Tshawb nrhiav Lub peev xwm ntawm Icelandic Seaweeds Extracts Ua Los ntawm Aqueous Pulsed Electric Fields-Assisted Extraction Rau Kev Siv Tshuaj pleev ib ce Part 2

Thov hu rauoscar.xiao@wecistanche.comyog xav paub ntxiv

2.5.4 ib. Hyaluronidase Inhibition Activity

Tag nrho cov seaweed extracts exhibited ho siab heevanti-hyaluronidasekev ua haujlwm (Table 4), uas qhia cov txiaj ntsig sib piv rau cov tshuaj tannic acid (ib tus paub zoo inhibitor ntawm hyaluronidase). Tshwj xeeb, A.esculenta extracts pom 100 feem pua ​​​​ntawm inhibition rau txhua txoj kev kuaj. Ntxiv mus, U. Lactuca extracts nthuav tawm cov dej num siab tshaj 90 feem pua ​​​​ntawm inhibition, qhov twg inhibition ntawm cov extracts uas tsim los ntawm PEF (96.8 feem pua) thiab kev sib xyaw ntawm PEF plus HW (97.3 feem pua) yog siab tshaj qhov inhibition tsim los ntawm cov tsoos dej kub. txoj kev 93.4 feem pua ​​)(p<0.05). all="" p.palmaria="" extracts="" exhibited="" similar="" activities="" (p=""><0.05), the="" inhibition="" of="" the="" extracts="" produced="" by="" pef="" was="" (91.9="" %)="" and="" the="" combination="" of="" pef+="" hw="" (89.5%)="" and="" the="" traditional="" hot="" water="" method="">

Thov nias ntawm no kom paub ntxiv

Lwm tus kws sau ntawv kuj tau piav qhia txog kev tiv thaiv hyaluronidase kev ua ntawm cov seaweeds sib txawv, tshwj xeeb tshaj yog rau cov extracts nplua nuj nyob hauvphlorotanninslos ntawm xim av algae [73,74]. Txawm li cas los xij, rau qhov zoo tshaj plaws ntawm peb txoj kev paub, qhov no yog thawj zaug uas hyaluronidase inhibitory kev ua ub no ntawm P. palmitate thiab U.lactuca extracts uas tsim los ntawm PEF tau tshaj tawm.

Cistanche tuaj yeem txhim kho kev tiv thaiv

Hyaluronic acid yog ib qho tseem ceeb ntawm cov dermis, qhov uas nws koom nrog hauv kev kho cov ntaub so ntswg, nws tawg nrog kev laus, ua rau wrinkles thiab poob ntawm daim tawv nqaij firmness. Hauv qhov kev nkag siab no, hyaluronidase inhibitors nce qib hyaluronic acid ntawm dermal extracellular matrix rau kev txhim kho cov tsos ntawm kev laus.daim tawv nqaij ntawm lub ntsej muag[13]. Yog li, cov txiaj ntsig ntawm txoj kev tshawb fawb no tuaj yeem qhib txoj hauv kev tshiab rau kev siv lub ntuj hyaluronidase inhibitors los ntawm cov peev txheej algae uas muaj peev xwm siv tau hauv cov khoom siv tshuaj pleev ib ce.

Hauv cov ntsiab lus, cov ntaub ntawv sau cia tau ua rau peb txiav txim siab tias A.esculenta extracts nthuav tawm tag nrho cov haujlwm inhibitory zoo dua li P.palmaria thiab U. Lactuca ntawm cov enzymes kuaj. Yog li ntawd, nws yog qhov zoo tshaj plaws seaweed hom nrog zoo heevanti-enzymatickev ua ub no thiab yog li ntawd nws tau raug xaiv rau kev kawm ntxiv hauv peb chav kuaj. Txawm hais tias crude extracts los ntawm A.esculenta zoo nkaus li yog cov neeg sib tw zoo hauv kev sim vitro, kev tshawb fawb ntxiv yuav tsum tau ua kom paub meej tias tus kheej ntawm cov metabolites lub luag haujlwm rau cov teebmeem lom.

2.6.Correlations ntawm Chemical Compounds thiab Bioactive Properties

Cov txiaj ntsig tau los ntawm kev txheeb xyuas cov ntsiab lus tseem ceeb (PCA), pom tias qhov sib cais tseem ceeb ntawm pawg tau txhais los ntawm PC1 thiab PC2, uas suav txog 71.9 feem pua ​​​​ntawm thiab 14.5 feem pua ​​​​ntawm qhov sib txawv ntawm cov ntaub ntawv, raws li (Daim duab 2). A.esculenta extracts tau muaj cov ntsiab lus siab dua ntawm flavonoids thiab phenolic tebchaw, inhibitory cuam tshuam rau enzymes (collagenase, tyrosinase thiab elastase), thiab DPPH thiab FRAP qhov tseem ceeb, dua li lwm hom, P. palmata thiab Ul. Lactuca. Ntawm qhov tod tes, A.esculenta muaj cov ntsiab lus carbohydrate qis, tshwj xeeb tshaj yog piv rau P. palmitate (uas nyob rau sab nraud ntawm PC1). Qhov sib txawv ntawm cov ntaub ntawv raws li PC2 feem ntau cuam tshuam nrog ABTS thiabhyaluronidaseinhibition. Raws li tau hais los ntawm qhov chaw nyob ntawm daim phiaj, P. palmata muaj kev sib raug zoo rau ABTS thaum U. Lactuca muaj feem cuam tshuam nroghyaluronidaseinhibition cuam tshuam, piv rau ob hom no.

