Isipekitire y’iyinjizwa ry’urumuri n’ubushobozi bwo kurwinjiza kuri mole by’ubururu bwa metilene: amakuru ajyana n’imimerere yapimiwemo

Isipekitire y’ubururu bwa metilene isa n’iyoroshye kugeza igihe ugerageje kongera gukoresha umubare uyikomokaho. Laboratwari imwe itanga koefisiyenti yo kuzimya urumuri kuri mole iri hafi ya 82,000. Indi itanga 67,000. Iya gatatu itanga irenga 100,000. Iyo mibare uko ari itatu ishobora gusobanura neza ibipimo bya buri laboratwari, kandi nta n’umwe ari agaciro kadahinduka muri buri mimerere y’iryo rangi.

Urwo rujijo ruva ku kuvanga ibitekerezo bibiri bitandukanye. Igipimo cy’urumuri rwinjijwe gipimwa ku muti runaka uri mu gakoresho runaka ko gupimiramo. Ubushobozi bwo kwinjiza urumuri kuri mole, bukunze kwandikwa nka epsilon, ni igipimo gikoreshwa mu cyitegererezo cy’imibare gisobanura uwo muti. Iyo uburyo molekile z’ubururu bwa metilene zishyira hamwe buhindutse, icyo cyitegererezo na cyo kirahinduka, bityo icyo gipimo kikahinduka. Uburyo bwiza ni ugufata buri koefisiyenti nk’agaciro gaherekejwe n’ibisobanuro: uburebure bw’umuraba, umushongeshaji, pH cyangwa umuti utuma pH idahinduka, ubushyuhe, intera ya konsantrasiyo n’uburebure bw’inzira y’urumuri bigomba kujyana n’uwo mubare.

Agace ka monomeri mu mazi arimo irangi rike

Mu muti w’amazi arimo irangi rike, agace kiganje ka katiyoni y’ubururu bwa metilene kaba hafi ya 664 kugeza kuri 665 nm; bitewe n’ibikoresho n’uburyo bwo kubigenzura, gakunze kuvugwa ko kari hagati ya 664 na 668 nm. Ako gace ni ko gakwiye gukoreshwa mu kumenya ingano iyo irangi riri mu muti ari rike bihagije ku buryo igice kinini cyaryo kiri nka monomeri.

Indangagaciro eshatu zo mu nyandiko z’ubushakashatsi zerekana impamvu imimerere y’igipimo iruta umubare uvugwa wonyine. Abashakashatsi ku ibumba bakoresheje ingero z’amazi ziri munsi ya hafi 7 micromolar mu dukoresho two gupimiramo twa 1 cm batangaje epsilon ya hafi 82,000 M-1 cm-1 kuri 665 nm. Ako gaciro gafite ibisobanuro birambuye bidasanzwe byerekana ko ari ubuhaname bw’umurongo w’igenzura wabonetse mu igerageza ku ngero z’amazi arimo irangi rike cyane. Nyuma yaho, igenzura ry’ibanze mu mazi yakuwemo iyoni, ryakoresheje 2.34 kugeza kuri 37.52 micromolar n’inzira y’urumuri ya 1 cm, ryatanze hafi 67,100 M-1 cm-1 kuri 665 nm ku irangi ryose. Ako gaciro kari hasi ni ko kari kitezwe, si ukuvuguruzanya, kuko igice cyo hejuru cy’iyo ntera y’igenzura kiba kimaze kugira dimeri zitari nke. Agaciro ka gatatu, hafi 73,460 M-1 cm-1 kuri 664 nm, kavuye mu bushakashatsi kuri pH 6.86 na 293 K bwatandukanyije isipekitire yapimwe mo ibice bya monomeri na dimeri, nk’uko byavunaguye mu isesengura riherutse gusuzuma ubushakashatsi kuri fotofiziki y’ubururu bwa metilene. Uwo mubare ni agaciro k’igice cya monomeri cyatandukanyijwe n’ikindi hakoreshejwe imibare, si ubuhaname bwapimwe butaziguye hagati y’igipimo cy’urumuri rwinjijwe n’irangi ryose.

