Daim ntawv thov ntawm Hibiscus Cannabinus L. (kenaf) Nplooj Extract Raws li daim tawv nqaij Whitening Thiab Anti-aging Agents hauv Natural Cosmetic Prototype

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

Yan Yi Sim & Kar Lin Nyam

TSAB NTAWV

Hauv Malaysia, kev lag luam pov tseg los ntawm covHibiscus cannabinusL. (kenaf) kev lag luam, tshwj xeeb tshaj yog tawm, ua rau muaj kev sib tw hauv kev ruaj ntseg. Kev ua haujlwm lom neeg ntawm kenaf nplooj tau tshaj tawm los ntawm ob peb txoj kev tshawb fawb yav dhau los, nws yog, yog li ntawd, ntseeg tau tias kenaf nplooj extract (KLE) tuaj yeem siv los ua cov khoom siv hauv cov tshuaj pleev ib ce. Yog li, lub hom phiaj ntawm txoj kev tshawb no yog los tsim cov tshuaj pleev ib ce ntuj uas muaj KLE thiab tshuaj xyuas nws cov yam ntxwv ntawm lub cev, cov khoom siv microbiological, kev ruaj khov, kev ua haujlwm lom (antioxidant, antityrosinase, thiabanti-aging), nyob rau hauv vitro cytotoxicity (ntawm tib neeg dermal fibroblast thiab B16F10 melanoma hlwb) thiab melanogenesis assay (intracellular tyrosinase kev ua si thiab txo cov melanin ntau lawm). Cov txiaj ntsig tau pom tias KLE lotion (KLEL) npaj los ntawm 15 feem pua ​​​​kenaf noob roj (KSO) (F2) nrog 0.1 feem pua ​​w/w KLE yields qhov zoo tshaj plaws lub cev thiab microbiological stability, tsis muaj toxicity ntawm tib neeg hlwb. KLEL kuj tau nthuav tawm cov ntsiab lus antioxidant txog 1.84 ± 0.07 mg caffeic acid sib npaug (CAE) / g thiab 21.62 ± 0.76 mg catechin hydrate sib npaug (CHE) / g rau tag nrho cov ntsiab lus phenolic (TPC ) thiab tag nrho cov ntsiab lus flavonoid (TFC), feem. Tsis tas li ntawd, KLEL nthuav tawm lub peev xwm ntawm antityrosinase ntawm inhibition ntawm mycophenolate (30.28 ± 3.90%) thiab diphenols (11.40 ± 0.29 feem pua) tsim, thiab qhia tawm, thawj zaug,anti-agingcov khoom los ntawm inhibiting collagenase (36.41 ± {2}}.54 feem pua) thiab elastase (23.13 ± 1.56 feem pua) cov dej num. Hais txog melanogenesis inhibitory kev ua ub no, KLEL nthuav tawm cov txiaj ntsig zoo hauv kev ua haujlwm ntawm cov cellular tyrosinase thiab cov ntsiab lus melanin ntawm B16F10 hlwb. Zuag qhia tag nrho, cov txiaj ntsig los ntawm txoj kev tshawb fawb no tau cog lus zoo heev rau kev txhim kho cov qauv tshuaj pleev ib ce uas siv cov nplooj kenaf.

suab puam cistanche zaj tshuaj ntsuab: anti-aging

1. Taw qhia

Tsis ntev los no, cov qauv kev lag luam ncig uas tuaj yeem txhim kho kev ruaj khov ntawm kev tswj cov khib nyiab los ntawm kev txo qis kev tsim khoom pov tseg thiab tswj kom muaj nuj nqis mus ntev, thaum txo qis qhov tsis zoo ntawm cov peev txheej tsis txaus thiab kev puas tsuaj ib puag ncig tau txais txiaj ntsig rau pej xeem (Morseletto, 2020). Feem ntau, ib qho tseem ceeb ntawm cov khoom lag luam nrog tus nqi qis nyiaj txiag yog tsim los ntawm kev lag luam qoob loo txhua xyoo.Hibiscus cannabinusL. KR9 (kenaf) kev lag luam, yog ib qho ntawm cov kev ua liaj ua teb pab txhawb rau Malaysia tus GDP (cov khoom lag luam hauv tsev) nrog cov khoom hnyav ntawm kenaf qhuav qia tau nce los ntawm 7.1 (000 ton) hauv xyoo 2013 txog 7.6 ({{7}) } ton) nyob rau hauv 2014, thiab lub ntiaj teb no kenaf kev ua lag luam npaj yuav hla US $ 854 lab los ntawm 2025 (Abdelrhman li al., 2016). Txawm li cas los xij, qhov ntau ntawm cov khoom lag luam raug tsim tawm, xws li kenaf noob thiab nplooj, uas ua rau muaj teeb meem sustainability.

