Cheebtsam Analysis Thiab Antioxidant Activity ntawm Polysaccharides Los ntawm Cistanche Deserticola

Lus Cim: Hauv daim ntawv no, polysaccharides tau muab rho tawm los ntawmCistanche deserticola, Cistanche deserticolacrudepolysaccharideCLP yog purified los ntawm DEAE cellulose kem chromatography, thiab nruab nrab qab zib CLPand acidic qab zib CLP2 ntawm Cistanche deserticola yog tau, Cov ntsiab lus ntawm tag nrho cov suab thaj, uronic acid thiab protein nyob rau hauvCistanche deserticola crude polysaccharideCLP, Cistanche deserticola nruab nrab qab zib CLPl thiabCistanche deserticola acidic qab zibCLP2 yog txiav txim los ntawm UV spectral scanning, thiab qhov hnyav molecular thiab monosaccharide muaj pes tsawg leeg ntawm polysaccharides raug txiav txim. Nyob rau tib lub sijhawm, DPPH radical scavenging muaj peev xwm thiab tag nrho cov antioxidant muaj peev xwm ntawm Cistanche deserticola crude polysaccharide CLP, Cistanche deserticola nruab nrab qab zib CLP1 thiab Cistanche deserticola acidic qab zib Cl.P2 raug soj ntsuam. Cov ntaub ntawv ntsuas tau qhia tias cov khoom ntawm Cistanche deserticola crud polysaccharide CLP, Cistanche deserticola nruab nrab qab zib CLP1 thiab Cistanche deserticola acidic qab zib CLP2 zoo ib yam, tab sis muaj qhov sib txawv loj hauv lawv cov ntsiab lus, Ntau qhov elution peaks tuaj yeem tau txais thaum Cistanche deserticola nruab nrab qab zib CLP1 thiab Cistanche. deserticola acidic qab zib CLP2 yog sib cais thiab purified los ntawm DEAE cellulose chromatographic kem, thiab molecular phaus faib yog 0~10 000 Da, qhia tias lub ntsiab Cheebtsam ntawm Cistanche deserticola polysaccharides yog me me molecular qab zib. Kev txiav txim siab monosaccharide tshwm sim ntawm Cistanche deserticola nruab nrab qab zib CLPl qhia tias monosaccharides hauv Cistanche deserticola nruab nrab qab zib CLP1 feem ntau yog mannose, galacturonic acid, qabzib, galactose, thiab arabinose, Qhov kev txiav txim siab monosaccharides ntawm Cistanche deserticola monosaccharides qhia tias cistanche deserticolaic acids monosaccharides qhia tau hais tias cov monosaccharides tshwm sim ntawm Cistanche deserticolatic acids. acidic qab zib ClP2 feem ntau yog rhamnose, galacturonic acid, qabzib thiab galactose, Cov txiaj ntsig ntawm DPPH radical scavenging sim thiab tag nrho cov antioxidant muaj peev xwm sim ntawm polysaccharides los ntawm Cistanche deserticola qhia tias cov antioxidant muaj peev xwm ntawm Cistanche deserticola nruab nrab qab zibCLPl yog siab dua li ntawm Cistanche deserticola crude. CLP thiab Cistanche deserticola acidic qab zib CLP2.

Ntsiab lus:Cistanche deserticola; polysaccharides; kev tshuaj ntsuam xyuas;kev ua haujlwm antioxidant; absorbance.

Tau txais Cistanche Tubulosa Extract Rau Lub Hom Phiaj Zoo

Cistancheyog ib tsob nroj herbaceous parasitic uas muaj coob tusphenolic glycosidessbiosynthesis,phenylethanoid glycosides, qab zib, thiab cawv. Cov glycans feem ntau yog nyob rau hauv daim ntawv ntawm polysaccharides. Cistanche polysaccharide muaj zog tiv thaiv kab mob ua haujlwm tswj kev ua haujlwm [1-2].

Raws li cov metabolites nruab nrab hauv tib neeg lub neej kev ua ub no, cov dawb radicals nyob rau hauv lub xeev tsis ruaj khov thiab muaj zog ntawm oxidative kev ua si, uas yuav ua rau lom macromolecules, yog li ntawd, kev siv cov natural antioxidant yog li ntawd, kev siv tej yam ntuj tso tshuaj tua kab mob kom tshem tawm cov dawb radicals. ua lub ntsiab lus tseem ceeb ntawm kev tshawb fawb txog zaub mov tam sim no. Cov ntsiab lus ntawm polysaccharideCistanchemuaj zog antioxidant muaj peev xwm, uas tuaj yeem txhawb kev tshem tawm ntawm cov dawb radicals hauv tib neeg lub cev. Cov polysaccharide muaj pes tsawg leeg ntawm Cistanches muaj zog antioxidant muaj peev xwm thiab tuaj yeem txhawb kev tshem tawm ntawm cov dawb radicals hauv tib neeg lub cev [3].