Icy’ingenzi mu bikorwa: hitamo koefisiyenti ifite imimerere ihuye n’ingero zawe, cyangwa ukore igenzura ryawe bwite. Ntukoreshe 82,000 mu isuzuma rya 30 micromolar rikoresha umuti wa fosifate utuma pH idahinduka ngo wibwire ko isano y’umurongo ugororotse ikomeza kubahirizwa.

Agatugu ku burebure bugufi bw’umuraba ni uruvange

Uko konsantrasiyo yiyongera, agatugu kari hafi ya 600 kugeza kuri 615 nm karakura ugereranyije n’impinga ya 664 nm. Ubushakashatsi bwa vuba bukunze gushyira uruhare rwa dimeri yo mu bwoko bwa H hafi ya 605 kugeza kuri 610 nm. Isesengura rusange ry’isipekitire z’ubururu bwa metilene mu mazi ritandukanya impinga ya dimeri kuri 607 nm n’impinga y’itsinda ririmo molekile nyinshi kuri 600 nm, hamwe n’ibice bya rezonansi bya monomeri kuri 664 na 650 nm. Inyandiko yaryo yose iboneka ku rupapuro rufunguye rwa PMID 33251415.

Ako gatugu ntigakwiye kwitwa igipimo cy’urumuri rwinjijwe na dimeri yonyine. Imiterere ya monomeri ijyanye no kunyeganyega kwa molekile n’inzego z’ingufu z’elekitironi na yo igira uruhare muri ako gace, bityo uburebure bw’agatugu bupimwe bukaba ari igiteranyo cy’ibyo bice. Mu guhuza icyitegererezo cy’imibare n’ibipimo byose, byagaragaye ko icyitegererezo cya monomeri na dimeri kitashoboye gusobanura intera yose ya 1.1 micromolar kugeza kuri 3.4 millimolar, mu gihe kongeramo tetrameri byatanze uguhuza kwiza kurusha ibindi, kandi ikimenyetso cya tetrameri cyari kimaze kugaragara no munsi ya 34 micromolar. Ubushobozi bw’ibice byatandukanyijwe muri iryo sesengura bwari bwinshi: hafi 107,800 M-1 cm-1 kuri 664 nm ku gice nyamukuru cya monomeri, hafi 101,400 kuri dimeri kuri 607 nm, na hafi 138,500 kuri tetrameri kuri 600 nm, mu mazi yakuwemo iyoni atarimo umuti utuma pH idahinduka, arimo sodium chloride kuva kuri 0 kugeza kuri 0.15 M, ku bushyuhe kuva kuri 282 kugeza kuri 333 K. Ibyo ni ibice byatandukanyijwe hakoreshejwe icyitegererezo cy’imibare, bityo ntibigomba gusimbura ubuhaname bwa Beer Lambert bwabonetse mu igerageza.

Ubukana bwo kwishyira hamwe kwa molekile zikabyara dimeri bwapimwe butaziguye. Kuri pH 6.86 na 293 K, ubushakashatsi bumwe bwabonye agaciro kadahinduka ko kurema dimeri kari hafi ya 3,900 M-1. Iyo ngano isobanura impamvu umuti wa 5 micromolar ukoreshwa mu kazi ugira imyitwarire hafi y’umurongo ugororotse, mu gihe umuti w’ibanze wa 50 micromolar utayigira. Ikigenzura cy’ingirakamaro mu gutegura uburyo bwo gupima ni ikigereranyo cy’igipimo cy’urumuri rwinjijwe hafi ya 610 nm ku gipimo cy’urumuri rwinjijwe hafi ya 665 nm. Iyo icyo kigereranyo cyiyongera hamwe na konsantrasiyo, epsilon kuri 664 nm iba itagisobanura ubwoko bumwe bwa molekile budahinduka.