Kenaf nplooj, accounting rau ib tug tseem ceeb npaum li cas ntawm tag nrho cov by-products, yog tsawg kawg nkaus tus yam ntxwv thiab muaj nuj nqis ntawm tag nrho cov by-products generated. Vim yog txoj hauv kev ncig, nws yog ib qho tseem ceeb uas yuav tsum tau rov siv dua lossis rov qab cov nplooj kenaf rau hauv cov khoom muaj nqis ntxiv vim nws muaj ntau yam txiaj ntsig hauv kev lag luam, ib puag ncig, thiab kev sib raug zoo (Coderoni thiab Perito, 2019). Kenaf nplooj muaj cov nplua nuj ntawm bioactive compounds xws li chlorogenic acid, caffeic pab, kaempferol, thiab catechin hydrate raws li pov thawj los ntawm kev tshawb fawb ua los ntawm Kho thiab al. (2019); Sim and Nyam (2019), and Haw et al. (2020). Txawm li cas los xij, nws feem ntau yog destined rau kev tsim cov khoom lag luam qis: cov khoom noj fiber ntau thiab tsiaj pub (Lim li al., 2020).

Lo lus "rov qab mus rau qhov xwm txheej" tau siv dav hauv kev tshawb fawb thiab kev tsim kho kom zoo nkauj, vim tias kev siv cov extracts ntawm botanical keeb kwm tau ua rau muaj kev lees paub zoo los ntawm cov neeg siv khoom. Raws li kev tshawb fawb ua los ntawm Sim et al. (2019), kenaf nplooj extract (KLE) pom tau hais tias cog lus antioxidant thiab antityrosinase zog, muaj peev xwm yuav siv tau raws li tus nqi-ntxiv cov khoom xyaw rau hauv txoj kev loj hlob ntawm cov khoom kom zoo nkauj. Nws yog ib qho tseem ceeb los tsim kom muaj kev ruaj ntseg thiab ruaj ntseg formulations uas muaj cov KLE raws li nws muaj ntau cov polyphenol tebchaw uas ua rau pom ntawm daim tawv nqaij.ntxuav hniav dawbthiabanti-agingkhoom. Muaj ntau yam yuav tsum tau coj mus rau hauv tus account thaum tsim cov tshuaj pleev ib ce tshiab, xws li hom formulation, lub hom phiaj ntawm kev siv, thiab muaj peev xwm sib cuam tshuam ntawm cov khoom xyaw uas siv rau hauv cov formulations, uas tag nrho, ua rau muaj kev xav tau kev ruaj ntseg thiab kev nyab xeeb kev tshawb fawb. (Garbossa thiab Maia Campos, 2016).

Kev tshawb fawb txog kev ruaj ntseg, uas suav nrog kev kawm ntawm lub cev, tshuaj, thiab cov yam ntxwv ntawm microbial, yog qhov tsim nyog rau kev tsim cov tshuaj pleev ib ce tom qab kev txhim kho. Tsis tas li ntawd, txhawm rau tiv thaiv kom tsis txhob muaj kev phiv los yog kev tsis haum tshuaj, cov khoom siv tshuaj pleev ib ce kuj tseem yuav tsum tau sim los ntawm kev siv tshuaj tua kab mob hauv vitro cytotoxicity ntawm tib neeg cov tawv nqaij ntawm tes. Yog li, lub hom phiaj ntawm txoj haujlwm no yog los tsim cov tshuaj pleev ib ce ntuj uas muaj KLE (KLE lotion- KLEL). Tom qab ntawd, KLEL tau raug tshuaj xyuas rau nws cov yam ntxwv ntawm lub cev, cov khoom siv microbiological, kev ruaj ntseg cia, hauv vitro cytotoxicity (ntawm tib neeg dermal fibroblast thiab B16F10 melanoma hlwb), thiab kev ntsuam xyuas melanogenesis (kev ua haujlwm ntawm intracellular tyrosinase thiab txo melanin ntau lawm). Txoj kev tshawb no kuj tau tso cai rau peb txiav txim siab, thawj zaug, tsis yog tsuas yog cov tshuaj tiv thaiv antioxidant thiab tiv thaiv tyrosinase ntawm KLEL tab sis kuj lawv cov kev ua haujlwm inhibitory ntawm collagenase thiab elastase.

cistanche txiaj ntsig: Anti-aging

2. Cov ntaub ntawv thiab cov txheej txheem

2.1. Cov khoom siv cog thiab tshuaj

TshiabHibiscus cannabinusL. KR9 (kenaf) tawm 90 hnub tom qab sowing thiab noob tau los ntawm Lembaga Kenaf & Tembakau Negara (LKTN, Malaysia). Span 20 thiab Tween 80 tau yuav los ntawm Sigma Aldrich (Munich, Lub Tebchaws Yelemees), Cosmedia® ACE, Iscaguard® PEG, thiab Emulgade® SE-PF tau los ntawm BASF (Malaysia), thiab glycerine tau yuav los ntawm Croda International Plc. (United Kingdom). Tib neeg dermal fibroblast (NHDF) kab ntawm tes tau muab los ntawm Lonza (Basel, Switzerland) thiab nas melanoma (B16F10) tau yuav los ntawm ATCC (Manassas, VA, USA). Dulbecco's Modified Eagle Medium (DMEM) tau yuav los ntawm Nacalai Tesque (Nyiv). Tag nrho lwm cov tshuaj siv tau yog qib analytical reagent (Belgium, Lub teb chaws Yelemees, Malaysia). Cov dej ntshiab ntshiab (Millipore, USA) tau siv thoob plaws hauv kev tshuaj xyuas.

