Spekitiramu Yekutorwa Kwechiedza neMethylene Blue neKoefishendi Yekudzimwa Kwechiedza Pamole

Spekitiramu yemethylene blue inoita seyakapusa kusvikira waedza kushandisazve nhamba inobva mairi. Imwe rabhoritari inotaura koefishendi yekudzimwa kwechiedza pamole iri pedyo ne82,000. Imwe inotaura 67,000. Yechitatu inotaura inopfuura 100,000. Dzose dziri nhatu dzinogona kutsanangura nemazvo zviyero zvadzo, asi hapana yadzo iri nhamba isingachinji yedhayi mumamiriro ose.

Kuvhiringidzika kunobva pakusanganisa pfungwa mbiri dzakasiyana. Mwero wekutorwa kwechiedza chiyero chinoitwa pamusanganiswa wakati uri musero rakati. Kugona kutora chiedza pamole, kunowanzonyorwa seepsilon, iparamita iri mumuenzaniso wemasvomhu wemusanganiswa iwoyo. Kana methylene blue ikachinja mamiriro ekubatana kwemamorekuru ayo, muenzaniso wacho unoshanduka, saka paramita yacho inoshandukawo. Tsika inobatsira ndeyekubata koefishendi imwe neimwe senhamba ine mamiriro ayo akanyorwa: urefu hwesaisai, chinonyungudutsa, pH kana buffer, tembiricha, chiyero cheuwandu hwechinhu pamwero wemvura, uye kureba kwenzira yechiedza zvinofambidzana nenhamba yacho.

Bhandi remonomera mumvura ine musanganiswa wakareruka

Mumvura ine methylene blue yakanyungudutswa pamwero wakaderera, bhandi guru rekationi yemethylene blue riri pedyo ne664 kusvika 665 nm, uye rinowanzonzi riri pakati pe664 ne668 nm zvichienderana nemudziyo nesarudzo dzekugadzirisa zviyero. Bhandi iroro ndiro rakakodzera kuyera huwandu kana musanganiswa wakareruka zvakakwana kuti dhayi yakawanda ive mumamorekuru ega ega emonomera.

Nhamba nhatu dziri mumabhuku ets research dzinoratidza kuti nei mamiriro achikosha kupfuura nhamba inotanga kutaurwa. Vatsvakurudzi vevhu veclay vakachengeta sampuli dzemvura dziri pasi peanenge 7 micromolar mumasero e1 cm vakataura epsilon yeanenge 82,000 M-1 cm-1 pa665 nm. Nhamba iyoyo ine zvinyorwa zvinonyatsoitsigira sekutenuka kwemutsetse wekugadzirisa zviyero kwakawanikwa nekuyera sampuli dzemvura dzakareruka zvikuru. Gare gare, kugadzirisa zviyero kwekutanga mumvura yakabviswa maioni, kuchishandisa 2.34 kusvika 37.52 micromolar nenzira yechiedza ye1 cm, kwakapa anenge 67,100 M-1 cm-1 pa665 nm padhayi yose. Mhedzisiro yakaderera iyi inotarisirwa, kwete inopesana neyekutanga, nokuti hafu yepamusoro yechiyero chakashandiswa kugadzirisa zviyero inotova nedimera yakawanda zvinooneka. Nhamba yechitatu, inenge 73,460 M-1 cm-1 pa664 nm, inobva mutsvakurudzo yakaitwa papH 6.86 ne293 K yakapatsanura spekitiramu yakaonekwa kuita zvikamu zvemonomera nedimera, sezvakapfupikiswa muongororo ichangoburwa inotsoropodza fizikisi yemethylene blue ine chekuita nechiedza. Nhamba iyoyo ndeyechikamu chemonomera chakabviswa pamusanganiswa nemasvomhu, kwete kutenuka kwemutsetse kunobva pakuyera mwero wekutorwa kwechiedza uchienzaniswa neuwandu hwedhayi yose.

Pfungwa yekushandisa ndeiyi: sarudza koefishendi ine mamiriro anoenderana nesampuli dzako, kana kuti gadzirisa zviyero zvako pachako. Usangoshandisa 82,000 muyedzo we30 micromolar uri muphophate buffer uchifunga kuti hukama hwemutsetse wakatwasuka huchiri kushanda.