Kev sib raug zoo siab thiab tseem ceeb ntawm TPC, TFC, DPPH, FRAP, thiab inhibitory cuam tshuam ntawm collagenase, elastase, thiabtyrosinasetau pom los ntawm Pearson correlation tsom xam (Table 5).

Daim duab 2. Kev tsom xam cov ntsiab lus tseem ceeb (PCA) biplot ntawm cov khoom thauj khoom (dub lub voj voog) thiab cov qhab nia rau cov algae rho tawm (P. palmata: xim av dots (sab saud); U. Lactuca: ntsuab daim duab peb sab (feem ntau nyob rau sab laug sab sauv); A . esculenta: xiav squares (qis dua ntawm daim phiaj).

Qhov no tau pom zoo nrog cov kev tshawb fawb yav dhau los, qhia tias phenolic tebchaw (xws li flavonoids) yog lub luag haujlwm tseem ceeb rau kev ua haujlwm antioxidant ntawm ntau cov nroj tsuag hiav txwv [75-77]. Kev ua haujlwm siab antioxidant ntawm cov extracts los ntawm cov xim av macroalgae tau cuam tshuam rau ib pawg ntawm polyphenols, phlorotannins, thiab lawv cov qauv molecular tshwj xeeb. Phlorotannis los ntawm cov algae xim av tau tshaj tawm tias muaj txog yim qhov sib cuam tshuam phenol rings uas ua raws li cov cuab yeej hluav taws xob [78,79]. Nws tau xav tias ABTs yuav cuam tshuam nrog TPC, lwm yam tshuaj tiv thaiv antioxidant. Cov laj thawj muaj peev xwm yuav yog tias cov txheej txheem yog raws li qhov sib txawv ntawm cov tshuaj tiv thaiv thiab qhov reactivity txawv ob qho tib si hais txog lub sijhawm thiab ntau yam ntawm cov khoom. Piv txwv li, ABTS reagent reacts nrog ntau qhov dav ntawmantioxidantstshaj DPPH radical [80]. Ntawm qhov tod tes, ib qho ntawm cov kev txwv uas tau hais rau ABTS yog cov tshuaj tiv thaiv ntev thiab lub sij hawm cov tshuaj tiv thaiv dav dav yuav tsis tso cai mus txog qhov kawg.

Cov txiaj ntsig tau qhia tias muaj kev sib raug zoo ntawm TPC thiab TFC rau qhov inhibitory kev ua ntawm collagenase, elastase, thiab tyrosinase ({{0}}}93-0.99), qhov kev sib raug zoo rau inhibition. ntawm hyaluronidase tsis muaj zog (r=0.42 thiab 0.54, feem). Qhov no qhia tau hais tias lwm yam khoom siv tuaj yeem ua rau inhibitory nyhuv ntawm cov extracts. Lwm cov kev tshawb fawb tau qhia tias polysaccharides muaj hyaluronidase-inhibitory kev ua, piv txwv li alginic acid nyob rau hauv cov xim av algae [81,82]. Cov kev tshawb fawb ntxiv ntawm cov tshuaj muaj pes tsawg leeg ntawm macroalgae hom rau cov teebmeem ntawm cov tshuaj sib cais ntawm cov enzyme yog xav tau los ntsuas qhov kev koom tes ntawm txhua yam tshuaj lom neeg xws li hauv txoj kev tshawb no tsom mus rau crude extracts.

Cov kev tshawb pom tau nyob rau hauv kev sib raug zoo nrog cov kev tshawb fawb yav dhau los, hais tias cov tshuaj muaj pes tsawg leeg thiab qib ntawm bioactivity ntawm cov extracts sib txawv heev ntawm peb kab (liab, ntsuab, thiab xim av algae) thiab ntawm ntau hom sib txawv ntawm tib lub phylum thiab cuam tshuam los ntawm hnub nyoog. thiab hom ntaub so ntswg. Tsis tas li ntawd, muaj pes tsawg leeg thiab cov yam ntxwv nyob ntawm ntau yam ib puag ncig cuam tshuam rau kev faib thiab kev loj hlob ntawm macroalgae. Piv txwv li, lub teeb (UV-radiation), qhov kub thiab txias, muaj cov khoom noj khoom haus, raug huab cua, dej txav, yoj raug, thiab ntsev. Qhov ntsuas kub tau piav qhia tias yog qhov tseem ceeb uas muaj qhov muaj zog tshaj plaws ntawm cov xim tsim thiab cov khoom noj khoom haus concentration, salinity, thiab UV hluav taws xob raws li cov yam ntxwv cuam tshuam rau cov concentration ntawm TPC [83].

