Te tūāwhiorangi ngongo me te kaha ngongo ā-mora o te puru metilīni: he tohutoro whai tapanga āhuatanga

He māmā te āhua o te tūāwhiorangi puru metilīni, kia whakamātau rā anō koe ki te whakamahi anō i tētahi tau nō reira. Ka pūrongo tētahi taiwhanga i te whakarea tinei ā-mora tata ki te 82,000. Ka pūrongo tētahi atu i te 67,000. Ka pūrongo te tuatoru i tētahi uara nui ake i te 100,000. Ka tika pea ēnei e toru hei whakaahuatanga mō ā rātou ake inenga, engari kāore tētahi o ēnei i te pūmau whānui mō taua waikano.

Ko te pūtake o te rangirua ko te whakaranu i ngā ariā motuhake e rua. Ko te ngongotanga he inenga ka mahia ki tētahi wairewa motuhake i roto i tētahi pūtau motuhake. Ko te kaha ngongo ā-mora, e tuhia nuitia ana hei epsilon, he tawhā i roto i tētahi tauira o taua wairewa. Ina rerekē te āhua honohono o te puru metilīni, ka rerekē te tauira, nō reira ka rerekē hoki te tawhā. Ko te tikanga whai hua, me titiro ki ia whakarea hei uara whai tapanga: me haere tahi te roangaru, te whakarewa, te pH, te pūreirei rānei, te pāmahana, te awhe kukū, me te roa ara me taua tau.

Te pae waetahi i te wairewa wai waimeha

I te wairewa wai waimeha, kei te takiwā o te 664 ki te 665 nm te pae matua o te katote tōrunga puru metilīni. Ka whakahuatia nuitia te awhe 664 ki te 668 nm, e ai ki ngā taputapu me ngā kōwhiringa whakatika ine. Ko taua pae te kōwhiringa tika mō te whakatau rahinga ina tino waimeha te wairewa, kia noho te nuinga o te waikano hei waetahi.

E toru ngā uara i ngā tuhinga rangahau e whakaatu ana he nui ake te hiranga o ngā āhuatanga i tō te tau matua. Ko ngā kairangahau uku i pupuri i ā rātou tauira wai ki raro iho i te takiwā o te 7 micromolar i roto i ngā pūtau 1 cm, i pūrongo i te epsilon tata ki te 82,000 M-1 cm-1 i te 665 nm. He tino pai ngā tuhinga taunaki mō taua uara hei rōnaki whakatika ine i takea mai i ngā whakamātautau mō ngā tauira wai tino waimeha. Nā tētahi whakatika ine tuatahi i te wai kua tangohia ōna katote i muri mai, i whakamahi i te 2.34 ki te 37.52 micromolar me te ara 1 cm, i puta ai te uara tata ki te 67,100 M-1 cm-1 i te 665 nm mō te waikano katoa. E tūmanakohia ana taua hua iti iho; ehara i te whakahē, nā te mea he nui kē te waerua i te haurua runga o taua awhe whakatika ine. Ko te uara tuatoru, tata ki te 73,460 M-1 cm-1 i te 664 nm, nō ngā mahi i te pH 6.86 me te 293 K i wehewehe i te tūāwhiorangi i kitea ki ngā wāhanga waetahi me ngā wāhanga waerua, e whakarāpopototia ana i tētahi arotake arohaehae nō nā tata nei mō te ahupūngao māramatanga o te puru metilīni. Ko taua tau he wāhanga waetahi kua wehea mā te tātari, ehara i te rōnaki tōtika o te ngongotanga ki te waikano katoa.

Ko te aronga mō te mahi: kōwhiria te whakarea e hāngai ana ōna āhuatanga ki ō tauira, whakatikaia rānei tāu ake ine. Kaua e whakamahi i te 82,000 mō tētahi whakamātautau i roto i te pūreirei pākawa 30 micromolar, me te whakapae ka mau tonu te hononga rārangi.

He ranunga te pokohiwi roangaru poto

Ka piki te kukū, ka tipu tētahi pokohiwi tata ki te 600 ki te 615 nm, ina whakatauritea ki te tihi 664 nm. I ngā mahi o ēnei rā, ka whakanohoia te takoha o te waerua momo H ki te takiwā o te 605 ki te 610 nm. Ka wehewehe te tātari tūāwhiorangi whānui o te puru metilīni i te wai i tētahi mōrahi waerua i te 607 nm me tētahi mōrahi o te kāhui nui ake i te 600 nm, me ngā wāhanga hāngai-oro waetahi i te 664 me te 650 nm. E wātea ana te tuhinga katoa mā te pūkete uru tuwhera mō PMID 33251415.

