enzymolysis pre-treatment by CEase contributed in enhancing the efficiency of Lp.EO release.The results of the current study found that Lp.EO which isolated by using US/MIC under optimal conditions could be used as anti-free radical and antibacterial in the biological systems.For La.EO isolation,US/MIC assisted preceded by enzymolysis treatment was used to isolate essential oil(EO)of La.Response Surface Methodology was employed for the same purpose.The yield% of La essential oil(La.EO)obtained from combined enzymes treatment(cellulase and hemicellulase)was higher than that obtained by a single enzyme treatment.Chemical composition analysis of La.EO isolated was carried by using GC-MS.Morphological structure of La tissues was observed using SEM.The anti-free radical activity was assayed using DPPH· and ABTS·+ methods.The isolation conditions consisted of the concentration of enzymes(12.5mg)with a range of(5-20mg),sonication time(52.5min)with a range of(15-90 min)and microwave power(275W)with a range of(500-50W).The antipathogenic bacterial activity results showed that the US/MIC assisted extracts were the most effective against Gram-positive and Gram-negative strains.The results of the current work also demonstrated that the uses of integrated US/MIC could induce the acoustic cavitation and rupture of plant cells.This facilitated the flow of liquid phase into the glandular trichomes in plant cells and enhanced the absorption from the plant matrix.Thus,enhanced the efficiency of bioactive extractions of aromatic plant and ensured a high selective release and component purity based on cavitation phenomenon.The bioactive components isolated from Lp and La could be used as food additives in food industries to prevent the oxidation and bacterial spoilage.This work also found that the La.EO was successfully encapsulated with SFo as a carrier material and WPI-coarse emulsion as a wall material.