2.2. Cov txheej txheem

2.2.1. Qhuav ntawm kenaf nplooj

Cov nplooj kenaf tau ntxuav nrog dej ntshiab huv, qhuav, thiab tom qab ntawd khaws cia ntawm − 80 ◦C thaum hmo ntuj. Tom qab ntawd, cov qauv no tau qhuav hauv lub tshuab ziab khaub ncaws (Christ, Lub teb chaws Yelemees), hauv qab 0.0004 bar rau 48 h, hauv av, lub tshuab nqus tsev ntim, thiab khaws cia ntawm -20 degree.

2.2.2. Kev npaj ntawm ib nrab purified kenaf nplooj extract (KLE)

Lub pulsed ultrasonic-pab extraction (PUAE) ntawm KLE tau ua raws li Sim li al. (2019). Cov qauv raug ntsuas nrog cov kuab tshuaj rho tawm (ethanol) hauv qhov sib piv ntawm 1:10. Tom qab ntawd, qhov sib tov tau raug rau lub zog ultrasonic-pab extraction (Sartorius, Lub teb chaws Yelemees) ntawm 50 feem pua ​​​​sonication amplitude, 1 min mem tes ntev lub sij hawm, thiab 1 min mem tes ncua sij hawm nrog kub tswj ntawm 18-22 ± 3 degree (peb lub voj voog. ). Tom qab ntawd, cov extract tau lim thiab concentrated hauv lub tshuab nqus tsev evaporator (Buchi, Switzerland). Kev ua kom huv ib nrab ntawm KLE tau ua raws li Seabra li al. (2010) nrog kev hloov kho me ntsis. Cov hmoov nplej tau siv rau hauv ib kem uas muaj silica gel thiab eluted nrog cov kuab tshuaj gradient ntawm n-hexane- ethyl acetate (100:00, 90:10, 80:20, 50:50 , 20:80, 10:90, 00:100) thiab ethyl acetate methanol (100:00, 90:10, 80:20, 50:50, 20:80, 10:90 , 00:100). Cov purified ib nrab KLE tau concentrated rau dryness los ntawm kev siv ib tug teb evaporator thiab khaws cia ntawm -20 degree rau yav tom ntej siv.

2.2.3. Cov kuab tshuaj extraction ntawm kenaf noob roj (KSO)

KSO tau muab rho tawm raws li Chew et al. (2015). Kenaf cov noob tau muab tso rau hauv cov hmoov zoo siv lub grinder (Panasonic, Nyiv) thiab ntxiv nrog hexane hauv qhov piv ntawm 1: 5. Cov roj tau muab rho tawm los ntawm kev siv Soxhlet extractor ntawm 60 ◦C rau 3 h, thiab cov hexane raug tshem tawm los ntawm kev siv lub evaporator teb. Tom qab yaug nrog nitrogen, KSO tau khaws cia ntawm -20 degree rau kev siv yav tom ntej (Chew et al., 2015).

2.2.4. Kev sim tsim cov tshuaj pleev ib ce uas muaj KLE

Thaum xub thawj, cov tshuaj pleev ib ce (tsis muaj KLE) tau ua kom zoo dua los ntawm kev siv 4 qhov sib txawv ntawm kenaf noob roj (KSO) (10 feem pua, 15 feem pua, 20 feem pua, thiab 25 feem pua ​​). Tom qab ntawd cov tshuaj pleev ib ce tau muab piv rau ntau yam tsis zoo: qhov tshwm sim ntawm lub cev, tsw, homogeneity, pH, viscosity, centrifugation, thiab khov-thaw kev ntsuam xyuas, los xaiv qhov zoo tshaj plaws lotion puag formulation rau kev kawm ntxiv. Hais txog cov txiaj ntsig los ntawm cov kev ntsuas saum toj no, qhov tsim nyog tshaj plaws tshuaj pleev ib ce (15 feem pua ​​​​KSO) raug xaiv rau kev koom ua ke nrog KLE (0.1 feem pua ​​w/w) (KLEL). Luv luv, tsis yog-polar theem (Emulgade® SE PF, Span 20, thiab KSO) thiab polar theem (dej, Tween 80, thiab glycerine) cov khoom xyaw tau ua kom sov txog 75 ± 2 degree. Tom qab ntawd, cov theem uas tsis yog-polar tau ntxiv poob los ntawm kev poob mus rau lub ncov qaumteb qabteb theem nrog ceev nrooj nplawm siv lub tshuab nqus hlau nplaum (Thermo Fisher Scientific, USA) ntawm 350 rpm los ua thawj emulsion, ua raws li high-shear homogenization siv IKA T25 digital ULTRA -TURRAX® (IKA Laboratory Equipment, Lub teb chaws Yelemees) ntawm 3200 rpm rau 3 min thiab ultrasonic homogenization ntawm 40 feem pua ​​​​amplitude rau 3 min. Cosmedia® ACE, Iscaguard® PEG, thiab KLE tau ntxiv thiab sib xyaw ntawm O / W emulsion ntawm chav tsev kub. KSO lotion base tsis muaj KLE ua haujlwm tswj (KSOL). Thaum cov tshuaj pleev ib ce ntxiv nrog kojic acid (0.1 feem pua ​​​​w / w) (KAL) tau ua haujlwm zoo rau kev tiv thaiv tyrosinase kev ua haujlwm thiab kev tshuaj xyuas melanogenesis. Cov tshuaj pleev ib ce ntxiv nrog catechin hydrate (0.1 feem pua ​​w / w) (CHL) tau txais txiaj ntsig zoo rau kev tiv thaiv collagenase thiab kev ua haujlwm antielastase