Kev faib tawm ntawm ntau hom macroalgae sib txawv nrog dej tob. Cov hauj lwm siab dua ntawm ntug dej hauv thaj chaw intertidal lossis littoral muaj kev ntxhov siab ntau dua li cov tsiaj loj hlob nyob rau ntawd, yuav tsum tiv taus ntau yam kev hloov pauv ntawm cov kab mob abiotic vim muaj kev hloov pauv tidal. Piv txwv li, cov huab cua qhuav ntawm huab cua, siab hnub ci irradiances (thaum dej ntws tsawg), kev hloov hauv salinity thiab kub thiab, nyob rau hauv tej yam kev mob ntawm huab cua tsis tshua muaj kub, nrog rau freezing. Hauv qab cov dej tsis tshua muaj, qhov tob zuj zus tuaj yeem ua rau txo qis heev hauv lub teeb siv thiab tsawg dua rau irradiance.

Algae loj hlob nyob rau hauv tidal ntau yam muaj qis rhiab heev rau UV Radiation thiab rov qab sai dua los ntawm hnub ci kev nyuaj siab. Whereas algae loj hlob nyob rau hauv sublittoral cheeb tsam yog rhiab heev rau UV hluav taws xob thiab muaj qis rov qab los ntawm hnub ci kev nyuaj siab [84]. Nyob rau tib lub sijhawm, cov kab dej muab kev tiv thaiv. Nyob rau hauv txoj kev tshawb no, qhov raug tshav ntuj yog qhov muaj zog dua rau P. palmata, piv rau lwm hom. Lwm cov kev tshawb fawb tau pom tias qhov tsim ntawm MAAs ncaj qha cuam tshuam rau lub hnub ci [85], tiv thaiv cov kab mob tiv thaiv UV-A thiab UV-B hluav taws xob. Tsis tas li ntawd, nws tau pom tias qhov tshwj xeeb ntawm MAAs tau txo qis nrog nce kev sau qhov tob. Kelps xws li A.esculenta, paub tias loj hlob nyob rau sab sauv sublittoral cheeb tsam tab sis kuj txuas mus rau qhov qis tshaj intertidal tsuas yog saum cov dej qis. Lub ntsiab lus ntawm cov dej kem muab kev tiv thaiv muaj zog dua rau P.palmata. Tsis tas li ntawd, cov yam ntxwv morphological txawv, cov hniav ntawm A.esculenta yog tuab dua piv rau lwm hom. UI. Lactuca, loj hlob nyob rau hauv lub intertidal thiab sublittoral yog muaj peev xwm mus photosynthesize thiab loj hlob nyob rau hauv tsawg heev irradiances. Kev cuam tshuam rau UVB lub teeb tau hais kom nrawm rov qab cov duab hluav taws xob tsis zoo ntawm U. Lactuca los ntawm qhov tsis zoo ntawm UVA lub teeb. Nws yog me me, yooj yim hauv cov qauv, thiab luv-nyob (3 lub hlis) dua ob qho tib si A.esculenta (5-7 vear) thiab P. palmata uas muaj kev loj hlob tshiab txhua xyoo.

Hauv cov ntsiab lus, cov kev xav tau tuaj yeem kos tau tias qhov sib txawv tseem ceeb hauv cov khoom ntawm cov khoom rho tawm yog qhov hloov pauv hauv lub neej ncua sijhawm, cov yam ntxwv morphological, thiab kev loj hlob ntawm hom algae.

3. Cov ntaub ntawv thiab cov txheej txheem 3.1.Cov ntaub ntawv

Icelandic seaweeds U.lactuca (ntsuab algae), A.esculenta (xim av algae), thiab P. palmitate (liab algae) tau muab los ntawm Icelandic Blue mussels thiab Seaweed, uas tau sau seaweeds hauv Breidafjordur (West-Iceland). Tom qab sau cov seaweeds tau qhuav (txog li 90 feem pua ​​​​ntawm cov khoom qhuav), milled, thiab xa lub tshuab nqus tsev ntim. Cov qauv raug khaws cia rau hauv qhov chaw qhuav thiab tsaus ntawm chav tsev kub kom txog thaum siv.

Tyrosinase los ntawm nceb, L-3, 4-dihydroxyphenylalanine (L-DOPA), elastase los ntawm porcine pancreas, ascorbic acid, N-Succinyl-Ala-Ala-Ala-p-nitroanilide (AAAPVN), hyaluronidase los ntawm bovine testes, quercetin, -tocopherol, tannic acid, 2, 2-diphenyl-1-picrylhydrazyl (DPPH), 24, 6-Tripyridyl-s-Triazine (TPTZ), Trolox, Folin-Ciocalteu reagent, gallic acid, thiab collagenase kev ua si colorimetric kuaj cov khoom (MAK293) tau yuav los ntawm Sigma-Aldrich Co.(St. Louis, MO, USA). Hyaluronic acid sodium ntsev tau yuav los ntawm MakingCosmetics (Redmond, WA, USA). Tag nrho lwm cov tshuaj thiab cov tshuaj reagents siv yog qib ntsuas thiab tau txais los ntawm VWR International, LLC. Deionized dej (ElixEssential, Merck, Darmstadt, Lub teb chaws Yelemees) tau siv rau kev rho tawm thiab npaj cov kev daws teeb meem dej.

