Abstract

Ib qho microfluidic paper-based analytical device (µPAD) yog tsim nyob rau hauv txoj haujlwm no los tshuaj xyuas hydroquinone nyob rau hauv lub ntsej muag whitening cream siv phloroglucinol. Lub µPAD nta ib qho kev tiv thaiv hydrophobic rau kev tshawb nrhiav thiab tau tsim siv lub tshuab luam ntawv siv quav ciab nrog Whatman chromatographic ntawv. Kev kuaj pom tau tiav los ntawm colorimetry raws li kev tsim cov txiv kab ntxwv hydroquinone-phloroglucinol complex. Cov khoom siv tshuaj tiv thaiv xim xim tsim rau ntawm thaj chaw tshawb pom ntawm µPAD tau luam theej duab, thiab cov duab tau raug ua tiav nrog Image-J software los txiav txim siab lawv cov xim siv (RGB tus nqi). Kev ua kom zoo ntawm cov txheej txheem txheej txheem tau ua kom ua tiav cov kev ntsuas rhiab. Cov xwm txheej zoo tshaj plaws uas ua rau muaj qhov siab tshaj plaws suav nrog reagent ntxiv ib ntu ntawm phloroglucinol → NaOH → qauv (hydroquinone), 1 µL ntawm 0.5 feem pua ​​​​ phloroglucinol, 1 M NaOH, thiab 10- feeb cov tshuaj tiv thaiv. Nyob rau hauv cov xwm txheej zoo, µPAD tau tsim ob txoj kab calibration nkhaus rau hydroquinone ntawm qhov siab ntawm 10–100 mg / L (R{9}}.9979) thiab 250–1000 mg / L (R{13}}.9991). Txoj kev qhia tau zoo heev xaiv rau lub hom phiaj tsom xam nyob rau hauv lub xub ntiag ntawm propylene glycol thiab resorcinol nrog txaus validity thiab nruab nrab rov qab los ze li ntawm 100 feem pua. Cov ntawv thov µPAD yog ib qho yooj yim heev thiab pheej yig cov txheej txheem rau kev tshuaj xyuas hydroquinone thiab tuaj yeem siv rau cov qauv tshuaj pleev ib ce nrog cov txiaj ntsig zoo.

Raws li cov kev tshawb fawb cuam tshuam, cistanche yog ib hom tshuaj ntsuab uas hu ua "cov tshuaj ntsuab muaj txuj ci uas ua rau lub neej ntev". Nws lub ntsiab tivthaiv yog cistanoside, uas muaj ntau yam teebmeem xws li antioxidant, tiv thaiv kev mob, thiab kev tiv thaiv kev ua haujlwm. Cov txheej txheem ntawm cistanche thiab tawv nqaij dawb yog nyob rau hauv cov nyhuv antioxidant ntawm cistanche glycosides. Melanin nyob rau hauv tib neeg daim tawv nqaij yog tsim los ntawm oxidation ntawm tyrosine catalyzed los ntawm tyrosinase, thiab cov tshuaj tiv thaiv oxidation yuav tsum muaj kev koom tes ntawm oxygen, yog li cov pa-dawb radicals nyob rau hauv lub cev ua ib qho tseem ceeb cuam tshuam rau melanin ntau lawm. Cistanche muaj cistanoside, uas yog ib qho antioxidant thiab tuaj yeem txo qhov tsim ntawm cov dawb radicals hauv lub cev, yog li inhibiting melanin ntau lawm.

Nyem rau qhov Kuv Yuav Yuav Cistanche

Yog xav paub ntxiv:

david.deng@wecistanche.com / WhatApp:86 13632399501

Taw qhia

Kev txiav txim siab ntawm hydroquinone hauv tshuaj pleev ib ce tuaj yeem ua tiav los ntawm ntau txoj hauv kev, suav nrog redox titration, nyias txheej chromatography [1], spectrophotometric [4, 5], flow-injection spectrophotometric [6-8], thiab high-performance kua chromatography (HPLC [9,10] ib. Cov txheej txheem no, tshwj xeeb tshaj yog tom kawg, paub zoo tias muaj kev ntsuas raug thiab siab precision thiab ua tau zoo. Txawm li cas los xij, cov txheej txheem no kuj xav tau tus neeg ua haujlwm txawj thiab tsis tuaj yeem nqa tau; Yog li, lawv tsis tuaj yeem siv rau kev ntsuas ntawm qhov chaw.