2.2.5. Physicochemical tsom xam

2.2.5.1. Cov tsos, tsw, thiab homogeneity.

Raws li Hanifah thiab Jufri (2018), cov tshuaj pleev ib ce tau tshuaj xyuas rau lawv cov xim, tsw thiab homogeneity, thiab theem sib cais.

2.2.5.2. pH. Cov txheej txheem daws teeb meem tau siv los ntsuas pH meter electrode ua ntej ntsuas.

Cov pH ntawm cov qauv tau txiav txim siab nrog pH meter (Mettler Toledo, Switzerland) ntawm chav tsev kub.

2.2.5.3. Xim.

Xim yuav raug ntsuas los ntawm kev siv lub colorimeter (Hunter Lab, United States). Cov txiaj ntsig yuav raug qhia raws li qhov chaw xim L * (lightness), a * (ntsuab), b * (daj), thiab tag nrho cov xim sib txawv (ΔE). Tag nrho cov xim sib txawv (ΔE) rau txhua qhov qauv tau suav los ntawm kev siv kab zauv raws li nram no: ̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅ (l * 0) 2 ntxiv (B * - B * 0) 2 √

2.2.5.4 ib. viscosity.

Cov viscosity ntawm cov qauv raug soj ntsuam hauv Brook teb viscometer siv LV-64 spindle raws li Gyawali li al. (2016) nrog kev hloov kho me ntsis. Cov qauv tau muab tso ncaj qha rau hauv qhov ntxaiv nrog 100 rpm kev sib hloov thiab qhov viscosity (cP) tau ntsuas.

2.2.5.5. Kev nthuav dav.

Kev nthuav dav ntawm cov qauv yog txiav txim siab los ntawm txoj kev sib npaug phaj (Gyawali li al., 2016). Cov qauv raug hnyav (1 g) thiab muab tso rau ntawm ib lub iav slides (20 × 20 cm2). Tom qab ntawd, lwm tus swb tau muab tso rau saum tus qauv thiab 100 g qhov hnyav tau muab tso rau ntawm tus swb sab saud, kom ntseeg tau tias cov qauv raug nias sib npaug. Tom qab 1 min, qhov hnyav raug tshem tawm, thiab ntsuas qhov sib kis (cm) .

2.2.5.6. Kev ntsuas centrifugation.

Cov qauv raug thauj mus rau hauv lub raj centrifugation thiab tom qab ntawd centrifuged ntawm 3750 rpm rau 30 min (Eppendorf 5417R, USA). Qhov kev ntsuam xyuas centrifugation no (sib npaug rau 1- xyoo lub ntiajteb txawj nqus) txiav txim siab qhov ruaj khov ntawm cov qauv (Hiola li al., 2018).

2.2.5.7 ib. Kev ntsuas khov-thaw.

Kev tshawb fawb khov-thaw tau ua tiav raws li cov txheej txheem dhau los nrog qee qhov kev hloov kho (Krongrawa li al., 2018). Txhua tus qauv tau hloov pauv ntawm qhov kub thiab txias, 4 ± 1 ◦C (24 h) thiab kub kub, 45 ± 1 ◦C (24 h) nrog 75% ± 2% RH ( txheeb ze humidity) rau 6 cycles hauv airtight iav ntim. Lub viscosity ntawm cov qauv tau txiav txim siab tom qab lub voj voog kub ntxhov.

2.2.6. Kev txiav txim siab ntawm cov ntsiab lus antioxidant

2.2.6.1. Tag nrho cov ntsiab lus phenolic (TPC).

Tag nrho cov ntsiab lus phenolic tau txiav txim siab raws li Lim et al. (2007). Cov qauv (10 mg / mL) tau sib xyaw nrog 10 feem pua ​​​​Folin-Ciocalteu reagent thiab 7.5 feem pua ​​(w/w) Na2CO3, ua raws li 30 min incubation hauv maub. Qhov nqus tau raug coj los ntawm 765 nm los ntawm kev siv UV-vis spectrophotometer (Secoman, Fabkis). Qhov sib npaug ntawm caffeic acid yog y=0.0269 ntxiv rau 9.69x (r2=0.999) (Sim et al., 2019). Cov txiaj ntsig tau qhia tias yog mg ntawm caffeic acid sib npaug (CAE) / g qauv.

