Kawm Txog Tus Qauv Zoo Ntawm Cistanche Deserticola Gouqi Jujuepian

PU Hongli, ZHOU Bo, WEI Shuchang (Lub Tsev Kawm Ntawv Pharmacy, Gansu University of suav tshuaj, Lanzhou, Gansu, 730000, Suav teb)

Abstract: Lub Hom Phiaj Tsim kom muaj kev tswj xyuas zoo ntawm Ｒoucongrong Gouqi JujuepianＲoucongrong Ｒoucongrong ( Cistanches Herba ) thiab Gouqizi ( Lycii Fructus ) Ｒoucongrong Gouqi Jujuepian Ｒoucongrong Ｒoucongrong Ｒoucongrong ( Cistanches Herba ) thiab Gouqizi ( Lycii Fructus ). ib Hypersil BDS C18(25{2}}．0 mm × 4.6 mm,5 μm) kem ntawm 25 degree .Ib qho isocratic elution nrog methanol thiab 0.1 feem pua ​​formic acid tau siv ntawm tus nqi ntws ntawm 1 mL / min .Ib qho aliquot ntawm 20 μL ntawm cov qauv tshuaj tau txhaj rau hauv kem thiab qhov kuaj pom wavelength yog 330 nm. Cov txiaj ntsig TLC cov yam ntxwv ntawm oucongrong ( Cistanches Herba) thiab Gouqizi (Lycii Fruc tus) tau pom tseeb nrog qhov muaj zog tshwj xeeb .Cov kab ntau ntawm echinacoside thiab verbascoside yog 41．8 txog 4180 ug / mL (r=0 {17}}) thiab 40 txog 400 ug / mL (r=0. 999 9) . Qhov nruab nrab cov qauv rov qab yog 99. 58 feem pua ​​( ＲSD=0.85 feem pua ​​) thiab 99.31 feem pua ​​( ＲSD= 1.13 feem pua ​​) Ｒ Cov txheej txheem qhia hauv daim ntawv no tuaj yeem siv rau kev tshawb fawb thiab ntsuas qhov zoo ntawm Ｒoucongrong Gouqi Jujuepian.

Cov lus tseem ceeb: cistanche deserticola Gouqi Jujuepian; HPLC; TLC; echinacoside; verbascoside; tus qauv zoo

Cistanche deserticola thiab goji Berry

Cistanchedeserticolathiab goji chewable ntsiav tshuaj yog tsim los ntawm Cistanche thiab goji berries, uas muaj kev noj qab haus huv zog xws li ncua kev qaug zog thiab ua kom qaug zog tshem tawm.. Lawv pab tswj lub cev ua haujlwm, txhim kho kev tiv thaiv kab mob, thiab tshem tawm cov kab mob hauv lub cev.Tam sim no, vim muaj sijhawm ua haujlwm ntev thiab ua haujlwm siab, muaj ntau tus neeg nyob hauv qee qhov kev lag luam hauv Suav teb uas qaug zog thiab noj qab haus huv [1-3]. Cov pab pawg kev lag luam no yog cov pej xeem tsim nyog rau cov zaub mov noj qab haus huv no. Peb tau tsim nws ua cov ntsiav tshuaj uas tuaj yeem noj tau raws sijhawm txawm tias tsis muaj dej. Tom qab cov ntsiav tshuaj tau chewed, nws yooj yim rau nqos thiab tuaj yeem ua kom cov tshuaj yaj yeeb sai, txhim kho cov nyhuv kho [4]. Tsab ntawv xov xwm no kawm txog cov qauv zoo ntawm cov ntsiav tshuaj chewable, thiab cov txheej txheem tau txais txiaj ntsig zoo thiab kev tshuaj xyuas ntau tuaj yeem cuam tshuam qhov zoo ntawm cov ntsiav tshuaj chewable thiab ua raws li cov qauv kev tswj hwm zoo rau kev tsim.

