Kawm Txog Kev Txhim Kho Ntawm Kev Siv Tshuab Extraction Thiab Kev Ua Haujlwm Antioxidant Ntawm Polysaccharides Los Ntawm Yunnan Coffea Arabica Paj

Abstract: Kom optimize lub extraction technology thiab kawm covantioxidantkev ua ntawmpolysaccharideslos ntawm lub paj kas fes (ACP), ultrasonic kub, ultrasonic lub sij hawm, piv ntawm cov kua-khoom, ultrasonic zog, qauv immersion lub sij hawm thiab ntim feem pua ​​ntawm ethanol tau soj ntsuam los ntawm kev siv polysaccharide extraction yield raws li qhia. Tom qab ntawd ultrasonic kub, lub sij hawm ultrasonic thiab lub zog ultrasonic tau siv los ua feem ntau cuam tshuam, lub tshuab rho tawm tau zoo los ntawm cov lus teb nto.AntioxidantKev ua haujlwm ntawm ACP tau tshawb xyuas los ntawm DPPH · thiab ABTS ntxiv rau · kev tshem tawm cov teebmeem, thiab FRAP kev soj ntsuam. Cov txiaj ntsig tau pom tias qhov kev tshem tawm ultrasonic zoo tshaj plaws yog: Ultrasonic kub 69.5 degree, ultrasonic lub sij hawm 93 min, ultrasonic zog 175 W, piv ntawm cov kua-khoom 10: 1 mL / g, qauv immersion lub sij hawm 3{{ 17}} min, thiab ntim feem pua ​​​​ntawm ethanol 80 feem pua. Raws li cov txiaj ntsig ntawm polysaccharides yog 2.292 feem pua. Cov txiaj ntsig tau qhia tias tus nqi ntawm IC50 raws li DPPH · scavenging nyhuv ntawm ACP yog 3.844 mg·mL−1, ABTS plus · scavenging kev ua si yog 0.921 mmol Trolox/g ACP. FRAP qhov tseem ceeb ntawm ACP los ntawm FRAP kev soj ntsuam yog 0.0565 mmol Fe2 ntxiv / g ACP, uas qhia tias ACP tsis muaj zog.antioxidantkev ua si. Txoj kev tshawb no yuav muab lub hauv paus theoretical rau kev siv dav dav thiab kev loj hlob ntawm kas fes los ntawm cov khoom lag luam.

Ntsiab lus:kas fes pajpolysaccharide extraction;kev ua haujlwm antioxidant

Kas fesyog ib tsob nroj ntawm Rubiaceae (Coffea) genus, feem ntau faib nyob rau hauv lub teb chaws xws li South America, Central America, Africa thiab Asia, thiab yog zus nyob rau hauv ntau tshaj 80 lub teb chaws thoob ntiaj teb [1]. Raws li "Suav Materia Medica", kas fes muaj qhov cuam tshuam ntawm kev ua kom lub cev, diuretic thiab plab, thiab nws yog siv rau kev puas hlwb qaug zog thiab tsis qab los noj mov. Nws yog feem ntau siv raws li ib tug refreshing, diuretic thiab mob plab. Kev tshawb fawb niaj hnub no tau pom tias kas fes muaj ntau yam khoom xyaw xws li alkaloids, phenolic acids, flavonoids, thiab terpenes, uas muaj ntau yam tshuaj siv tshuaj xws li kev tiv thaiv lub siab, kev tiv thaiv neuroprotection,antioxidant, thiab antidiabetic [2-3]. kuv lub teb chawskas fesKev cog qoob loo yog ua los ntawm cov kas fes me me, thiab ntau dua 99 feem pua ​​​​ntawm nws tau muab faib rau hauv Yunnan. Yunnan me-grain kas fes yog nplua nuj nyob rau hauv cov tshuaj, ntxiv rau caffeine, chlorogenic acid, trigonelline thiab lwm yam khoom, nws kuj muaj ascarosides.

I~II[4], paniculoside VI[4], cofaryloside I[4], villanovane I[4], caffarolides A~H[5], caffruenol AB[6], caffruones AD[6] and caffruolide AB[7] thiab qee qhov tshiab terpenoids. Ntawm lawv, caffarolides C, D thiab F tau lees paub tias muaj qee yam kev ua haujlwm ntawm kev ua kom platelet aggregation hauv vitro [5]; caffruenol AB thiab caffruolide AB muaj cov nyhuv ntawm inhibiting lipopolysaccharide-induced NO ntau lawm hauv 264.7 macrophages [7]. Nrog rau kev kawm tob ntawm kas fes, tus nqi ntxiv ntawm kas fes tseem nce ntxiv. Nyob rau hauv xyoo tas los no, kas fes los ntawm cov khoom lag luam muaj nplua nuj nyob hauv phenolic acids, flavonoids, terpenes, alkaloids thiab lwm yam khoom siv roj ntsha.

