Chemical Constituents Ntawm nplooj ntawm Cistanche

Abstract: Cistanchenplooj tau ntev tau siv los ua tshuaj raukhoischemic mob stroke, mob angina, thiabntshav siabthiab raws li kev noj qab haus huv haus thiabtshuaj pleev ib ce rauanti-aging. Txoj kev tshawb no tsom kom cais tawm ntau lub tebchaw raws li qhov ua tau los ntawm ethanol extractntawm persimmon nplooj los txheeb xyuas cov biologically active compounds. Cov nyhuv antioxidativeethyl acetate txheej los ntawm ethanol extract ntawm persimmon nplooj tau pom los ntawm2,2-diphenyl-1-picrylhydrazyl (DPPH) assay thiab online-performance kua chromatography-2,20 -casino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (HPLC-ABTS) kev tshuaj xyuas. Ib qho tshiab flavonoid,kaempferol -3-O- -D-2"-coumaroylgalactoside (1), thiab ib qho chaw tshiab, kaempferol-3-O- -D-2"-feruloylglucoside (3) raug cais tawm ntawm ethyl acetate txheej, nrog rau 25 yav dhau lospaub cov tebchaw, suav nrogkaum plaub flavonoids, ib ionone, ob coumarins, xya triterpenoids,thiab ib qho acetophenone. Kev txhais cov ntaub ntawv spectrometric thiab spectroscopic txiav txim lawv cov qauv. Txhua qhov sib xyaw ua ke tau raug soj ntsuam sai heev los ntawm kev siv onlineHPLC-ABTSsoj ntsuam, thiab ntawm no, compounds4–8, 11, 13, 15, thiab16 qhia meej meejAntioxidative teebmeem. Covtus nqi ntawm cov tebchaw no yog {{0}}.3–0.65 feem pua ntawm cov extract.

Ntsiab lus:Diospyros kaki; persimmon nplooj; flavonoid; antioxidant

Flavonoid (8)

Nyem qhov no kom tau txais Cistanche nplua nuj nyob rau hauv Flavonoids Rau Cosmetic Anti-Aging

1. Taw qhia

Diospyros kakiThunb. (persimmon) belongs rau tsev neeg ntawm Ebenaceae thiab davmuab faib rau hauv Kauslim, Tuam Tshoj, thiab Nyiv. Nws cov txiv hmab txiv ntoo yog noj tshiab los yog qhuav, thaum nplooj muajntev tau siv los ua cov tshuaj ib txwm siv los kho ischemic stroke, angina, kub siab,atherosclerosis, thiab kab mob sib kis [1]. Tsis tas li ntawd, nws cov nplooj tau sivraws li kev noj qab haus huv dej haus thiab tshuaj pleev ib ce vim lawv anti-aging zog thiab muaj peev xwmpab tiv thaiv cov cholesterol thiab melanin txuam nrog [1]. Kev tshawb fawb tsis ntev los no tau qhiahais tias cov extracts ntawm persimmon nplooj muaj ntau yam khoom lom,suav nrog radical scavenging, neuroprotection, thrombosis inhibition, anti-atherosclerosis,thiab anti-allergy [2–6]. Kev tshawb nrhiav phytochemical yav dhau los qhia tias ntau yamhom ntawmflavonoidsthiab terpenoids yog cov ntsiab lus tseem ceeb [7], thiab ntau tannins,Naphthoquinones, coumarins, ionones, thiab fatty acids kuj tau tshaj tawm [8–12]. Reactive oxygen hom (ROS) yog reactive molecules tsim nyob rau hauv biological systems,thiab qhov sib npaug ntawm tiam thiab tshem tawm ntawm ROS yog tswj tau zoo nyob rau hauv ib txwm muajcellular physiology [13]. Txawm li cas los xij, ntau dhau ntawm ROS ua rau oxidative puas tsuaj,thiab nyob rau hauv lem, kev laus thiab muaj hnub nyoog cov kab mob xws li mob cancer, ntshav qab zib, thiab Parkinson'skab mob [14]. Yog li, nrhiav pom cov tshuaj antioxidants xws li flavonoids thiab phenolic tebchawtuaj yeem yog lub tswv yim zoo rau kev kho cov kab mob no.