Suav cov khoom siv tshuaj xws li cistanche deserticola

1. Ntsuas thiab ntsuas tshuaj

1.1 Cov cuab yeej

Agilent1260 high-performance kua chromatograph (Agilent, USA), UV Bluestar B ultraviolet-pom spectrophotometer (Beijing Laibetai Technology Co., Ltd.), WFH-203 (ZF-1) peb-txoj kev ultraviolet analyzer ( Shanghai Jingke Industry Co., Ltd.), BT125D 1/100000 hluav taws xob tshuav nyiaj li cas (Sartorius Scientific Instrument Co., Ltd.), DZF-6090 lub tshuab nqus tsev ziab (Shanghai New Scientific Instrument Co., Ltd.), SY{ {10}}T ultrasonic tu tshuab (Shanghai Shengyuan Ultrasonic Instrument Co., Ltd.), polyamide zaj duab xis (Zhejiang Taizhou Luqiao Sijia Biochemical Plastic Factory).

1.2 Tshuaj thiab reagent:

Echinacetincov khoom siv (batch tooj 111670-201505), poolside reference substance (batch number 111530-201411), Cistanche deserticola reference substance (batch number 121101-200402), and Lycium barbarum reference substance (batch tooj {{3}) }}) tag nrho tau txais los ntawm Tuam Tshoj lub koom haum rau Kev Tswj Xyuas Khoom Noj thiab Tshuaj. Cistanche deserticola ma thiab Lycium barbarum chewable ntsiav tshuaj yog tus kheej tsim los ntawm Tuam Tshoj Tshuaj Laboratory ntawm Tsev Kawm Ntawv Pharmacy, Gansu University of Traditional suav tshuaj, thiab methanol yog chromatographically ntshiab, Tag nrho lwm cov reagents yog analytical ntshiab.

Echinoside, cov tshuaj tseem ceeb ntawm Cistanche deserticola

2 Cov txheej txheem thiab cov txiaj ntsig

2.1 Nyias txheej chromatography (TLC) qhia

2.1.1 TLC kev txheeb xyuas ntawm Cistanche deserticola

Siv ib qho tsim nyog ntawm Cistanche deserticola thiab goji txiv hmab txiv ntoo chewable ntsiav tshuaj thiab grind lawv mus rau hauv hmoov. Noj 1.5 g ntawm hmoov zoo thiab muab tso rau hauv ib lub khob conical. Ntxiv 20 mL ntawm methanol thiab kho nrog ultrasound (lub zog 250 W, zaus 40 kHz) rau 15 feeb. Txias thiab lim. Concentrate lub filtrate kom ze li ntawm qhuav, thiab yaj cov residue nyob rau hauv 5 mL ntawm methanol raws li cov tshuaj ntsuam xyuas. Noj 1 g ntawm cov tshuaj siv tshuaj ntsuab Cistanche deserticola thiab npaj cov tshuaj ntawm cov tshuaj siv tshuaj siv tib txoj kev. Noj 0.5 g ntawm cov qauv tsis zoo yam tsis muaj Cistanche deserticola, thiab npaj cov tshuaj tiv thaiv tsis zoo siv tib txoj kev raws li kev sim tshuaj. Siv 0.5 txhua los ntawm kev tshuaj ntsuam xyuas, kev tswj tsis zoo, thiab siv cov tshuaj siv tshuaj μ L. Muab cov qauv ntawm tib lub polyamide zaj duab xis, tsim nrog methanol glacial acetic acid dej (2: 1: 7) raws li tus neeg sawv cev tsim, tshem tawm, cua qhuav. , thiab tshuaj xyuas nyob rau hauv lub teeb ultraviolet (365 nm). Hauv chromatogram ntawm cov qauv kuaj, muaj 4 qhov chaw nrog tib xim ntawm cov haujlwm sib txuas ntawm cov khoom siv tshuaj chromatogram, thiab kev tswj tsis zoo tsis muaj kev cuam tshuam. Thov saib daim duab 1 kom paub meej.