Cov khoom xyaw nquag, uas tuaj yeem siv los ua cov khoom siv ntuj tsim thiab ruaj khov ntawm cov khoom xyaw nquag xws liantioxidants, kev tiv thaiv lub siab thiab kev tiv thaiv paj hlwb, tau ua rau kev tshawb fawb ntawm kas fes los ntawm cov khoom lag luam muaj kev txhawj xeeb ntau dua los ntawm cov kws tshawb fawb [8-10]. Campa et al. qhia tias nplooj kas fes muaj phenolic tebchaw [11]; Chen tau tshuaj xyuas cov khoom siv tshuaj lom neeg nplua nuj ntawm alkaloids, flavonoids, phenolic acids, terpenes, thiab lwm yam hauv kas fes nplooj thiab lawv cov haujlwm tshuaj xws li antioxidant, tiv thaiv kab mob, thiab tshuaj tua kab mob. [12], thiab kawm txog cov teebmeem ntawm kas fes nplooj ua txoj kev thiab nplooj hnub nyoog ntawm nws cov tshuaj muaj pes tsawg leeg thiab kev ua haujlwm [13].

Tsis tas li ntawd, Fu Xiaoping et al. [14-15] pom tias cov kua txiv hmab txiv ntoo ntawm Yunnan me me kas fes tev muaj qee yam kev tiv thaiv thiab rov ua haujlwm rau tib neeg lub cev nqaij daim tawv endothelial hlwb puas lawm, thiab tseem muaj peev xwm.antioxidantcov teebmeem, thiab pom tias lub ntsiab paj Cov cyanidins yog cyanidin-3-glucoside thiab cyanidin-3-rutinoside.

Cov paj kas fes feem ntau muab pov tseg raws li cov khoom lag luam loj hauv kev lag luam kas fes. Txawm li cas los xij, cov kev tshawb fawb uas twb muaj lawm tau pom tias paj kas fes yog nplua nuj nyob hauv cov tshuaj lom neeg. Stashenko thiab al. [16] siv GC-MS los tshuaj xyuas cov khoom tsis zoo thiab cov khoom tsis zoo hauv cov paj kas fes me me, thiab cov txiaj ntsig tau txiav txim siab tag nrho ntawm 150 lub tebchaw, nrog rau cov ntsiab lus ntawm n-pentadecane. Qhov siab tshaj plaws, ua raws li geraniol. Tsis tas li ntawd, Nguyen et al. [17] tau kawm txog cov khoom xyaw nquag hauv paj kas fes thiab pom tias paj kas fes muaj cov ntsiab lus siab ntawm phenolic tebchaw, yog li paj kas fes tuaj yeem siv los ua cov khoom siv raw rau kev tau txais ntuj.antioxidantcov khoom xyaw nquag. Tsis tas li ntawd, kas fes paj kuj muaj caffeine thiab trigonelline. Caffeine yog txuam nrog txo kev pheej hmoo ntawm cov kab mob neurodegenerative [18-19]

Trigonelline tuaj yeem tiv thaiv kev mob ntshav qab zib thiab lub raum puas, thiab tseem muaj txiaj ntsig ntawm kev kho cov kab mob neurodegenerative [20-21]. Pinheiro et al. [22] txheeb xyuas cov ntsiab lus ntawm plaub yam tseem ceeb ntawm trigonelline, chlorogenic acid, gallic acid thiab caffeine hauv kas fes paj nyob rau hauv ntau txoj kev ziab thiab rho tawm los ntawm HPLC, ntawm cov caffeine thiab trigonelline muaj cov ntsiab lus siab tshaj plaws; Kev ua haujlwm antioxidant tau soj ntsuam los ntawm ABTS thiab DPPH kev sim, uas tau lees paub tias kas fes paj muaj cov haujlwm antioxidant thiab tuaj yeem siv los ua cov khoom siv raw rau kev haus tshuaj yej. Tam sim no, muaj ob peb cov ntaub ntawv tshawb fawb txog paj kas fes, tab sis nws tuaj yeem pom los ntawm cov ntawv tshaj tawm uas twb muaj lawm tias paj kas fes muaj cov ntawv thov dav dav raws li qhov muaj peev xwm ntawm bioactive compounds.

Polysaccharidesyog macromolecular compounds uas muaj ntau tshaj 10 monosaccharides khi los ntawm glycosidic bonds, thiab dav pom nyob rau hauv cov tsiaj, nroj tsuag thiab kab mob. Polysaccharides yog structurally complex, nrog sib txawv conformations thiab txheeb ze molecular masses, raws li zoo raws li theem nrab cov qauv ntawm intrachain thiab interchain hydrogen bonds. Cov kev tshawb fawb niaj hnub no tau pom tias polysaccharides muaj cov tshuaj siv tshuaj xws li antioxidant [23-24], tiv thaiv kev laus [25], tiv thaiv kab mob [26], tiv thaiv kab mob [27], thiab tiv thaiv qog [28]. Kev ua haujlwm lom neeg ntawm polysaccharides muaj feem xyuam rau nws cov purity, tshuaj qauv, solubility, thiab lwm yam. Nyob rau hauv xyoo tas los no, cov khoom siv roj ntsha ntawm polysaccharides tau dhau los ua kev tshawb fawb hotspot ntawm cov tshuaj ntuj, thiab nws kuj yog ib txoj hauv kev tshawb nrhiav cov tshuaj tshiab thiab tsim cov khoom noj khoom haus. . Yog li ntawd, polysaccharides ua lub luag haujlwm tseem ceeb hauv kev kho mob thiab zaub mov. kuv lub teb chaws Yunnan yog qhov chaw tsim khoom tseem ceeb rau kev cog kas fes, thiab paj kas fes muaj peev xwm txhim kho tus nqi, tab sis muaj qee qhov kev tshawb fawb txog kev loj hlob ntawm Yunnan kas fes paj, thiab lub peev xwm ntawm kas fes paj tsis tau tapping. Yog li ntawd, daim ntawv no siv Yunnan me me-grain kas fes paj los ua cov khoom tshawb fawb los ua kev tshawb fawb ntawm nws cov active polysaccharides, aiming mus tshawb fawb tob txog kev siv cov nqi ntawm Yunnan me-grain kas fes.