3.2. Kev sim tsim

Ib qho kev tsim qauv tsim tau siv los ntsuas qhov cuam tshuam ntawm Icelandicseaweedspecies (U.lactuca, A.esculenta, P. palmata) thiab kev kho cov dej kub (HW, 95 degree C), PEF-pab extraction (PEF) thiab kev sib xyaw ua ke ntawm ob qho tib si cov tswv yim (PEF ntxiv rau HW), ntawm cov extract muaj pes tsawg leeg thiab bioactivity (Table 6). Cov extraction yog nqa tawm nyob rau hauv triplicate rau txhua pab pawg neeg thiab txhua txhua extract replicate yog soj ntsuam nyob rau hauv triplicate.

3.3. Extraction ntawm Bioactives los ntawm Icelandic Seaweeds

Kev siv cov macroalgal biomass nyob rau ntau theem tau txhawb cov kws tshawb fawb los tshawb nrhiav ntau dua eco-phooj ywg, siv tau zoo, thiab cov txiaj ntsig zoo extraction, raws li cov txheej txheem rho tawm ntsuab. Hauv kev ua haujlwm no, PEF-pab extraction tau raug soj ntsuam raws li txoj kev tshiab thiab ntsuab los tsim cov khoom siv tau zoo, thaum cov dej kub ib txwm siv los sib piv. Ntxiv mus, cov nyhuv ntawm kev sib xyaw ua ke ntawm ob txoj kev, PEF kev kho mob ntawm macroalgae ua raws li cov tsoos dej kub extraction, ntawm bioactive rov qab tau kawm. Vim tias qhov xav tau electroporation tsim nyob rau hauv cell membranes tom qab kev kho mob lub cev, cov nram qab no extraction nrog dej kub yuav ntxiv pab txhawb kev tso tawm ntawm intracellular cov ntaub ntawv [86], nce lub extraction yield. Ib lub sij hawm tom qab kev kho mob yog xav tau rau cov ntaub ntawv kom diffuse tawm ntawm lub hlwb [87,88], thiab nyob rau hauv qhov kev sim no, cov ncua kev kawm ntawv tos ib hmos kom txog rau thaum lub sib cais ntawm cov kua (extract) los ntawm lub pulp.

Hais txog kev rho tawm nruab nrab, cov dej distilled tau siv los tsim cov seaweed extracts kom kov yeej cov kev txwv txog kev siv tshuaj lom thiab cov kuab tshuaj organic. Dej tau ua pov thawj los ua cov kuab tshuaj zoo rau kev tshem tawm ntau cov tshuaj bioactive los ntawm seaweed [46,{1}}] thiab yog ib puag ncig zoo. Tsis tas li ntawd, dej feem ntau yog siv rau PEF-pab extraction vim nws yog ib tug zoo conductor rau hluav taws xob.

3.3.1. Cov txheej txheem rho tawm

Rau txhua qhov sib txawv hauv txhua pab pawg, seaweeds (15 g) tau soaked thaum hmo ntuj ntawm chav tsev kub (22 degree) hauv dej deionized (300 mL). Tom qab ntawd, qhov kev ncua raug kho nrog PEF (PEF), rhuab (HW), los yog ob qho tib si PEF-kho thiab rhuab (PEF ntxiv rau HW). Cov kev tshem tawm tau khaws cia rau hmo ntuj hauv lub tub yees tom qab los ntawm kev pom nrog cov ntawv ntxhib (20 μm) lim ntawv. Tom qab ntawd cov filtrates (extracts) tau khaws cia ntawm 4 degree kom txog rau thaum lawv cov kev tshuaj ntsuam.

Lub tshuab hluav taws xob hluav taws xob-pab extraction tau ua los ntawm kev siv lub tshuab hluav taws xob mem tes ua hauv tsev. Nws muaj FuGHCK-200-2000 capacitor (Fu.G.Elektronik GmbH, Rosenheim, Lub teb chaws Yelemees) thiab qhov sib txawv (18.5 kV OG75, Perkin-Elmer Optoelectronics, GMBH, Wiese-baden, Lub teb chaws Yelemees). Cov khoom siv PEF tau tsim cov khoom lag luam exponential decay pulses nrog qhov dav ntawm 0.96 peb thiab amplitude ntawm 18 kV. Lub plexiglass kho chamber nrog qhov ntev (L × H × W) 20 × 8 × 2.5 cm, nrog qhov luv tshaj plaws nyob nruab nrab ntawm cov phaj electrodes tau siv los kho cov kev ncua nrog 8 kV / cm hluav taws xob teb ntawm 1.2 Hz rau 10 min. HW-extracts tau npaj los ntawm cua sov lub ncua kev kawm ntawv nyob rau hauv ib tug beaker nyob rau hauv ib tug thermostatic da dej thiab khaws cia ntawm 95 degree rau 45 feeb. Rau kev sib xyaw ua ke ntawm cov hluav taws xob hluav taws xob thiab kev kho cua sov, cov kev ncua tau raug kho PEF thiab muab tso rau hauv lub beaker, rhuab hauv dej da dej, thiab khaws cia ntawm 95 degree rau 45 min.

