Isihloko
Ukuhlonza i-methylene blue elabhorethri: amaspekhthramu e-infrared nezindlela ezihambisanayo
Cabanga ngamalabhorethri amathathu athola impushana efanayo eluhlaza okwesibhakabhaka. Eyokuqala iqopha ispekhthramu ye-infrared bese imemezela ukuthi iyi-methylene blue. Eyesibili yenza ikhromathografi bese ibika u-0.4% we-Azure B. Eyesithathu isebenzisa ukuphambuka kwemisebe ye-X kumakristalu bese isho uhlobo lwe-hydrate. Zontathu zilungile, kodwa ayikho ephendule eminye imibuzo emibili. Ukudideka kuqala lapho umphumela owodwa ufundwa sengathi uphendula yomithathu imibuzo.
Lesi sihloko sihlukanisa leyo mibuzo. Ukuhlonza nge-infrared kuqinisekisa ubunjalo bemolekhuli ngokuqhathanisa nesilinganiso esiyinkomba. Ikhromathografi iqinisekisa ukwakheka ngokwalokho okuqukethwe. Ukuphambuka kwemisebe kuqinisekisa uhlobo lwesimo esiqinile. Ukuphumelela ekuqhathaniseni ubunjalo akusona isitifiketi sobumsulwa.
Ukuhlonza nge-infrared kuphendula kahle umbuzo owodwa
Ukuhlolwa kobunjalo be-methylene blue okusencwadini esemthethweni yamazinga emithi kuwukuqhathanisa ispekhthramu nesilinganiso esiyinkomba, hhayi uhlu lweziqongo okufanele zibe khona. Ngaphansi kokubuka kuqala umbhalo wamazinga we-USP we-methylene blue, ukuhlolwa kokuhlonza A kubhekisela ekuhlaziyeni nge-infrared kusetshenziswa izinto zokubona ze-ATR noma ithebhulethi le-potassium bromide. Ispekhthramu yesampula kufanele ihambisane nespekhthramu yesilinganiso esiyinkomba se-USP, isampula nesilinganiso esiyinkomba kulungiswe futhi kuqoshwe ngaphansi kwezimo ezilinganayo. Alukho uhlu olungaguquki lwezinombolo zamagagasi zokuhlonza olunquma umphumela. Ukuqhathanisa amaspekhthramu ngokuwabeka phezu kwelinye yilokho okuwunqumayo.
Lokhu kukhethwa kwendlela kubonisa indlela amaspekhthramu e-FTIR e-methylene blue aziphatha ngayo. Izici eziqine kakhulu zisendaweni yespekhthramu efana nesigxivizo somunwe. Ukuhlotshaniswa kwamabhendi nokudlidliza okubikwe ezincwadini zesayensi kubeka ngokungaguquguquki ibhendi eyinhloko yesiyingi esinamabhondi ahlangene eduze kuka-1590 kuya ku-1600 cm-1, lapho izinhlobo zokweluleka kwamabhondi e-aromatic phakathi kwekhabhoni nekhabhoni kanye nekhabhoni nenayitrojeni zihlangana khona. Amanye amabhendi eduze kuka-1490 no-1390 cm-1 anezici zokuguquka komumo wesiyingi, zamabhondi ekhabhoni nenayitrojeni, nezamabhondi ekhabhoni nehayidrojeni. Amabhendi eduze kuka-1355 no-1330 cm-1 abandakanya amabhondi ekhabhoni nenayitrojeni e-dimethylamino kanye nokuguquka komumo wamaqembu e-methyl. Izici eziseduze kuka-1225 kuya ku-1250 cm-1 zihlobene nohlaka lwe-heterocyclic, kanti amabhendi eduze kuka-1170 kuya ku-1130 naku-1066 cm-1 anezici zamabhondi ekhabhoni nesibabule, nokugoba kwe-C-H ye-aromatic eduze kuka-880 cm-1. Izinhlobo zokweluleka zivela ezinombolweni zamagagasi eziphakeme: i-C-H ye-aromatic eduze kuka-3040 cm-1 kanye ne-C-H ye-N-methyl phakathi kuka-2900 no-3000 cm-1 cishe. Ukuhlotshaniswa okushicilelwe kuyahluka ngemininingwane ngoba uhlaka lwe-phenothiazinium ludlidliza njengohlelo olunezingxenye ezixhumene, ngakho phatha noma yiliphi ithebula elilodwa lokuhlobana njengokubikiwe, hhayi njengokuphela kwencazelo engenzeka.