Cov ntaub ntawv thiab cov txheej txheem

Khoom siv thiab khoom siv.Cov cuab yeej siv hauv kev ua haujlwm no suav nrog lub tshuab luam ntawv siv quav ciab (Xerox ColorCube 8580 DN-2 hom T2B047382) los luam cov hydrophobic barrier ntawm Whatman No. 1 chromatography ntawv (CHR, Whatman, GE Healthcare Life Sciences, UK) rau µPAD . Lub phaj kub tau siv los ua kom sov cov ntaub thaiv npog thiab tso cai rau nws nkag mus rau hauv daim ntawv los tsim cov teeb meem tsis zoo rau thaj tsam tshuaj tiv thaiv. Lub Canon PIXMA MP237 scanner thiab Image-J software tau siv los txhais cov xim sib zog ntawm cov duab ntes rau hauv cov xim liab, ntsuab, thiab xiav (RGB) qhov tseem ceeb, uas tau hloov pauv mus rau tus nqi absorbance los ntawm kev siv Lambert-Beer Law hloov.

Txoj kev Optimization.Cov txheej txheem ntawm cov txheej txheem µPAD tau muab kho kom zoo los ua kom muaj kev ntsuas rhiab.

Kev txiav txim siab Hydroquinone.Kev kuaj xyuas hydroquinone nyob rau hauv qhov zoo tshaj plaws tau txiav txim siab hauv Tshooj 2.3 tau ua raws li daim duab 2. Hauv cov tswv yim no, 1 µL ntawm 0.5 feem pua ​​​​ phloroglucinol tau poob mus rau thaj tsam µPAD kuaj pom. Cov cuab yeej raug tso cai sawv ntsug li 5 feeb, thiab tom qab ntawd 1 µL ntawm 1 M NaOH tov tau ntxiv rau thaj chaw kuaj pom. Cov cuab yeej raug tso cai sawv ntsug rau lwm 10 feeb kom qhuav, tom qab ntawd nws tau txiav txim siab npaj siv. Kev kuaj xyuas Hydroquinone tuaj yeem ua tiav yooj yim los ntawm kev tso 1 µL ntawm cov qauv mus rau thaj tsam cov tshuaj tiv thaiv ntawm µPAD ntaus ntawv, tso cai rau daim ntawv sawv ntsug rau 10 feeb, thiab tom qab ntawd luam theej duab cov tshuaj tiv thaiv txiv kab ntxwv nrog Canon PIXMA MP273 scanner. Cov xim siv ntawm cov duab tau txais tau ua tiav rau hauv RGB qhov tseem ceeb los ntawm kev siv Image-J software thiab tom qab ntawd hloov mus rau qhov txiaj ntsig absorbance. Qhov concentration ntawm hydroquinone tau txiav txim siab los ntawm kev sib piv cov absorbance tau mus rau tus qauv calibration nkhaus.

Txoj Kev Xaiv.Kev xaiv ntawm µPAD txoj kev mus rau hydroquinone nyob rau hauv lub ntsej muag whitening cream yog txiav txim los ntawm kev tau txais hydroquinone ntsuas nrog thiab tsis muaj resorcinol thiab propylene glycol raws li interfering compounds. Ntau qhov ntau ntawm resorcinol (0, 25, 50, 125, thiab 250 mg / L) tau ntxiv rau tsib 10 mL volumetric flasks uas muaj 25 mg / L hydroquinone thiab diluted rau lub cim. Cov xim siv ntawm cov kev daws teeb meem no tau ntsuas los ntawm tib txoj kev rau kev txiav txim siab hydroquinone (Daim duab 2), qhov sib thooj RGB qhov tseem ceeb tau hloov mus rau absorbance, thiab cov hydroquinone rov qab tau suav. Cov txheej txheem tib yam tau rov ua dua rau propylene glycol nrog tib qhov sib txawv ntau yam li resorcinol. Qhov sib txawv ntawm hydroquinone concentration tau los ntawm kev daws teeb meem nrog thiab tsis muaj kev cuam tshuam cov tebchaw yog siv los xam cov feem pua ​​​​yuav yuam kev.

