HPLC Fingerprinting Thiab Spectrum-antitumor Effect Relationship For Discrimination Ntawm Mylabris Phalerata Pallas Thiab Mylabris Cichorii Linnaeus

Hu rau: emily.li@wecistanche.com

Jian-Yong Zhang, Qi-Hong Chen, Xian Pei1, Rong Yan, Can-Can Duan, Yun Liu4, Xiao-Fei Li

1 Lub Tsev Kawm Ntawv Pharmacy, Zunyi Medical University, Zunyi, Suav teb,

2 Lub Chaw Haujlwm Tseem Ceeb ntawm Cov Tshuaj Kho Mob ntawm Lub Tsev Haujlwm Saib Xyuas Kev Kawm thiab Kev Tshawb Fawb Kev Tshawb Fawb Thoob Ntiaj Teb ntawm Ethnomedicine ntawm Ministry of Education, Zunyi Medical University, Zunyi, Guizhou, Suav.

3 Lub Tsev Kawm Ntawv Qib Siab Kev Kho Mob, Zunyi Medical University, Zunyi, Tuam Tshoj, 4Guizhou Provincial College-based Key Lab rau qog tiv thaiv thiab kho mob nrog cov tshuaj tshwj xeeb, Zunyi Medical University, Zunyi, Suav.

Nyem qhov no kom tau txais cov ntaub ntawv ntau ntxiv txog Cistanche anti-tumor

Qhov tseem ceeb

Txoj kev tshawb no muab ib txoj kev siv tau, ntseeg tau, thiab ua tau zoo dua HPLC txoj kev rau kev ntxub ntxaug ob hom Mylabris. Thiab, HPLC fingerprinting thiab spectrum-cov teebmeem antitumortau koom ua ke thiab peb qhov kev sib txawv tseem ceeb tau pom rau tus cim tshiab ntawm Mylabris.

Abstract

Lub Hom Phiaj: Kev ntsuam xyuas kev ntxub ntxaug ntawm ob hom Mylabris raws li HPLC ntiv tes thiab spectrum-antitumor nyhuvkev sib raug zoo. Txoj Kev: Hauv txoj kev tshawb no, ib qho yooj yim thiab ua tau zoo ua kua chromatography (HPLC) txoj kev sib koom ua ke nrog kev tshuaj ntsuam chemometric thiab spectrum-antitumor nyhuvKev sib raug zoo tau tsim los rau kev ntxub ntxaug ntawm ob hom Mylabris: Mylabris phalerata Pallas (MP) thiab Mylabris cichorii Linnaeus (MC). Cov txiaj ntsig: Hauv kev txheeb xyuas tus ntiv tes, 14 tus yam ntxwv siab tshaj plaws tau raug xaiv los ntsuas qhov sib txawv ntawm MP thiab MC siv qhov zoo sib xws thiab cov qauv kev ntsuas kev paub siv PCA thiab OPLS-DA. HPLC chromatograms ntawm cov qauv los ntawm 10 cheeb tsam ntawm Tuam Tshoj tau pom qhov sib txawv ntawm MP thiab MC, thiab 7 cov cim tshuaj lom neeg tau pom. Hauv spectrum-antitumor nyhuvKev txheeb xyuas kev sib raug zoo, 4 cov cim kev ua si tau ua lub luag haujlwm tseem ceeb hauv kev txo qis IC50 thiab tej zaum yuav yog cov tshuaj tiv thaiv kab mob ntawm Mylabris los ntawm kev txheeb xyuas grey kev sib raug zoo thiab kev txheeb xyuas ntau yam kab rov tav. Kev tshuaj ntsuam chemometric ua ke nrog cov txiaj ntsig kev sib raug zoo ntawm cov txiaj ntsig tau qhia tias peaks 2 (cytosine), 4 (tsis paub), thiab 14 (tsis paub) yog cov cim tseem ceeb sib txawv rau qhov txawv ntawm ob hom Mylabris.