Cov tshuaj tseem ceeb ntawm Cistanche deserticola

Nyem qhov no mus saib Cistanche cov khoom

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2.1.2 TLC kev txheeb xyuas ntawm goji berries

Siv ib qho tsim nyog ntawm Cistanche deserticola thiab goji txiv hmab txiv ntoo chewable ntsiav tshuaj, grind lawv mus rau hauv hmoov, hnyav 2.5g ntawm hmoov zoo rau hauv ib lub raj centrifuge, yaj nyob rau hauv 35 mL dej, centrifuge, thiab extract lub supernatant nrog 15 mL ntawm ethyl acetate. Siv ib txheej ntawm ethyl acetate thiab mloog zoo rau 1 mL raws li cov tshuaj ntsuam xyuas. Noj 0.5 g ntawm goji txiv hmab txiv ntoo raws li cov ntaub ntawv siv, txiav mus rau hauv me me pieces, ntxiv 35 mL dej, tshav kub, thiab boil rau 15 feeb, cia nws txias, centrifuge, thiab rho tawm supernatant nrog 15 mL. ntawm ethyl acetate. Siv ib txheej ntawm ethyl acetate thiab mloog zoo rau 1 mL raws li cov ntaub ntawv siv tshuaj. Noj 2g ntawm cov qauv tsis zoo yam tsis muaj goji berries thiab npaj cov tshuaj tiv thaiv tsis zoo los ntawm kev npaj cov tshuaj ntsuam xyuas. Siv 2 cov qauv ntawm cov tshuaj ntsuam xyuas, 2 cov qauv ntawm cov kev tswj tsis zoo, thiab 2 cov qauv ntawm cov tshuaj siv tshuaj μ L. Cov qauv kuaj ntawm tib lub polyamide zaj duab xis, tsim nrog ethyl acetate chloroform formic acid (3: 2: 1) raws li tus neeg sawv cev tsim, tshem tawm, cua qhuav, thiab tshuaj xyuas nyob rau hauv lub teeb ultraviolet (365 nm). Nyob rau hauv chromatogram ntawm cov qauv kuaj, muaj ib qho chaw nrog tib xim ntawm qhov chaw sib txuas nrog cov ntaub ntawv siv tshuaj, thiab kev tswj tsis zoo tsis muaj kev cuam tshuam. Thov saib daim duab 2 kom paub meej.

1. Siv cov khoom siv tshuaj; 2, 3, kuaj qauv; 4. Kev tswj tsis zoo.

Daim duab 1 TLC daim duab ntawm Cistanche deserticola

1. Siv cov khoom siv tshuaj; 2. Kev tswj tsis zoo; 3. Cov qauv kuaj.

Daim duab 2 TLC daim duab ntawm goji berries

2.2 Kev Ntsuam Xyuas Ntawm Glycosides los ntawm Fructus Moringa thiab Echinacetin

2.2.1 Kev npaj cov kev siv tshuaj Precisely luj tsim nyog tus nqi ntawm echinacea thiab muab cov ntaub ntawv siv, muab tso rau hauv ib tug xim av volumetric flask, thiab ntxiv 50 feem pua ​​methanol los npaj ib tug tov reference tov uas muaj 0.836 mg ntawm echinacea thiab 0.799 mg ntawm poolside ib milliliter thiab tau txais [5].

2.2.2 Kev npaj cov tshuaj ntsuam xyuas

Noj cov ntsiav tshuaj Cistanche deserticola medlar chewable, zom rau hauv cov hmoov (dhau los ntawm 60 mesh sieve), hnyav 1.4 g meej, muab tso rau hauv 50 mL xim av volumetric hwj, ntxiv 50 feem pua ​​​​methanol rau lub teev. , co nws, hnyav, tsau rau 30 min, kho nws nrog ultrasound (lub zog 250 W, zaus 40 kHz) rau 40 min, txias nws, hnyav, siv 50 feem pua ​​​​methanol los ua kom txo qis, co nws nce, sawv ntsug, coj tus supernatant, siv 0.45 μ Lim nrog microporous membrane thiab coj cov filtrate tas mus li kom tau [6].