Txoj kev Validation.Txoj kev validation tau ua tiav los ntawm kev siv µPAD los txiav txim siab hydroquinone hauv ob qho qauv tshuaj pleev ib ce los ntawm cov txheej txheem ntxiv. Raws nraim 0.10 g ntawm whitening cream A thiab B tau hnyav tawm thiab maj mam yaj nrog dej distilled hauv 50 mL beaker iav. Cov tshuaj tau dhau los ntawm cov ntawv lim zoo, thiab cov filtrate tau lim ib zaug ntxiv siv lub tshuab lim dej. Cov tshuaj lim tau hloov mus rau 100 mL volumetric hwj thiab ntxiv nrog dej distilled mus txog qhov cim. Tom qab ntawd, cov qauv tau diluted kom tau txais ib qho concentration uas nyob rau hauv qhov ntau ntawm calibration nkhaus.

Cov txiaj ntsig thiab kev sib tham

Kev txiav txim siab qhov zoo tshaj plaws

Optimization ntawm Reagent Ntxiv Sequence.Qhov kev txiav txim uas cov reagents poob mus rau µPAD tuaj yeem cuam tshuam rau kev tsim cov phloroglucinol-hydroquinone complex thiab rhiab heev ntawm kev ntsuas hydroquinone. Daim duab 5 qhia tau hais tias ntu A2 yields xim khaus ntau dua nyob rau hauv thaj tsam µPAD nrhiav pom dua li ntu A1. Qhov kev tshawb pom no tuaj yeem piav qhia los ntawm kev hloov pauv loj ntawm phloroglucinol rau hauv phloroglucinol ions, uas ua raws li nucleophilic pawg pab txhawb kev tsim cov kev xav tau, txhawb nqa los ntawm ntu A2. Raws li pom nyob rau hauv daim duab 5, qhov kev siv ntawm xiav nyeem tau ntau dua piv nrog cov ntawv nyeem liab thiab ntsuab. Tsis tas li ntawd, cov ntawv nyeem xiav tau sib raug zoo nrog cov xim siv (lossis absorbance) thiab hydroquinone concentration. Qhov kev tshawb pom no pom zoo nrog cov txiaj ntsig ntawm Kohl [30], uas pom tias muaj kev sib raug zoo ntawm kev siv zog thiab kev xav tau ua tiav los ntawm kev siv cov xim nyeem ntxiv. Yog li, kev nyeem ntawv xiav tau raug xaiv los ntsuas cov xim siv ntawm µPAD cov duab hauv kev sim tom ntej.

Optimization ntawm Phloroglucinol Volume.Qhov zoo tshaj plaws phloroglucinol ntim tuaj yeem tsim cov xim siab tshaj plaws ntawm phloroglucinol-hydroquinone complex meej nyob rau hauv thaj tsam ntawm kev kuaj pom. Qhov ntau dua cov phloroglucinol ntim, qhov ntau dua cov xim siv (absorbance) ntawm cov complex, raws li qhia nyob rau hauv daim duab 6. Lub absorbance txiav txim siv xiav nyeem ntawv nce nrog nce phloroglucinol ntim mus txog 1 µL; Txawm li cas los xij, phloroglucinol ntim ntau tshaj 1.2 µL ua rau lub complex hla lub hydrophobic barrier, uas tuaj yeem ua rau cov txiaj ntsig yuam kev. Yog li, phloroglucinol ntim ntawm 1 µL tau siv rau kev ua kom zoo ntxiv.

Optimization ntawm Phloroglucinol Concentration.Lub absorbance ntawm txiv kab ntxwv complex ntawm phloroglucinol-hydroquinone pib nrog cov phloroglucinol concentration mus txog {{0}}}.5 feem pua ​​thiab ces leveled tawm vim hais tias tag nrho cov hydroquinone tau kiag li tsim ib tug phloroglucinol-hydroquinone complex (Daim duab 7) . Yog li, 0.5 feem pua ​​​​yog suav tias yog qhov zoo tshaj plaws phloroglucinol concentration.