2.2.6.2. Tag nrho cov ntsiab lus flavonoid (TFC).

Tag nrho cov ntsiab lus flavonoid tau soj ntsuam raws li Ogbunugafo li al. (2011). Feem ntau, cov qauv (10 mg/mL) yog tov nrog 15 feem pua ​​NaNO2, 10 feem pua ​​AlCl3 tov, ultra-ntshiab dej, thiab 1 M NaOH. Qhov nqus tau raug ntsuas tam sim ntawm 510 nm hauv kev sib piv nrog tus qauv npaj siv catechin hydrate (0.02- 0.1 mg / mL). Qhov sib npaug ntawm catechin hydrate yog y=0.004 ntxiv rau 3.1x (r{17}}.999) (Sim et al., 2019). Tag nrho cov ntsiab lus flavonoid (TFC) tau qhia raws li milligrams ntawm catechin hydrate sib npaug (CHE) / g qauv.

cistanche extract cov txiaj ntsig: tiv thaiv kev laus

2.2.7. Anticollagenase kev ua haujlwm

Kev ua haujlwm los tiv thaiv collagenase raws li kev ua kom tsis muaj zog ntawm collagenase thiab hluavtaws substrate (FALGPA- N-(3-[2-}Furyl]- acryloyl)-Leu-Gly-Pro-Ala) ntawm 345 nm hauv muaj collagenase inhibitors tau ua raws li Barrantes thiab Guinea (2003) nrog qee qhov kev hloov kho. Lub 0.25 units / mL collagenase (40 μL) muab los ntawm Clostridium histolyticum tau tso cai rau hnov ​​​​mob nrog 10 μL kuaj cov qauv thiab 20 μL 50 mM tricine tsis (pH 7.5, nrog 100 mM CaCl2 thiab 5 mM NaCl15) hauv qhov tsaus ntuj. Tom qab ua ntej incubation, tus nqi ntawm 40 μL ntawm 2 mM FALGPA tov tau ntxiv rau txhua qhov dej. Txhua tus qauv tau nrog ib qho khoob uas muaj tag nrho cov khoom tshwj tsis yog FALGPA thiab qhov nqus tau raug txiav txim siab tom qab incubated rau 20 min.

2.2.8 ib. Bacteriological txee lub neej tsom xam ntawm KLEL

Cov txheej txheem siv rau tag nrho aerobic microbial suav (TAMC) thiab tag nrho cov poov xab thiab pwm suav (TYMC) ntawm KLEL tau hloov kho los ntawm FDA bacteriological analytical phau ntawv (BAM) (Huang li al., 2017). Cov qauv (1 g) yog tov nrog 1 mL sterile Tween 80 thiab ntim tau kho nrog sterile peptone dej kom tau ib tug tiav dilution series los ntawm 10− 1 mus rau 10− 3. Rau lub TAMC, cov qauv tau inoculated rau nutrient agar siv a Txoj kev sib kis ntawm cov phaj, thiab tom qab ntawd cov phaj tau incubated ntawm 30 ± 2 ◦C rau 48 h. Nyob rau hauv TYMC rooj plaub, cov qauv tau inoculated rau daim hlau ntawm qos yaj ywm dextrose agar siv txoj kev sib kis, thiab cov phaj tau tom qab incubated ntawm 30 ± 2 ◦C rau 7 hnub. Cov phiajcim tau kuaj xyuas kev loj hlob ntawm microbial tom qab lub sijhawm incubation.

2.3. Kev txheeb cais

Tag nrho cov txiaj ntsig tau raug tshuaj xyuas siv Minitab 16.2.1 cov ntaub ntawv pov thawj pob (Minitab Inc., Pennsylvania, Tebchaws USA), Ib Txoj Kev Ntsuam Xyuas ntawm qhov sib txawv (ANOVA) tau ua, ua raws li Tukey qhov kev sim los txiav txim siab qhov sib txawv tseem ceeb (p < 0="" .05).="" txhais="" tau="" tias="" ±="" tus="" qauv="" sib="" txawv="" (sd)="" (n="3)" tau="" nthuav="" tawm="" rau="" kev="" txheeb="" xyuas="" cov="" ntaub="">