Mu ntera ya konsantrasiyo yiganjemo ubwoko bubiri bwa molekile bushobora guhindukamo, isipekitire zishobora guhurira ku ngingo ya izosibesitiki, yabonetse mu igerageza hafi ya 625 nm mu ruhererekane rw’imiti y’amazi. Ingingo isobanutse idahindura aho iri ihuje no kuba hari ubwoko bubiri bwiganje. Iyo iyo ngingo yimuka cyangwa ikazimira uko konsantrasiyo yiyongera, bifate nk’ikimenyetso cy’uko haje amatsinda arimo molekile nyinshi cyangwa ko uburinganire bw’imimerere bwahindutse, aho kubifata nk’ikosa ry’igikoresho.

Umushongeshaji, pH n’ubushyuhe bihindura isipekitire

Amazi yorohereza molekile kwishyira hamwe. Kongeramo dimethyl sulfoxide bigabanya ikorwa rya dimeri, bikimura impinga z’iyinjizwa ry’urumuri n’iza fuloresansi ku burebure burebure bw’umuraba, kandi bikongera umusaruro wa kwantumu wa fuloresansi hafi incuro enye, nk’uko bivugwa mu bushakashatsi ku mishongeshaji y’ubururu bwa metilene mu mvange z’amazi na DMSO, bwanditswe nka PMID 30933781. Ubwo bushakashatsi bufata isipekitire zo mu mazi nk’izisobanurwa n’uburinganire hagati ya monomeri na dimeri, aho monomeri yinjiza urumuri hafi ya 650 kugeza kuri 670 nm, dimeri ikarwinjiza hafi ya 580 kugeza kuri 615 nm, kandi bukura indangagaciro zidahinduka zo kurema dimeri muri isipekitire zapimwe kuri 293.15 K.

Alukoro zihindura aho impinga iri n’ubushobozi bugaragara bwo kwinjiza urumuri, ariko ntizivanaho kwishyira hamwe kwa molekile. Mu igereranya ry’ibanze ryakorewe kuri 31.3 micromolar no ku bushyuhe bwo mu cyumba, ryatangajwe mu bushakashatsi ku mishongeshaji y’ubururu bwa metilene, impinga ku burebure burebure bw’umuraba yari kuri 665 nm mu mazi ariko kuri 655 nm muri methanol, kandi indangagaciro zigaragara ku muti wose zari hafi 37,100 mu mazi na 60,800 muri methanol. Irindi genzura ryakorewe muri ethanol itarimo amazi kuri 298 K ryatanze impinga kuri 655 nm, na epsilon ya hafi 68,000.

Impinduka zisanzwe za pH zimura isipekitire ku rugero ruto kurusha uko amabwiriza menshi y’ibikorwa abitekereza. Isesengura ryo gusuzuma ubushakashatsi ryavuzwe haruguru rivuga ko kuva hafi ya pH 2 kugeza kuri pH 12, impinga yagumye hafi ya 665 nm, ubushobozi bwo kwinjiza urumuri bugahinduka gato gusa kugeza kuri pH 11. Ubwoko bw’umuti utuma pH idahinduka n’imbaraga za iyoni bishobora guhindura ibisubizo ku rugero rungana n’urwa pH yanditswe, bityo andika umuti n’umunyu byakoreshejwe koko, aho kwandika gusa igipimo cyasomwe ku gipimisho cya pH. Aside ikomeye cyangwa alikali ikomeye ni ikibazo cyihariye kuko irangi rishobora guhinduka mu buryo bwa shimi aho kwishyira hamwe mu buryo bushobora gusubira inyuma. Ubushyuhe n’umunyu na byo bihindura uburinganire, ari yo mpamvu ubushakashatsi bwo mu mazi bwo mu 2020 bwahinduye byombi ku bushake.

