Chemical Properties Of A Mannoglucan Los Ntawm Cistanche Deserticola

Hu rau: Audrey Hu Whatsapp / hp: 0086 13880143964 Email:audrey.hu@wecistanche.com

1. Khoom siv

Qhov ncauj pastes rau nees los ntawm ob lub tuam txhab sib txawv tau siv nrog 10 mg / g (A) lossis 20 mg / g (B) ntawm ivermectin, feem. Qhov kev ntsuam xyuas ntau rau ivermectin tau ua thiab cov ntsiab lus yog 104.4- 107.7 feem pua ​​​​ntawm cov lus tshaj tawm.

2. Kev kuaj kev puas tsuaj

Qhov kev xeem tau ua nrog Ph. Eur. paddle apparatus (Pharma Test, Heinberg, Lub teb chaws Yelemees), ntawm agitations ntawm 75 rpm lossis 50 rpm, ntawm 37 -0.5 C. Sodium lauryl sulfate (BDH Chemicals, Poole, England) 0.5 feem pua ​​(w /v) aqueous tov tau siv los ua qhov nruab nrab dissolution (900 ml). Cov muab tshuaj txhuam (4 g ntawm A los yog 2 g B) tau muab tso rau hauv lub nkoj nrog ib tug syringe nrog ib tug cannula (A) los yog xa rov qab cov qauv ntawm qhov chaw ntawm dissolution nruab nrab (B). Tom qab lub sij hawm ncua sij hawm (15, 30, 45, 60 min, thiab 2 h), ib qho piv txwv ntawm kev sib cais nruab nrab yog rho tawm (2 ml), lim los ntawm lub khob lim, thiab tshuaj xyuas.

3. Kev tshuaj xyuas HPLC

Kev txiav txim siab ntawm ivermectin tau ua nrog kev hloov kho HPLC raws li tau piav qhia hauv Ph. Eur. Cov tshuaj ntsuam xyuas tau diluted nrog methanol (1: 10). Lub HPLC system muaj ib kab octadecyl (250 -4 hli; 5 hli; Lichrospher RP-18; Merck), ib qho kev sib koom ua ke D-12500 A, tus ntes UV-Vis L{{7} }, twj L-6200A (Merck-Hitachi, Darmstadt, Germany). Kev sib xyaw ntawm acetonitrile: methanol: dej (64: 24: 16) tau siv los ua lub sijhawm txawb nrog tus nqi ntws ntawm 1.9 ml / min. Qhov ntim ntawm cov qauv txhaj tshuaj yog 20 ml thiab kuaj pom ntawm 254 nm.

4. Kev tshawb fawb txog Solubility

Ib qho kev nkag tau ntawm cov khoom raug shaken nyob rau hauv nruj kaw iav flasks nrog ib tug haum hnyav (Table 1) rau 20 h ntawm 37 0.1 C. Cov raug tshem tawm tau lim thiab cov filtrate, tom qab dilution hauv methanol, tau soj ntsuam los ntawm HPLC .

5. Kev ruaj ntseg hauv cov kua qaub

Cov kev daws teeb meem ntawm ivermectin hauv HCl {{0}}.1 M tau muab cia rau hauv cov iav kaw ntom ntom ntawm 20 C thiab 37 C. Chromatograms ntawm cov tshuaj tau sau tseg ntawm t ¼ 0 thiab tom qab 1, 2 thiab 6 h. Thaj tsam ntawm ivermectin (lub sijhawm khaws cia 10.5 min) thiab qhov siab ntxiv tau suav.

Cov ntaub ntawv

FDA Qhia rau kev lag luam: SUPAC-SS nonsterile semisolid dosage forms. Kev hloov pauv thiab kev pom zoo tom qab: chemistry, kev tsim khoom, thiab kev tswj hwm; Hauv vitro tso tawm kev sim thiab hauv vivo bioequivalence cov ntaub ntawv. Tsib Hlis 1997. FIP cov lus qhia rau kev ntsuas kev sib cais ntawm cov khoom siv qhov ncauj (1997) Dissolute. Tshuab. 4:5–14. Siewert M, Dressman J, Brown C, Shah VP (2003) FIP/AAPS cov lus qhia rau kev sib cais / hauv vitro tso tawm kev sim cov ntawv tshiab / tshwj xeeb ntau npaum li cas, AAPS PharmSciTech, 4 (1) tsab xov xwm 7 Modern Research Center for Traditional Chinese Medicine, School ntawm Pharmaceutical Sciences, Peking University Health Science Center, Beijing, Suav

Xiang mei Wu, Pengfei Tu Tau txais Lub Ob Hlis 13, 2004, tau txais lub Plaub Hlis 4, 2004, Prof. Dr. Pengfei Tu, Niaj hnub nimno Research Center for Traditional Chinese Medicine, Tsev Kawm Ntawv Pharmaceutical Sciences, Peking University Health Science Center, No. 38, Xueyuan Road , Haidian District, Beijing 100083, Suav teb Pengfeitu@bjmu.edu.cn Pharmazie 59: 815–816 (2004)

Ib tug tshiab mannoglucan los ntawmCistanche deserticolalos ntawm Tuam Tshoj yog characterized. Lub compound yog lub luag hauj lwm rau ib tug me me stimulation ntawm mitogen-induced T thiab B lymphocyte proliferation.