Kev xaus: Txoj kev siv tau, muaj kev ntseeg siab, thiab muaj txiaj ntsig zoo rau kev ntxub ntxaug MP thiab MC, thiab yuav muab txoj hauv kev tshiab rau kev tswj xyuas cov tshuaj tua kab kom zoo.

Keywords: HPLC, Fingerprinting, Spectrum-antitumor nyhuv, Mylabris, Kev ntxub ntxaug

Keeb kwm

Mylabris, hu ua ban mao nyob rau hauv Suav teb, tau siv los ua tshuaj suav tshuaj (TCM) rau kev kho mob furuncle, tob hauv paus rwj, plab plab, thiab yog ib qho tseem ceeb.antitumortus neeg sawv cev. Tsis tas li ntawd, nws yog dav siv nyob rau hauv cov teb chaws Europe raws li pej xeem tshuaj [1]. Cov khoom tseem ceeb ntawm Mylabris yog cantharidin, uas yog qhov ua tau zootshuaj tiv thaiv qogcompound [2]. Cov kev tshawb fawb tshuaj niaj hnub no tau pom tias Mylabris muaj ntau yam dej num, thiab muaj txiaj ntsig zoo rau kev kho mob qog nqaij hlav vim yog nws ob qho tib si.tshuaj tiv thaiv kab mobcov khoom thiab muaj peev xwm nce cov leukocytes [3-4]. Tam sim no, Mylabris tau siv dav hauv qee cov tshuaj tiv thaiv kab mob qog noj ntshav TCM hauv Suav teb, xws li Aidi txhaj thiab Compound txwv tsis pub mao capsule.

Raws li Pharmacopoeia ntawm Tuam Txhab Tuam Txhab Tuam Tshoj (2015 version), Mylabris txhais tau tias yog lub cev qhuav ntawm Mylabris phalerata Pallas (MP) thiab Mylabris cichorii Linnaeus (MC), uas txawv ntawm lawv cov yam ntxwv ntawm botanical, cov pej xeem dynamics, thiab ecology [ 5-6]. Mylabris kuj yog ib qho tshuaj lom heev. Txij li cov tshuaj muaj nyob hauv Suav teb feem ntau yog sib xyaw ntawm MP thiab MC, nws yog ib qho tsim nyog los txheeb xyuas cov tshuaj thiab kev ua haujlwm sib txawv ntawm ob hom.

Txij li TCM's muaj ntau cov khoom xyaw uas paub thiab tsis paub, kev txheeb xyuas qhov profile ntawm lawv cov khoom siv tshuaj ua rau muaj kev sib tw tseem ceeb. High-performance liquid chromatography (HPLC) fingerprinting plays lub luag hauj lwm tseem ceeb hauv kev tswj kom zoo (QC) ntawm TCM vim nws tag nrho cov yam ntxwv profile ntawm feem ntau cov tebchaw nyob rau hauv ib tug complex system. Tsis tas li ntawd, kev ntsuam xyuas ntiv tes ntawm TCMs muaj qhov zoo rov ua dua thiab ruaj khov rau hom kev txheeb xyuas thiab kev soj ntsuam zoo, thiab tau txais los ntawm World Health Organization, Lub Xeev Food and Drug Administration ntawm Tuam Tshoj (SFDA), thiab European Medicines Evaluation Agency (EMEA). Tsis ntev los no, kev tshuaj ntsuam chemometric combining HPLC fingerprinting tau raug tsim los rau QC thiab kev ntxub ntxaug ntau yam ntuj tsim [7]. Txawm li cas los xij, nws tsis muaj txiaj ntsig rau kev ntxub ntxaug qhov sib txawv ntawm TCMs vim nws tsis tuaj yeem cuam tshuam qhov sib txawv ntawm cov khoom sib txawv, uas tseem ceeb dua rau kev siv tshuaj kho mob. Txog tam sim no, spectrum-antitumor nyhuvCov txheej txheem kev sib raug zoo tau siv los txheeb xyuas cov khoom siv bioactive ntawm TCMs [8]. HPLC fingerprinting ua ke nrog kev tshuaj ntsuam chemometric thiab spectrum-antitumor effect kev sib raug zoo yuav zoo dua qhia qhov txawv ntawm ntau qhov chaw ntawm TCMs.

