Ispektram Yokufunxa ye-Methylene Blue neKhowefishiyenti Yokucinywa Kokukhanya Ngokwemole

Ispektram ye-methylene blue ibonakala ilula de uzame ukuphinda usebenzise inani elivela kuyo. Enye ilabhoratri inika ingxelo yekhowefishiyenti yokucinywa kokukhanya ngokwemole ekufutshane ne-82,000. Enye inika i-67,000. Eyesithathu inika ngaphezu kwe-100,000. Zontathu zinokuba ziinkcazelo ezichanekileyo zemilinganiselo yazo, kodwa akukho nanye kuzo elixabiso elingaguqukiyo lale dayi kuzo zonke iimeko.

Oku kubhideka kuvela ekuxutyweni kweekhonsepthi ezimbini ezahlukileyo. Umlinganiselo wokufunxwa kokukhanya ngumlinganiselo owenziwa kwisisombululo esithile esikwiseli ethile yokulinganisa. Umlinganiselo wokufunxa ngokwemole, odla ngokubhalwa njenge-epsilon, yiparamitha kwimodeli yeso sisombululo. Xa i-methylene blue itshintsha indlela iimolekyuli zayo ezidityana ngayo, imodeli iyatshintsha, ngoko neparamitha iyatshintsha. Umkhwa oluncedo kukuphatha ikhowefishiyenti nganye njengexabiso elineenkcukacha ezichaziweyo: ubude bamaza, isinyibilikisi, i-pH okanye isithinteli se-pH, iqondo lobushushu, uluhlu loxinaniso nobude bendlela yokukhanya kufuneka zihambe kunye nenani.

Ibhanti yemonomer kumanzi anoxinaniso oluphantsi

Kwisisombululo samanzi esinoxinaniso oluphantsi, ibhanti ephambili yekheyishini ye-methylene blue ikufutshane ne-664 ukuya kwi-665 nm, yaye idla ngokuchazwa ngaphakathi kwe-664 ukuya kwi-668 nm ngokuxhomekeke kwizixhobo nakukhetho lokulinganisa. Loo bhanti yeyona ifanelekileyo ekumiseleni ubungakanani xa uxinaniso lwesisombululo luphantsi ngokwaneleyo kangangokuba uninzi lwedayi lukwimo yemonomer.

Amaxabiso amathathu akuncwadi lophando abonisa ukuba kutheni iimeko zibaluleke ngaphezu kwenani eliphambili. Abaphandi bodongwe ababegcina iisampulu zamanzi zingaphantsi malunga ne-7 micromolar kwiiseli eziyi-1 cm banika ingxelo ye-epsilon emalunga ne-82,000 M-1 cm-1 kwi-665 nm. Elo xabiso libhalwe ngokucacileyo ngokukodwa njengethambeka lokulinganisa elifunyenwe ngovavanyo kwiisampulu zamanzi ezinoxinaniso oluphantsi kakhulu. Ulinganiso oluphambili lwamva emanzini asuswe ii-ion, olusebenzisa i-2.34 ukuya kwi-37.52 micromolar nendlela yokukhanya eyi-1 cm, lwanika malunga ne-67,100 M-1 cm-1 kwi-665 nm kwidayi iyonke. Isiphumo esisezantsi silindelekile, asiphikisani nesinye, kuba isiqingatha esingaphezulu solo luhlu lokulinganisa sele sinomlinganiselo obonakalayo wedimer. Ixabiso lesithathu, elimalunga ne-73,460 M-1 cm-1 kwi-664 nm, livela kuphando olwenziwa kwi-pH 6.86 nakwi-293 K olwahlula ispektram eboniweyo yaba ngamacandelo emonomer nedimer, njengoko kushwankathelwe kuphononongo lwamva oluhlalutya nzulu ifotofiziksi ye-methylene blue. Elo nani licandelo lemonomer elahlulwe ngokuhlalutya ispektram, hayi ithambeka elithe ngqo lomlinganiselo wokufunxwa kokukhanya xa kuthelekiswa nedayi iyonke.

Ingongoma esebenzayo: khetha ikhowefishiyenti eneemeko ezihambelana neesampulu zakho, okanye wenze olwakho ulinganiso. Musa ukusebenzisa i-82,000 kuvavanyo lwesisombululo esine-phosphate njengesithinteli se-pH esiyi-30 micromolar uze ucinge ukuba ulwalamano lusengumgca othe ngqo.