Hauv daim ntawv no, lubpolysaccharideyield tau siv los ua qhov ntsuas ntsuas txhawm rau txhawm rau txhim kho cov txheej txheem polysaccharide extraction thiab antioxidant peev xwm ntawm kas fes paj sau los ntawm Baoshan City, Yunnan Province, muab cov ntaub ntawv yooj yim rau kev txhim kho ntxiv ntawm cov khoom siv roj ntsha polysaccharides. Thiab muab kev siv rau kev kawm Yunnan me nplej kas fes thiab txhim kho nws cov nqi ntxiv.

1 Cov ntaub ntawv thiab cov txheej txheem

1.1 Cov khoom siv thiab cov cuab yeej siv

Kas fespaj Baoshan City, Yunnan Province; Anhydrous Cawv Tianjin Chemical Reagent Co., Ltd.; Anthrone purity 98.0 feem pua ​​, Sinopharm Chemical Reagent Co., Ltd.; concentrated sulfuric acid, hydrochloric acid Chongqing Chuandong tshuaj Co., Ltd.; 1,1-diphenyl-2- Trinitrophenylhydrazine (DPPH), 2,2'-Diaza-bis-3-ethylbenzothiazoline{{10}}sulfonic acid (ABTS), 2, 4,6-Tripyridyltriazine (TPTZ), Rutin purity 98.0 feem pua ​​, Shanghai Ruiyong Biotechnology Co., Ltd.; dej-soluble vitamin E purity 98.0 feem pua, Hefei Bomei Biotechnology Co., Ltd.; ferric chloride hexahydrate analytically ntshiab, Western

Long Science Co., Ltd.; Potassium Persulfate Analytical Qib, Tianjin Damao Chemical Reagent Factory; PBS tsis, sodium acetate tsis- Xiamen Haibiao Technology Co., Ltd.

Nyem qhov no kom paub ntau ntxiv txog Rich inpolysaccharide

Cistanche instant kas fes khoom

FA2104N Electronic Balance, 722-Sspectrophotometer Shanghai Qinghua Technology Co., Ltd.; KQ-250DB Ultrasonic Instrument, SHZ-D (Ⅲ) Circulating Water Vacuum Pump Gongyi Yuhua Instrument Co., Ltd.; Xuman 1000Y Multifunctional Grinder Yongkang City Boou Hardware Khoom Co., Ltd.; 800 Fais Centrifuge Jintan Fuhua Instrument Co., Ltd.; VFD-3000 Lub tshuab nqus tsev nqus tsev vacuum Beijing Bo Yikang Experimental Instrument Co., Ltd.

1.2 Kev sim txoj kev

1.2.1 Extraction ntawm kas fes pajpolysaccharidesXa mus rau txoj kev ntawm Zheng Tingting li al. [29] nrog kev hloov kho. Cov paj kas fes sau los ntawm Baoshan City, Yunnan Province tau qhuav hauv qhov ntxoov ntxoo ntawm chav tsev kub, pulverized nrog lub pulverizer thiab dhau los ntawm 80-mesh sieve siv. Nyias 2.0 g ntawm kas fes paj hmoov, ntxiv 20.0 mL dej ntshiab, soak rau 30 feeb

Ultrasonic ntawm 100 W rau 30 min ntawm 40 degree, txias rau chav sov, thiab cov lim tau khaws cia tom qab nqus pom. Ethanol tau ntxiv rau cov lim mus rau qhov concentration ntawm 80 feem pua ​​​​rau nag lossis daus thiab tso cai rau sawv ntsug rau 12 teev. Tom qab centrifugation ntawm 4000 r / min rau 10 min, lub supernatant raug muab pov tseg, thiab precipitate tau yaj nyob rau hauv dej thiab lyophilized ntawm -80 degree kom tau crude polysaccharide.

Npaj 5 mg·mL−1 kas fes paj polysaccharide tov thiab tso tseg.