Raws li ib feem ntawm peb txoj haujlwm txuas ntxiv mus nrhiav cov tshuaj lom neeg lom neeg [15], uaAntioxidative nyhuv ntawm ethanol (EtOH) extract thiab hnyav partitions los ntawm persimmonnplooj raug soj ntsuam siv 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay thiab online ua haujlwm siabkua chromatography-2,20 -casino-bis (3-ethylbenzothiazoline-6-sulfonic acid)(HPLC-ABTS). Kev tshawb fawb phytochemical ntawm nplooj persimmon coj mus rau kev sib caisntawm ib qho tshiab flavonoid (1), ib tug tshiab natural compound (3), thiab25 ua ntejpaub tebchaw. Daim ntawv thov ntawm spectroscopic thiab spectrometric txoj kev characterizes cov qauv. Txhua qhov sib xyaw ua ke tau raug tshuaj xyuas sai rau lawvAntioxidative teebmeem siv online HPLC-ABTS. Tsis tas li ntawd, qhov kev ntsuam xyuas ntau ntawmtag nrho cov sib cais cais tau ua nyob rau hauv txoj kev tshawb no.

2. Cov txiaj ntsig thiab kev sib tham

2.1. Antioxidative nyhuv ntawm Persimmon nplooj

Cov nyhuv antioxidative ntawm ethanol (EtOH) extract ntawm persimmon nplooj raug soj ntsuamrau kev ntsuam xyuas ua ntej los ntawm DPPH (Daim duab1A). Cov 0.125 mg/mL ntawm covextract scavenged kwv yees li 8 0 feem pua ntawm DPPH radical, thaum 0.025 mg / mL ntawm ascorbicacid ua 94 feem pua ntawm cov radical. Kev ntsuam xyuas HPLC-ABTS online tau ua tiav saixyuas kom muaj kev ntseeg siab ntawm cov txiaj ntsig no (Daim duab1D). Gallic acid thiab Trolox tau siv los ua sab hauvqauv. Lub chromatogram ntawm 734 nm (tsis zoo ncov) qhia tias kwv yees licuaj qhov chaw tuaj yeem muaj cov haujlwm tiv thaiv oxidative. Gallic acid thiab Trolox (6-hydroxy- 2,5,7,8-tetramethyl chroman-2-carboxylicacid) tau siv los ua cov qauv sab hauv los xyuas kom meej qhovkev ntseeg tau ntawm cov txiaj ntsig. Nws tau raug xam tias feem ntau ntawm cov ncov no yog flavonoid derivativesxws li flavonoid glycoside thiab flavanol, raws li kev soj ntsuam dereplication tau ualos ntawm kev sib piv ultraviolet (UV) thiab huab hwm coj spectra ntawm cov tebchaw nrog cov luam tawmcov ntaub ntawv. Bioassay-guided fractionation qhia tias cov tshuaj tiv thaiv oxidative no yogmuaj ntau nyob rau hauv ethyl acetate (EtOAc) txheej, thaum dej (H2O) txheej pom tsis muaj zogkev ua haujlwm (daim duab1B,C).

cistanche for dpph scavenging

Daim duab 1. DPPH scavenging teebmeem ntawm EtOH extract (A), EtOAc txheej (B), thiab HO txheej (C) HhO txheej ntawm persimmon nplooj: (D) online ABTS-HPLC chromatogram ntawm EtOH extract.

cistanche research for dpph scavenging

2.2. Kev tshawb nrhiav Phytochemical

Txhawm rau txheeb xyuas cov tshuaj tiv thaiv oxidative, ntau yam chromatographic thiab spectroscopiccov txheej txheem tau nqa tawm rau kev cais thiab cov yam ntxwv ntawm tus yam ntxwvtshuaj sib xyaw. Ib qho tshiab flavonoid (1) thiab ib tug tshiab natural compound (3) tau muab los ntawm covethyl acetate txheej ntawm ethanol extract, ua ke nrog 25 cov lus qhia yav dhau los,uas yog kaempferol-3-O- -2"-coumaroylglucoside (2) [16], (ntxiv )-catechin (4) [17], hyperoside(5) [17isoquercitrin (6) [18], quercetin -3-O- -2"-galloyylgalactoside (7) [19], quercetin -3-O- -2"-galloyylglucoside (8) [20trifolin (9) [18, astragalin (10) [18], kaempferol-3-O- - 2"-galloylgalactoside (11) [21], kaempferol-3-O- - arabinoside (12) [22], kaempferol-3-O- - 2"-galloyylglucoside (13) [23], quercetin -3-O- -2"-coumaroylglucoside (14) [24], quercetin(15) [15], kaempferol (16) [25], (6S,9S) - roseoside (17) [26], scopoletin (18) [27], umbelliferone(19) [28], 1-(2,4-dihydroxy-6-methylphenyl) ethanone (20) [29], barbinervic acid (21) [30], diospyric acid B (22) [7], rotungenic acid (23) [31pomolic acid (24) [32], siresinolic acid(25) [33oleanolic acid, uas yog,26) [25thiab ursolic acid (27) [25] los ntawm kev siv spectroscopic thiabspectrometric thiab lub cev cov ntaub ntawv sib piv nrog cov ntaub ntawv luam tawm thiab kuj nrog nyiastxheej chromatography (TLC) tsom xam (Figures2 thiab3). Ntawm no, compounds2, 16, 17, 19, 20, thiab24 tau thawj zaug cais tawm ntawmD. kev.