Cistanche deserticolaYC Ma yog ib qho holoparasite loj hlob ntawm cov hauv paus hniav ntawm cov nroj tsuag dicotyledonous Haloxylon ammodendron Bunge uas yog dav faib nyob rau sab qaum teb hnub poob ntawm Tuam Tshoj. Raws li nws yog ib qho tseem ceeb tonic nyob rau hauv cov tshuaj oriental, muaj ntau yam active Cheebtsam thiab pharmacological kev ua ub no (Karasawa li al. 1986; Yoshizawa li al. 1990). Cov khoom no tuaj yeem cuam tshuam nrog qee yam ntawm cov khoom siv polysaccharideCistancheuas tseem tsis tau tshawb nrhiav. Mitogenic pectic polysaccharides thiab neutral polysaccharides tau raug cais tawm thiab cov yam ntxwv los ntawmCistanche deserticolalos ntawm Mongolia (Radna et al. 1996; Anna et al. 1997; Bozena et al. 1999). Ntawm no peb tshaj tawm cov tshuaj lom neeg ntawm mannoglucan tshiab los ntawmCistanche deserticolalos ntawm Tuam Tshoj, uas yog lub luag hauj lwm rau me me stimulation ntawm mitogen-induced T thiab B lymphocyte proliferation. Tom qab stems ntawmCistanche deserticolaYC Ma tau muab rho tawm nrog 95 feem pua ​​​​ethanol, dej txias distilled yog siv los rho tawm cov seem. Cov dej txias extract tau precipitated nrog EtOH, cov protein raug tshem tawm thiab dialyzed, tau cov crude polysaccharide CCDP. Lub crude polysaccharide yog fractionated ntawm DEAE kem thiab Sephadex G-150 kem kom them tau cov protein-dawb polysaccharide (CDP-6), uas muaj cov piam thaj (89.2 feem pua) thiab mannose (1{{58}) }.4 feem pua ​​), thiab qhov nruab nrab Mr tau kwv yees yog 6.8 -104. Cov polysaccharide CDP-6 qhia [a 20 D 42 (c 1, H2O). Cov kev teeb tsa tiag tiag ntawm cov suab thaj tau txiav txim siab los ntawm GC ntawm TMS (– )-2-butyl glycosides. CDP-6 yog methylated (Needs et al. 1993), hydrolyzed, hloov mus rau alditol acetates, thiab soj ntsuam los ntawm GC thiab GC-MS (Table). Cov txiaj ntsig no ua ke nrog cov ntaub ntawv molar piv los ntawm kev tshuaj ntsuam methylation tau pom tias CDP-6 muaj feem ntau 1,6- txuas Glcp thiab qaum 1,6- txuas mus rau ib qho me me ceg ntawm O{{ 24}} ntawm mannose. Kev tshuaj xyuas methylation ntawm ob lub polymers (CDP-6-1, CDP-6-2) los ntawm ib nrab hydrolysis tau lees paub tias CDP-6 muaj cov pob txha uas muaj 1,6-txuas ntxiv cov glucosyl residues thiab 1,6- txuas mannosyl residues, uas tau hloov pauv ntawm O-3 ntawm mannose los ntawm ceg uas muaj lub davhlau ya nyob twg, 1,6- txuas glucosyl residues, thiab 1,3,{{38} }linked mannosyl residues. Raws li molar ratios thiab cov ntaub ntawv (Bozena li al. 1999; Bacon et al. 1996), 13C NMR spectrum ntawm CDP-6 pom cov teeb liab vim yog cov pa roj carbon monoxide ntawm a- d-Glcp (C{{45). }} ntawm 98.4 ppm), bd-Map (C-1 ntawm 104.3–104.9 ppm) thiab ad-Map (C-1 ntawm 100.0 ppm). Lub 1 H NMR spectrum ntawm CDP-6-2 hauv D2O pom cov cim proton sib raug rau ad-Glcp (H-1 ntawm 4.94–4.95 ppm) thiab a- d-Map (H-1 ntawm 4.96–5.00 ppm). Lub 13C NMR spectrum ntawm CDP-6-2 hauv D2O pom cov cim proton sib raug rau aD-Glcp (C-1 ntawm 98.6 ppm) thiab ad-Map (C-1 ntawm 98.4ppm). Mannoglucans tau tshaj tawm tias muaj 1, 4- txuas glucopyrannan thiab 1, 4- txuas mannopyrannan qaum qaum (Radjabi Nassab li al. 1984; Tanabe et al. 2000). CDP-6 muaj qee yam qauv sib txawv los ntawm nws qhov -1, 6-txuas Glcp thiab b-1,6- txuas daim ntawv qhia qaum, thiab nws tau hloov los ntawm mannoglucan ceg (Fig.). Yuav kom tshawb xyuas kev siv ntawmCistanche deserticolaHauv cov tshuaj pej xeem, cov txiaj ntsig ntawm CDP-6 ntawm T thiab B cell systems raug soj ntsuam. Nws tau pom tias yuav ua rau mob me me mitogen-induced T thiab B lymphocyte proliferation.