Tsis ntev los no qee txoj hauv kev tau tshaj tawm rau kev txiav txim siab cov ntsiab lus ntawm cantharidin, uas tuaj yeem siv los txiav txim siab qhov zoo ntawm Mylabris [9]. Qee qhov chromatographic ntiv tes luam ntawv kuj tau thov rau QC [10-11]. Tsuas yog ib txoj kev tshawb fawb tau siv ib txoj hauv kev uas cuam tshuam nrog gradient elution ntawm cov dej-soluble compounds ntawm Mylabris raws li HPLC. Lub caij no, tsuas yog kaum tus qauv tau siv, thiab tsis muaj MC cov qauv [12]. Rau qhov zoo tshaj plaws ntawm peb txoj kev paub, kev ntxub ntxaug MP thiab MC raws li tshuaj lom neeg muaj pes tsawg leeg thiab kev ua haujlwm tsis tau kawm ua ntej.

Txoj kev tshawb no tsom los tsim ib txoj hauv kev yooj yim thiab ua tau zoo los cais ob hom Mylabris siv HPLC ntiv tes ua ke nrog kev tshuaj ntsuam xyuas thiab spectrum-antitumornyhuvkev sib raug zoo, uas yuav muab lub hauv paus rau QC ntawm Mylabris. Txoj kev no tseem tuaj yeem siv rau kev txheeb xyuas QC cim ntawm lwm TCMs.

Cov ntaub ntawv thiab cov txheej txheem

Nyob rau hauv tag nrho, 20 pawg ntawm Mylabris phalerata Pallas (MP) thiab Mylabris cichorii Linnaeus (MC) tau sau los ntawm ntau lub xeev ntawm Tuam Tshoj (Table 1) thiab txheeb xyuas los ntawm peb tus kws tshaj lij Xiao-Fei Li. Cov ntawv pov thawj tau muab tso rau hauv peb lub chaw kuaj mob.

Cov qauv siv cytosine, uridine, guanosine, thiab adenosine tau yuav los ntawm ChengduPush Biotechnology Co., Ltd. (Chengdu, Suav). HPLC qib acetonitrile thiab methanol tau yuav los ntawm Thermofisher Scientific (Fairlawn, NJ, USA). Watson cov dej ntshiab tau siv thoob plaws hauv txoj kev kawm. Lwm cov tshuaj thiab cov kuab tshuaj yog cov qib tshuaj ntsuam xyuas thiab tau txais los ntawm Aladdin Reagent (Shanghai, Tuam Tshoj), Chengdu Kelong Reagent (Chengdu, Tuam Tshoj) thiab Sinopharm Chemical Reagent (Beijing, Suav)

Apparatus thiab chromatographic tej yam kev mob

Kev tshuaj xyuas HPLC tau ua los ntawm Agilent 1260 Infinity HPLC system ntawm 30 degree ntawm Phenomenex Synergy Polar-RP80 (4.6 mm × 250 mm, 5 μm) rau kev luam ntiv tes. Linear gradient elution nrog eluents A (dej / glacial acetic acid, 100: 1, v / v) thiab B (methanol) tau siv rau kev sib cais. Txoj haujlwm gradient tau tsim raws li hauv qab no: 0-1 min, 3.0-4.6 feem pua ​​B; 1-9 min, 4.6-6.8 feem pua ​​B; 9-25 min, 6.8-51.0 feem pua ​​B; 25-30 min, 51.0-100 feem pua ​​. Tus nqi ntws ntawm cov theem mobile yog 1.0 mL / min. Lub chromatogram tau soj ntsuam ntawm 254 nm. Tom qab 15 feeb sib npaug, 10 μL ntawm cov qauv siv rau kev txhaj tshuaj.

