Cov khoom xyaw muaj txiaj ntsig ntawm Cistanche: Kev Sib cais thiab Kev Ua kom huv ntawm Phenylethanoid Glycosides
Mar 14, 2022
Hu rau:joanna.jia@wecistanche.com
Kev rho tawm thiab ua kom huv ntawm Phenylethanoid Glycosides Los ntawm Cistanche Deserticola Los Ntawm Kev Kub Ceev Counter-tam sim no Chromatography
Li a,b, Rong Tsao b,*, Raymond Yang b, Chunming Liu a,
J. Christopher Young b, Honghui Zhu b
Department of Chemistry, Changchun Normal University, Changchun 130032, Suav
b Guelph Food Research Center, Agriculture thiab Agri-Food Canada, 93 Stone Road West, Guelph, Ontario, Canada N1G 5C9
Tau txais 31 Tsib Hlis 2007; tau txais hauv daim ntawv kho dua 16 Lub Kaum Hli 2007; tau txais 29 Lub Kaum Hli 2007
Abstract:
Tsibphenylethanoid glycosides(PhGs),echinacoside cov tshuaj, cistanoside A, acteoside, isoacteoside, thiab 20-acetylacteoside, raug cais thiab ntxuav los ntawmCistanche deserticolarau thawj zaug los ntawm high-speed counter-tam sim no chromatography (HSCCC) siv ob lub tshuab biphasic, ib qho muaj ethyl acetate-ethanol-dej (5:0.5:4.5, v/v/v) thiab lwm yam ntawm ethyl acetate-n-butanol-ethanol-dej ({{10}}.5:{14}}.5:0.1:1, v/v/v/v). Tag nrho ntawm 28.5 mg ntawm echinacoside, 18.4 mg ntawmcistanoside A, 14,6 mg vacteoside, 30.1 mg ntawm isoacteoside thiab 25.2 mg ntawm 20-acetylacteoside tau purified los ntawm 1412 mg ntawm n-butanol extract ntawmCistanche deserticola, txhua ntawm ntau dua 92.5 feem pua purity raws li kev txiav txim los ntawm HPLC. Cov qauv raug txheeb xyuas los ntawm lawv lub sijhawm khaws cia, UV, LC-ESI-MS hauv hom tsis zoo ion, thiab tau lees paub los ntawm NMR kev sim. Tus yam ntxwv LC-ESI-MSn fragmentation qauv ntawm tsib lub tebchaw yog tham, thiab pom tias yog ib qho tshwj xeeb thiab muaj txiaj ntsig zoo rau kev txheeb xyuas cov qauv ntawm PhGs los ntawm cov nroj tsuag tshuaj tseem ceeb no.
Crown Copyright © 2007 Luam tawm los ntawm Elsevier Ltd. All rights reserved.
Ntsiab lus:Cistanche deserticola; Phenylethanoid;Echinacoside; Cistanoside A;Acteoside; Isoacteoside; 20-Acetylacteoside; HSCCC; LC-ESI-MS; NMR
1. Taw qhia
Cistanche deserticolaYC Ma yog ib lub npe nrov suav tshuaj ntsuab thiab tau siv dav hauv kev npaj tshuaj zoo ib yam li niaj hnub no cov khoom xyaw ua haujlwm lossis cov khoom noj khoom haus (Wong, Li, Cheng, & Chen, 2006). Cov khoom tseem ceeb bioactive hauv C. deserticola yogphenylethanoidglycosides (PhGs), suav nrogechinacoside cov tshuaj, acteoside, isoacteoside, 20-acetylacteoside, thiabcistanoside A(Kobayashi, Karasawa, & Miyase, 1984; Kobayashi li al., 1987). PhGs tau muab faib rau hauv lub nceeg vaj cog thiab tau kawm ntau yam rau lawv cov haujlwm lom neeg xws li hepatoprotective (Xiong li al., 1998), anti-inflammatory, antinociceptive kev ua haujlwm (Schapoval et al., 1998), thiab cov haujlwm antioxidant (Cheng. , Wei, Guo, Ni, & Liu, 2005; He, Lau, Xu, Li, & But, 2000; Li, Wang, & Wang, 1997; Li, Wang, Zheng, Liu, & Jia, 1993; Wang, Jiang , Wu, & Wang, 2001; Xiong, Kadota, Tani, & Namba, 1996), txhim kho kev sib deev (Xie & Wu, 1993; Zong, He, Wu, & Chen, 1996), thiab sedative effect (Lu, 1998) .