1.2.2 Kev npaj ntawm polysaccharide tus qauv nkhaus thiab kev txiav txim siab ntawm cov ntsiab lus polysaccharide hauv kas fes paj Cov piam thaj tus qauv nkhaus tau kos nrog siv rau txoj kev anthrone-sulfuric acid [30]. Npaj 0.0, 25. }, 150.0, thiab 200.0 ug/mL qabzib cov qauv kev daws teeb meem, feem. Ua kom raug pipette 1.00 mL ntawm cov txheej txheem cov piam thaj saum toj no nrog ntau qhov sib txawv, ntxiv 1.00 mL dej ntshiab, thiab muab tso rau hauv 25 mL stoppered test tube. Ntxiv 5.0 mL ntawm 2.1 mg·mL-1 anthrone-sulfuric acid tov, co zoo, txias nyob rau hauv ib tug dej khov da dej, kub nyob rau hauv boiling dej da dej rau 7 min, thiab sai sai txias rau chav tsev kub nyob rau hauv ib tug dej khov da dej. Siv cov dej deionized ua kev tswj tsis pub muaj,

Absorbance A tau ntsuas colorimetrically ntawm 625 nm. Noj cov ntsiab lus anhydrous qabzib raws li abscissa (0.0, 25.0, 50.0, 1{{10 }}}0.0, 150.0, 200.0 ug), thiab tus nqi absorbance raws li kev tswj hwm, kos tus qauv nkhaus, thiab qhov sib npaug ntawm tus qauv nkhaus yog: Y=0.0051X−0.0092 ( R2=0.9970) (qhov twg: Y yog tus nqi nqus, X yog cov piam thaj, ug).

Precisely pipette ib lub ntim ntawm 5 mg / mL kas fes paj polysaccharide tov npaj saum toj no rau hauv 25 mL xeem raj nrog ib tug stopper, thiab ntxiv dej ntshiab kom txog li 2.00 mL. Ntxiv 5.0 mL ntawm 2.1 mg·mL−1 anthrone-sulfuric acid tov, co zoo, txias nyob rau hauv ib tug ice-dej da dej, tshav kub nyob rau hauv boiling dej da dej rau 7 min, thiab sai sai txias rau chav tsev kub nyob rau hauv lub chav tsev kub. da dej dej khov. Absorbance A tau ntsuas colorimetrically ntawm 625 nm siv dej deionized raws li kev tswj tsis pub muaj. Lub paj kas fes polysaccharide yield raug xam raws li cov piam thaj tus qauv nkhaus sib npaug, thiab txhua tus qauv tau rov ua dua 3 zaug. Cov txiaj ntsig tau nthuav tawm raws li tus nqi nruab nrab, thiab cov qauv kev suav tau raws li hauv qab no:

Kas fes paj polysaccharide tawm los (%) =X × m1 × 10−35 × V × ms × 100

Hauv cov mis: X yog cov ntsiab lus polysaccharide hauv V ntim ntawm kas fes paj polysaccharide tov, ug; V yog qhov ntsuas ntim ntawm cov tshuaj polysaccharide, mL; 5 yog qhov concentration ntawm cov tshuaj kas fes paj polysaccharide npaj, 5 mg·mL−1; m1 yog lub paj kas fes khov-qhuav tag nrho ntawm polysaccharide, g; ms yog qhov loj ntawm kas fes paj qauv, g.

1.2.3 Ib-qhov kev sim

Nyob rau hauv tus txheej txheem ntawm ultrasonic-pab extraction ntawm kas fes paj polysaccharide, lub tseem ceeb yam tseem ceeb cuam tshuam lub yield ntawm polysaccharide mas muaj xws li ultrasonic kub, ultrasonic sij hawm, kua-khoom siv piv, ultrasonic zog, soaking lub sij hawm thiab dej cawv los nag concentration. Tsib theem ntawm ultrasonic kub tau xaiv: 40, 50, 60, 70, thiab 80 degree; tsib theem ntawm ultrasonic lub sij hawm raug xaiv: 30, 60, 90, 120, thiab 150 min; 1, 30: 1 mL / g tsib theem; ultrasonic zog xaiv tsib theem ntawm 100, 125, 150, 175, 200 W; immersion lub sij hawm xaiv tsib theem ntawm 30, 60, 90, 120, 150 min; ethanol concentration xaiv tsib theem 75 feem pua, 80 feem pua, 85 feem pua, 90 feem pua, 95 feem pua ​​tsib theem, feem, ib qho kev sim. Thaum lim rau ib qho ntawm cov kev txwv

, lwm yam yog: ultrasonic kub 40 degree, ultrasonic lub sij hawm 30 min,

Ua kua rau cov khoom sib piv 10: 1 mL / g, ultrasonic zog 100 W, soaking sij hawm 30 min

thiab dej cawv nag lossis daus concentration ntawm 80 feem pua.

1.2.4 Teb nto optimization xyaum ua raws li lub Box-Benhnken sim tsim txoj cai, nrog rau cov tawm los ntawm kas fes paj polysaccharide raws li cov lus teb kuj sib txawv, peb yam uas muaj lub zog loj tshaj plaws ntawm cov qoob loo ntawm kas fes paj polysaccharide raug xaiv los ntawm ib qho tseem ceeb. kuaj tau, raws li qhia nyob rau hauv Table 1. Optimization ntawm ultrasonic kub nrog cov tawm los ntawm kas fes paj polysaccharide raws li ib tug Performance index degree, ultrasonic lub sij hawm thiab ultrasonic zog.