Kev npaj ua qauv

Cov qauv qhuav tau raug muab tso rau hauv cov hmoov, thiab 2.5 g ntawm txhua cov qauv hmoov tau muab rho tawm ob zaug nrog 50 mL ntawm 75 feem pua ​​​​ethanol los ntawm reflux rau 1.5 teev txhua zaus. Cov qauv muab rho tawm yog tov thiab concentrated nyob rau hauv txo siab mus rau 20 mL. Tom qab ntawd cov sib tov yog precipitated nrog 80 mL dej rau 24 teev nyob rau hauv 4 degree. Tom qab ntawd, cov extract tau centrifuged ntawm 3000 g min{11}} rau 10 min kom cais cov supernatant. Lub supernatant yog concentrated nyob rau hauv txo siab mus rau 25 mL. Thaum kawg, tus supernatant tau lim los ntawm 0.22 μm Millipore zaj duab xis ua ntej HPLC tsom xam.

Methodology validation

Txoj kev HPLC tau raug lees paub rau qhov tseeb, rov tsim dua tshiab, thiab ruaj khov (0 h, 2 h, 4 h, 8 h, 12 h, 24 h). Qhov kev lees paub tau kwv yees raws li lub sijhawm khaws cia (RT) thiab thaj chaw siab tshaj (PA). Thaum kawg, chemical fingerprints ntawm 20 pawg ntawm Mylabris tau tsim los txheeb xyuas cov yam ntxwv ntawm kev sib xyaw ua ke ntawm ntau lub tebchaw. Qee cov txheej txheem chemometric tau qhia txog cov tshuaj ntiv tes rau kev tshuaj xyuas.

Pattern recognition tsom xam

Qhov sib txawv ntawm ob hom ntawm thaj chaw ncov tau raug tshuaj xyuas rau qhov tseem ceeb ntawm cov ntaub ntawv siv cov tub ntxhais kawm tsis muaj kev sib tw t-test. P-tus nqi tsawg dua 0.05 tau suav tias yog qhov tseem ceeb. Raws li cov txheej txheem classic tsis saib xyuas, kev tsom xam cov ntsiab lus tseem ceeb (PCA) tau siv dav rau kev txheeb xyuas cov ntaub ntawv txheeb xyuas. Hloov chaw ntawm kev siv ntau qhov sib txawv, PCA yuav siv qee lub PCs me me yam tsis tau poob ntau cov ntaub ntawv, thiab cov qhab nia tau pom zoo rau kev sib cais dawb ntawm kev soj ntsuam. Nyob rau hauv txoj kev tshawb no, PCA tau ua nyob rau ntawm qhov chaw siab tshaj plaws ntawm txhua qhov chaw hauv HPLC cov ntiv tes siv SIMCA-P ntxiv rau 14.0 software (Umetrics, Umea, Sweden) txhawm rau nrhiav kev ntxub ntxaug ntawm ob hom Mylabris.

Rau kev ntxub ntxaug ntau dua ntawm ob hom Mylabris, ib txoj hauv kev saib xyuas ntawm orthogonal ib feem tsawg squares cais cais (OPLS-DA) tau siv los tshuaj xyuas qhov sib txawv ntawm qhov chaw siab tshaj plaws ntawm txhua yam hauv HPLC ntiv tes. Tus nqi VIP yog qhov hnyav ntawm cov squares ntawm OPLS qhov hnyav, cuam tshuam txog kev sib raug zoo ntawm txhua tus X sib txawv rau tus qauv. Qhov sib txawv nrog VIP> 1, ua ke nrog S-plot tau suav tias yog qhov cuam tshuam rau kev sib cais ntawm cov qauv hauv cov qhab nia tau tsim los ntawm OPLS-DA tsom xam. Tom qab ntawd, cov yam ntxwv tshuaj cim rau kev ntxub ntxaug tau txais.