Kaua taua pokohiwi e tapaina hei ngongotanga waerua anake. Ka whai wāhi hoki te hanganga wiri-hiko o te waetahi i taua takiwā, nō reira he whakakikorua te teitei e inea ana. I kitea e tētahi whakaōritenga whānui kāore tētahi tauira waetahi me te waerua i hāngai puta noa i te awhe 1.1 micromolar ki te 3.4 millimolar. Engari, ko te tāpiri i te waewhā i whakaputa i te whakaōritenga pai rawa, ā, i taea kē te kite i te tohu waewhā i raro iho i te 34 micromolar. He nui ngā kaha o ngā wāhanga i whakaōritea: tata ki te 107,800 M-1 cm-1 i te 664 nm mō te wāhanga waetahi matua, tata ki te 101,400 mō te waerua i te 607 nm, me te tata ki te 138,500 mō te waewhā i te 600 nm, i roto i te wai kua tangohia ōna katote, kāore he pūreirei, me te konutai pūhaumāota mai i te 0 ki te 0.15 M me ngā pāmahana mai i te 282 ki te 333 K. He wāhanga ēnei i wehea mā te tauira, nō reira kaua e whakamahia hei whakakapi mō tētahi rōnaki Beer Lambert i takea mai i ngā whakamātautau.

Kua whakatauria tōtikatia te kaha o te hanga waerua. I te pH 6.86 me te 293 K, i whiwhi tētahi rangahau i te pūmau hanga waerua tata ki te 3,900 M-1. Nā taua rahinga e whakamārama ana he aha te wairewa mahi 5 micromolar ka whai whanonga tata ki te rārangi, engari kāore te wairewa pūtake 50 micromolar e pērā. He arowhai whai hua i te wā e whakawhanakehia ana te tikanga, ko te ōwehenga o te ngongotanga tata ki te 610 nm ki te ngongotanga tata ki te 665 nm. Ina piki taua ōwehenga i te pikinga o te kukū, kua kore te epsilon i te 664 nm e whakaahua i tētahi momo matū kotahi e noho pūmau ana.

I roto i tētahi awhe kukū e noho matua ai tētahi takirua e huri ana tētahi ki tētahi, ka whai wāhi ngā tūāwhiorangi ki tētahi ira ōrite-ngongo; i ngā whakamātautau raupapa wairewa wai, kei te takiwā o te 625 nm tēnei. Ko tētahi ira pūmau, mārama hoki, e hāngai ana ki te noho matua o ngā momo matū e rua. Ina neke, ina ngaro rānei taua ira i te pikinga o te kukū, tirohia hei whakatūpato kua puta mai ngā kāhui nui ake, ngā taurite e neke ana rānei, kaua hei hapa taputapu.

Ka nekehia te tūāwhiorangi e te whakarewa, te pH, me te pāmahana

Ka hāpai te wai i te honohono. Mā te tāpiri i te dimethyl sulfoxide e pēhi te hanga waerua, e nuku ngā mōrahi ngongo me ngā mōrahi kōrekoreko ki ngā roangaru roa ake, ā, e whakapiki te putanga ā-kuanta kōrekoreko kia tata ki te whā whakareatanga, e ai ki te rangahau whakarewa mō te puru metilīni i te wai me ngā ranunga DMSO, kua whakarārangitia hei PMID 30933781. I taua mahi, ka tirohia ngā tūāwhiorangi wairewa wai hei taurite waetahi-waerua, me te ngongotanga waetahi tata ki te 650 ki te 670 nm me te ngongotanga waerua tata ki te 580 ki te 615 nm. Ka tātai hoki taua mahi i ngā pūmau hanga waerua mai i ngā tūāwhiorangi i te 293.15 K.

Ka nekehia e ngā waipiro te tūnga me te kaha e kitea ana, engari kāore e whakakorehia te whakahuihui. I tētahi whakataurite tuatahi i te 31.3 micromolar me te pāmahana rūma, i pūrongotia i tētahi rangahau whakarewa puru metilīni, ko te mōrahi roangaru roa ko te 665 nm i te wai, engari ko te 655 nm i te methanol. Ko ngā uara e kitea ana mō te wairewa katoa, kei te takiwā o te 37,100 i te wai me te 60,800 i te methanol. Nā tētahi whakamana motuhake i te ethanol kore-wai i te 298 K i puta ai te mōrahi i te 655 nm me te epsilon tata ki te 68,000.