Raws li qhov tseem ceeb ntawm bioactivities saum toj no, ntau cov ntsiab lus ntshiab yog qhov xav tau ceev ceev raws li cov qauv siv thiab rau ntau yam hauv vitro thiab hauv vivo kev tshawb fawb txog kev siv tshuaj suav tshuaj. Cov kev siv tau zoo rau kev sib cais, kev huv, thiab kev tsim cov qauv ntawm PhGs, yog li tsim nyog. Txawm li cas los xij, kev ua haujlwm zoo li no feem ntau yuav tsum tau siv ntau yam chromato-graphic cov kauj ruam rau cov qauv ntxuav thiab cais tawm (Gross, Lahloub, Anklin, Schulten, & Sticher, 1988; Nishimura, Sasaki, Inagaki, Chin, & Mitsuhashi, 1991; Ravn, Nishibe. , Sasahara, & Li, 1990; Shoyama, Matsumoto, & Nishioka, 1987), uas feem ntau ua rau cov nqi rov qab qis vim yog qhov tsis zoo ntawm PhGs adsorptions mus rau cov khoom txhawb nqa thaum sib cais (Lei li al., 2001). Nyob rau hauv sib piv, high-speed counter-tam sim no chromatography (HSCCC) tau dhau los ua lwm txoj hauv kev zoo rau cov txheej txheem chromatographic rau kev sib cais ntawm qee cov PhGs los ntawm cov nroj tsuag rho tawm (Lei li al., 2001; Li et al., 2005). Lei et al. ntse sib caisacteosidethiab 20-acetylacteoside los ntawm Cistanches salsa (CA Mey.) G. Beck los ntawm kev siv HSCCC (Lei et al., 2001). Cov neeg sau ntawv ntawm daim ntawv no yav dhau los tau tshaj tawm txog kev sib cais ntawmacteosidethiab isoacteoside los ntawm Plantago psyllium L. los ntawm HSCCC (Li et al., 2005). Txawm li cas los xij, tsis muaj daim ntawv tshaj tawm tau tshaj tawm txog kev sib cais thiab kev huv ntawm ntau PhGs los ntawmCistanche deserticolasiv HSCCC. Vim tias tsis muaj cov qauv, LC-MS txoj kev tau tsim thiab siv los ua cov cuab yeej tshuaj ntsuam xyuas muaj zog rau kev ua tus cwj pwm sai thiab kev txheeb xyuas ntawm qee cov PhGs hauv cov nroj tsuag extract (Li et al., 2005; Wang et al., 2000).
Hauv daim ntawv no, peb qhia txog HSCCC txoj kev tsim los rau kev npaj echinacoside,cistanoside A, acteoside, isoacteoside thiab 20-acetylacteoside los ntawmCistanche deserticola. Cov cwj pwm thiab kev tsom xam ntawm tsib PhGs sib cais tau ua tiav los ntawm kev siv LC ua ke nrog hauv kab ESI pawg spectrometry thiab NMR kev sim. Lub sijhawm khaws cia, qhov hnyav molecular, thiab cov yam ntxwv ntawm cov ions ntawm tsib PhGs tau nthuav tawm thiab sib tham hauv daim ntawv no. Cov qauv ntawm tsib PhGs tau txheeb xyuas hauv qhov kev tshawb xyuas no tau qhia hauv daim duab 1.
echinacoside cov tshuajhauv cistanche
2. Kev sim
2.1. Tshuaj thiab reagents
Acteosidetau yuav los ntawm Sigma-Aldrich (Oak-Ville, ON), echinacoside tau yuav los ntawm ChromaDex (Santa Ana, CA). Isoacteoside raug cais los ntawm P. psyllium L. (Li et al., 2005).Cistanche deserticolatau yuav los ntawm Beijing TongRenTang Medicinal Store (Suav teb). Tag nrho cov kuab tshuaj yog qib HPLC thiab yuav los ntawm Caledon Laboratories Ltd. (Georgetown, ON).