3. Cov txiaj ntsig thiab kev sib tham

3.1. Optimization ntawm lotion puag formulation

Raws li Hiola et al. (2018), kev ua kom zoo ntawm cov tshuaj pleev ib ce (pre-formulation) muaj lub luag haujlwm tseem ceeb kom tau txais cov qauv zoo thiab ruaj khov. Yog li ntawd, cov tshuaj pleev ib ce tau ua kom zoo dua los ntawm kev siv 4 feem pua ​​​​ntawm KSO raws li qhia hauv Table 1. Lawv tau soj ntsuam los ntawm cov tsos, tsw, homogeneity, lub cev ruaj khov, pH, kis tau, viscosity, thiab khov-thaw tsom xam (cov ntaub ntawv ntxiv-S1 thiab S2). Tag nrho cov tshuaj pleev ib ce npaj los ntawm kev siv KSO muaj lub teeb daj daj rau cov xim daj daj, nrog cov ntxhiab tsw zoo (Chu thiab Nyam, 2020). Raws li cov txiaj ntsig tau txais, tag nrho cov tshuaj pleev xim rau cov qauv tsim 'pH qhov tseem ceeb nyob rau hauv thaj tsam ntawm daim tawv nqaij pH (4-6), thiab qhov no yog qhov tseem ceeb kom txo qis cov tshuaj tsis haum thiab ua kom muaj kev ruaj ntseg ntawm cov tshuaj pleev ib ce nyob rau lub sij hawm cia (Chu thiab Nyam). , 2020). Tag nrho cov tshuaj pleev ib ce qhia pom qhov poob qis tom qab kev tshuaj xyuas khov-thaw, tab sis tseem nyob hauv thaj tsam rau cov tshuaj pleev zoo (500-5000 cP) (Kusuma li al., 2017). Rau qhov kis tau tus mob, qhov siab dua ntawm KSO, qhov me dua qhov kis tau tus kab mob vim muaj viscosity siab. Qhov loj dua qhov sib kis tau yooj yim, qhov yooj yim ntawm cov tshuaj pleev ib ce tuaj yeem siv rau ntawm daim tawv nqaij. Rau qhov kev ntsuam xyuas kev ruaj ntseg los ntawm kev tsis kam mus rau centrifugation, F2 thiab F3 pom tias tsis muaj theem sib cais, qhia tias ob qho tib si formulations ruaj khov rau 1 xyoo. Txawm li cas los xij, F3 tsis tuaj yeem nchuav tau yooj yim thiab muab cov tsos zoo li cov tshuaj nplaum vim nws muaj viscosity siab dua F2. Yog li ntawd, F2 tau raug xaiv los ua cov tshuaj pleev qhov zoo tshaj plaws los koom nrog KLE.

Dab tsi yog cistanche siv rau: anti-aging

3.2. Kev ntsuam xyuas ntawm KLE lotion (KLE) formulation

3.2.1. Physicochemical tsom xam

Raws li cov txiaj ntsig tau tshwm sim hauv Table 2, cov tshuaj pleev ib ce ntxiv nrog KLE pom cov xim mis nyuj nrog cov ntxhiab tsw qab ntxiag thiab tsis muaj theem sib cais. KLEL pom tias muaj pH ntau dua li kev tswj hwm, qhia tias KLE ntxiv rau hauv cov tshuaj pleev ib ce tuaj yeem ua rau nce pH tus nqi. Cov pH qhov tseem ceeb ntawm cov qauv yog txhua yam sib xws nrog cov tawv nqaij pH ntau thiab muaj kev nyab xeeb rau daim tawv nqaij. Tsis muaj qhov sib txawv tseem ceeb hauv viscosity thiab kev sib kis ntawm KLEL thiab kev tswj hwm, uas qhia tias ntxiv KLE rau hauv emulsion tsis cuam tshuam rau viscosity thiab kis tau. Cov kev hloov pauv tseem ceeb tau pom nyob rau hauv L *, a *, thiab b * qhov tseem ceeb ntawm KLEL piv nrog kev tswj hwm, uas ua pov thawj tias cov xim ntawm cov tshuaj pleev ib ce cuam tshuam los ntawm cov xim ntsuab ntsuab ntawm nplooj kenaf. KLEL muaj qhov ntsuab tshaj plaws rau a* tus nqi (-2.24 ± 0.03) thiab yellowest rau b* tus nqi (15.54 ± 0.{{13} }1). Tag nrho cov xim sib txawv (ΔE) ntawm KLEL piv nrog kev tswj yog qhov tseem ceeb ntawm p＜0.05, nrog rau tus nqi ntawm 6.82 ± 0.04 rau KLEL. Cov kev tshawb fawb yav dhau los suav hais tias ΔE=2 yog qhov pib rau kev ntxub ntxaug (Zhou et al., 2009). Yog li, cov txiaj ntsig ntawm ΔE tau ua pov thawj tias qhov sib ntxiv ntawm KLE tuaj yeem cuam tshuam cov xim ntawm cov tshuaj pleev ib ce.

3.2.2. Antioxidant cov ntsiab lus tsom xam

Table 3 nthuav qhia tus nqi tag nrho phenolic (TPC) thiab cov ntsiab lus flavonoid (TFC). Cov txiaj ntsig tau pom tau qhia tias cov tshuaj pleev xim hauv qab nrog KLE muaj TPC ntau dua (1.84 ± 0.07 mgCAE/g) dua li kev tswj hwm (1.64 ± 0.0 6 mgCAE/g). Thaum, rau TFC, KLEL kuj tau pom qhov tseem ceeb hauv TFC (21.62 ± 0.76 mgCHE / g) piv nrog kev tswj hwm (19.42 ± 0.27 mgCHE / g). Nyob rau tib lub sijhawm ntawm qee cov khoom cog qoob loo, suav nrog KLE ntawm cov kab mob dermatological tseem nthuav tawm cov ntsiab lus antioxidant siab, qhia txog nws qhov muaj peev xwm ntawm cov tshuaj polyphenol hauv kev lag luam tshuaj pleev ib ce (Adhikari li al., 2019; Rodrigues li al., 2014. ). Raws li kev tshawb fawb los ntawm Haw et al. (2020), kenaf nplooj yog nplua nuj nyob rau hauv ntau hom polyphenol tebchaw xws li caffeic acid, tannic acid, catechin, thiab chlorogenic acid.