Igxalaba elikubude bamaza obufutshane ngumxube

Njengoko uxinaniso lunyuka, igxalaba elikufutshane ne-600 ukuya kwi-615 nm liyakhula xa lithelekiswa nencopho ye-664 nm. Uphando lwale mihla ludla ngokubeka igalelo ledimer yohlobo lwe-H kufutshane ne-605 ukuya kwi-610 nm. Uhlalutyo oluhlangeneyo lweespektram ze-methylene blue emanzini lwahlula incopho yedimer kwi-607 nm nencopho yeqela elikhulu leemolekyuli kwi-600 nm, kunye namacandelo eresonansi yemonomer kwi-664 nakwi-650 nm. Umbhalo walo opheleleyo uyafumaneka kwirekhodi efikeleleka simahla ye-PMID 33251415.

Elo gxalaba akufanele lichazwe njengomlinganiselo wokufunxwa kokukhanya yidimer yodwa. Ulwakhiwo lwemonomer olunxulumene nokungcangcazela neenguqu ze-elektroni nalo lunegalelo kwakuloo mmandla, ngoko ukuphakama okulinganisiweyo kukudibana kwamagalelo. Olunye ulungelelaniso lwemodeli kuyo yonke idatha lwafumanisa ukuba imodeli yemonomer nedimer ayiphumelelanga kulo lonke uluhlu lwe-1.1 micromolar ukuya kwi-3.4 millimolar, ngoxa ukongeza itetramer kwanika olona lungelelwaniso lufanelekileyo, umqondiso wetetramer sele ubonakala nangaphantsi kwe-34 micromolar. Amandla amacandelo alungelelanisiweyo ayemakhulu: malunga ne-107,800 M-1 cm-1 kwi-664 nm kwicandelo eliphambili lemonomer, malunga ne-101,400 kwidimer kwi-607 nm, kunye ne-138,500 kwitetramer kwi-600 nm, kumanzi asuswe ii-ion angenasithinteli se-pH, ane-sodium chloride ukusuka kwi-0 ukuya kwi-0.15 M namaqondo obushushu ukusuka kwi-282 ukuya kwi-333 K. La ngamacandelo ahlukaniswe yimodeli, ngoko akufanele asetyenziswe endaweni yethambeka le-Beer Lambert elifunyenwe ngovavanyo.

Amandla okwakheka kwedimer alinganiswe ngokuthe ngqo. Kwi-pH 6.86 nakwi-293 K, olunye uphando lwafumana ixabiso elingaguqukiyo lokwakheka kwedimer elikufutshane ne-3,900 M-1. Obo bukhulu buchaza ukuba kutheni isisombululo sokusebenza esiyi-5 micromolar siziphatha phantse ngokolwalamano lomgca othe ngqo, ngoxa isisombululo esingundoqo esiyi-50 micromolar singakwenzi oko. Uvavanyo oluluncedo ngexesha lokuphuhlisa indlela ngumlinganiselo phakathi kokufunxwa kokukhanya kufutshane ne-610 nm nokufunxwa kokukhanya kufutshane ne-665 nm. Xa loo mlinganiselo unyuka kunye noxinaniso, i-epsilon kwi-664 nm ayisachazi uhlobo olunye olungaguqukiyo.

Kuluhlu loxinaniso olulawulwa ikakhulu ziintlobo ezimbini eziguqukela kwenye nenye, iispektram zinokwabelana ngenqaku le-isosbestic, elifunyenwe ngovavanyo kufutshane ne-625 nm kuthotho lwezisombululo zamanzi. Inqaku elicacileyo elihlala kwindawo enye liyahambelana nobukho beentlobo ezimbini eziphambili. Xa elo nqaku lisuka okanye linyamalala njengoko uxinaniso lunyuka, lithathe njengesilumkiso sokuba kungene amaqela amakhulu eemolekyuli okanye iimeko zolungelelwano eziguqukayo, hayi njengempazamo yesixhobo.