Imiphumela emibili yokulungiswa kwesampula idinga ukunakwa ngoba ishintsha amaspekhthramu e-IR e-methylene blue ngaphandle kokushintsha udayi. Okokuqala, into enamanzi esakhiweni sayo ibonisa ibhendi ebanzi ye-O-H ezungeze u-3300 kuya ku-3500 cm-1 kanye nokugoba kwamanzi eduze kuka-1630 kuya ku-1640 cm-1, futhi izinhlobo ezihlukene ze-hydrate zisusa amabhendi endaweni yesigxivizo somunwe ngezinga elingalinganiswa. Okwesibili, isisekelo sethebhulethi le-KBr simunca umswakama: umswakama omunyiwe ukhiqiza amabhendi kulezo zindawo zamanzi ncamashi futhi ungabonakala sengathi ungamanzi e-hydrate avela kusampula. Ukuhlola nge-ATR kugwema isisekelo se-KBr, kodwa amandla amabhendi e-ATR ayashintsha ngokujula kokungena kwemisebe okuncike enombolweni yegagasi, ngakho ispekhthramu ye-ATR nespekhthramu yokudlula kwemisebe yempushana efanayo azikwazi ukubekwa ngokuqondile phezu kwelinye ukuze kuqhathaniswe. Umthetho osebenzayo ulandela imfuneko yencwadi esemthethweni yamazinga: qhathanisa isampula nesilinganiso esiyinkomba okuqoshwe ngendlela efanayo, futhi bhala ukuthi iyiphi esetshenzisiwe. Ukuze uthole ikhemistri ye-cation, usawoti, nama-hydrate okuyisisekelo sala maspekhthramu, bheka inkomba yesakhiwo nezakhiwo ze-methylene blue.
Ithebula lokukhetha indlela yokuhlaziya
Leli thebula liyithuluzi lokukhetha. Funda umugqa ovundlile ukuze uthole ukuthi indlela iphendula muphi umbuzo, ukuthi umphumela ophumelelayo uqinisekisa ini, nokuthi yini engakaqinisekiswa.
| Indlela | Umbuzo ewuphendulayo | Lokho okuqinisekiswa umphumela ophumelelayo | Lokho engakwazi ukukuqinisekisa |
|---|---|---|---|
| I-FTIR iqhathaniswa nesilinganiso esiyinkomba | Ingabe le nto iyi-methylene blue? | Ubunjalo bemolekhuli obuhambisana nespekhthramu esiyinkomba | Ubumsulwa, izinhlobo namanani okungcola, uhlobo lwe-hydrate ngokwayo, noma ukugxila |
| Ukuhlolwa kwenani nokungcola okuphilayo nge-HPLC-UV (ikholomu ye-phenyl, 246 nm) | Ingakanani i-methylene blue futhi ingakanani i-Azure B? | Ubunjalo ngekhromathografi ngokuhambisana kwesikhathi sokugcinwa; inani lento kanye namanani okungcola okubaluliwe | Uhlobo lwesimo esiqinile; ubunjalo beziqongo ezingalindelekile ngaphandle kwezilinganiso eziyinkomba |
| I-LC-MS/MS enezilinganiso eziyinkomba ezifanele eziqinisekisiwe | Yibaphi odayi abakhishwe amaqembu e-methyl abakhona? | Ukuhlukanisa nokuhlonza ochungechungeni lwe-MB, Azure A, Azure B, Azure C, ne-thionine | Ukuhambisana nencwadi esemthethweni yamazinga; lokhu kuyindlela eyengeziwe yezinga locwaningo, hhayi ukuhlolwa okuvamile kombhalo wamazinga |
| I-XRD yempushana | Iyiphi i-hydrate eyikristalu ekhona? | Isigaba sesimo esiqinile, izingxube zezigaba ezingaphezu komkhawulo wokutholwa, nezinga lokwakheka kwamakristalu | Ubunjalo bemolekhuli bokungcola okungenasakhiwo sekristalu; ukugxila kwesixazululo |
| Ukumuncwa kwemisebe ye-UV-Vis | Ingabe isixazululo simunca imisebe njengoba kulindelekile? | Izindawo zokumunca okuphezulu eziyizici zento nokuziphatha okuncike ekugxileni | Ubunjalo noma ubumsulwa ngokwayo |
| Ukuhlonza i-chloride nokulahleka kwesisindo lapho komiswa (okwesekelayo) | Ingabe i-ion ephikisayo ikhona; singakanani isisindo sezinto ezihwamukayo? | Uhlobo lukasawoti nesisindo esilahleka ekomisweni ngaphansi kwezimo ezichaziwe | Ukuthi iyiphi imolekhuli enikeza umbala |
Kungani ukufana kwe-infrared kungesona isitifiketi sobumsulwa
Ukufana kobunjalo kukhipha amathuba okuthi kube udayi ongafanele. Akubaluli konke okusesampuleni. Imikhawulo emithathu ibaluleke kakhulu.