Pfudzi riri paurefu hwesaisai hupfupi musanganiswa

Sezvo uwandu hwechinhu pamwero wemvura huchikwira, pfudzi riri pedyo ne600 kusvika 615 nm rinokura kana richienzaniswa nenhongonya ye664 nm. Tsvakurudzo dzemazuva ano dzinowanzoisa chikamu chinobva kudimera yerudzi rweH pedyo ne605 kusvika 610 nm. Ongororo yespekitiramu yemethylene blue mumvura yose pamwe chete inopatsanura nhongonya yedimera iri pa607 nm nenhongonya yemapoka makuru emamorekuru iri pa600 nm, pamwe nezvikamu zverezonensi yemon om era zviri pa664 ne650 nm. Chinyorwa chayo chose chinowanikwa kuburikidza nerekodhi rinowanikwa pachena rePMID 33251415.

Pfudzi iroro harifaniri kunzi rinongova kutorwa kwechiedza nedimera chete. Chimiro chemonomera chine chekuita nekudengenyeka kwemamorekuru nemamiriro emaerekitironi chinopawo chikamu munzvimbo imwe cheteyo, saka kureba kunoyerwa kubatanidzwa kwezvikamu zvakasiyana. Kumwe kuenzanisa muenzaniso nemiyero yose kwakawana kuti muenzaniso une monomera nedimera wakatadza kuenderana nechiyero chose che1.1 micromolar kusvika 3.4 millimolar, asi kuwedzera tetramera kwakapa kuenderana kwakanakisisa, chiratidzo chetetramera chichitoonekwa pasi pe34 micromolar. Masimba ezvikamu zvakawanikwa nekuenzanisa aive makuru: anenge 107,800 M-1 cm-1 pa664 nm pachikamu chikuru chemonomera, anenge 101,400 padimera pa607 nm, uye anenge 138,500 patetramera pa600 nm, mumvura yakabviswa maioni isina buffer, ine sodium chloride kubva pa0 kusvika 0.15 M uye tembiricha kubva pa282 kusvika 333 K. Izvi zvikamu zvakapatsanurwa nemuenzaniso wemasvomhu, saka hazvifaniri kutsiva kutenuka kwemutsetse weBeer Lambert kwakawanikwa nekuyera chaiko.

Simba rekuumbwa kwedimera rakayerwa zvakananga. PapH 6.86 ne293 K, imwe tsvakurudzo yakawana konstanti yekuumbwa kwedimera iri pedyo ne3,900 M-1. Kukura kwenhamba iyoyo kunotsanangura kuti nei musanganiswa wekushandisa we5 micromolar uchitevera hukama hunenge hwemutsetse wakatwasuka asi musanganiswa wekuchengetera we50 micromolar usingadaro. Chimwe chinhu chinobatsira kuongorora pakugadzira nzira yekuyera ireshiyo yemwero wekutorwa kwechiedza pedyo ne610 nm nemwero wacho pedyo ne665 nm. Kana reshiyo iyoyo ichikwira pamwe neuwandu hwechinhu pamwero wemvura, epsilon pa664 nm inenge isisatsananguri rudzi rumwe chete rusingachinji.

Muchikamu cheuwandu hwechinhu pamwero wemvura chinotongwa nemhando mbiri dzinoshandukirana, maspekitiramu anogona kugoverana poindi yeisosbestic, inowanikwa mumiedzo pedyo ne625 nm mumisanganiswa yemvura yakatevedzana. Poindi yakajeka inoramba panzvimbo imwe chete inoenderana nekuvapo kwemhando mbiri huru. Kana poindi yacho ichitsvedza kana kunyangarika apo uwandu hwechinhu pamwero wemvura huchikwira, ibate seyambiro yekuti mapoka makuru emamorekuru kana kuenzana kuri kuchinja kwapinda, kwete sekukanganisa kwemudziyo.