Hauv daim ntawv no, lubpolysaccharidesnyob rau hauv Cistanche stannic tau muab rho tawm thiab cais los ntawm high-efficiency gel permeation. Qhov hnyav molecular ntawm polysaccharides hauv Cistanche tau txiav txim siab los ntawm kev ua haujlwm siab gel permeation chromatography. Qhov hnyav molecular ntawm polysaccharides ntawm Cistanche tau txiav txim siab los ntawm kev ua tau zoo ntawm cov gel permeation chromatography, thiab muaj pes tsawg leeg ntawm polysaccharides kuj yog qhov hnyav molecular ntawm polysaccharides ntawm Cistanche tau txiav txim siab los ntawm kev ua haujlwm siab gel permeation chromatography, thiab cov muaj pes tsawg leeg ntawm polysaccharides yog. soj ntsuam.

Kev ua haujlwm antioxidant ntawm Cistanche tau txheeb xyuas los ntawm kev txiav txim siab dawb radical scavenging zog thiab tag nrho cov antioxidant muaj peev xwm ntawm cov khoom siv polysaccharide. Peb kuj tau txiav txim siab lub peev xwm dawb radical scavenging thiab tag nrho cov antioxidant muaj peev xwm ntawm polysaccharide Cheebtsam ntawm Cistanch, thiab muab lub hauv paus theoretical rau kev txhim kho thiab kev siv ntawm Cistanche thiab nws cov nqi ntxiv.

1 Kev npaj thiab rho tawm cov tshuaj polysaccharide los ntawm Cistanche

Nqa 2.5 kg ntawm cistanche, hlais nws, ntxiv 12.5L ntawm distilled dej thiab boil rau 3h. Tom qab boiling rau 3h, cov lim tau sau los ntawm kev pom thiab ntxiv rau 75 feem pua ​​​​ethanol, thiab sab laug rau sawv ntsug rau 24 teev. Tom qab boiling, cov filtrate tau sau los ntawm kev pom, ntxiv rau 75 feem pua ​​​​ethanol thiab sab laug rau sawv ntsug rau 24h, centrifuged ntawm 4000r / min rau 30min, thiab cov precipitate tau sau. Lub precipitate tau sau, khov thiab qhuav kom tau Cistanche crude polysaccharide CLP [4]. Lub precipitate tau khov-qhuav kom tau cov crude polysaccharide CLP ntawm Cistanche [4]. DEAE cellulose tau noj, tsau thiab o, degassed Nws tau thauj mus rau DEAE cellulose anion txauv kem thiab sib npaug nrog dej distilled thiab NaCl tov. Dej thiab NaCl tov tau siv rau kev sib npaug. Qhov hnyav 100 mg ntawm crude polysaccharide ntawm Cistanche tubulosa CLP yog yaj nyob rau hauv 10 mL ntawm distilled dej, thiab cov eluate tau sau tom qab elution nrog dej distilled.

Cov eluate tau sau tom qab elution nrog dej distilled thiab khov-qhuav los ntawm concentration kom tau cov qab zib nruab nrab CLP1 ntawm Cistanche; Cov eluate tau sau tom qab elution nrog NaCl tov, thiab tom qab ntawd concentrated thiab khov-qhuav tom qab lim dej kom tau Cistanche acidic qab zib CLP1. Tom qab elution nrog NaCl tov, cov eluate tau sau thiab concentrated los ntawm kev lim ntshav thiab khov-dried kom tau Cistanche acidic qab zib CLP2 [5].

Cov piam thaj nruab nrab CLP1 thiab acidic qab zib CLP2 ntawm Cistanche tubulosa tau npaj ntawm qhov concentration ntawm 1 mg / kg, feem. Cov tshuaj polysaccharide tau npaj ntawm qhov concentration ntawm 1 mg / mL, thiab cov polysaccharides tau txheeb xyuas los ntawm UV spectroscopy siv lub spectrophotometer ntawm 210-600 nm. Lub UV spectra tau raug soj ntsuam siv lub spectrophotometer ntawm 210-600 nm, thiab thaum lub ncov tshwm sim ntawm 260-280 nm Thaum lub ncov tshwm sim ntawm 260-280 nm, cov tshuaj tau pov thawj muaj protein ntau thiab nucleic acid. [6].