2.2. Kev npaj ua qauv
Cistanche deserticola(20 g) tau muab rho tawm tsib zaug ntawm chav tsev kub rau 12 teev txhua nrog 100 mL ntawm 80 feem pua aqueous ethanol. Txhua zaus cov khoom sib tov tau lim los ntawm Whatman No.1 daim ntawv lim (Whatman International Ltd., Maidstone, England). Cov filtrate ua ke tau concentrated rau 100 mL hauv vacuo ntawm <40 degree.="" cov="" txiaj="" ntsig="" aqueous="" daws="" tau="" defatted="" ob="" zaug,="" txhua="" tus="" nrog="" 100="" ml="" ntawm="" hexane,="" thiab="" tom="" qab="" ntawd="" muab="" rho="" tawm="" ua="" ntu="" zus="" rau="" tsib="" zaug,="" txhua="" tus="" nrog="" 100="" ml="" n-butanol.="" cov="" txheej="" n-butanol="" tau="" sib="" xyaw="" ua="" ke="" thiab="" ua="" rau="" kom="" qhuav="" hauv="" lub="" tshuab="" nqus="" tsev="" ntawm=""><40 degree,="" uas="" tau="" txais="" 2.2="" g="" ntawm="" n-butanol="" extract.="" cov="" extract="" tau="" khaws="" cia="" ntawm="" -20="" degree="" ua="" ntej="" hsccc="" sib="">
2.3. HSCCC kev cais tawm
Kev npaj HSCCC tau ua tiav hauv Model CCC {{0}}} high-speed counter-tam sim no chromatography (Pharma-Tech Research, Baltimore, Maryland USA). Cov cuab yeej no muaj peb qhov kev npaj coils, txuas nrog hauv series (tag nrho ntim, 325 mL). Lub kiv puag ncig ceev ntawm lub cuab yeej tuaj yeem tswj hwm ntawm 0 thiab 2000 rpm. Lub HSCCC system tau nruab nrog lub twj tso kua mis HPLC (Pharma-Tech Research, Baltimore, Maryland, USA), tus qauv 450 UV ntes (Alltech, USA), tus qauv L 120 E flat-bed recorder (Linseis Inc., Princeton Jct, Teb Chaws Asmeskas), cov khoom siv feem ntau (Advantec MFS Inc., USA) thiab cov qauv txhaj tshuaj valve nrog 10-mL qauv voj.
Ib qho kev sib xyaw ntawm ethyl acetate-ethanol-dej (5:0.5:4.5, v/v/v) tau shaken hnyav nyob rau hauv ib qho kev sib cais thiab cia sawv ntsug thiab sib cais ntawm chav tsev kub. Ob theem tau siv hauv HSCCC tom qab lawv mus txog qhov sib npaug. Tag nrho cov kab coiled yog thawj zaug nrog cov txheej sab saud, uas ua haujlwm raws li theem nyob ruaj khov. Cov txheej txheem qis (theem txawb) tau muab tso rau hauv lub taub hau-kawg ntawm kem ntawm tus nqi ntws ntawm 1.5 mL / min. Kev sib hloov ceev tau teem rau ntawm 105{20}} rpm. Ib qho piv txwv (ca. 230 mg txhua lub sijhawm) tau daws hauv 8 ml ntawm kev sib xyaw ntawm ethyl acetate-ethanol-dej (5: 0.5: 4.5, v / v / v) tau ntim rau hauv qhov kev txhaj tshuaj valve tom qab lub kaw lus mus txog hydrodynamic. kev sib npaug. Qhov no biphasic hnyav system raug xaiv raws li kev faib tawm coefficient (K), uas yog 0.87, 1.11, thiab 1.32 rauacteoside, isoacteoside, thiab 20 acetylacteoside, feem. Tus nqi K yog qhov sib piv ntawm cov concentrations nyob rau sab saum toj thiab hauv qab cov khaubncaws sab nraud povtseg ntawm tib lub compound raws li tau txiav txim los ntawm HPLC (Fig. 2). Lub qhov hluav taws xob los ntawm qhov hluav taws xob ntawm kem tau saib xyuas tsis tu ncua los ntawm UV ntes ntawm 254 nm thiab sau rau hauv cov raj kuaj nrog cov khoom siv feem ntawm 4 min rau txhua lub raj.