Isinyibilikisi, i-pH neqondo lobushushu zitshintsha ispektram

Amanzi akhuthaza ukudibana kweemolekyuli. Ukongeza i-dimethyl sulfoxide kunciphisa ukwakheka kwedimer, kususa iincopho zokufunxa nezefluoresensi ziye kubude bamaza obude ngakumbi, yaye kwandisa isivuno sekhwantam sefluoresensi malunga namaxesha amane, ngokutsho kophando lwezinyibilikisi ze-methylene blue emanzini nakwimixube ye-DMSO, olubhaliswe njenge-PMID 30933781. Olo phando luphatha iispektram zamanzi njengolungelelwano phakathi kwemonomer nedimer, apho ukufunxwa kokukhanya y imonomer kukufutshane ne-650 ukuya kwi-670 nm nokwedimer kukufutshane ne-580 ukuya kwi-615 nm, yaye lufumana amaxabiso angaguqukiyo okwakheka kwedimer kwiispektram kwi-293.15 K.

Ii-alcohol zitshintsha indawo namandla abonakalayo, kodwa azikuphelisi ukudibana kweemolekyuli. Kolunye uthelekiso oluphambili kwi-31.3 micromolar nakwiqondo lobushushu legumbi, oluchazwe kuphononongo lwezinyibilikisi ze-methylene blue, incopho ekubude bamaza obude yayiyi-665 nm emanzini kodwa iyi-655 nm kwi-methanol, kunye namaxabiso abonakalayo esisombululo siphela akufutshane ne-37,100 emanzini ne-60,800 kwi-methanol. Uqinisekiso olwahlukileyo kwi-ethanol engenamanzi kwi-298 K lwanika incopho kwi-655 nm ene-epsilon ekufutshane ne-68,000.

Utshintsho oluqhelekileyo lwe-pH lususa ispektram kancinci kunokuba iinkqubo ezininzi zicinga. Uphononongo oluhlalutya nzulu olukhankanywe ngasentla lunika ingxelo yokuba ukusuka malunga ne-pH 2 ukuya kwi-pH 12, incopho yahlala kufutshane ne-665 nm, kukho utshintsho oluncinane kuphela kumandla ukuya kwi-pH 11. Uhlobo lwesithinteli se-pH namandla e-ion zinokutshintsha iziphumo kangangoko i-pH ebhaliweyo inokwenjenjalo, ngoko bhala esona sithinteli se-pH netyuwa zisetyenzisiweyo, hayi kuphela ufundo lwemitha ye-pH. Iasidi eyomeleleyo okanye ialkali eyomeleleyo yimeko eyahlukileyo kuba idayi inokutshintsha ngokwemichiza endaweni yokudibana okunokubuyiselwa umva. Iqondo lobushushu netyuwa nazo zitshintsha ulungelelwano, yiyo loo nto uphando lwamanzi luka-2020 lwazitshintsha zombini ngabom.

Ukulinganisa ukuze inani lihlale lisebenza

Umsebenzi wokumisela ubungakanani uxhomekeke kulwalamano lwe-Beer Lambert, apho umlinganiselo wokufunxwa kokukhanya ulingana ne-epsilon ephindaphindwe ngoxinaniso nangobude bendlela yokukhanya, kunye nobungqina bokuba uluhlu lolwalamano lomgca othe ngqo luyasebenza ngokwenene phantsi kweemeko zakho.

Sebenzisa iseli eyi-1 cm kumsebenzi okuxinaniso oluphantsi lwe-micromolar. Xa i-epsilon ikufutshane ne-82,000, i-5 micromolar inika umlinganiselo wokufunxwa kokukhanya okufutshane ne-0.41, kwaye i-10 micromolar inika malunga ne-0.82. Kumlinganiselo ogxile kwimonomer kumanzi angenazo ezinye izongezo, ukuhlala kufutshane okanye ngaphantsi malunga ne-5 ukuya kwi-7 micromolar kukhuselekile ngakumbi kunokwenza ulinganiso olufikelela kumashumi e-micromolar. Kwizisombululo ezingundoqo ezinoxinaniso oluphezulu, nciphisa ubude bendlela yokukhanya endaweni yokwamkela umlinganiselo wokufunxwa kokukhanya ongaphezulu kakhulu kwe-1.

Misela ufundo olungenadayi usebenzisa umxube ofanayo ncam: umgangatho ofanayo wamanzi, umlinganiselo ofanayo wesinyibilikisi esongezelelweyo, isithinteli se-pH neetyuwa ezifanayo, kodwa kungabikho dayi. Lungisa izisombululo zokulinganisa kuloo mxube mnye nakwiqondo lobushushu elifanayo. Ngexesha lophuhliso, skena lonke uluhlu lokukhanya okubonakalayo, ngokufanelekileyo malunga ne-550 ukuya kwi-750 nm, endaweni yokufunda ubude bamaza obunye kuphela. Landela imilo, hayi ukuphakama kuphela. Ukuba ukuxutywa ngokulandelelana ukuze kuncitshiswe uxinaniso kutshintsha imilo elungelelanisiweyo, inkqubo ixhomekeke kuxinaniso ngokwemichiza. Incopho engaguqukiyo nomlinganiselo ongaguqukiyo we-610 ukuya kwi-665 nm bubungqina obunamandla ngakumbi boluhlu olwamkelekileyo kunexabiso eliphezulu lika-R ophindwe ngesikwere lodwa.