Cov tshuaj tseem ceeb ntawm Cistanche deserticola

2.2.3 Kev npaj cov tshuaj tiv thaiv tsis zoo

Noj ib qho tsim nyog tus nqi ntawm cov qauv tsis zoo yam tsis muaj Cistanche deserticola, zom rau hauv cov hmoov (sieved los ntawm 60 mesh sieve), raug hnyav 1.4 g, thiab npaj cov kev tswj tsis zoo raws li txoj hauv kev "2.2.2".

2.2.4 Chromatographic tej yam kev mob

Cov kab chromatographic yog Hypersil BDS C18 (250). raws li theem mobile B. Gradient elution yog ua raws li Table 1; Flow tus nqi 1 mL / min; Qhov kub ntawm qhov kub yog 25 degrees. Raws li cov txheej txheem chromatographic saum toj no, rho tawm 20 feem pua ​​​​ntawm cov kev siv tshuaj, 20 feem pua ​​​​ntawm cov tshuaj ntsuam xyuas, thiab 20 feem pua ​​​​ntawm cov kev tswj tsis zoo, raws li μ Txhaj L qauv, thiab sau cov chromatogram, raws li qhia hauv daim duab 3. Cov txiaj ntsig qhia tau tias lwm yam khoom tsis cuam tshuam nrog kev txiav txim siab ntawm poolside thiab echinacea, thiab kev sib cais ntawm poolside thiab echinacea yog qhov zoo.

Table 1 Gradient elution tej yam kev mob rau methanol (A) -0.1 feem pua ​​formic acid (B) mobile theem

2.2.5 Kev tshuaj xyuas kev sib raug zoo

Qhov tseeb ntsuas {{0}.5, 1.0, 2.0, 3.0, 4.0, thiab 5. {31}} mL ntawm kev siv tshuaj sib xyaw ntawm echinacea thiab poolside, muab tso rau hauv 10 mL xim av volumetric flasks, dilute lawv nrog 50 feem pua ​​​​methanol rau qhov teev, thiab txiav txim siab thaj chaw ncov raws li cov xwm txheej chromatographic hauv "2.2.4" . Linear regression tau ua tiav siv qhov chaw siab tshaj qhov tseem ceeb raws li kev tswj hwm (Y) thiab txhaj tshuaj raws li abscissa (X). Cov kev sib npaug regression rau echinacoside thiab poolside raug xam raws li Y=1493.8X-8.0748 (r=0.999 9) thiab Y=1895.8X{{ 25}}.486 (r=0.999 9), raws. Cov txiaj ntsig tau pom tias cov anthocyanins ntawm Echinacea thiab Moringa nyob nruab nrab ntawm 41.8-418.0 thiab 40-400, raws li μ Muaj kev sib raug zoo ntawm cov kab hauv g / mL.

2.2.6 Precision xeem

Ntsuas ntsuas qhov sib xyaw ua ke ntawm echinacea glycoside thiab pas dej ua ke 20 μ L. Txhaj 5 cov qauv sib law liag thiab sau thaj chaw ncov. Cov txiaj ntsig tau pom tias RSDs ntawm pineal thiab poolside yog 0.62 feem pua ​​​​thiab 0.51 feem pua ​​​​, raws li, qhia tau hais tias qhov zoo ntawm qhov ntsuas.

2.2.7 Stability test

Siv cov khoom siv tib yam ntawm cov tshuaj ntsuam xyuas thiab txhaj tshuaj ntawm 0, 4, 8, 16, thiab 32 teev raws li, sau qhov chaw ncov. Cov txiaj ntsig tau pom tias RSD ntawm pineal thiab pas dej ua ke yog 1.30 feem pua ​​​​thiab 1.35 feem pua ​​​​, raws li qhia tau hais tias cov tshuaj ntsuam xyuas tau ruaj khov hauv 32 teev.