2.4. Cov xwm txheej LC
static auto-sampler thiab photodiode array detector (DAD) tau siv rau kev tsom xam ntawm PhGs hauv n-butanol extract ntawmCistanche deserticolathiab feem ntau sau los ntawm HSCCC sib cais. Kev sib cais tau ua nyob rau hauv Phenomenex ODS-C18 kem (250 × 4.6 mm, 5 lm) nrog C18 guard kem. Cov theem binary mobile muaj xws li acetonitrile (cov kuab tshuaj A) thiab cov dej uas muaj 2 feem pua acetic acid (hloov B). Tag nrho cov kuab tshuaj tau lim los ntawm a
{{0}}.45 lm lim ua ntej siv. Tus nqi ntws mus tas li ntawm 1.0 mL/min rau tag nrho lub sijhawm khiav ntawm 25 min. Lub kaw lus tau khiav nrog qhov kev pab cuam gradient: 0–20 min: 90 feem pua B txog 60 feem pua B; 20–22 min: 60 feem pua B rau 0 feem pua B; thiab 22– 25 feeb, 0 feem pua B mus rau 90 feem pua B. Cov qauv txhaj tshuaj yog 10 lL. Peaks ntawm kev txaus siab tau saib xyuas ntawm 320 nm los ntawm DAD detector.
2.5. LC-ESI-MS kev sim
Kev sim LC-MS tau ua tiav siv Finnigan LCQ DECA ion trap mass spectrometer (Thermo Finnigan, San Jose, CA, USA) nruab nrog lub tshuab hluav taws xob ionization (ESI) qhov chaw. Cov qauv no tau soj ntsuam nyob rau hauv tib lub chromatographic mob. Tus qauv tsis zoo tau siv los sau cov ntaub ntawv. Lub sheath gas thiab auxiliary flow-rates tau teem rau ntawm 96 thiab 7 (arbitrary unit), feem. Lub capillary voltage tau teem rau ntawm 29 V thiab nws qhov kub thiab txias tau tswj ntawm 350 degree. Lub qhov hluav taws xob nkag ntawm lub lens tau kho ntawm 40 V thiab ntau lub RF amplitude tau teeb tsa ntawm 540 V. ESI koob voltage tau tswj ntawm 4.5 kV. Lub raj lens offset yog 16 V, lub lens multipole 1 offset yog 8.20 V thiab multipole lens 2 offset yog 10.5 V. Lub electron multiplier voltage yog teem rau 980 V rau ion detection.
2.6. NMR rau kev txheeb xyuas
NMR spectra raug kaw rau ntawm Bruker Avance-600 spectrometer (Bruker BioSpin Ltd., Milton, Canada). Tsuas yog cov tebchaw 2 thiab 5 (tsis muaj qauv) tau raug rau kev sim NMR. Cov qauv tau yaj hauv CD3OD.
3. Cov txiaj ntsig thiab kev sib tham
3.1. HSCCC sib cais
N-butanol extract ntawmCistanche deserticolathiab cov feem cuam tshuam rau txhua qhov siab tshaj plaws uas raug cais los ntawm HSCCC tau txheeb xyuas los ntawm HPLC, thiab cov txiaj ntsig tau muab rau hauv daim duab 2. Tsib lub ntsiab lus tseem ceeb (peaks 1–5) tau sib cais thiab kuaj nrog lub sijhawm tuav ntawm 9.2 min, 10.7 min, 12.3 min. ,
13.3 min thiab 16.2 min.