3.3.2. Conductivity, pH, thiab ntsuas kub

Cov hluav taws xob conductivity thiab pH ntawm seaweed suspensions tau ntsuas tom qab soaking thiab tom qab kev kho mob extraction, nyob rau hauv chav tsev kub, siv ib tug pH-meter (Orion StarTM A215 pH / Conductivity Benchtop Meter, Thermo Scientific, Waltham, MA, USA) nruab nrog ib tug conductivity sensor thiab pH / ARC triode ua ke electrode. Tsis tas li ntawd, qhov kub thiab txias hloov vim kev kho mob raug kaw.

3.4. Spectral Profiles ntawm Seaweed Extracts

UV-VIS absorption spectra ntawm cov seaweed sib txawv tau ntsuas rau qhov ntau ntawm 200 txog 450 nm siv ob lub teeb Thermo Scientific Evolution 350 UV-Vis Spectrophotometer (Thermo Fisher Scientific, Waltham, MA, USA) nrog 1 cm quartz. Peb scans tau ua rau txhua seaweed extract. 3.5. Kev txiav txim siab ntawm Tag Nrho Cov Ntsiab Lus Polyphenolic

Tag nrho cov ntsiab lus phenolic (TPC) hauv seaweed extracts tau txiav txim siab los ntawm kev siv Folin-Ciocalteu reagent tom qab ib qho kev hloov kho me ntsis piav qhia los ntawm Zhang [92] siv Multiskan Sky Microplate Spectrophotometer (Thermo Fisher Scientific, Waltham, MA, USA). 20 μL ntawm seaweed extract lossis serial standard tov yog tov nrog 100 μL ntawm Folin-Ciocalteu reagent (10 feem pua ​​​​ntawm cov dej distilled). tau muab tso rau ntawm chav tsev kub thiab qhov tsaus ntuj rau 30 min. Kev nqus tau ntsuas ntawm lub wavelength ntawm 760 nm. Cov dej distilled tau siv los ua qhov tsis muaj, tus qauv nkhaus ntawm gallic acid yog siv los txiav txim siab tag nrho cov ntsiab lus phenolic thiab qhia tias yog ug ntawm gallic acid sib npaug. (GAE) ib gram ntawm cov khoom qhuav (ug GAE / g dw).

3.6. Kev txiav txim siab ntawm tag nrho cov ntsiab lus Flavonoid

Tag nrho cov ntsiab lus flavonoid (TFC) hauv seaweed extracts tau txiav txim siab los ntawm txoj kev piav qhia los ntawm Kamtekar 【93】 thiab hloov mus rau 96-zoo microplates. Luv luv, ib ntim ntawm 25 μL ntawm seaweed extract los yog serial txheem tov yog tov nrog 100 μL ntawm sodium nitrite (0.375 feem pua ​​w/v). Tom qab 5 min, 25 μL ntawm txhuas chloride (3 feem pua ​​w / v) tau ntxiv rau qhov sib tov thiab incubated rau 6 min ntawm chav tsev kub. Tom qab ntawd, 100 l ntawm sodium hydroxide (2 feem pua ​​​​w / v) tau ntxiv rau qhov sib tov thiab sib xyaw. Tam sim ntawd, absorbance tau ntsuas ntawm lub wavelength ntawm 510 nm. Cov dej distilled thiab ethanol tau siv ua qhov khoob. Ib qho qauv nkhaus ntawm quercetin (solved hauv ethanol) tau siv los txiav txim siab tag nrho cov ntsiab lus phenolic thiab qhia tias yog ug ntawm quercetin equivalents (QE) ib gram ntawm cov khoom qhuav (ug QE / g ua). 3.7. Kev txiav txim siab ntawm cov ntsiab lus Carbohydrate

Cov ntsiab lus ntawm cov piam thaj dawb tau ntsuas raws li txoj kev piav qhia los ntawm [94], nrog kev hloov kho me ntsis. Ib 50 μL ntawm phenol tov (4 feem pua) thiab 250 μL ntawm sulfuric acid (96 feem pua) tau ntxiv rau 100μL ntawm cov qauv lossis cov qauv tshuaj. Tom qab 10 min ntawm incubation ntawm chav tsev kub, lub absorbance ntawm cov sib tov tau nyeem ntawm 490 nm. Ib qho qauv nkhaus ntawm cov piam thaj tau siv los txiav txim siab tag nrho cov ntsiab lus carbohydrate thiab qhia tias yog mg ntawm cov piam thaj sib npaug (GluE) ib gram ntawm cov khoom qhuav (mg GluE / g dw).