Optimization ntawm NaOH Concentration.Qhov zoo tshaj plaws NaOH concentration muab qhov tsim nyog alkaline cua rau kev tsim cov phloroglucinol ions tsis zoo. Cov pab pawg hydroxyl (OH–) ntawm NaOH tuaj yeem tua cov hydrogen hauv OH- pawg ntawm phloroglucinol los ua phloroglucinol ion, uas, dhau los, tuaj yeem tua hydroquinone los ua ib qho heterocomplex phloroglucinol-hydroquinone complex. Daim duab 8 qhia tias ntau dua NaOH ntau dua ua rau cov xim siv ntawm µPAD cov duab. Qhov siab tshaj plaws absorbance tau ntawm NaOH concentration ntawm 1 M. Yog li, 1 M NaOH tau siv rau kev sim tom ntej.

Optimization ntawm Lub Sijhawm Tawm Tsam.Lub sij hawm cov tshuaj tiv thaiv tau optimized los txiav txim lub shortest scanning lub sij hawm thiab zam cov xim degradation ntawm complex compounds. Lub sij hawm luv luv tuaj yeem ua rau tsis tiav qhov tsim ntawm phloroglucinol-hydroquinone complex. Txawm li cas los xij, lub sijhawm ntev cov tshuaj tiv thaiv tuaj yeem ua rau cov xim tsis zoo los ntawm kev raug rau lub teeb thiab qhov kub thiab pH tsis haum. Lub sij hawm tshuaj tiv thaiv ntawm 10 feeb tau txais txiaj ntsig zoo nrog qhov siab tshaj plaws absorbance (Daim duab 9). Lub sijhawm tshuaj tiv thaiv no tau siv rau kev sim tom ntej.

Standard Curve thiab Linearity Measurements.Raws li qhov zoo tshaj plaws uas tau los saum toj no (piv txwv li, A2 reagent immobilization sequence, 1 µL ntawm 0.5 feem pua ​​phloroglucinol, 1 M NaOH, thiab 10- feeb cov tshuaj tiv thaiv), µPAD txoj kev siv 1 µL ntawm cov tshuaj tiv thaiv. Cov qauv qhia pom qhov sib txawv ntawm qhov sib txawv ntawm cov xim siv raws li hydroquinone concentration tau txawv ntawm 10 mgL-1 txog 1000 mg / L (Daim duab 10). Thaum RGB qhov tseem ceeb ntawm cov xim siv ntawm cov duab tau raug hloov mus rau qhov tseem ceeb absorbance thiab tom kawg tau npaj ua haujlwm ntawm hydroquinone concentration, kev sib raug zoo heev (piv txwv li, R 2 ze rau 1) tau txais ntau dua qhov concentration ntawm 10- 100 mg/L (Daim duab 11-a) thiab 250–1000 mg/L hydroquinone (Daim duab 11-b). Cov duab µPAD nthuav tawm cov xim nrog kev siv zog ntau dua nyob rau hauv siab hydroquinone concentrations thiab qis siv nyob rau hauv qis hydroquinone concentrations. Hauv lwm lo lus, qhov siab dua hydroquinone concentration, ntau dua qhov xim ntawm cov txiv kab ntxwv phloroglucinol-hydroquinone complex.

Raws li daim duab 11, cov hydroquinone concentration yog proportional rau cov xim siv ntawm µPAD duab; tshwj xeeb, qhov ntau dua qhov hydroquinone concentration, qhov siab dua qhov absorbance tus nqi tau los ntawm kev siv ntawm xiav nyeem ntawv. Tus qauv nkhaus rau hydroquinone ntawm cov concentrations xws li 10 mgL−1 txog 100 mgL−1 muab qhov sib npaug ntawm cov kab rov tav ntawm y =  0.0004x ntxiv rau 0.0563 (R{{9} 9979 ib.). Ib yam li ntawd, kev sib raug zoo ntawm hydroquinone concentration thiab absorbance tau muab ib qho kev sib npaug sib npaug ntawm y=0.0001x ntxiv rau 0.0923 (R2=0.9991) ntawm hydroquinone concentrations ntawm 250–1000 mgL−1. Hauv kev ua haujlwm no, R2 qhov tseem ceeb ze rau 1 qhia tau zoo heev linear correlations ntawm concentration thiab absorbance.