2 Kev txiav txim siab ntawm polysaccharide tshuaj muaj pes tsawg leeg ntawm Cistanche

Txoj kev phenol-sulfuric acid tau siv los txiav txim siab tag nrho cov ntsiab lus qab zib hauv Cistanche cov qauv. Glucose daws ntawm 0.1 mg/mL, 0.2, 0.4, 0.6, 0.8 thiab 1 mL raug ntsuas nyob rau hauv cov iav kuaj cov raj, thiab cov dej distilled tau ntxiv los kho qhov ntim rau 1 mL. Tom qab ntawd 0.5 mL ntawm phenol reagent thiab 2.5 mL ntawm cov concentrated sulfuric acid nrog 6 feem pua ​​​​ concentration tau ntxiv rau hauv cov raj kuaj, shaken zoo thiab txias rau chav tsev kub. Lub absorbance yog ntsuas ntawm 490 nm siv lub spectrophotometer, thiab tus qauv nkhaus tau npaj nrog cov ntsiab lus qab zib raws li kab rov tav kev sib koom tes thiab absorbance raws li txoj kab ntsug, thiab qhov sib npaug regression tau xam [7-8].

Cov ntsiab lus protein ntawm Cistanche cov qauv tau txiav txim siab siv Komas Brilliant Blue txoj kev. Cov ntsiab lus protein ntawm cov qauv tau ntsuas los ntawm spectrophotometer ntawm 495 nm thiab cov ntsiab lus ntawm cov protein raug suav raws li tus qauv nkhaus sib npaug [9].

Cov ntsiab lus ntawm glyoxalate hauv cov qauv Cistanche tau txiav txim siab siv txoj kev m-hydroxyphenyl. Qhov sib txawv ntawm cov qauv thiab 0.4 mL ntawm cov qauv tov nrog ib qho concentration ntawm 0.1 mg / mL tau ntxiv rau lub raj kuaj nrog 2.5 mL ntawm concentrated sulfuric acid, shaken

Qhov concentration ntawm m-hydroxybiphenyl thiab 0.5 feem pua ​​​​sodium hydroxide tov tau ntxiv rau hauv lub raj kuaj, shaken zoo thiab sab laug rau 30 min. Lub absorbance ntawm 525 nm tau ntsuas los ntawm spectrophotometer, thiab tus nqi ntawm glyoxalate hauv cov qauv raug xam raws li tus qauv nkhaus sib npaug [10-11]. Qhov txheeb ze molecular hnyav ntawm polysaccharides ntawm Cistanche tau txiav txim siab los ntawm kev ua haujlwm siab gel chromatography, thiab cov ntsiab lus ntawm monosaccharides ntawm Cistanche tau txiav txim siab los ntawm kev ua haujlwm siab ua kua chromatography. Cov monosaccharide muaj pes tsawg leeg ntawm Cistanche tau txiav txim siab los ntawm kev ua haujlwm siab ua kua chromatography [12].

3 Cov txiaj ntsig thiab kev tshuaj xyuas ntawm polysaccharide muaj pes tsawg leeg ntawm Cistanche Tubulosa

Los ntawm daim duab 1, nws tuaj yeem pom tias qhov nruab nrab qab zib CLP1 ntawm Cistanche Tubulosa tau pom qhov pom tseeb qhov siab tshaj plaws ntawm 280nm, thaum lub acidic qab zib CLP2 ntawm Cistanche Tubulosa tsis muaj qhov pom tseeb qhov siab siab ntawm 260-280nm, tab sis UV absorption spectrum tau siab dua. Txij li cov tryptophan thiab tyrosine muaj nyob rau hauv cov proteins muaj qhov nqus loj dua rau UV ntawm 280 nm, nws tuaj yeem txiav txim siab tias Cistanche nruab nrab qab zib CLP1 muaj glycoproteins, tab sis nws tsis tuaj yeem txiav txim seb Cistanche acidic qab zib CLP2 muaj nucleic acids thiab cov proteins.

Nws tsis tuaj yeem txiav txim siab seb cov piam thaj acidic CLP2 ntawm Cistanche puas muaj nucleic acids thiab cov proteins.

DEAE cellulose tsis zoo ion pauv chromatography kem tsom xam tuaj yeem adsorb ionic tshuaj thiab impurities ntawm kem kom ua tiav cov khoom sib cais ntawm polysaccharide, thiab tom qab sib xyaw thiab ziab cov dej distilled eluate, tag nrho ntawm 34.68 mg ntawm Cistanche nruab nrab qab zib CLP1 tau txais, thiab tom qab ntawd. siv cov tshuaj NaCl kom elute thiab mloog zoo thiab qhuav, tag nrho ntawm 16.52 mg ntawm Cistanche acidic qab zib CLP2 tau. Tus qauv nkhaus sib npaug ntawm tag nrho cov ntsiab lus qab zib, cov ntsiab lus protein thiab glyoxylate tus qauv nkhaus sib npaug. Kev ntsuas tag nrho cov piam thaj, protein thiab glyoxylate ntawm Cistanche tau txais. Tag nrho cov suab thaj, protein thiab glucuronic acid cov ntsiab lus ntawm Cistanche tau ntsuas thiab qhia hauv Table 1.