1.2.5 Antioxidant muaj peev xwm kuaj

1.2.5.1 DPPH dawb radical scavenging thwmsim Lub DPPH dawb radical scavenging thwmsim tau ua raws li piav nyob rau hauv Ref. [31]. Noj 3.9 mL ntawm 0. Cov tshuaj tiv thaiv tau nqa tawm ntawm chav tsev kub rau 30 min hauv qhov tsaus ntuj, thiab qhov ntsuas qhov nqus tau ntsuas ntawm 515 nm. Siv rutin raws li kev tswj kom zoo, DPPH dawb radical scavenging tus nqi raug xam raws li: I feem pua ​​=[(A0–As)/A0] × 100 (qhov twg: Raws li yog qhov absorbance ntawm cov qauv tov; A0 yog qhov absorbance ntawm cov tshuaj tsis muaj qauv), thiab cov kev ua antioxidant yog qhia raws li 50 feem pua ​​inhibition tus nqi (IC50).

1.2.5.2 ABTS ntxiv rau kev sim dawb radical scavenging

ABTS plus radical scavenging thwmsim tau ua siv txoj kev piav qhia hauv Ref. [32]. Noj 2 mL ntawm cov tshuaj polysaccharide thiab ntxiv rau 2 mL ntawm ABTS ntxiv rau cov tshuaj dawb radical raws li, tom qab sib xyaw ua ke, hnov ​​​​mob hauv chav sov li 6 min, thiab ntsuas qhov nqus UV ntawm 734 nm, thiab rutin yog qhov zoo tswj. Cov mis laij lej ntawm ABTS ntxiv rau cov dawb radical scavenging muaj peev xwm yog raws li nram no: I(%)=[(A0–As)/A0] ×10{ {12}} (qhov twg Raws li yog qhov nqus ntawm cov qauv kev daws teeb meem; A0 yog qhov nqus ntawm cov tshuaj tsis muaj qauv) Tus qauv nkhaus yog txiav txim siab los ntawm kev ntsuas Drawing ntawm Trolox tus qauv kev daws teeb meem nrog ntau qhov sib txawv (I feem pua ​​=0.0247C −0.0046, R2=0.9937), the ABTS anti-

Oxidative kev ua haujlwm yog qhia raws li mmol Trolox/g.

1.2.5.3 FRAP txoj kev

Txoj kev tau piav qhia hauv Ref. [33] tau siv rau kev txiav txim siab ntawm FRAP antioxidant muaj peev xwm. Noj 5.0 mL ntawm TPTZ, 5.0 mL ntawm 20 mmol/L FeCl3 thiab 50 mL ntawm sodium acetate buffer solution (300

mmol / L, pH 3.6) los npaj FRAP ua haujlwm daws; 100 μL qauv yog tov nrog 300 μL dej thiab 3.0 mL FRAP ua hauj lwm tov, muab tso rau hauv ib da dej ntawm 37 degree rau 30 min; Lub absorbance yog ntsuas ntawm 595 nm. Tus qauv nkhaus tau npaj nrog FeSO4 raws li cov khoom siv txheem (A=0.572C0.008, R2=0.9974), thiab rutin tau siv los ua qhov zoo tswj, raws li tus qauv nkhaus

Xam qhov txo hwj chim hauv mmol FeSO4/g polysaccharide.

1.3 Kev ua cov ntaub ntawv

Txhua qhov kev sim tau rov ua dua peb zaug thiab qhov nruab nrab tus nqi raug coj mus. DesignExpert 8.

2 Cov txiaj ntsig thiab kev txheeb xyuas

2.1 Cov txiaj ntsig kev sim ib leeg

Cov txiaj ntsig ntawm qhov kev sim ib zaug tau pom hauv daim duab. Cov txiaj ntsig ntawm qhov ntsuas kub ntawm ultrasonic ntawm cov qoob loo ntawm kas fes paj polysaccharide: qhov ntsuas kub ntawm ultrasonic yog 40-80 degree , thiab polysaccharide yield yog 1.0048 feem pua ​​-1.7982 feem pua. Hauv qhov ntau ntawm 40-70 degree, cov txiaj ntsig ntawm kas fes paj polysaccharide maj mam nce nrog qhov nce ntawm ultrasonic kub, mus txog qhov siab tshaj plaws ntawm 70 degree , thiab pib txo qis tom qab 70 degree . Qhov no tej zaum yuav yog vim qhov txo qis polysaccharide tawm los vim kev puas tsuaj ntawm cov qauv ntawm kas fes paj polysaccharides nyob rau hauv qhov kub thiab txias, uas tau zoo sib xws nyob rau hauv cov ntaub ntawv [29,34-35]. Qhov kub sonication tau xaiv los ua 70 degree .

Cov nyhuv ntawm ultrasonic lub sij hawm ntawm yield ntawm kas fes paj polysaccharide: lub sij hawm ultrasonic yog 30-150 min, lub polysaccharide yield yog 1.0369 feem pua ​​-1.5853 feem pua, cov polysaccharide yield tau nce nrog nce lub sij hawm ultrasonic, thiab mus txog qhov siab tshaj plaws ntawm 90 min, , cov tawm los pib txo nrog qhov nce ntawm sonication lub sij hawm. Qhov no yog vim lub sij hawm luv luv ultrasonic extraction tsis haum rau tag nrho cov dissolution ntawm polysaccharides, thaum lub sij hawm ntev ultrasonic extraction yuav degrade polysaccharides thiab ua rau ib tug txo nyob rau hauv yield, uas kuj qhia nyob rau hauv cov ntaub ntawv [29,34-35]. Yog li ntawd, lub sij hawm sonication raug xaiv los ua 90 min.