3.8. Antioxidant Properties ntawm Seaweeds Extracts

3.8.1.2,2 Diphenyl{5}}picrylhydrazyl (DPPH) Dawb Radical Scavenging Assay

Kev ua haujlwm antioxidant (DPPH) ntawm seaweed extracts tau txiav txim siab raws li txoj kev piav qhia yav dhau los 【94】 nrog qee qhov kev hloov kho. Luv luv, 200 μL ntawm 10.825 ×10-5M DPPH tov tau ntxiv rau 100 μLof qauv (1∶1 hauv methanol) hauv 96- zoo phaj. Tib lub ntim ntawm DPPH tau tov nrog 50μL tus qauv ntxiv rau 50 μL methanol. Tom qab ntawd cov qauv thiab cov qauv tau muab tso rau hauv qhov chaw tsaus ntawm chav sov li 30 min. Lub absorbance yog ntsuas ntawm lub wavelength ntawm 517 nm. Cov dej distilled tau siv los ua qhov khoob. Lub peev xwm txhawm rau txhawm rau DPPH radical tau suav nrog kev sib npaug hauv qab no:

qhov twg A tswj yog qhov nqus ntawm kev tswj (DPPH tov tsis muaj qauv), tus qauv A yog qhov nqus ntawm cov qauv kuaj (DPPH tov ntxiv rau cov qauv kuaj), tus qauv A yog qhov nqus ntawm cov qauv nkaus xwb (piv txwv tsis muaj DPPH tov. ) thiab Amethanol dawb paug yog qhov nqus ntawm methanol nkaus xwb. Kev lag luam antioxidants (ascorbic acid, gallic acid, thiab -tocopherol) tau siv los ua kev tswj xyuas zoo.

3.8.2 ib. Ferric Ion Txo Antioxidant Power (FRAP) Kev Tshawb Fawb

Kev ua FRAP tau ntsuas raws li txoj kev ntawm Benzie thiab Strain[95]. Luv luv, acetate buffer (300 mM, pH3.6), 2,4, 6-tripyridyl-s-triazine (TPTZ) 10 mM hauv 40 mM HCl, thiab FeCl3·6H2O (20 mM) tau sib xyaw hauv qhov sib piv ntawm 10: 1: 1 kom tau txais kev ua haujlwm FRAP reagent. Cov tshuaj tiv thaiv sib xyaw yog incubated ntawm 37C rau 10 min. Ib qho piv txwv A50μL los ntawm txhua qhov extract tau tov nrog 150μL ntawm kev ua haujlwm FRAP tov rau 8 min ntawm chav tsev kub. Lub absorbance ntawm cov khoom xim, Ferrous-TPTZ tau ntsuas ntawm lub wavelength ntawm 593 nm. FRAP qhov tseem ceeb ntawm seaweeds rho tawm tau qhia raws li μM ntawm Trolox equivalents (TE) ib gram ntawm cov khoom qhuav.

3.8.3.2,2 Azino-bis(3-ethylbenzothiazoline-6-sulfonic Acid)(ABTS) Assay

Kev tsom xam tau ua tiav siv ABTS decolorization raws tu qauv [76] nrog qee qhov kev hloov kho. Ib qho ABTS radical cation (ABTS ntxiv) yog tsim los ntawm reacting ABTS (66 mg) nrog 10 mL ntawm poov tshuaj persulphate tov (2.45 mM).Cov sib tov tau tso rau hauv qhov tsaus ntuj ntawm chav tsev kub rau 12-16 h ua ntej siv. Cov tshuaj ABTS ntxiv tau diluted nrog dej mus rau qhov nqus ntawm 0.700 ntawm 734 nm. Cov tshuaj tiv thaiv sib xyaw (200 ul) raug xa mus rau lub microplate, 50 ul ntawm cov qauv ntxiv thiab tom qab ntawd 150 ul ntawm cov tshuaj reagent. Lub phaj tau shaken rau 10 s ntawm nruab nrab ceev, thiab lub absorbance yog ntsuas ntawm 734 nm tom qab 5 min ntawm incubation ntawm chav tsev kub. Ib qho qauv nkhaus tau npaj los ntawm kev npaj qhov inhibition ntawm A734nm ntawm Trolox qauv raws li kev ua haujlwm ntawm lawv cov concentrations. Tus nqi Trolox sib npaug antioxidant muaj peev xwm (TEAC) tus nqi ntawm cov qauv tau suav nrog qhov sib npaug tau los ntawm cov kab rov tav ntawm tus qauv nkhaus hloov ntawm A734nm qhov tseem ceeb rau txhua tus qauv:

Cov tshuaj tiv thaiv antioxidant tau qhia nyob rau hauv cov nqe lus ntawm TEACconcentration, umol / g qhuav hnyav algae.