Spectrum-antitumor effect kev txheeb xyuas

Tshawb nrhiav qhovtshuaj tiv thaiv qogCov nyhuv peak cheeb tsam rau Mylabris cov qauv ua ke nrog cov ntaub ntawv ntiv tes, thiab sib piv qhov sib txawv ntawm ob hom, kev sib raug zoo ntawm lub ncov ntawm cov ntiv tes thiabtiv thaiv-qogcov teebmeem tau kawm. Ob txoj hauv kev sib xyaw ua ke grey kev txheeb xyuas kev txheeb xyuas thiab kev txheeb xyuas ntau hom kab rov tav kab rov tav (MLRA) tau siv los ntawm SPSS19.0 (IBM, USA) thiab GM6.0 mos (Grey Systems Theory Institution, NUAA, Tuam Tshoj). Covtiv thaiv-qogkuaj tau ua los ntawm peb cov txheej txheem luam tawm [12], tib neeg hepatocellular carcinoma cell kab HepG2 hlwb tau khaws cia hauv RPMI1640 nruab nrab ntxiv nrog 10 feem pua ​​​​fetal bovine serum. Cov hlwb tau loj hlob nyob rau hauv ib qho chaw humidified uas muaj 5 feem pua ​​CO2 ntawm 37 C. Txoj kev SRB tau siv los ntsuas qhov kev loj hlob rautiv thaiv-qogkev ua si, tom qab ntawd IC50 ntawm HepG2 hlwb raug xam, thaum kawg, IC50 thiab qhov chaw siab tshaj plaws tau siv rau kev tshuaj ntsuam chemometric. Tom qab ntawd, cov cim kev ua yeeb yam ntawm Mylabris tau txais.

Cov txiaj ntsig thiab kev sib tham

Optimization ntawm extract tej yam kev mob

Rau kom txaus extraction ntawm cov dej-solut compounds los ntawm Mylabris, lub extraction system yog optimized. Ultrasonic thiab refluxing pom tias cov tebchaw no tuaj yeem ua kom zoo los ntawm refluxing.

Optimization ntawm HPLC txoj kev

Txhawm rau kom tau txais qhov siab tshaj plaws chromatographic peaks los piav qhia tag nrho cov yam ntxwv ntawm cov tshuaj ntsuab, muaj pes tsawg leeg ntawm chromatographic kem, theem mobile, thiab kev tshawb nrhiav wavelengths (200, 254, 265, thiab 278 nm) tau tshawb xyuas. Cov txiaj ntsig tau pom tias Phenimenex Synergy Polar-RP80 kem, dej (muaj 1 feem pua ​​​​glacial acetic acid) thiab methanol, thiab 254 nm yog qhov zoo tshaj plaws rau Mylabris HPLC tsom xam (Daim duab 1), cov qauv ntawm 4 tus qauv tshuaj muaj nyob rau hauv daim duab 2. .

Txoj kev validation

Nyob rau hauv txoj kev tshawb no, 14 peaks tau zoo sib cais thiab siv ua "peaks". Lub precision, repeatability, thiab stability yog raws li nyob rau hauv lub sij hawm tuav (RT) thiab ncov cheeb tsam (PA). Tus nqi RSD ntawm RT thiab PA rau qhov tseeb (n=6) tsis tshaj 0.2 thiab 4 feem pua; RSD qhov tseem ceeb ntawm RA thiab PA rau kev rov ua dua (n=6) yog qis dua 0.2 thiab 4 feem pua, raws li, thiab RSD tus nqi rau kev ruaj ntseg (0-24 h) tsawg dua { {10}}.4 thiab 5 feem pua ​​​​, feem, uas qhia tias cov qauv ruaj khov hauv 24 teev. Cov txiaj ntsig no qhia tau hais tias qhov zoo ntawm cov qauv kawm thiab HPLC txoj kev tau ruaj khov thiab tswj tau zoo.