Cov nyhuv ntawm cov kua-kua piv ntawm cov txiaj ntsig ntawm kas fes paj polysaccharide: cov nyhuv ntawm cov kua-kua piv ntawm cov txiaj ntsig polysaccharide yog me me. Cov txiaj ntsig ntawm polysaccharide tau nce ntxiv nrog qhov nce ntawm cov kua-rau-cov khoom sib piv, thiab mus txog qhov siab tshaj plaws ntawm 25: 1 mL / g. Tom qab 25: 1 mL / g, cov txiaj ntsig tau txo qis nrog qhov nce ntawm cov kua-rau-cov khoom sib piv. Cov kuab tshuaj tsawg dua yuav ua rau tsis txaus ntawm cov kuab tshuaj polysaccharide, uas ua rau qis qis ntawm polysaccharide; ntau cov kuab tshuaj yuav yaj polysaccharide thiab ua rau nws nyuaj rau precipitate tawm, thiab nyob rau tib lub sij hawm, cov tawm los yuav raug txo vim yog nqus ntawm ultrasonic hluav taws xob los ntawm cov hnyav. 29,34–35] kuj muaj cov lus qhia zoo sib xws. Xav tias qhov sib piv ntawm cov kua-rau-cov ntaub ntawv muaj qhov cuam tshuam me ntsis ntawm cov khoom lag luam, thiaj li txuag tau tus nqi ntawm cov reagents, cov kua-rau-cov ntaub ntawv piv tau raug xaiv li 10: 1 mL / g.

Cov nyhuv ntawm lub zog ultrasonic ntawm cov txiaj ntsig ntawm kas fes paj polysaccharide: lub zog ultrasonic raug xaiv raws li 100-200 W, thiab cov txiaj ntsig polysaccharide yog 1.1185 feem pua ​​-1.8583 feem pua. Tom qab W, qhov tawm los poob nrog qhov nce ntawm lub zog ultrasonic. Qhov nce ntawm lub zog ultrasonic tuaj yeem ua kom puas hlwb thiab cov ntaub so ntswg kom yaj polysaccharides hauv cov kuab tshuaj, yog li ua kom lub zog ultrasonic muaj txiaj ntsig zoo rau nag lossis daus ntawm polysaccharides; Txawm li cas los xij, cov nyhuv fragmentation thiab thermal nyhuv uas ua los ntawm cov ultrasonic tsis loj dua yuav ua rau kom qhov kev sib cais ntawm impurities hauv kas fes paj. , cov nyhuv thermal yuav rhuav tshem cov khoom siv polysaccharide thiab ua rau cov txiaj ntsig ntawm polysaccharide txo qis, uas kuj tau tshaj tawm hauv cov ntaub ntawv [36-37]. Yog li ntawd, lub zog ultrasonic raug xaiv los ua 175 W.

Cov nyhuv ntawm lub sij hawm soaking ntawm cov qoob loo ntawm kas fes paj polysaccharide: cov nyhuv ntawm soaking lub sij hawm ntawm yield ntawm polysaccharide yog me me, lub sij hawm soaking yog 30-150 min, thiab cov polysaccharide yield yog 1.1827 feem pua ​​-1. 4609% ua. Hauv qhov ntau ntawm 30-90 min, cov txiaj ntsig polysaccharide tau nce ntxiv nrog rau qhov nce ntawm lub sijhawm so, thiab mus txog qhov siab tshaj plaws ntawm 90 min. Tom qab 90 min, nrog rau qhov nce ntawm lub sij hawm soaking, cov tawm los txo me ntsis thiab tended kom tiaj tus. Ntev lub sij hawm ntub dej tuaj yeem pab txhawb cov nag lossis daus ntawm polysaccharides thaum lub sij hawm ultrasonication thiab txo lub zog noj. Tab sis kev ntub dej ntev dhau tsis tuaj yeem ua rau muaj txiaj ntsig ntau dua, thiab kev ntub dej ntev dhau kuj tseem ua rau lwm cov khoom tso tawm thiab cuam tshuam rau polysaccharide yield. Qhov no zoo ib yam li tau tshaj tawm hauv [38]. Xav tias qhov cuam tshuam ntawm lub sij hawm soaking me me, thiaj li yuav txuag tau lub sij hawm, lub sij hawm soaking tau xaiv li 30 min.