1. Cov khoom siv sib xyaw; 2. Cov qauv kuaj; 3. Kev tswj tsis zoo;

A. Echinacetin; B. Maorui paj glycoside.

Daim duab 3 HPLC Daim duab ntawm Echinacetin thiab Fructus Moringae Glycoside

2.2.8 Repeatability test

Nqa 5 cov qauv los ntawm tib lub batch thiab npaj thiab ntsuas lawv ua ntu zus raws li txoj kev npaj ntawm cov tshuaj ntsuam xyuas. Cov txiaj ntsig tau pom tias RSD ntawm pineal thiab pas dej ua ke yog 0.82 feem pua ​​​​thiab 0.75 feem pua ​​​​, raws li, qhia tau hais tias qhov rov ua tau zoo ntawm txoj kev no.

2.2.9 Sample recovery test

Siv 5 cov qauv uas paub txog cov ntsiab lus ntawm anthocyanin thiab anthocyanin, ntxiv rau cov qauv tswj ntawm anthocyanin thiab anthocyanin, npaj cov tshuaj ntsuam xyuas raws li txoj hauv kev "2.2.2", thiab txiav txim siab tus nqi rov qab raws li cov xwm txheej chromatographic hauv "2.2. .4". Cov txiaj ntsig tau pom tias qhov nruab nrab rov qab los ntawm echinacea glycoside thiab poolside yog 99.58 feem pua ​​​​thiab 99.31 feem pua, feem, nrog RSD ntawm 0.85 feem pua ​​thiab 1.13 feem pua. Thov mus saib cov lus 2 thiab 3 kom paub meej.

2.2.10 Kev txiav txim cov ntsiab lus piv txwv

Siv cov txheej txheem kev txiav txim siab saum toj no los txiav txim siab cov ntsiab lus ntawm echinacoside thiab poolside hauv peb pawg ntawm cov qauv. Cov txiaj ntsig tau pom nyob rau hauv Table 4.

Table 2 Cov txiaj ntsig ntawm qhov rov qab ntsuas ntsuas rau echinacea glycoside ntxiv

Table 3 Cov txiaj ntsig ntawm kev ntsuas tus nqi rov qab rau qhov sib ntxiv ntawm Maorui paj glycosides

Table 4 Cov ntsiab lus kev txiav txim siab ntawm peb pawg ntawm cov qauv

3 Kev sib tham

Vim tias Cistanche deserticola yog cov tshuaj tseem ceeb hauv cov tshuaj ntawm Cistanche deserticola thiab Lycium barbarum chewable ntsiav tshuaj, nws suav rau feem ntau ntawm cov qauv. Yog li ntawd, kev soj ntsuam zoo thiab qhov ntau tau ua rau Cistanche deserticola, thiab kev soj ntsuam zoo tau ua rau Lycium barbarum noob. Cov txheej txheem siv yog yooj yim, rhiab heev, thiab muaj zog tshwj xeeb, uas tuaj yeem ua lub luag haujlwm tswj tau zoo.

Cistanche ntxiv nyob ze kuv

Hauv kev tsom xam kom muaj nuj nqis, kom ua tiav qhov kev sib cais zoo ntawm echinacoside thiab poolside, cov qauv hmoov yog thawj zaug soaked hauv 50 feem pua ​​​​methanol rau 30 feeb, thiab tom qab ntawd raug kho ultrasonic. Txhawm rau txiav txim siab cov ntsiab lus ntawm echinacoside thiab poolside hauv cov ntsiav tshuaj chewable, lub sij hawm rho tawm ultrasonic tau tshawb xyuas. Piv nrog rau cov qauv muab rho tawm rau 30, 40, thiab 50 feeb, nws tau pom tias echinacoside thiab pas dej ua ke tuaj yeem muab tshem tawm tag nrho tom qab 40 feeb.. Txoj kev tsim hauv kab lus no tuaj yeem tswj tau qhov zoo ntawm cov ntsiav tshuaj chewable.

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