Bhala imo yereyijenti. Imveliso ethengiswayo inokuba yi-methylthioninium chloride engenamanzi okanye ihayidreyithi efana netrihydrate, ngoko ubunzima ngokwemole obusetyenziswa ekubaleni isisombululo esingundoqo kufuneka buhambelane noko kukwibhotile yokwenene. Eli nyathelo lilodwa lichaza inxalenye yokungavumelani ngoxinaniso ngokwemole olubhaliweyo phakathi kweelabhoratri. Uqwalaselo olufanayo luyasebenza xa iziphumo zisetyenziswa kwizibalo zovavanyo lobungakanani, xa iispektram zixhasa ukujongwa kozinzo lokwakheka komxube, naxa iinkcukacha zokungcola zifuna uphononongo lobunyulu nemigaqo ebekiweyo.

Isishwankathelo esineemeko ezichaziweyo

Sebenzisa le theyibhile njengethempleyithi yokuqala yerekhodi yelabhoratri, hayi njengomthombo wamaxabiso angaguqukiyo anokusetyenziswa endaweni yelinye nelinye.

Uhlobo nobude bamaza	Iimeko	I-epsilon okanye indlela yokuziphatha echaziweyo
Imonomer, 665 nm	Amanzi, ngaphantsi malunga ne-7 micromolar, 1 cm, i-pH ayichazwanga	Malunga ne-82,000 M-1 cm-1, ithambeka ledayi iyonke elifunyenwe ngovavanyo
Icandelo lemonomer, 664 nm	Amanzi, pH 6.86, 293 K, lahlulwe kwidimer ngohlalutyo	Malunga ne-73,460 M-1 cm-1, icandelo lemonomer kuphela
Idayi iyonke, 665 nm	Amanzi asuswe ii-ion, 2.34 ukuya kwi-37.52 micromolar, 1 cm	Malunga ne-67,100 M-1 cm-1, ithambeka lokulinganisa kuluhlu apho ukudibana kweemolekyuli kwenzeka ngokuyinxenye
Iresonansi yemonomer, 664 nm	Amanzi asuswe ii-ion angenasithinteli se-pH, uthotho lwe-1.1 micromolar ukuya kwi-3.4 millimolar, NaCl 0 ukuya kwi-0.15 M, 282 ukuya kwi-333 K	Malunga ne-107,800 M-1 cm-1, icandelo elahlulwe yimodeli
Idimer, 607 nm	Uthotho olufanayo nolo lungasentla	Malunga ne-101,400 M-1 cm-1, icandelo elahlulwe yimodeli
Itetramer, 600 nm	Uthotho olufanayo nolo lungasentla	Malunga ne-138,500 M-1 cm-1, icandelo elahlulwe yimodeli
Idayi iyonke, 665 nm emanzini xa kuthelekiswa ne-655 nm kwi-methanol	31.3 micromolar, iqondo lobushushu legumbi	Malunga ne-37,100 emanzini, malunga ne-60,800 kwi-methanol, amaxabiso abonakalayo kolo xinaniso
Ulungelelwano phakathi kwemonomer nedimer	Imixube yamanzi ne-DMSO, 293.15 K	I-DMSO inciphisa idimer, iincopho zisuka ziye kubude bamaza obude ngakumbi, isivuno sokukhutshwa kokukhanya siyenyuka

Ngoko ke, ubude bamaza okufunxwa kokukhanya yi-methylene blue yimpendulo emfutshane enengcaciso ende esezantsi. Chaza incopho oyilinganisileyo, kwisisombululo osilinganisileyo, kuxinaniso olulinganisileyo. Loo mkhwa uthintela ixabiso elinye lesisombululo samanzi esinoxinaniso oluphantsi kufutshane ne-664 nm ekubeni lijike libe lixabiso elingaguqukiyo elingeyonyani kuzo zonke iimeko.