Okokuqala, odayi abahlobene eduze babukeka befana ku-infrared. I-Azure B ihluka ku-methylene blue ngeqembu elilodwa kuphela le-methyl, ngakho ispekhthramu yayo ifana kakhulu neye-methylene blue. Ingxube equkethe amaphesenti ambalwa e-Azure B isahambisana kahle nespekhthramu esiyinkomba ye-methylene blue lapho ebekwa phezu kwelinye. Ukuthola lokho kungcola kuwumsebenzi wekhromathografi, ochazwe kumhlahlandlela wokungcola kwe-methylene blue nezindlela zokuhlola.
Okwesibili, ukuhlaziya nge-infrared kuyindlela ehlola isampula ngobuningi bayo futhi enekhono elilinganiselwe lokuthola izingxenye ezikhona ngamanani amancane. Amanani amancane odayi abahlobene, izincibilikisi ezisele, noma izinsalela ezingaphili afihleka ngaphansi kwamabhendi amakhulu. Ukungabikho kweziqongo ezengeziwe kuwubufakazi obubuthakathaka bokungabikho kokungcola.
Okwesithathu, isilinganiso esiyinkomba silawula konke ukuqhathanisa. Isilinganiso esiyinkomba esonakele, esinamaspekhthramu ashintshile ngenxa yamanzi esakhiweni, noma esiphathwe kabi sishintsha isisekelo sokwahlulela. Kwenza okufanayo nokuhlanganisa izindlela ezingahambisani, njengokwahlulela ispekhthramu yesampula ye-ATR ngokuyiqhathanisa nesilinganiso esiyinkomba se-KBr. Qopha iqoqo lesilinganiso esiyinkomba, indlela, kanye nokulungiswa kwesampula kanye nesinqumo ngasinye.
Izingxube zengeza isixwayiso sokugcina. Ispekhthramu efana kakhulu nesiyinkomba kodwa enamahlombe emabhendini noma amandla angajwayelekile ayiwona umphumela ophumelelayo odinga nje amanothi angezansi. Iwuphawu lokuthi isampula kufanele ihlukaniswe ngaphambi kokuyihlonza.
Izindlela ezihambisanayo ekusebenzeni
I-HPLC ye-methylene blue inikeza impendulo ngokwakheka ngokwalokho okuqukethwe, okuyinto i-infrared engakwazi ukuyenza. Indlela yamanje yencwadi esemthethweni yamazinga isebenzisa ikholomu yohlobo lwe-phenyl enengxube yamanzi ne-acetonitrile eshintsha ngokuhamba kwesikhathi, equkethe i-trifluoroacetic acid, futhi ithola izinto ku-246 nm, hhayi ebhendini yokukhanya okubonakalayo. Ubunjalo buvela ekuhambisaneni kwesikhathi sokugcinwa nesilinganiso esiyinkomba; ukuqhathanisa okufanayo kunikeza nesilinganiso senani. Ukuhlolwa kokungcola okuphilayo kuhlonza i-Azure B kusetshenziswa into yayo eyinkomba, ngesikhathi sokugcinwa esiqhathanisiwe esiseduze kuka-0.8 uma i-methylene blue iku-1.0, futhi kudinga ukuhlukaniseka okuphezulu phakathi kweziqongo ezimbili. Qaphela ukukhethwa kobude begagasi: uchungechunge lokususwa kwamaqembu e-methyl lumunca imisebe ngokuya ngokuya ngasohlangothini olubomvu, kusukela ku-thionine eduze kuka-600 nm kudlule ku-Azure C, Azure A, ne-Azure B kuze kufike ku-methylene blue eduze kuka-670 nm. Ngakho umtshina omiswe eduze kuka-660 nm uphendula ngokungalingani kulo lonke uchungechunge futhi ubika amanani aphansi kunawoqobo kwezinye izinhlobo ezihlobene. Ukuhlola ngokugcwele lonke uchungechunge, lapho kudingeka, kufanele kwenziwe ngendlela ye-C18 LC-MS/MS enezilinganiso eziyinkomba ezifanele eziqinisekisiwe zento ngayinye ehlaziywayo.