Kev ua tiav HSCCC kev sib cais feem ntau yog nyob ntawm qhov tsim nyog ob-theem hnyav qhov system uas muab qhov zoo tagnrho faib coefficient (K) nyob ib ncig ntawm 1 rau qhov xav tau. Xws li cov txheej txheem biphasic kuj tseem yuav ua rau lub sijhawm luv luv (Chen, Games, & Jones, 2003; Foucault & Chevolot, 1998; Oka, Oka, & Ito, 1991). Hauv peb qhov kev sim, peb xaiv plaub lub tshuab hnyav raws li kev solubility ntawm lub hom phiaj sib txuas hauvCistanche deserticola. HPLC tau siv los ntsuas qhov concentration hauv txhua theem, los ntawm qhov K qhov tseem ceeb ntawm lub hom phiaj sib txuas tau suav. Ob lub tshuab, ethyl acetate- n-butanol-ethanol-dej (4:0.6:0.6:5, v/v/v/v) thiab ethyl acetate-dej (1: 1, v/v), tau siv yav dhau los hauv HSCCC los caisacteosidethiab 20-acetylacteoside los ntawm C. salsa,acteoside, thiab isoacteoside los ntawm P. Psyllium, raws li (Lei li al., 2001; Li et al., 2005). Txawm hais tias thawj qhov system muaj lub sijhawm luv luv, nws muaj kev ua haujlwm tsis zoo hauv kev sib cais PhGs ntawmCistanche deserticola, vim qhov qis K qhov tseem ceeb rau cov tebchaw 1 thiab 2, thiab siab K qhov tseem ceeb rau cov tebchaw 3-5. Cov txiaj ntsig K tau qis heev rau cov tebchaw 1-4 hauv qhov system thib ob, tab sis siab heev rau compound 5 (Table 1). Ib qho kev hloov kho uas muaj ethyl acetate–ethanol-dej (5:{{10}}.5:4.5, v/v/v), muab qhov zoo tagnrho K tus nqi rau cov tebchaw 3–5 ntawm 0.87, 1.11, thiab 1.32, raws li, thiab ua rau muaj kev sib cais zoo ntawm peb lub tebchaw (Fig. 3A thiab B). Qhov no, txawm li cas los xij, tsim
me me heev tus nqi K rau cov tebchaw 1 thiab 2, ua rau ob lub tebchaw sib xyaw ua ke nyob ze ntawm qhov hnyav pem hauv ntej (feem 1 hauv daim duab 3A). Ib qho kev hloov kho ntxiv rau cov kab ke (ethyl acetate–n-butanol-ethanol-water (0.5:0.5:0.1:1, v/v/v/ v) elevated tus nqi K rau compound 1 thiab 2 rau 0.52 thiab 0.92, raws li, thiab coj mus ua kom tiav kev sib cais (Fig. 3C). Fig. 3A qhia HSCCC sib cais ntawm ib qauv muaj 230 mg ntawm n-butanol extract ntawmCistanche deserticolasiv ethyl acetate-ethanol-dej (5:0.5:4.5, v/v/v). Cov seem uas tau lees paub los ntawm HPLC kom muaj cov ntsiab lus 3, 4, lossis 5 tsuas yog sib xyaw ua ke, thiab cov uas muaj cov khoom sib xyaw 3 thiab 4 tau muab sib xyaw, khov-qhuav, thiab rov raug rau HSCCC rau kev sib cais ntxiv (Fig. 3B). Qhov ob-kauj ruam HSCCC sib cais tau piav qhia saum toj no tau muab tag nrho ntawm 14.6 mg, 30.1 mg, thiab 25.2 mg ntawm cov tshuaj 3-5 los ntawm 1412 mg n-butanol extract. Fig. 3C qhia HSCCC kev sib cais ntawm cov qauv uas muaj cov khoom sib txuas 1 thiab 2 (feem 1 hauv qhov kev sib cais thawj zaug) siv ethyl acetate-n-butanol-ethanol-water (0.5:0.5 :0.1:1, v/v/v/v). Tag nrho ntawm 28.5 thiab 18.4 mg ntawm cov tshuaj 1 thiab 2 tau txais. Cov chromatographic purities ntawm lub khov-qhuav cov tebchaw 1-5 yog tshaj 92.5 feem pua, uas tau siv ncaj qha rau LC-ESI-MS thiab NMR tshuaj ntsuam xyuas.