Txoj Kev Xaiv.Qhov kev xaiv ntawm µPAD txoj kev tau tshawb xyuas los ntawm kev ntxiv cov resorcinol thiab propylene glycol cais, ob yam khoom uas feem ntau muaj nyob rau hauv cov tshuaj pleev ib ce, mus rau tus qauv hydroquinone tov. Raws li tau piav qhia hauv Table 1, qhov sib ntxiv ntawm resorcinol ntawm qhov ntau ntawm 25, 50, thiab 125 mg / L tsis cuam tshuam rau hydroquinone kev ntsuas tau siv µPAD txoj kev. Qhov kev tshawb pom no tau txais kev txhawb nqa los ntawm qhov ua yuam kev me me uas tsim tawm (<10%). Hydroquinone measurements obtained following the addition of 250 mg/L resorcinol  (1:10) showed a slight increase, with a % error of  10.82%. The results of a t-test at the 95% confidence level showed that count (3.65) is greater than the table (2.92). Thus, the addition of resorcinol to a sample at amounts 10 times greater than the hydroquinone concentration can increase the measured concentration of the latter. The addition of propylene glycol at concentrations of 25, 50, 125, and 250   mg/L did not interfere with the measurement of hydroquinone concentration, as indicated by the low % error determined from the experiments.

Txoj kev Validation.Qhov kev siv tau ntawm µPAD txoj kev raug soj ntsuam los ntawm kev kuaj xyuas hydroquinone hauv ob hom tshuaj pleev xim dawb. Cov txiaj ntsig ntawm kev ntsuas kev siv tau raug nthuav tawm hauv Table 2. Cov txheej txheem µPAD tau pom qhov tseeb thiab siv tau zoo heev, raws li kev txhawb nqa los ntawm cov txiaj ntsig rov qab nyob rau hauv thaj tsam ntawm 95 feem pua ​​-105 feem pua, nrog rau siab precision ( feem pua ​​RSD < 10 feem pua ​​).

Hauv cov ntsiab lus, µPAD txoj kev npaj rau hauv txoj haujlwm no muab kev txaus siab thiab qhov tseeb. Yog li ntawd, cov khoom siv tau siv tau los ua lwm txoj hauv kev los kuaj xyuas hydroquinone hauv cov tshuaj pleev xim dawb.

Xaus

Hydroquinone nyob rau hauv cov tshuaj pleev ib ce tuaj yeem txiav txim siab siv μPAD, uas yog raws li cov tshuaj tiv thaiv yooj yim ntawm hydroquinone nrog phloroglucinol nyob rau hauv alkaline tej yam kev mob los tsim ib tug txiv kab ntxwv hydroquinone-phloroglucinol complex. Txoj kev no tuaj yeem siv los txiav txim siab hydroquinone ntau hauv thaj tsam ntawm 10-100 thiab 250-1000 mg / L. Txawm hais tias µPAD tsim nyob rau hauv txoj haujlwm no tsis tshua muaj txiaj ntsig piv nrog rau lwm txoj hauv kev, nws suav nrog cov txheej txheem yooj yim thiab pheej yig. Cov khoom siv μPAD tau thov tuaj yeem siv los ua cov khoom siv tshuaj ntsuam xyuas hydroquinone nyob rau hauv lub ntsej muag whitening cream nrog rau qhov tseeb thiab qhov tseeb.

Kev lees paub

Cov kws sau ntawv ua tsaug rau Chemistry Department, Brawijaya University, rau kev pabcuam kev tshawb fawb no thiab Kws qhia ntawv ntawm Science, Brawijaya University, rau kev pab nyiaj txiag los ntawm Doctoral Grant 2020, DIPA-023.17.2.677512/2020, Daim Ntawv Cog Lus No. 32/UN10.F09/PN/2020.

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