Ukuphambuka kwemisebe empushaneni kuphendula umbuzo wesimo esiqinile lezi zindlela ezimbili ezingenhla eziwuqeda noma eziwuziba: ukuncibilikisa impushana ukuze kwenziwe ikhromathografi kususa imininingwane yamakristalu, futhi ukudlidliza kwamamolekhuli kubika ngamabhondi esikhundleni sokuhleleka kwawo ezindaweni ezinde. Amaphethini okuphambuka kwemisebe ahlukanisa izigaba ze-hydrate nezinga lokwakheka kwamakristalu, futhi i-methylene blue idinga lokho kuhlolwa ngokucophelela ngaphezu kwezinto eziningi. Umsebenzi wezimo eziqinile ka-Rager nozakwabo wahlonza izakhiwo ezinhlanu ezihlukene ze-hydrate, kuhlanganise ne-pentahydrate, ama-dihydrate amabili, i-monohydrate, ne-hydrate ephakathi, futhi awutholanga i-trihydrate yangempela phakathi kwezinhlobo ezachazwa. Ukuphathwa elabhorethri kubalulekile lapha: umswakama nezinga lokushisa kushintsha ukuzinza kwe-hydrate, futhi ukugaya ngamandla kungashintsha isigaba noma kunciphise izinga lokwakheka kwamakristalu, ngakho lawula futhi uqophe umswakama ohlobene nezinga lokushisa, bese ugcina ukugaya kukuncane ngangokunokwenzeka. Ukubhalwa kohlobo lwe-hydrate kumalebula nomthelela wako esisindweni esikaliwe kuchazwe kabanzi ezinhlotsheni ze-hydrate ze-methylene blue nokubhalwa kwazo kumalebula.
Ukumuncwa kwemisebe ye-UV-Vis kudlala indima yokwesekela. Izindawo ezaziwayo zokumunca okuphezulu ziqinisekisa ukuziphatha okulindelekile kwesixazululo futhi zivumela umsebenzi wokulinganisa ukugxila, kodwa isixazululo esiluhlaza okwesibhakabhaka esimunca imisebe ebudeni begagasi obulindelekile siyahambisana ne-methylene blue ngaphandle kokukufakazela, ngoba odayi abahlobene bamunca imisebe ebudeni bamagagasi obuseduze.
Okufanele ukubuze ngaphambi kokuthenga
Guqula lokhu kwehluka kube imibuzo yokuthenga. Buza ukuthi yiliphi iqoqo elahlolwa, iyiphi indlela eyaqinisekisa ubunjalo, ukuthi ukuhlolwa kokungcola nge-HPLC kwasebenzisa yini into eyinkomba ye-Azure B, nokuthi uhlobo lwe-hydrate lwahlolwa lwachazwa noma lwathathwa nje njengolwazi oluqinisekile. Ngaphezu kwakho konke, buza ukuthi isitifiketi sihlanganisa zonke izimfuneko noma ukuhlolwa okukhethiwe kuphela. Abathengisi abaningi abathi bahambisana nencwadi esemthethweni yamazinga bahlola izinsimbi ezisindayo kuphela, okushiya imibuzo yobunjalo, yenani lento, yokungcola okuphilayo, yezincibilikisi, neyam amagciwane ingaphenduliwe.
Blupreme ihlola zonke izimfuneko ze-USP, ezihlanganisa ubunjalo, ubumsulwa, ukungcola okuphilayo, izincibilikisi ezisele, ukungcola kwezakhi zamakhemikhali, izinsalela ngemva kokushiswa, imikhawulo yamagciwane, nama-endotoxin amagciwane, ngokubambisana nomkhiqizi we-methylene blue yezinga lemithi. Isitifiketi sokuhlaziya somphakathi seqoqo ngalinye sishicilelwa ekuhlolweni kwekhwalithi kwe-Blupreme, ukuze isisekelo sokulinganisa inani nemiphumela yokungcola kufundwe ngokuqondile esikhundleni sokucatshangelwa. Imiphumela eseduze kuka-100% isadinga ukuthi isisekelo sokubala kwayo sishiwo, njengoba umhlahlandlela wesilinganiso senani esingaphezu kwamaphesenti angu-100 uchaza. Ukuqhathanisa ubunjalo kuvula uphenyo. Irekhodi eliphelele liyaluvala.