cistanche research for dpph scavenging

Daim duab 3.HPLC chromatogram ntawm EtOH extract los ntawm persimmon nplooj

Compound 1 tau txais los ua cov hmoov daj, uas cov mis mos molecular tau tsim los ua C3olO3 raws li cov ntaub ntawv siab daws teeb meem loj spectrometry (HRMS). Cov spectrum tau nthuav tawm cov khoom nqus nqus ntawm 207 thiab 315 nm, qhia tias compound1 muaj flavonol qaum. Cov ntaub ntawv 'H nuclear magnetic resonance (NMR) cov ntaub ntawv (Table 1Figure S1) tau pom ib qho qauv ntawm coumaroylated flavonol glycoside, qhia ob pawg ntawm AA'BB'-hom teeb liab ({{10}}}{{11 }} (2H, d, J= 8.5 Hz, H-2' thiab H-6 ),6.87 (2H, d, J{{20}} .5 HzH-3' thiab H-5')) nyob rau hauv lub nplhaib B ntawm kaempferol nrog rau cov teeb liab (0 7.45 (2H, d, J=8). 5 HzH-2 thiab H{{3{{104}}}}"), 6.81 (2H, d, J=8.5 Hz, H-3 thiab H-5 feem pua ) nyob rau hauv lub nplhaib aromatic ntawm coumaroyl pawg. (1Hd, J=16. d, J=8.0 Hz, H-1) tau pom nyob rau hauv thaj av qab zib, qhia tias muaj B-configurated cyclic qab zib pawg. Lub 13C thiab distortionless txhim kho los ntawm polarization hloov NMR cov ntaub ntawv (Iable 1Daim duab S2) pom 30 resonances suav nrog ob trans-olefinic carbons, kaum aromatic carbons, thiab rau glucosyl moiety carbons, thiab kaum non-protonated carbons nrog rau ob carbonyl carbons. Tshwj xeeb, cov tshuaj hloov pauv ntawm C-2, C-3, thiab C-4 (8c 158.1, 134.9 thiab 179.2) yog cov cim qhia ntawm flavonol3-0-glycoside. Tsis tas li ntawd, carbonyl carbon teeb liab ntawm C-1" (8c 168.7) thiab ob lub trans-olefinic carbon signals ntawm C-2'" thiab C-3"(6 146.9 , 115.2) yog cov tshuaj lom neeg hloov pauv ntawm pawg coumaroyl. Cov paib carbon anomeric ntawm C-1(6C 100.4) thiab lwm cov cim rau glycosyl moiety ntawm C-2" rau C-6" ' (6c74.3, 73.4, 70.5, 77.4, thiab 62.0) tau pom. Cov ntaub ntawv ib-dimensional (1D) NMR no yog superimposable rau cov kaempferol-3-0-B-2"-coumaroylglucoside (2) (16]. Txawm li cas los xij, kev sib piv ceev faj ntawm 13C NMR cov ntaub ntawv ntawm ob lub tebchaw qhia tias cov compound 1 muaj galactose moiety, uas tau nthuav tawm ntxiv los ntawm nuclearOverhauser li spectroscopy (NOESY) NMR cov ntaub ntawv (Daim duab S6). Thaum lub NOES correlation ntawm H-2 thiab H-4 tau pom nyob rau hauv compound 2, tsis muaj kev sib raug zoo ntawm cov protons nyob rau hauv compound 1. Feem ntau, txhais 13C NMR thiab NOESYNMR cov ntaub ntawv yog ib txoj hauv kev zoo los txiav txim seb hom glycosyl moiety . Qhov chaw ntawm galactose moiety tau txiav txim siab los ntawm C-3 raws li kev hloov pauv ntawm C-2thiab C-4, raws li kev pov thawj ntxiv los ntawm heteronuclear ntau qhov sib raug zoo (HMBCbetween H{ {123}}" thiab C-3 (Daim duab S5). Txoj hauj lwm ntawm pawg coumaroyl tau pom tias yog nyob ntawm C-2" raws li kev hloov pauv hauv qhov chaw downfield (0 5.36 (1H, dd, J=10.0, 8.0 Hz, H{ {134}}")) thiab HMBC kev sib raug zoo ntawm H-2' thiab C-1. Raws li qhov tshwm sim, cov qauv ntawm compound 1 tau txiav txim siab ua kaempferol-3-0-B-2"-coumaroylgalactoside. Txawm hais tias compound 2 tau raug cais tawm los ntawm ntau qhov chaw, suav nrog Ouercus suber (16] thiab Alliumporrum (34), compound 1 tau cais tawm thiab cov qauv tsim thawj zaug.