Gupima ku buryo umubare ukomeza kuba ufite agaciro

Gupima ingano bishingira ku isano ya Beer Lambert: igipimo cy’urumuri rwinjijwe kingana na epsilon ikubwe na konsantrasiyo ikubwe n’uburebure bw’inzira y’urumuri, hamwe n’ibimenyetso by’uko intera ikurikiza umurongo ugororotse yubahirizwa koko mu mimerere ukoresha.

Koresha agikoresho ko gupimiramo ka 1 cm igihe ukora kuri konsantrasiyo ntoya zo mu rwego rwa micromolar. Iyo epsilon iri hafi ya 82,000, 5 micromolar itanga igipimo cy’urumuri rwinjijwe cya hafi 0.41, naho 10 micromolar igatanga hafi 0.82. Mu gupima monomeri mu mazi asanzwe, kuguma hafi cyangwa munsi ya hafi 5 kugeza kuri 7 micromolar ni byo byizewe kurusha gukora igenzura rigera cyane mu makumi ya micromolar. Ku miti y’ibanze ifite konsantrasiyo nyinshi, gabanya uburebure bw’inzira y’urumuri aho kwemera igipimo cy’urumuri rwinjijwe kirenze cyane 1.

Koresha icyitegererezo kitagira irangi gifite ibigize umuti bisa neza n’iby’urugero: amazi y’urwego rumwe, igice kingana cy’umushongeshaji wunganira, umuti umwe utuma pH idahinduka n’imyunyu imwe, ariko hatarimo irangi. Tegura imiti y’igenzura muri urwo ruvange rumwe no ku bushyuhe bumwe. Mu gutegura uburyo bwo gupima, genzura intera yose y’urumuri rugaragara, byaba byiza hafi ya 550 kugeza kuri 750 nm, aho gusoma uburebure bumwe gusa bw’umuraba. Kurikirana imiterere, atari uburebure bw’impinga gusa. Niba kugenda ugabanya konsantrasiyo mu ruhererekane bihindura imiterere ya isipekitire yashyizwe ku rugero rumwe, iyo sisitemu iterwa na konsantrasiyo mu buryo bwa shimi. Impinga iguma aho iri n’ikigereranyo cya 610 kuri 665 nm kidahinduka ni ibimenyetso bikomeye kurusha agaciro kanini ka R ku mpande ebyiri konyine mu kwemeza intera yemewe.

Andika imiterere y’ikinyabutabire ukoresha. Ibicuruzwa bishobora kuba methylthioninium chloride itarimo amazi cyangwa ubwoko burimo amazi, nka trihydrate, bityo uburemere kuri mole bukoreshwa mu kubara umuti w’ibanze bugomba guhura n’ibiri mu icupa koko. Iyi ntambwe yonyine isobanura igice cy’itandukaniro rya konsantrasiyo kuri mole zandikwa na laboratwari zitandukanye. Kwitonda nk’uko birakenewe iyo ibisubizo bikoreshwa mu mibare y’amasuzuma y’ingano, iyo isipekitire zifasha kugenzura ko imvange idahinduka, n’iyo imiterere y’imyanda isaba gusuzuma ubuziranenge n’ibisabwa ku gicuruzwa.

Incamake iherekejwe n’imimerere y’ibipimo

Koresha iyi mbonerahamwe nk’icyitegererezo cyo gutangiriraho wandika amakuru ya laboratwari, aho kuyifata nk’isoko y’indangagaciro zidahinduka zishobora gusimburana.

Bityo, uburebure bw’umuraba w’urumuri rwinjizwa n’ubururu bwa metilene ni igisubizo kigufi gisaba ibisobanuro birebire. Tanga impinga wapimye, mu muti wapimye, kuri konsantrasiyo wapimye. Uwo muco urinda agaciro kamwe ko hafi ya 664 nm kabonetse mu mazi arimo irangi rike guhinduka agaciro kadahinduka kavugwa ko gakora hose kandi atari byo.