He iti ake te neke o te tūāwhiorangi i ngā rerekētanga pH noa i tā te maha o ngā kawa e whakapae ana. E ai ki te arotake arohaehae i whakahuatia i runga ake nei, mai i te takiwā o te pH 2 ki te pH 12, i noho te mōrahi tata ki te 665 nm, ā, he iti noa ngā rerekētanga kaha tae noa ki te pH 11. Ka taea e te momo pūreirei me te kaha katote te whakarerekē i ngā hua kia rite ki tō te pH ingoa noa, nō reira tuhia te pūreirei me te tote i whakamahia, kaua ko te pānui mita pH anake. He take motuhake te waikawa kaha, te kawakore kaha rānei, nā te mea ka pā pea he panoni matū ki te waikano, kaua ko te honohono ka taea te whakahoki. Ka nekehia hoki te taurite e te pāmahana me te tote; koia te take i āta whakarerekētia ai ēnei e rua i te rangahau wai o 2020.

Te ine kia mau tonu ai te mana o te tau

Ko te tūāpapa o te mahi whakatau rahinga ko te hononga Beer Lambert: ka ōrite te ngongotanga ki te epsilon whakareatia ki te kukū, whakareatia ki te roa ara. Me whai taunakitanga hoki ka mau tonu te awhe rārangi i raro i ō āhuatanga.

Whakamahia he pūtau 1 cm mō ngā mahi kukū micromolar iti. Ina tata te epsilon ki te 82,000, ka puta te ngongotanga tata ki te 0.41 i te 5 micromolar, ā, ka puta te tata ki te 0.82 i te 10 micromolar. Mō te ine e aro ana ki te waetahi i te wai noa, he haumaru ake te noho tata ki te 5 ki te 7 micromolar, ki raro iho rānei, i te whakatika ine ki roto rawa i ngā tekau micromolar. Mō ngā wairewa pūtake kukū, whakapotohia te ara, kaua e whakaae ki te ngongotanga kei runga noa atu i te 1.

Whakatakotoria te ine pātea mā te huinga wairewa ōrite pū: kia ōrite te kōeke wai, te hautanga o te whakarewa hoa, te pūreirei me ngā tote, engari kia kore te waikano. Whakaritea ngā wairewa whakatika ine i taua huinga wairewa anō, i taua pāmahana anō. Matawaia te awhe katoa o te māramatanga kitea i te wā whakawhanake, ko te mea pai ko te takiwā o te 550 ki te 750 nm, kaua e pānui i te roangaru kotahi anake. Aroturukihia te āhua, kaua ko te teitei anake. Ki te panoni te āhua kua whakatauritea e te waimehatanga ā-raupapa, ka whakawhirinaki te pūnaha ki te kukū i te taha matū. He taunakitanga kaha ake te mōrahi pūmau me te ōwehenga pūmau o te 610 ki te 665 nm mō tētahi awhe e tika ana, i tētahi R pūrua teitei anake.

Tuhia te āhua o te tauhohe matū. Ko te matū e hokona ana he methylthioninium chloride kore-wai pea, he matū whai wai rānei pērā i te trihydrate. Nō reira, me hāngai te papatipu ā-mora e whakamahia ana mō ngā tātai wairewa pūtake ki te mea kei roto tonu i te pounamu. Mā tēnei mahi anake e whakamārama tētahi wāhanga o ngā rerekētanga o ngā kukū ā-mora ingoa noa i waenga i ngā taiwhanga. Me pēnei anō te āta mahi ina whakamahia ngā hua i roto i ngā tātai whakamātautau whakatau rahinga, ina tautoko ngā tūāwhiorangi i ngā arowhai pūmautanga o te hanganga ranunga, ā, ina hiahiatia e ngā kōtaha poke he arotake i te parakore me ngā whakaritenga.

He whakarāpopototanga whai tapanga āhuatanga

Whakamahia tēnei ripanga hei tauira tīmatanga mō tētahi pūkete taiwhanga, kaua hei puna pūmau ka taea te whakawhitiwhiti.

Nō reira, he whakautu poto te roangaru ngongotanga o te puru metilīni, engari he kupu āpiti roa tōna. Pūrongotia te tihi i inea e koe, i roto i te wairewa i inea e koe, i te kukū i inea e koe. Mā taua tikanga e aukati te huri o tētahi uara wairewa wai waimeha tata ki te 664 nm hei pūmau whānui hē.