Cov nyhuv ntawm dej nag lossis daus concentration ntawm kas fes paj polysaccharide yield: cawv nag lossis daus concentration muaj me ntsis cuam tshuam rau polysaccharide yield, cawv nag lossis daus concentration yog 75 feem pua ​​~ 95 feem pua, polysaccharide yield yog 0.9703 feem pua ​​~ 1.2806 feem pua. Cov txiaj ntsig ntawm polysaccharide nce nrog qhov nce ntawm ethanol concentration, thiab mus txog qhov siab tshaj plaws ntawm 85 feem pua. Tom qab 85 feem pua, cov txiaj ntsig tau poob nrog qhov nce ntawm ethanol concentration. Kev rho tawm dej thiab dej cawv los nag yog siv cov polysaccharide insoluble hauv cawv kom nws tawm los. Thaum tus nqi ntawm ethanol ntxiv nce, cov polysaccharide yog insoluble hauv ethanol thiab precipitates tawm, thiab tawm los nce. Thaum cov dej cawv nag lossis daus ntau tshaj 85 feem pua, cov txiaj ntsig ntawm polysaccharide tsis tuaj yeem txhim kho, tab sis cov tshuaj reagents yog nkim. Qhov no zoo ib yam li tau tshaj tawm hauv [38]. Txhawm rau ua kom yooj yim rau kev ua haujlwm, daim ntawv no siv txoj hauv kev ncaj qha ntxiv ethanol los kho cov cawv concentration rau nag lossis daus. Nyob rau tib lub sijhawm, txij li cov txiaj ntsig ntawm 80 feem pua ​​​​thiab 85 feem pua ​​​​polysaccharide tsis txawv ntau, nws tuaj yeem txuag cov reagents thiab txo cov khib nyiab. Yog li, cov dej cawv nag lossis daus concentration tau raug xaiv los ua 80 feem pua.

2.2 Teb qhov ntsuas qhov ntsuas

2.2.1 Teb qhov ntsuas qhov ntsuas kub Ultrasonic, lub sijhawm ultrasonic thiab lub zog ultrasonic muaj kev cuam tshuam zoo. Yog li ntawd, nyob rau hauv lub hauv paus ntawm cov saum toj no ib qho kev sim, cov lus teb nto txoj kev yog optimized rau peb tej yam kev mob ntawm ultrasonic kub, ultrasonic lub sij hawm thiab ultrasonic zog. Cov txiaj ntsig tau qhia hauv Table 2..

Noj cov polysaccharide yield (Y) raws li cov lus teb Performance index, ib tug regression qauv tau tsim nrog peb yam ntawm ultrasonic kub, ultrasonic lub sij hawm thiab ultrasonic zog, thiab lub quadratic regression sib npaug tau txais:

Y{{0}}.29−0.067A plus 0.{{10}}54B−{14}}. 0.34 AB + 0.083 AC + 0.011 BC - 0.40A2

−0.19B2−0.27C2

2.2.2 Kev ntsuas qhov tseem ceeb ntawm qhov sib txawv

Cov txiaj ntsig kev xeem tau pom nyob rau hauv Table 3.

Raws li cov txiaj ntsig ntawm kev txheeb xyuas qhov sib txawv hauv Table 3, tag nrho cov qauv yog qhov tseem ceeb (P<0.0001), and the model reached a very significant level, indicating that the difference between different factors was significant; according to the absolute value of the linear coefficient of the regression equation, it can be seen that each factor has a significant effect on the total polysaccharide yield. The order of influence is: A>B>C, that is, ultrasonic temperature> ultrasonic time> ultrasonic power. Lack of fit item P=0.5764>0.05, qhov tsis zoo ntawm cov khoom xeem tsis yog qhov tseem ceeb, qhia tias yam tsis paub muaj feem cuam tshuam me ntsis rau cov txiaj ntsig kev xeem, thiab cov khoom seem feem ntau yog tshwm sim los ntawm qhov yuam kev random, qhia tias kev xaiv tus qauv tsim nyog thiab raug. . Lub zog ntawm AB tseem ceeb heev (P<0.05), and the influence of A2, B2, and C2 was extremely significant (P<0.01). In the whole model, the adjustment coefficient R2Adj=0.9277 in the model, indicating that 92.77% of the response value changes can be carried out through the model. Explanation, the coefficient of determination R2 = 0.9684, indicating that the model is highly reliable, and the model fits well with the experiment, and this model can be used for analysis and prediction [39−42].

2.2.3 Teb nto thiab contours

Cov lus teb saum npoo daim duab ntawm kev sib cuam tshuam ntawm ntau yam ntawm cov txiaj ntsig ntawm kas fes paj polysaccharide yog qhia hauv daim duab 2. Kev sib cuam tshuam ntawm ultrasonic kub thiab lub sij hawm ultrasonic pom tias kev sib cuam tshuam ntawm ob yog qhov tseem ceeb; Thaum lub ntsuas kub ntawm ultrasonic tseem tsis hloov, cov txiaj ntsig ntawm kas fes paj polysaccharides thawj zaug nce thiab tom qab ntawd txo qis nrog qhov nce ntawm lub sijhawm ultrasonic; thaum lub sij hawm ultrasonic tseem tsis hloov, kas fes Cov txiaj ntsig ntawm paj polysaccharide thawj zaug nce thiab tom qab ntawd txo qis nrog qhov nce ntawm ultrasonic kub. Los ntawm kev sib cuam tshuam ntawm ultrasonic kub thiab lub zog ultrasonic, nws tuaj yeem pom tias thaum lub ntsuas kub ntawm ultrasonic tsis tas li, cov txiaj ntsig ntawm kas fes paj polysaccharide thawj zaug nce thiab tom qab ntawd txo qis nrog qhov nce ntawm lub zog ultrasonic; Thaum lub zog ultrasonic tseem tsis hloov pauv, cov txiaj ntsig ntawm kas fes paj polysaccharide nce nrog lub ultrasonic yoj. Qhov kub nce ua ntej nce thiab tom qab ntawd txo qis. Los ntawm kev sib cuam tshuam ntawm lub sij hawm ultrasonic thiab lub zog ultrasonic, nws tuaj yeem pom tau tias thaum lub sij hawm ultrasonic yog tas li, cov txiaj ntsig ntawm kas fes paj polysaccharide thawj zaug nce thiab tom qab ntawd txo qis nrog qhov nce ntawm lub zog ultrasonic; Thaum lub zog ultrasonic tsis tu ncua, cov txiaj ntsig ntawm kas fes paj polysaccharide nce nrog qhov nce ntawm lub zog ultrasonic. Lub sij hawm nce ua ntej thiab tom qab ntawd txo.