3.2. Kev txheeb xyuas tus qauv los ntawm LC-ESI-MS thiab NMR
Kev txheeb xyuas qhov tseeb ntawm cov tebchaw 1, 3, thiab 4 tau ua tiav los ntawm cov sijhawm khaws cia thiab UV spectral cov ntaub ntawv nrog cov qhabnias echinacoside,acteoside, thiab isoacteoside (Daim duab 2). Cov tshuaj 2 thiab 5 tsis paub, txawm li cas los xij, UV spectra ntawm tag nrho tsib lub tebchaw tau zoo sib xws, qhia txog cov yam ntxwv zoo sib xws.
Txhawm rau tshawb xyuas cov qauv ntawm tsib lub tebchaw no, kev sim LC-ESI-MSn tau sim thiab cov txiaj ntsig tau tshwm sim hauv daim duab 4 thiab Table 2. Cov tshuaj muaj feem xyuam rau cov peaks (1–5) hauv daim duab 2 tau nthuav tawm cov ions hnyav deprotonated molecular ions. [MH] — ntawm m/z 785, 799, 623, 623, thiab 665, ntsig txog, hauv hom tsis zoo. Dimeric [2M H] — ions kuj tau pom rau peaks 1–4 hauv daim duab 2. Cov no tau lees paub qhov hnyav molecular ntawm peaks 1–5 yog 786, 800, 624, 624, thiab 666, raws li. Cov ntaub ntawv LC–MSN (Table 2) muab cov ntaub ntawv muaj txiaj ntsig zoo rau tsib PhGs, xws li kev poob nruab nrab ntawm
Cov txheej txheem sim: kwv yees li 1 mg ntawm txhua tus qauv raug ntsuas hauv 10 mL xeem raj rau hauv uas 1 mL ntawm txhua theem ntawm pre-equilibrated ob-theem hnyav system tau ntxiv. Lub raj ntsuas tau capped thiab shaken rau 1 min, thiab tso cai rau sawv ntsug kom txog thaum nws sib cais tag nrho. Ib qho aliquot ntawm 100 lL ntawm txhua txheej tau muab tshem tawm thiab evaporated nyias mus rau dryness hauv vacuo ntawm<40 °c.="" the="" residue="" was="" dissolved="" in="" 10="" ll="" methanol="" and="" analyzed="" by="" hplc="" for="" determining="" the="" partition="" coefficient="" (k)="" of="" compounds="" 1–5.="" the="" k="" value="" was="" expressed="" as="" the="" peak="" area="" of="" the="" target="" compound="" in="" the="" upper="" phase="" divided="" by="" that="" in="" the="" lower="">
caffeoyl moiety (162), qabzib moiety (162), rhamnose moiety (146), CH2 radical (14), thiab COCH2 pawg (42).
LC-ESI-MS ntawm ncov 1 yog qhia hauv daim duab 4A. Lub deprotonated molecular ion [MH] — ntawm m/z 785 nrog ib tug siab abundance thiab ib tug dimeric deprotonated molecular
ion [2M H] — ntawm m / z 1571 tau pom nyob rau hauv hom tsis zoo, qhia txog qhov hnyav molecular ntawm 786, uas yog tib yam li echinacoside. Kev tshawb nrhiav ntxiv hauv LC-MS2 qhov kev sim ntawm m/z 785 ions yielded ib tug ntxhais tseem ceeb ion ntawm m/z 623 (Fig. 4B) ua ncaj qha los ntawm m/z 785 los ntawm kev poob ntawm caffeoyl moiety los yog hexose moiety li [M 162 H]— ib. LC-MS3 spectrum ntawm m/z 623 tau nthuav tawm ob lub ions loj ntawm m/z 477 thiab 461, thiab ob qho me me ntawm m/z 315 thiab 179 (Fig. 3C, Table 2). Qhov loj qhov sib txawv ntawm m / z 623 thiab cov fragment ions m / z 477 thiab 461 yog 146 thiab 162, raws li qhov poob ntawm ib chav rhamnose thiab cov piam thaj moiety lossis caf-feel moiety [M 162 H] -. Lub m / z 623 ions kuj poob caffeoyl moiety thiab rhamnose moiety los tsim m / z 315 ions. Lub ion ntawm m / z 179 tau tsim los ntawm kev sib cais ntawm caffeoyl moiety, nrog rau cov nqi tsis zoo uas tseem tshuav ntawm cov caffeoyl moiety. LC-ESI-MSN kev sim ntawm cov qhabnias echinacoside pom cov qauv sib cais. Yog li ntawd, Peak 1 tau lees paub tias yog echinacoside.