3.2.3. Antioxidant kev ua haujlwm tsom xam

Lub peev xwm dawb radical scavenging ntawm cov tshuaj pleev ib ce tau tshawb xyuas los ntawm kev siv DPPH thiab ABTS assay. Table 3 qhia tias KLEL pom tau tias siab dua DPPH (1.16 ± 0.18 mgTEAC/g) thiab ABTS (0.50 ± 0.0 4 mgTEAC/g) radical scavenging kev ua piv nrog rau kev tswj. DPPH thiab ABTS kev soj ntsuam kev tshawb pom tau pom cov qauv zoo ib yam li hauv TPC thiab TFC, qhov uas nws tuaj yeem txhim kho cov tshuaj tiv thaiv kab mob antioxidant los ntawm kev ntxiv KLE rau hauv cov tshuaj pleev ib ce, cov kev tshawb pom no tau pom tias KLE muaj cov dawb radical scavengers uas thaum siv rau hauv cov tshuaj pleev ib ce, tuaj yeem ua si. Lub luag haujlwm tseem ceeb yog thawj cov tshuaj tiv thaiv antioxidant hauv kev tshem tawm cov dawb radical uas ua rau tawv nqaij laus. Tsis tas li ntawd, KLEL (0.69 ± 0.20 mgTEAC/g) kuj pom tau tias muaj peev xwm txo qis ferric ion ntau tshaj qhov kev tswj (0.46 ± 0.09 mgTEAC/g), pom zoo nrog DPPH thiab ABTS. Yuav tsum muaj cov khoom noj tsis tu ncua ntawm cov tshuaj tiv thaiv antioxidant los ntawm lwm qhov chaw xws li tshuaj pleev ib ce los kho cov tawv nqaij tiv thaiv kab mob tiv thaiv ROS uas ua rau cov tawv nqaij laus (Działo li al., 2016). Tsis tas li ntawd, cov tshuaj pleev ib ce nrog cov tshuaj tua kab mob radical thiab txo lub zog tuaj yeem siv los ua daim tawv nqaijntxuav hniav dawbTus neeg sawv cev los ntawm kev tswj hwm UV-induced melanin ntau lawm (Działo li al., 2016).

3.2.4. Enzyme inhibitory kev ua haujlwm

Cov tshuaj pleev ib ce tau sim tawm tsam tyrosinase, tooj liab-muaj metalloprotein enzyme koom nrog hauv melanin biosynthesis, rau kev ntsuam xyuas hauv vitro antityrosinase kev ua. Table 5 qhia tau hais tias KLEL (30.28 ± 3.9{18}} feem pua ​​) tsis muaj qhov sib txawv tseem ceeb hauv kev tiv thaiv tyrosinase nrog kev tswj zoo (34.05 ± 4.60%) thaum siv L-tyrosine ua substrate . Txawm li cas los xij, TAG NRHO (11.40 ± 0.29 feem pua) tau pom tias muaj kev ua haujlwm antityrosinase qis dua li kev tswj hwm zoo (15.01 ± 0.84%), thaum siv L-DOPA ua substrate. Qhov no qhia tau tias KLEL feem ntau inhibited mycophenolate es tsis yog diphenols. Hauv vitroanti-agingCov khoom raug soj ntsuam los ntawm collagenase thiab elastase inhibitory kev ua haujlwm. Ib qho enzyme xws li collagenase thiab elastase tuaj yeem degrade collagen thiab elastin, uas yog lub luag haujlwm rau kev ncaj ncees ntawm daim tawv nqaij thiab elasticity (Jesumani li al., 2019). Yog li, qhov inhibition ntawm elastase thiab collagenase kev ua ub no yuav txo qhov degradation ntawm elastin thiab collagen yog li tiv thaiv wrinkle tsim, ib qho ntawm cov cim tseem ceeb ntawm kev laus. Raws li pom nyob rau hauv Table 5, KLEL qhia tau zoo kawg nkaus anti collagenase (36.41 ± 0.54 feem pua ​​) thiab antielastase (23.13 ± 1.56 feem pua) kev ua ub no nyob rau hauv kev sib piv nrog rau kev tswj zoo (CHL). KSOL kuj tau qhia txog antityrosinase (18.82 ± 0.17 feem pua ​​(L-tyrosine as substrate); 8.48 ± {22}}.29 feem pua ​​(L-DOPA as substrate), anti collagenase (18.84 ± {{26). }}.63 feem pua ​​​​) thiab antielastase (5.78 ± 0.59 feem pua ​​) kev ua haujlwm, tab sis qis dua TAG NRHO. Qhov no yuav qhia tau tias kev sib xyaw ntawm KLE thiab KSO pom tau tias muaj kev sib koom ua ke nrog kev ua haujlwm zoo enzyme inhibitory. Pascoal li al. (2015) thiab Chew et al. (2016), uas KSO yog nplua nuj nyob rau hauv tocopherols, thaum KLE yog nplua nuj nyob rau hauv quercetin thiab kaempferol derivatives.ntxuav hniav dawbthiabanti-agingcov khoom, nrog rau kev txaus siab rau kev txo cov tawv nqaij hyperpigmentation thiab wrinkles ntau lawm (Lin li al., 2007; Keen thiab Hassan, 2016).