3.9. Anti-Enzymatic Activities ntawm Seaweeds Extracts 3.9.1. Collagenase Inhibition Assay

Ib qho kev ua haujlwm collagenase colorimetric assay kit (MAK293), yuav los ntawm Sigma-Aldrich, tau siv los txiav txim qhov collagenase inhibition ntawm seaweeds extracts. Cov khoom siv ntsuas collagenase kev ua haujlwm siv hluavtaws peptide (FALGPA) uas ua raws li cov qauv collagen. Cov txheej txheem tau ua raws li cov lus qhia ntawm cov khoom siv.

3.9.2 ib. Elastase Inhibition Assay

Lub elastase inhibition ntawm seaweeds rho tawm tau tshawb xyuas hauv TRIS tsis kho nrog cov kev hloov kho raws li tau piav qhia ua ntej 【96】. Luv luv, 100μL ntawm 0.1 M TRISbuffer tov (pH8.0), 25 μL ntawm elastase (1 U / mL hauv TRISbuffer) thiab 25μL cov qauv rho tawm tau sib xyaw thiab ua rau 15 feeb ntawm 30 C ua ntej ntxiv cov substrate los pib cov tshuaj tiv thaiv. Tom qab lub sij hawm incubation, 50 uL ntawm 2 mM AAAPVN tov tau ntxiv. Tom qab ntawd, qhov absorbance ntawm 420nm tau saib xyuas rau 20 min siv lub microplate nyeem ntawv nyob rau hauv qhov kub thiab txias ntawm 30 C. Thaum kawg, elastase inhibition raug xam nyob rau hauv feem pua ​​​​ntawm kev siv qhov sib npaug:

qhov twg Abs tswj yog qhov absorbance ntawm qhov kev ntsuam xyuas siv qhov tsis hloov ntawm inhibitor (piv txwv) thiab Abssampleis lub absorbance ntawm cov qauv rho tawm. Quercetin tau siv los ua kev tswj xyuas zoo. Tris tsis yog siv los ua qhov khoob.

3.9.3 ib. Tyrosinase Inhibition Assay

Tyrosinase inhibitory assay tau ua raws li cov txheej txheem yav dhau los tau piav qhia los ntawm 【66】 siv L-DOPA ua substrate. Ib 20 μL ntawm cov qauv, 10 μL ntawm nceb tyrosinase tov (50 U / mL hauv phosphate tsis) thiab 80 μL ntawm phosphate tsis (pH =6.8) tau tov rau hauv microplate thiab pre-incubated ntawm 37 degree. rau 5 min. Tom qab ntawd, 90 μL ntawm L-DOPA (2 mg / mL) tau ntxiv. Qhov tsim ntawm dopachrome tau saib xyuas tam sim ntawd rau 20 min ntawm 475 nm hauv microplate nyeem ntawv hauv qab qhov kub thiab txias ntawm 37 degree. Qhov feem pua ​​inhibition ntawm tyrosinase enzyme tau suav nrog kev sib npaug:

qhov twg Abs tswj yog qhov absorbance ntawm qhov kev ntsuam xyuas siv qhov tsis hloov ntawm inhibitor (piv txwv) thiab Abs qauv yog qhov nqus ntawm cov qauv rho tawm. Quercetin tau siv los ua kev tswj xyuas zoo. Phosphate tsis yog siv los ua qhov khoob.

3.9.4 ib. Hyaluronidase Inhibition Assay

Hyaluronidase inhibitory kev ua haujlwm tau ntsuas raws li yav dhau los tau piav qhia los ntawm [66] nrog qee qhov kev hloov kho. Ib ntim ntawm 100 ulof type-1-Sbovine testes hyaluronidase (21{26}}0 U/mL) yaj hauv 0.1 M acetate buffer (pH 3.5) yog tov nrog 100 μL ntawm extract thiab incubated ntawm 37 degree rau 20 min. Ib lub ntim ntawm 200 μL ntawm 6mM ntawm calcium chloride tau ntxiv rau cov tshuaj tiv thaiv sib tov, thiab tom qab ntawd cov sib tov tau incubated ntawm 37 degree rau 20 min. Qhov Ca2 plus activated hyaluronidase no tau kho nrog 250 μL ntawm sodium hyaluronate (1.2 mg / mL) yaj hauv 0.1 M acetate tsis (pH 3.5) thiab tom qab ntawd incubated hauv dej da dej ntawm 37 degree rau 40min.A 50 μL ntawm sodium hydroxide. thiab 100 μL ntawm 0.2 M sodium borate tau ntxiv rau cov tshuaj tiv thaiv sib tov, thiab tom qab ntawd incubated nyob rau hauv ib lub boiling dej da dej rau 5 min. Tom qab txias rau chav sov, 250 μL ntawm p-dimethylaminobenzaldehvde (DAMB) tov tau ntxiv rau cov tshuaj tiv thaiv sib tov. Cov tshuaj DAMB tau npaj los ntawm dissolving 0.25 g ntawm DAMB hauv 21.88 mL ntawm 100 feem pua ​​​​acetic acid thiab 3.12 ml ntawm 10N hydrochloric acid. Cov pab pawg tswj tau raug kho nrog 100μL ntawm 5 feem pua ​​​​ntawm cov dej es tsis txhob rho tawm. Lub absorbance tau ntsuas ntawm lub wavelength ntawm 585 nm tom qab 45 min. Qhov feem pua ​​​​enzyme inhibition tau suav nrog kev sib npaug hauv qab no:

qhov twg Abs tswj yog qhov absorbance ntawm qhov kev ntsuam xyuas siv qhov tsis hloov ntawm inhibitor (piv txwv) thiab Abs qauv yog qhov nqus ntawm cov qauv rho tawm. Tannic acid yog siv los ua tus qauv siv.

3.10. Kev txheeb cais

Qhov nruab nrab ntawm kev tsom xam triplicate ntawm txhua qhov kev rho tawm tau suav thiab siv los nrhiav cov txiaj ntsig nruab nrab thiab cov qauv sib txawv rau txhua pawg (n=3). General linear qauv (GLM) rau cov khoom ruaj khov tau siv los ntsuas cov teebmeem tseem ceeb thiab kev sib cuam tshuam ntawm ob txoj hauv kev ntawm cov kev sim (hom thiab cov txheej txheem rho tawm) ntawm qhov ntsuas qhov sib txawv. Tsis tas li ntawd, ANOVA thiab Tukey-Kramer xeem tau siv los txheeb xyuas qhov tseem ceeb (p<0.05)differences between="" the="" groups.="" pearson="" correlation="" was="" used="" to="" evaluate="" the="" linear="" relationship="" between="" the="" variables.="" principal="" component="" analysis(pca)was="" used="" to="" detect="" structure="" in="" the="" relationship="" between="" measured="" variables="" and="" experimental="" factors.="" the="" pca="" reduces="" voluminous="" data="" to="" a="" small="" set="" of="" linear="" combinations="" of="" related="" variables="" (i.e.,="" factors)="" based="" on="" patterns="" of="" correlation="" among="" the="" original="" variables.="" the="" resulting="" linear="" attribute="" combinations="" can="" be="" used="" for="" profiling="" specific="" product="" characteristics="" based="" on="" the="" variables="" studied.="" all="" statistical="" analyses="" were="" performed="" using="" ncss="" 2020="" statistical="" software="" (2020)="" (ncss,="" llc.,="" kaysville,="" ut,="">

4. Cov lus xaus

Cov txiaj ntsig ntawm qhov kev ntsuam xyuas thawj zaug no tau pom muaj peev xwm ntawm peb hom Icelandic seaweed los ntawm kev muab cov txiaj ntsig zoo ntawm ntau txoj hauv kev. Txoj hauv kev ntsuab tau tsim los ntawm kev siv cov av hluav taws xob aqueous pulsed tau nthuav tawm cov txiaj ntsig zoo sib xws rau kev rho tawm cov dej kub ib txwm muaj, uas qhia ntau yam zoo xws li nws qhov tsis muaj cua sov thiab lub sijhawm rho tawm luv dua (10 min vs. 45 min). Ntawm peb hom algal, cov xim av macroalgae A.esculenta tau pom cov ntsiab lus siab tshaj plaws ntawm TPC thiab TFC kuj nthuav tawm qhov muaj peev xwm loj tshaj plaws antioxidant Ntxiv mus, A. esculenta dej extracts nthuav tawm cov haujlwm inhibitory zoo dua li P. palmaria thiab U. Lactuca ntawm collagenase, elastase, tyrosinase, thiab hyaluronidase yog cov feem ntau cog lus seaweed hom nrog zoo heev anti-enzymatic kev ua ub no rau lawv siv nyob rau hauv daim tawv nqaij dawb, anti-aging thiab daim tawv nqaij noj qab haus huv. Interestingly, A. esculenta extracts uas tsim los ntawm PEF txoj kev qhia ib tug collagenase inhibition ntawm 91 feem pua, siab tshaj qhov inhibition kev ua qhia los ntawm cov tsoos dej kub extraction thiab txawm siab tshaj qhov inhibitor muab los ntawm cov khoom siv coj mus muag. Hauv kev xaus, peb txoj kev tshawb fawb ua ntej qhia tias Icelandic seaweed-raws li extracts, tshwj xeeb tshaj yog cov extracts los ntawm cov xim av macroalgae A.esculenta, uas ua los ntawm aqueous pulsed hluav taws xob teb-pab extraction yog muaj peev xwm ua tau cov khoom xyaw uas yuav siv tau raws li active compounds rau cosmetic thiab cosmetic formulations. nyob rau yav tom ntej.

Kab lus no yog muab rho tawm los ntawm Mar. Tshuaj 2021, 19, 662. https://doi.org/10.3390/md19120662 https://www.mdpi.com/journal/marinedrugs