Rau lub ncov 2, LC-ESI-MS tau pom m/z 799 raws li lub deprotonated molecular ion [MH]- thiab m/z 1599 raws li nws dimeric ion, qhia txog molecular mass ntawm 800. Thaum lub sij hawm MS2 thwmsim, lub m/z 799 ions tsim peb tus ntxhais
ions ntawm m/z 637, 623, thiab 475 (Table 1). Lub ion ntawm m / z 637 tau tsim ncaj qha los ntawm niam txiv ion ntawm m / z 799 dua vim qhov nruab nrab poob ntawm caféoyl [M-162-H]- lossis qabzib moiety [M-162-H]—. Lub ion ntawm m / z 623 tshwm sim los ntawm qhov poob ntawm CH2 radical. Lub ion ntawm m / z 475 tau tsim los ntawm qhov tsis muaj kev poob ntawm ob qho tib si caffeoyl moiety [M 162 H] - thiab cov piam thaj moiety los ntawm niam txiv ion. Qhov kev sim MS3 ntawm m / z 637 tau tsim peb ions ntawm m / z 619, 491, thiab 475, sib xws rau qhov poob ntawm ib qho dej, chav tsev rhamnose, thiab cov piam thaj moiety, raws li. Lub m / z 623 tus ntxhais ion tsim m / z 461 thiab 315 hauv MS3 txoj kev tshawb fawb, uas ua raws tib txoj kev sib cais raws li echinacoside raws li tau tham saum toj no. Cov ntaub ntawv LC-ESI-MSN tau txhawb nqa kev txheeb xyuas qhov tseeb rau qhov siab tshaj 2 raws licistanoside A.
Kev sim LC-ESI-MS kuj tau ua rau qhov siab tshaj
3 thiab 4 (tR 12.49 thiab 13.46 min hauv daim duab 2). Ob lub ncov pom tib yam [MH] - ion ntawm m / z 623 thiab dimer ntawm m / z 1247 nyob rau hauv hom tsis zoo (Table 1), qhia tias lawv muaj peev xwm isomers nrog tib qhov hnyav ntawm molecular.
624, ibacteosidethiab isoacteoside. Lub MS2 spectra ntawm [MH] — ions kuj tau pom ib tug ntxhais ion ntawm m/z 461, uas qhia txog qhov poob ntawm caffeoyl moiety los ntawm niam txiv ion m/z 623 (Table 1). Zoo sib xws MS3 spectra tau txais rau ob lub tebchaw. Rau lub ncov 3, MS3 spectrum ntawm ion ntawm m / z 461 tsim peb ions ntawm m / z 315, 161, thiab 135. Lub m / z 315 yog tsim tom qab poob rhamnose raws li tau tham ua ntej. Lub ion m/z 161 yog tsim los ntawm qhov cleavage ntawm caffeoyl moiety, ua raws li los ntawm ib tug ntxiv poob ntawm dej; tus nqi tseem nyob ntawm ib feem ntawm caffeoyl moiety. Cov ion ntawm m / z 135 [agly-cone 18 H] - tshwm sim los ntawm qhov sib cais ntawm glycosidic daim ntawv cog lus ntawm C1 txoj hauj lwm nrog ib qho dej ntxiv, tawm hauv tus nqi mus rau ntawm aglycone moiety. Lub MS3 spectrum ntawm ncov 4 ua raws tib txoj kev fragmentation li ncov 3 tshwj tsis yog rau cov ion uas ploj lawm ntawm m/z 135. Cov molecular ion thiab cov qauv fragmentation ntawm ob lub tebchaw no zoo ib yam nrog cov ntaub ntawv cov ntaub ntawv ntawmacteosidethiab isoacteoside (Wang li al., 2000), txawm hais tias ib qho ntxiv ion m/z 153 tau pom thiab
muab ua [aglycone H] — los ntawm Wang et al. (Wang et al., 2000). Cov ion no tej zaum yuav tsis ruaj khov thiab poob dej los muab m/z 135 hauv peb qhov kev sim. Raws li cov ntaub ntawv MS thiab lub sijhawm khaws cia sib koom ua ke ntawm qhov siab tshaj 3 thiab 4 nrog cov qauv, lawv raug txheeb xyuas raws liacteosidethiab isoacteoside, feem.