Kev ntsuam xyuas tus ntiv tes zoo sib xws

The fingerprint similarity analysis was used to evaluate the similarity of HPLC peaks. The similarities of MP and MC were calculated by the reference HPLC fingerprint, respectively. As shown in Table 1, except for two MP samples (from Guizhou2 and Guizhou5) with lower similarities (0.880 and 0.920, respectively), other samples of MP were >0.930. The similarities of MC samples were >{{0}}.921 tshwj tsis yog cov qauv MC (los ntawm Guizhou2 thiab Guizhou4), uas nws qhov zoo sib xws yog 0.888 thiab 0.887, raws li. Cov ntaub ntawv no tau qhia tias qhov zoo ntawm Mylabris hauv ib hom yog ruaj khov. Txawm li cas los xij, qhov zoo sib xws ntawm tsuas yog peb tus qauv MP piv nrog cov ntaub ntawv HPLC ntiv tes yog > 0.930; thiab qhov zoo sib xws ntawm tsuas yog ob qho qauv MC piv nrog qhov siv HPLC ntiv tes yog> 0.921. Cov txiaj ntsig no tau pom tias ob hom Mylabris tau hloov pauv ntau hauv qee thaj chaw ncov.

Kev ntxub ntxaug ntawm MP thiab MC los ntawm kev txheeb xyuas tus qauv paub

For global analysis of the difference, PCA was used to find the quality variation of the samples from the two species of Mylabris. Figure 3 shows that the two-dimensional PCA model was constructed by the first two PCs, which included approximately 61.8% of the original data. The score plot showed that the MP and MC samples could not be separated by PCA. To understand the differences between MP and MC, an OPLS-DA model was established. As shown in Figure 4, the Mylabris samples could be classified into two groups with R2X=0.502, R2Y=0.492, and Q2=0.0769 as compared to the PCA model. These results showed that the OPLS-DA model was more suitable than the PCA model for distinct separation of the test samples based on their different components. From the S-plot of OPLS-DA, peak markers including peaks 1, 2, 4, 7, 9, 12, and 14 between MP and MC could be found (Figure 5). Based on VIP>1, peaks 1, 2, 4, 7, 9, 12, thiab 14 tej zaum yuav yog qhov sib txawv tseem ceeb tshaj plaws hauv kev ntxub ntxaug ntawm ob hom (Daim duab 6). Cov pab pawg siab tshaj (1, 2, 4, 7, 9, 12, thiab 14) tuaj yeem ua lub luag haujlwm tseem ceeb hauv kev sib txawv ntawm MP thiab MC raws li cov cim tshuaj lom neeg.

Spectrum-effect kev sib raug zoo los ntawm grey kev txheeb xyuas kev txheeb xyuas

To further evaluate the relationship between the variations of normalized peak area and IC50, grey relational analysis (GRA) was performed. The influence rank by normalized peak area was P13>P14>P3>P2>P12>P4>P9>P1>P10>P7>P6>P8>P5> P11 as shown in Table 2. The results indicated that the top-6 peak including peaks 13, 14, 12, 2, 3, and 4 were the main influencing factors for the antitumor effect based on the standard of Relative Grey correlative degree (RGCE) >0.75.

Spectrum-effect kev sib raug zoo los ntawm MLRA

MLRA qauv yog tus qauv qauv tshaj plaws rau kev txiav cov kev sib raug zoo ntawm ob lossis ntau qhov sib txawv thiab cov lus teb uas tau tsim los ntawm cov qauv hauv qab no