Table 1.1H thiab13C NMR data of compounds1 thiab3 hauv methanol-d4

cistanche research for dpph scavenging

Compound3 raug cais raws li cov hmoov daj, thiab cov mis molecular tau tsimraws li C31H28O14 los ntawm kev txheeb xyuas cov ntaub ntawv HRMS. UV spectrum pom UVnqus ntawm 210 thiab 327 nm vim tib yam aglycone nrog cov tebchaw1 thiab2. Cov1H NMR data (Table1, Daim duab S10) zoo ib yam li cov compound2, tab sis compound3 muaj ib pawg feruloyl es tsis txhob ntawm coumaroyl pawg, raws li muaj pov thawj los ntawm muaj ib tugntxiv methoxy pawg (δH 3.91 (3H, s, 3000 - OCH3)). Tsis tas li ntawd, ib qho anomeric protonteeb liab (δH 5.64 (1H, d, J {{0}}.0 Hz, H-100 )) tau pom, qhia tias qhov sib txuas glycosylua a- kev teeb tsa, thiab downfield-shifted teeb liab (5.03 (1H, t,J = 8.5 Hz, H-200 )) tau pom nyob rau hauv cheeb tsam qab zib, raws li nrog compound1. Cov13C NMR data (Table1, Daim duabS11) pom 31 resonances suav nrog obtrans-olefifinic carbons, kaum quarternary carbons,kaum cov pa roj carbon monoxide, 6 glucosyl moiety carbons, thiab ib qho methoxy carbon, thiab kaum tsis-protonatedcarbons, suav nrog ob carbonyl carbons sib raug rau kaempferol, feruloyl,thiab pab pawg qabzib. Tshwj xeeb, carbon signals los ntawm C-200 rau C-600 (δC 75.8, 76.3, 71.5, 78.8 thiab 62.5) qhia tias muaj cov piam thaj moiety. Qhov chaw ntawm cov piam thaj moietythiab feruloyl pawg tau raug muab los ntawm qhov ntev HMBC kev sib raug zoo ntawm H-100 thiabC-3 (δC 134.8) thiab H-200 thiab C-1000 (δC 168.4) (Daim duab S14). Txoj hauj lwm ntawm ib qho ntxivmethoxy pawg tau txiav txim siab los ntawm qhov tseem ceeb correlation ntawm 3000 - OCH3 thiab C-3000 (δC 149.4). Cov txiaj ntsig saum toj no qhia txog cov qauv ntawm kev sib xyaw3 as kaempferol-3-O- - 2 "-feruloyl glucoside. Rau qhov zoo tshaj plaws ntawm peb kev paub, compound3 tsuas yog qhia raws li ib tugcov khoom ntawm hydrolysis ntawm 3-O- -(2-O-feruloyl)-glucosyl-7,40 -di-O- -glucosyl kaempferol, cais tawm ntawmAllium tuberosum[35]. Yog li ntawd, tus qauv ntawm3 twb elucidated li ib tug tshiabnatural compound.