Lub scanning nkhaus ntawm UV spectrum ntawm photometer los ntawm 210 mus rau 600 nm yog pom nyob rau hauv daim duab 1.

Los ntawm daim duab 1, nws tuaj yeem pom tias Cistanche neutrophilic qab zib CLP1 tau pom qhov pom tseeb ntawm qhov nqus ntawm 280 nm, thaum Cistanche acidic qab zib CLP2 pom qhov nqus siab siab ntawm 260-280 nm. thiab cov kua qaub qab zib CLP2 ntawm Cistanche Tubulosa pom qhov pom tseeb nqus ntawm 260-280 nm. Txawm hais tias tsis muaj qhov pom tseeb qhov siab tshaj plaws ntawm 260-280 nm, UV nqus spectrum siab dua. Txij li thaum tryptophan thiab tyrosine muaj nyob rau hauv cov proteins muaj ib tug loj dua tus nqi nqus ntawm 280 nm rau UV absorption tus nqi, nws yuav txiav txim tau hais tias Cistanche neutrophic qab zib CLP1 muaj glycoproteins, tab sis nws tsis muaj peev xwm txiav txim siab seb Cistanche acidic qab zib CLP2 muaj nucleic acids thiab proteins. . Lub xub ntiag ntawm nucleic acids thiab cov proteins hauv Cistanche CLP2 tsis tuaj yeem txiav txim siab.

DEAE cellulose tsis zoo ion pauv chromatography kem tsom xam tuaj yeem nqus cov khoom ionic thiab impurities rau hauv kem. Kev sib cais ntawm cov tshuaj polysaccharide tau ua tiav los ntawm adsorbing cov tshuaj ionic thiab impurities ntawm kab. Cov dej distilled eluate tau concentrated thiab qhuav kom tau txais cov suab thaj nruab nrab ntawm Cistanche Tom qab ua kom tiav thiab ziab cov dej distilled eluate, tag nrho ntawm 34.68 mg ntawm CLP1 tau txais, thiab tom qab concentrating thiab ziab nrog NaCl tov, acidic qab zib CLP1 tau txais. Cov kua qaub qab zib CLP2 ntawm Cistanche tau txais li 16.52 mg. Cov qauv nkhaus sib npaug ntawm tag nrho cov ntsiab lus qab zib, cov ntsiab lus protein thiab glyoxylate cov ntsiab lus ntawm Cistanche tau txais. thiab glyoxylate tus qauv nkhaus sib npaug. Kev ntsuas tag nrho cov piam thaj, protein thiab glyoxylate ntawm Cistanche tau txais. Tag nrho cov suab thaj, protein thiab glucuronic acid cov ntsiab lus ntawm Cistanche tau ntsuas thiab qhia hauv Table 1.

Table 1 Cov ntsiab lus ntawm tag nrho cov suab thaj, protein thiab uronic acid hauv Cistanche deserticola

Qhov hnyav molecular thiab qhov chaw siab tshaj ntawm tus qauv qauv tuaj yeem txiav txim siab, thiab qhov sib npaug regression tuaj yeem tau los ntawm kev hloov lub teeb emission teeb liab ntawm tus qauv coj mus kuaj rau hauv qhov sib npaug regression. Qhov hnyav molecular ntawm tus qauv yuav raug kuaj tau. Qhov hnyav molecular ntawm cov qauv yuav raug kuaj tau los ntawm kev hloov lub teeb emission teeb liab qhov tseem ceeb ntawm cov qauv coj mus kuaj rau hauv qhov sib npaug regression. Qhov nruab nrab molecular hnyav, qhov nruab nrab hnyav molecular hnyav thiab dispersion coefficient ntawm Cistanche Tubulosa CLP1 thiab Cistanche Tubulosa CLP2 tau txais. Cistanche nruab nrab qab zib CLP1 thiab Cistanche acidulous CLP2 tau txiav txim siab los ntawm kev ua haujlwm siab gel permeation chromatography. Cov txiaj ntsig ntawm lub sijhawm khaws cia thiab kev suav qhov hnyav molecular yog qhia hauv Table 2. Qhov piv ntawm qhov hnyav nruab nrab molecular hnyav rau tus lej nruab nrab molecular hnyav tau siv los qhia qhov dav ntawm qhov kev faib tawm molecular. Qhov piv ntawm qhov hnyav nruab nrab molecular hnyav rau tus lej nruab nrab molecular hnyav yog siv los qhia qhov dav ntawm molecular tis.

Nug ntxiv:

Email: wallence.suen@wecistanche.com

Whatsapp / Tel: ntxiv rau 86 15292862950