Qhov twg Y yog tus nqi kwv yees thiab sawv cev rau cov lus teb; Xn yog qhov sib txawv ntawm tus kheej, b{{0}} yog qhov cuam tshuam, thiab bn yog qhov sib txawv ntawm qhov sib txawv rau Xn. Hauv txoj kev tshawb no, MLRA tau siv los tsim kom muaj kev sib raug zoo ntawm tus ntiv tes-ua tau zoo ntawm cov txiaj ntsig ntawm thaj chaw ncov hauv HPLC cov ntiv tes thiab IC50 ntawm cov tshuaj tiv thaiv HepG2, thiab tom qab ntawd nrhiav cov tshuaj tiv thaiv kab mob. Kev sib koom ua ke ntawm cov ntaub ntawv tau pom los ntawm ib qho qauv MLRA, uas tsis tsim nyog los tshawb txog kev sib raug zoo ntawm Y (IC50) thiab X (PA). Tus qauv PCA MLRA tau siv los kawm txog kev sib raug zoo ntawm tus ntiv tes, thiab thawj rau lub PCs nrog rau qhov sib txawv ntawm qhov sib txawv: 91.068 feem pua ​​​​tau raug xaiv los tshuaj xyuas. Thaum kawg, qhov sib npaug hauv qab no tau tsim los raws li SPSS cov zis thiab cov PCs: IC50=1.115864 ntxiv （1.87268PA1- 699.722PA2 ntxiv rau 25.7138PA3 - 24.1528PA4 ntxiv rau 7.22878PA{{ 23}}.2114PA6 plus 2.95283PA7- 15.5305PA8 plus 13.0297PA9 plus 22.5683PA10 - 10.3462PA11 plus 123.762PA12 plus 31.0428PA.7}{42}. (R yog 0.682, P <>

Integrating Analysis ntawm Ob Mylabris Hom

Qhov kev sim tau pom tias HPLC ntiv tes tuaj yeem siv los cuam tshuam cov yam ntxwv ntawm Mylabris. Cov txheej txheem zoo sib xws, PCA, thiab OPLS-DA tau siv los nrhiav qhov sib txawv ntawm MC thiab MP. Cov pab pawg siab tshaj (1, 2, 4, 7, 9, 12, thiab 14) tau txhais tias yog cov cim tshuaj lom neeg.

Tsis tas li ntawd, ob qho tib si GRA thiab MLRA tau ua pov thawj los ua ib txoj hauv kev txaus siab rau kev ntxub ntxaug ntxiv. Kev sib xyaw ua ke ntawm GRA thiab MLRA kev soj ntsuam cov txiaj ntsig, qhov siab tshaj 2, 4, thiab 14 raws li cov cim kev ua si ntawm Mylabris yuav tsum muaj lub luag haujlwm rau cov nyhuv tiv thaiv qog, uas tej zaum yuav yog cov khoom siv tshuaj muaj zog ntawm Mylabris.

Cov cim tseem ceeb sib txawv tau raug txiav txim siab rau lub ncov nrog cov yam ntxwv ntawm cov tshuaj lom neeg thiab cov khoom siv cim npe. Yog li, ntawm MC thiab MP ntawm HPLC cov ntiv tes tom qab kev txheeb xyuas kev sib raug zoo ntawm kev sib raug zoo, lub ncov 2 (cytosine), 4 (tsis paub), thiab 14 (tsis paub) yog cov cim tseem ceeb rau qhov sib txawv ntawm MP thiab MC raws li qhia hauv daim duab 7.

Xaus

Qhov tseeb, hauv kev tshawb fawb tam sim no, ib txoj kev HPLC tau npaj rau kev tshuaj ntsuam ntiv tes ntawm MP thiab MC. Los ntawm kev sib txuas HPLC ntiv tes, PCA, thiab spectrum-zoo kev txheeb xyuas kev sib raug zoo xws li GAS thiab MLRA, cov khoom siv tshuaj thiab tshuaj ntawm ob hom Mylabris tuaj yeem raug cais tawm. Qhov no qhia tau hais tias cov tebchaw los ntawm cov peaks 2 (cytosine), 4 (tsis paub), thiab 14 (tsis paub) raws li cov cim tseem ceeb sib txawv ua lub luag haujlwm tseem ceeb hauv kev sib txawv ntawm MP thiab MC. Cov qauv ntawm peaks 4 thiab 14 yuav tsum tau txheeb xyuas los ntawm lwm yam thev naus laus zis. Cov txheej txheem ntawm HPLC ntiv tes ua ke nrog kev txheeb cais, tshuaj ntsuam xyuas tshuaj, thiab kev txheeb xyuas kev sib raug zoo-spectrum-zoo tau pom tias muaj txiaj ntsig zoo hauv kev tshawb pom cov cim cim lossis kev txhawb nqa QC ntawmtshuaj ntsuab.

Cov ntaub ntawv

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