Compound11 raug cais raws li cov hmoov daj. Cov1Cov ntaub ntawv H NMR (Daim duab S19)tso tawm ib txheej ntawm AA0 BB0 - hom teeb liab (δH 8.06 (2H, d, J {{0}}.0 Hz, H-20 , H-60 ), 6.87 (2H, d, J {{0}}.0 Hz, H-30, H-50 )) nyob rau hauv B nplhaib ntawm kaempferol thiab ib tug singlet teeb liab ntawmδH 7.02 (2H, s, H-3000 , H-7000 ) ntawm galloyl moiety nyob rau hauv cheeb tsam aromatic, uas yog ib tug yam ntxwv teeb liabntawm faib flavonol. Lub teeb liab anomeric proton (δH 5.78 (1H, d, J {{0}}.0 Hz, H-100 )) qhia tau hais tias glycosyl linkage yog a- kev teeb tsa. Tsis tas li ntawd, ib qho downfieldhloov pauv proton teeb liab (5.27 (1H, t,J = 9.5 Hz, H-200 )) qhia tias pawg galloyltau txuas ntawm pawg hydroxyl ntawm C-200 vim qhov kev hloov no tuaj yeem raug ntaus nqi raulub anisotropic cawv ntawm covO-galloyl moiety [21]. Cov13Cov ntaub ntawv C NMR (Daim duab S20)nthuav tawm 26 resonances, qhia tias faib flavonol glycoside. Cov pa roj carbon signalslos ntawm C-200 rau C-600 (δC 71.1, 72.7, 68.2, 76.0, thiab 60.1) qhia tias muaj cov galactosemooj. Yog li ntawd, cov qauv ntawm compound11 tau lees paub tias yog kaempferol-3-O- -200 - Galloylgalactoside. Txawm tias compound11 yav dhau los raug cais tawm los ntawm ntau qhov chaw,suav nrogD. kev[21,36], tsuas yog cov1H NMR thiab MS cov ntaub ntawv tau tshaj tawm yav dhau los. Yog li,tus13C NMR cov ntaub ntawv tau tshaj tawm thawj zaug hauv txoj kev tshawb no

Echinacoside in cistanche

2.3. Antioxidative kev ua ub no ntawm Isolated Compounds

Tag nrho cov tshuaj sib cais tau raug soj ntsuam rau lawv cov teebmeem antioxidative siv ceev ceevkab HPLC-ABTS tsom xam los txiav txim seb cov tebchaw twg pab rau cov tshuaj tiv thaiv oxidativecov nyhuv ntawm persimmon nplooj. Cov tshuaj4–8, 11, 13, 15, thiab16 pom muaj zog antioxidative activities (Daim duab4). Feem ntau ntawm cov tshuaj no yog kaempferol thiab quercetinderivatives, tab sis qee qhov derivatives (1–3, 9, 10, 12, thiab14) tsis qhia kev ua ub ua no. Covkev sib raug zoo-kev sib raug zoo tsis tau txiav txim siab tag nrho tab sis tau nthuav tawm ib nrab. Hauvtshwj xeeb, quercetin thiab kaempferol nrog galloyl moieties (7, 8, 11, thiab13) tau pommuaj cov haujlwm muaj zog, tab sis cov neeg uas muaj coumaroyl lossis feruloyl moieties (1–3 thiab16) tsis tauqhia tej kev ua ub no.

cistanche research for dpph scavenging

Daim duab 4.Radical scavenging kev ua ub no ntawm cov khoom sib cais (4–8, 11, 13, 15, thiab16) tau soj ntsuam los ntawm onlineHPLC-ABTS kev xeem.

2.4. Quantitative Analysis ntawm Isolated Compounds

Kev soj ntsuam kom muaj nuj nqis ntawm txhua qhov sib xyaw ua ke hauv EtOAc-soluble extracttau ua kom paub meej tias cov tshuaj tiv thaiv antioxidant ntawm cov nplooj persimmon tuaj yeem yogtshwm sim los ntawm cov active compounds. Ib txoj kev HPLC tshwj xeeb nrog diode array nrhiav thiabevaporative light scattering detector tau tsim los rau kev txiav txim siab ib txhijntawm 27 sib xyaw. Cov ntsiab lus ntawm tag nrho cov triterpenoids cais yog kwv yees li 5.9 feem puacov extract, thiab ntawm no, cov ntawm siaresinolic acid (25), oleanolic acid (26), thiab ursolicacid (27) suav rau ib feem tseem ceeb, raws li qhia hauv cov kev tshawb fawb yav dhau los [7]. CovCov ntsiab lus ntawm flavonoids yog kwv yees li 5.4 feem pua, thiab ntawm cov no, cov tshuaj nquagyog 3.2 feem pua (Table2). Tshwj xeeb, cov ntsiab lus ntawm kaempferol -3-O- -200 -galloylgalactoside(9), kaempferol-3-O- -2"-galloyylglucoside (10), isoquercitrin (13), thiab quercetin -3-O- -2"- galloylglucoside (15) tau ntau dua 0.3 feem pua . Txoj kev tsom xam tau txheeb xyuas siv ayooj yim validation txheej txheem los xyuas kom meej qhov tseem ceeb ntawm txoj kev, uas pom tau txaustshwj xeeb, linearity, raug, thiab precision.