Kev sim

1. Cov txheej txheem dav dav

Kev hloov kho qhov muag tau ntsuas nrog W22-1S tsis siv neeg polarimeter. IR spectra tau txiav txim siab ntawm Perkin-Elmer 599B spectrometer. GC tsom xam tau ua tiav ntawm Agilent HP6890N ntsuas, nruab nrog FID ntes. GC-MS tau ua ntawm Finnigan Trace GC-MS ntsuas. Homogeneity thiab Mr ntawm CDP-6 tau kwv yees los ntawm calibration nkhaus ntawm elution ntim ntawm tus qauv dextrans, uas tau ua rau

ib qho Agilent 1100 series apparatus nrog Shodex KS-805 column.1 H NMR thiab 13C NMR spectra tau kaw nrog INOVA-500 ntsuas. Cov ntsiab lus ntawm cov protein tau txheeb xyuas los ntawm txoj kev ntawm Bradford (Andre li al. 1975). Monosaccharides tau soj ntsuam raws li lawv cov alditol acetate tom qab lawv tau hydrolyzed (2 M TFA, 2 h, 110 C), txo thiab acetated. Cov hydrolysates tau txheeb xyuas siv TLC ntawm lub silica gel phaj uas muaj 5 feem pua ​​​​sodium dihydrogen phosphate hauv BuOH-EtOAc-isopropyl cawv-HOAC––H2Opyridine (3.5: 10: 6: 3.5: 3: 3) thiab cov suab thaj tau kuaj pom los ntawm kev txau nrog lossis cinol. -H2SO4. Qhov tshwm sim alditol acetates tau soj ntsuam los ntawm GLC siv HP -5 kab capillary (30 m 0.32 mm). CDP-6 yog methylated los ntawm Ciucanu txoj kev, ua raws li hydrolysis, thiab tau soj ntsuam raws li ib feem methylated alditol acetates los ntawm GC-MS. Tom qab ib nrab hydrolysis, cov khoom tau dialyzed thiab sib cais, tau txais ob lub polymers, CDP-6-1 thiab CDP-6-2.

2. Cov khoom siv cog

Cov nroj tsuag tau sau los ntawm Inner Mongolia Xeev hauv Suav teb thaum Lub Yim Hli 2001 thiab tau txheeb xyuas los ntawm Hu-Biao Chen, Department of Natural Medicine, School of Pharmaceutical Sciences, Peking University Health Science Center. Ib daim ntawv pov thawj [E-1-(9)] ntawm cov nroj tsuag no tau muab tso rau hauv Tsev Kawm Ntawv Pharmaceutical Sciences, Peking University. 3. Extraction, cais, thiab purification ntawm polysaccharide CDP-6 Lub qia ntawmCistanche deserticolaYC Ma (1500 g) tau muab rho tawm nrog 95 feem pua ​​​​ethanol (6 L) nyob rau hauv reflux. Cov qhuav ethanol-insoluble residue yog macerated peb zaug nrog dej txias distilled ib 12 teev. Cov dej txias extract tau lim thiab cov filtrate centrifuged. Tom qab nag lossis daus nrog plaub ntim ntawm EtOH ( stirring thiab sawv rau 24 h ntawm 4 C), cov khoom precipitate (35.5 g) tau txais. Cov protein nyob rau hauv precipitate raug tshem tawm los ntawm Savage txoj kev. Tom qab soluble hauv dej distilled, qhov seem (25.5 g) tau dialyzed thiab lyophilized (yield: 10.6 g). Lub crude polysaccharide (8 g) tau yaj hauv dej distilled (100 mL, 8 feem pua ​​w/v) thiab sib cais siv DEAE kem (70 5 cm, gradient ntawm 0–2 M NaCl, ib 1.5 L). Ib qho dej eluted feem (600 mg) tau txais. Ib qho piv txwv (250 mg) raug ntxuav los ntawm kem gel-permeation chromatography ntawm Sephadex G- 150 kab ({25}} cm, 1600 mL, 5 mL ib feem). Cov feem ntawm CDP-6 (1500–1600 mL) tau muab sib sau ua ke, dialyzed, thiab khov-qhuav. Tom qab raug tshem tawm los ntawm Sephadex G-25 (60 1 cm). CDP-6 (110 mg) tau txais thiab txiav txim siab ua ib qho siab tshaj plaws los ntawm HPSGC.

Kev lees paub:Tus sau ua tsaug rau Dr. Baomei Shao thiab Prof. Xiaoming Gao, Lub Tsev Kawm Ntawv ntawm Kev Kho Mob Kev Tshawb Fawb, Peking University Health Science Center, rau kev ntsuas cov tshuaj tiv thaiv kab mob ntawm CDP-6.