Cov ntaub ntawv LC–ESI-MS ntawm ncov 5 yog qhia nyob rau hauv Table 2. Ib tug deprotonated molecular ion [MH]— (m/z 665) yog tib tug ion pom nyob rau hauv qhov tsis zoo hom, implying ib molecular mass ntawm 666. Peb tus ntxhais ions tau pom ntawm m/z 623, 503, thiab 461 hauv MS2 sim (Table 2). Cov ntxhais ions ntawm m / z 623 thiab 503 tau tsim ncaj qha los ntawm niam txiv ion los ntawm kev poob ntawm pawg COCH2 thiab caffeoyl moiety, raws li. Lub ion ntawm m/z 461 los ntawm qhov poob ntawm caféoyl thiab COCH2 moiety [M 162 42 H] — los ntawm niam txiv ion. Hauv kev sim MS, m / z 623 tsim m / z 461, thiab m / z 503 yielded peb ions ntawm m / z 485, 461, thiab 315. Lub MS3 spectrum ntawm tus ntxhais ion m / z 461 muab ob ions ntawm 443. thiab 315. Los ntawm kev muab piv rau LC– MSN fragmentation qauv ntawm ncov 5 nrog rau lwm cov tebchaw qhia nyob rau hauv txoj kev tshawb no thiab nrog rau lwm yam qhia (Li et al., 2005; Wang et al., 2000), peb xaus lus tias ncov 5 yog structurally muaj feem xyuam nrog. rau acteoside nrog qhov sib txawv tsuas yog COCH3 chav nyob ntawm R3 txoj haujlwm. Yog li ntawd, ib qho kev paub txog tus kheej tau muab rau ncov 5 raws li 20-acetylacteoside (Fig. 1).
Cov qauv ntawm ob lub ntsiab lus uas tau txheeb xyuas qhov tseeb, ncov 2 (raws li cistanoside A) thiab ncov 5 (raws li 20-acetyl-acteoside) tau lees paub ntawm lawv cov qauv los ntawm 1H NMR. Cov tshuaj hloov pauv thiab sib txuas ua ke ntawm tag nrho cov protons hauv cov tebchaw 2 thiab 5, raws li qhia hauv Table 3., sib phim nrog cov ntaub ntawv qhia NMR raucistanoside Athiab 20-acetylacteoside, raws li (Kobayashi li al., 1984, 1987). 2D NMR thwmsim (ntev-ntev COSY, ROESY, thiab CH correlation) kuj tau ua nyob rau hauv txoj kev tshawb no thiab lawv tau qhia ntxiv txog qhov kev txheeb xyuas (cov ntaub ntawv tsis qhia).
phenylethanoidglycosides hauv cistanche
4. Cov lus xaus
Nyob rau hauv daim ntawv tam sim no, HSCCC tau ua tiav siv rau kev sib cais thiab purification ntawm echinacoside,cistanoside A, acteoside, isoacteoside, thiab 20-acetylacteoside los ntawm n-butanol extract ntawm C. deserticola. Yog li ntawd nws yog ib qho pov thawj txhais tau tias rau semi-npaj sib cais ntawm bioactive. Lub caij no, cov qauv ntawm tsib PhGs hauv Cistanche deserticola tau tshawb xyuas los ntawm LC-ESI-MSN; Qee cov yam ntxwv ntawm PhGs tau pom, uas tso cai rau peb los txiav txim siab ua haujlwm hauv cov qauv. Txoj kev LC-ESI-MSN yog qhov cuab yeej muaj zog rau kev txheeb xyuas sai ntawmphenylethanoidsthiab lawv cov glycosides hauvCistanche deserticolarho tawm, tshwj xeeb tshaj yog thaum pom los ntawm NMR cov ntaub ntawv.
Kev lees paub
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Cov ntaub ntawv
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