Yog li ntawd, siv cov polysaccharide yield raws li tus qauv kev ntsuam xyuas, qhov optimization ntawm cov lus teb nto txoj kev rau peb tej yam kev mob ntawm lub sij hawm ultrasonic, ultrasonic kub thiab ultrasonic zog yog: ultrasonic kub 69.56 degree, ultrasonic lub sij hawm 92.99 min thiab ultrasonic zog 174.01 W, nws kwv yees tias nyob rau hauv cov xwm txheej no 2.290 feem pua. Raws li qhov teeb meem tiag tiag, qhov kub ntawm ultrasonic ntawm 69.5 degree, lub sij hawm ultrasonic ntawm 93. 1 mL / g thiab ethanol concentration ntawm 80 feem pua ​​​​tau raug xaiv rau 4 qhov kev xeem sib npaug.

Qhov nruab nrab tawm los yog 2.292 feem pua ​​±0.061 feem pua ​​. Nws yog qhov pib ze rau qhov kev ntsuas qhov ntsuas tau los ntawm qhov kev sim, qhia tias muaj qhov sib haum zoo ntawm tus nqi kwv yees thiab tus nqi tiag tiag, yog li cov txheej txheem optimized tsis tau txais los ntawm qhov chaw teb hauv qhov kev tshawb fawb no yog qhov tseeb thiab ntseeg tau [43].

2.3 Kev sim cov txiaj ntsig ntawm antioxidant muaj peev xwm

DPPH xeem yog ib qho kev ntsuam xyuas zoo thiab rhiab rau cov nroj tsuag antioxidant muaj peev xwm. Lub peev xwm dawb radical scavenging ntawm cov qauv kuaj yog muaj feem xyuam rau nws cov peev xwm proton-dawb peev; ABTS xeem yog dav siv los kwv yees lub peev xwm antioxidant ntawm cov qauv cog, uas tuaj yeem kuaj cov qauv Antioxidant kev ua ntawm lipophilic thiab hydrophilic Cheebtsam hauv FRAP txoj kev; Lub peev xwm txo qis ntawm cov khoom ntuj raug ntsuas los ntawm kev txo Fe3 ntxiv -TPTZ rau Fe2 ntxiv -TPTZ [44−45]. Cov txiaj ntsig ntawm kev sim tshuaj tiv thaiv kab mob ntawm kas fes paj polysaccharides tau pom nyob rau hauv Table 4. Kas fes paj polysaccharides muaj qee yam kev ua haujlwm antioxidant tiv thaiv DPPH dawb radicals thiab ABTS ntxiv rau cov dawb radicals, tab sis lawv cov tshuaj tiv thaiv antioxidant qis dua li ntawm rutin.

3 Kev xaus

Hauv qhov kev sim no, Yunnan me-grain kas fes paj tau siv los ua raw khoom, thiab covpolysaccharidentawm Yunnan me-grainkas fespaj tau muab rho tawm los ntawm ultrasound. Nws tau pom tias lub sijhawm ultrasonic, qhov kub thiab txias ultrasonic muaj qhov cuam tshuam tseem ceeb ntawm kev rho tawm ntawm kas fes paj polysaccharides. Tom qab ntawd, lub sijhawm ultrasonic, qhov kub thiab txias ultrasonic tau ua kom zoo dua los ntawm qhov chaw teb, thiab cov txheej txheem zoo ntawm kas fes paj polysaccharide tau txiav txim siab raws li hauv qab no: ultrasonic kub 69.5 degree, ultrasonic lub sij hawm 93 min, ultrasonic zog 175 W, kua-khoom siv piv. 10: 1 mL / g, Lub sij hawm soaking yog 30 min, thiab ethanol concentration yog 80 feem pua. Raws li qhov xwm txheej no, cov txiaj ntsig ntawm polysaccharide yog 2.292% ± 0.061 feem pua. Txoj kev tuaj yeem txhim kho cov txiaj ntsig ntawm kas fes pajpolysaccharide, thaum shortening lub sij hawm rho tawm thiab txo tus nqi ntawm ethanol siv. Cov txiaj ntsig ntawm kev sim tshuaj tua kab mob antioxidant tau pom tias kas fes paj polysaccharides pom muaj peev xwm antioxidant tsis muaj zog. Txoj kev tshawb no yuav muab cov ntaub ntawv siv rau kev sib cais thiab kev ua kom huv ntawm kas fes pajpolysaccharidethiab kev tshawb fawb txog nws txoj haujlwm thiab kev ua haujlwm, thiab tseem yuav muab cov ntsiab lus theoretical thiab kev txhawb nqa rau kev txhim kho ntxiv thiab kev siv kas fes.

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