3.2.5. Kev ntsuam xyuas hauv vitro melanogenesis

Txhawm rau tshawb xyuas daim tawv nqaij ntxivntxuav hniav dawbCov khoom ntawm KLEL, B16F10 melanoma cell qauv tau siv los kawm txog cov nyhuv inhibitory ntawm melanogenesis. Rau kev ua haujlwm ntawm cellular tyrosinase, raws li qhia hauv daim duab 5a, KLEL (32.35 feem pua) ntawm 500 ug/mL pom tau tias muaj zog inhibition ntawm cellular tyrosinase kev ua ntau dua li KAL (14.85 feem pua) thiab KSOL (13.64 feem pua), thaum piv nrog a-MSH kho tswj cell. Rau cov ntsiab lus ntawm cov cellular thiab intracellular melanin, KLEL koob tshuaj-dependently txo cov ntsiab lus melanin ntawm B16F10 hlwb (Fig. 5b thiab c). TAG NRHO (500 ug / mL) tau pom tias muaj qhov sib piv inhibitory cuam tshuam rau cov ntsiab lus melanin extracellular (56.13 feem pua) (piv nrog MSH-kho cell cell) nrog KAL (54.53 feem pua). Tsis tas li ntawd, KLEL (36.52 feem pua) kuj pom tias muaj zog inhibitory nyhuv ntawm intracellular melanin cov ntsiab lus ntau dua KAL (7.98 feem pua). Thaum, KSOL kuj tau pom tias muaj kev cuam tshuam inhibitory ntawm cov ntsiab lus melanin extracellular (17.73 feem pua), tab sis tsis cuam tshuam rau intracellular melanin cov ntsiab lus. Piv nrog KAL, qhov muaj zog inhibitory cuam tshuam ntawm KLEL ntawm inhibition ntawm melanogenesis tuaj yeem qhia meej los ntawm kev ua haujlwm sib koom ua ke ntawm KSO thiab KLE, uas tuaj yeem txhim kho KLE daim tawv nqaij dawb lub zog hauv cov tshuaj pleev ib ce. Cov txiaj ntsig no tau ua pov thawj ntxiv tias KLE tuaj yeem siv los ua tus neeg sawv cev zoo nkauj ntawm daim tawv nqaij hauv cov qauv tshuaj pleev ib ce.

3.2.6. Hauv vitro cytotoxicity assay

Los ntawm kev ntsuam xyuas biocompatibility ntawm KLEL (MTT tsom xam), NHEF hlwb muaj peev xwm (tom qab 24, 48, 72 h) loj hlob nyob rau hauv muaj cov sib txawv KLEL concentrations (0.125- 2 mg / mL) tau khaws cia saum toj no 95 feem pua ​​(Fig. 4a). Qhov no tau qhia tias KLEL yuav tsis ua rau muaj kev phom sij rau tib neeg. Thaum B1610 melanoma hlwb tau siv dav hauv kev tshawb fawb rau daim tawv nqaijntxuav hniav dawbcov neeg sawv cev (Chatatikun et al., 2019; Lee et al., 2019). Ua ntej kev ntsuas lub peev xwm ntawm KLEL txhawm rau txo qis melanogenesis hauv B16F10 melanoma hlwb, cytotoxicity ntawm KLEL tau soj ntsuam. Raws li qhia hauv daim duab 4b, nws tau lees paub tias KLEL tsis muaj tshuaj lom rau B16F10 melanoma hlwb ntawm qhov concentration qis dua 500 ug / mL. Yog li, 32.5- 500 ug / mL KLEL tau siv rau cov kev sim hauv qab no.

4. Cov lus xaus

Txoj kev tshawb no tau ua pov thawj tias muaj txiaj ntsig zoo hauv cov qauv kev lag luam ua tiav, raws li nws nthuav tawm cov ntawv thov tshiab rau kenaf nplooj raws li cov khoom muaj nqis ntxiv nrog cov khoom tu tawv nqaij rau kev lag luam tshuaj pleev ib ce, uas yog antioxidant,anti-aging, thiab tiv thaiv melanogenic kev ua ub no. Cov kev tshawb pom tau nthuav tawm hauv txoj kev tshawb fawb kuj tau qhia tias cov tshuaj pleev ib ce npaj los ntawm 15 feem pua ​​​​KSO (F2) nrog 0.1 feem pua ​​KLE yields qhov zoo tshaj plaws lub cev thiab microbiological stability, tsis muaj toxicity ntawm tib neeg hlwb. Hauv kev txuas ntxiv, kev tshawb fawb ntxiv yuav tsum tau tsom mus rau KLEL microbial kev sib tw xeem thiab kev kho mob ua tau zoo.

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