Chinonyungudutsa, pH, netembiricha zvinoshandura spekitiramu

Mvura inofarira kubatana kwemamorekuru. Kuwedzera dimethyl sulfoxide kunoderedza kuumbwa kwedimera, kunofambisa nhongonya dzekutorwa kwechiedza nefluorescence kuenda kuurefu hwesaisai hwakareba, uye kunowedzera goho rekwantamu refluorescence zvakapetwa kanenge kana, maererano netsvakurudzo yezvinonyungudutsa methylene blue mumisanganiswa yemvura neDMSO, yakanyorwa pasi pePMID 30933781. Tsvakurudzo iyoyo inobata maspekitiramu emvura sekuenzana pakati pemonomera nedimera, monomera ichitora chiedza pedyo ne650 kusvika 670 nm uye dimera pedyo ne580 kusvika 615 nm, uye inoburitsa makonstanti ekuumbwa kwedimera kubva mumaspekitiramu pa293.15 K.

Maalcohol anochinja nzvimbo pamwe nesimba rinoratidzika, asi haabvisi kuungana kwemamorekuru. Mune kumwe kuenzanisa kwekutanga pa31.3 micromolar netembiricha yemumba, kwakataurwa mutsvakurudzo yezvinonyungudutsa methylene blue, nhongonya iri paurefu hwesaisai hwakareba yaive pa665 nm mumvura asi pa655 nm mumethanol, nenhamba dzinoratidzika dzemusanganiswa wose dziri pedyo ne37,100 mumvura ne60,800 mumethanol. Kumwe kusimbisa kwakasiyana muethanol isina mvura pa298 K kwakapa nhongonya pa655 nm neepsilon iri pedyo ne68,000.

Shanduko dzepH dzakajairika dzinofambisa spekitiramu zvishoma kupfuura zvinofungidzirwa nenzira zhinji dzakanyorwa dzekuita miedzo. Ongororo inotsoropodza yataurwa pamusoro inoti kubva paanenge pH 2 kusvika pH 12, nhongonya yakaramba iri pedyo ne665 nm, kuchingova neshanduko duku dzesimba kusvika papH 11. Rudzi rwebuffer nesimba remaioni zvinogona kushandura mhedzisiro zvakangofanana nepH yakatarwa, saka nyora buffer chaiyo nemunyu, kwete nhamba yemita yepH chete. Asidhi yakasimba kana arikari yakasimba inyaya yakasiyana nokuti dhayi inogona kuchinja pamakemikari pane kungobatana kwemamorekuru kunogona kudzokera shure. Tembiricha nemunyu zvinoshandurawo kuenzana uku, ndosaka tsvakurudzo yemvura ya2020 yakachinja zvose zviri zviviri nemaune.

Kuyera kuitira kuti nhamba irambe ichishanda

Basa rekuyera huwandu rinovakirwa pahukama hweBeer Lambert: mwero wekutorwa kwechiedza wakaenzana neepsilon yakapetwa neuwandu hwechinhu pamwero wemvura, yobva yapetwa nekureba kwenzira yechiedza, pamwe neuchapupu hwekuti hukama hwemutsetse wakatwasuka hunoshanda chaizvo muchikamu chakasarudzwa pasi pemamiriro ako.

Shandisa sero re1 cm pabasa reuwandu hwakaderera hwe micromolar. Kana epsilon iri pedyo ne82,000, 5 micromolar inopa mwero wekutorwa kwechiedza uri pedyo ne0.41 uye 10 micromolar inopa anenge 0.82. Pakuyera kwakanangana nemonomera mumvura isina zvimwe zvakawedzerwa, kugara pedyo kana pasi peanenge 5 kusvika 7 micromolar kwakachengeteka kupfuura kugadzirisa zviyero kusvika pamakumi mazhinji e micromolar. Pamisanganiswa yekuchengetera ine uwandu hwakakwirira, pfupisa nzira yechiedza pane kubvuma mwero wekutorwa kwechiedza uri kure pamusoro pe1.

Gadzira chiyero chisina dhayi uchishandisa musanganiswa wezvimwe zvinhu wakafanana chaizvo: mvura yemhando imwe chete, chikamu chakafanana chechimwe chinonyungudutsa, buffer nemunyu zvakafanana, asi pasina dhayi. Gadzirira misanganiswa yekugadzirisa zviyero mumusanganiswa iwoyo wezvimwe zvinhu uye patembiricha imwe chete. Paunenge uchigadzira nzira, ongorora chikamu chose chechiedza chinoonekwa, zvakanakisisa kubva paanenge 550 kusvika 750 nm, pane kungoverenga urefu hwesaisai humwe chete. Tevera chimiro, kwete kureba chete. Kana kuderedza uwandu zvishoma nezvishoma nekuwedzera chinonyungudutsa kuchichinja chimiro chakaiswa pachiyero chimwe chete, mamiriro emakemikari emusanganiswa anotsamira pauwandu hwechinhu pamwero wemvura. Nhongonya inoramba panzvimbo imwe chete nereshiyo yakagadzikana ye610 kusvika 665 nm zvinopa uchapupu hwakasimba hwechiyero chinogamuchirika kupfuura R yakakwidzwa kusimba repiri iri pamusoro yoga.

Nyora chimiro chereajenti. Chinhu chinotengeswa chinogona kuva methylthioninium chloride isina mvura kana hydrate yakadai setrihydrate, saka huremu pamole hunoshandiswa kuverenga misanganiswa yekuchengetera hunofanira kuenderana nezviri mubhodhoro chairo. Danho iri roga rinotsanangura chimwe chikamu chekusiyana kwemwero yemolar inotaurwa mumarabhoritari akasiyana. Kuchenjerera kwakafanana kunodiwa kana mhedzisiro ichishandiswa kuverenga huwandu mumiedzo, kana maspekitiramu achitsigira kuongorora kugadzikana kwemusanganiswa wakagadzirwa, uye kana kuongorora mhando dzezvisina kuchena kuchida kuongororwa kwekuchena nezvinodiwa zvakatarwa.

Pfupiso ine mamiriro akanyorwa

Shandisa tafura iyi semuenzaniso wekutangira kunyora rekodhi yerabhoritari, kwete setsime remakonstanti anogona kungotsivana.

Rudzi rwemamorekuru neurefu hwesaisai	Mamiriro	Epsilon kana maitiro akataurwa
Monomera, 665 nm	Mvura, pasi peanenge 7 micromolar, 1 cm, pH haina kutaurwa	Anenge 82,000 M-1 cm-1, kutenuka kwemutsetse wedhayi yose kwakawanikwa nekuyera
Chikamu chemonomera, 664 nm	Mvura, pH 6.86, 293 K, chakapatsanurwa kubva kudimera nemasvomhu	Anenge 73,460 M-1 cm-1, chikamu chemonomera chete
Dhayi yose, 665 nm	Mvura yakabviswa maioni, 2.34 kusvika 37.52 micromolar, 1 cm	Anenge 67,100 M-1 cm-1, kutenuka kwemutsetse wekugadzirisa zviyero muchikamu chine kumwe kuungana kwemamorekuru
Rezonensi yemon om era, 664 nm	Mvura yakabviswa maioni isina buffer, nhevedzano ye1.1 micromolar kusvika 3.4 millimolar, NaCl 0 kusvika 0.15 M, 282 kusvika 333 K	Anenge 107,800 M-1 cm-1, chikamu chakapatsanurwa nemuenzaniso wemasvomhu
Dimera, 607 nm	Nhevedzano imwe chete neiri pamusoro	Anenge 101,400 M-1 cm-1, chikamu chakapatsanurwa nemuenzaniso wemasvomhu
Tetramera, 600 nm	Nhevedzano imwe chete neiri pamusoro	Anenge 138,500 M-1 cm-1, chikamu chakapatsanurwa nemuenzaniso wemasvomhu
Dhayi yose, 665 nm mumvura ichienzaniswa ne655 nm mumethanol	31.3 micromolar, tembiricha yemumba	Anenge 37,100 mumvura, anenge 60,800 mumethanol, nhamba dzinoratidzika pauwandu ihwohwo
Kuenzana pakati pemonomera nedimera	Misanganiswa yemvura neDMSO, 293.15 K	DMSO inoderedza dimera, nhongonya dzinoenda kuurefu hwesaisai hwakareba, goho rekuburitswa kwechiedza rinokwira

Saka urefu hwesaisai hwekutorwa kwechiedza nemethylene blue imhinduro pfupi ine tsananguro refu inofanira kuiperekedza. Taura nhongonya yawakayera, mumusanganiswa wawakayera, pauwandu hwechinhu pamwero wemvura hwawakayera. Tsika iyoyo inodzivirira kuti nhamba imwe yemusanganiswa wakareruka wemvura iri pedyo ne664 nm isashandurwa kuita konstanti yenhema inonzi inoshanda mumamiriro ose.