Kumenya ubururu bwa metilene muri laboratwari: sipekitirumu z'imirasire ya infraruje n'uburyo buzunganira

Tekereza laboratwari eshatu zakiriye ifu imwe y'ubururu. Iya mbere ifata sipekitirumu y'imirasire ya infraruje maze ikemeza ko ari ubururu bwa metilene. Iya kabiri ikoresha kromatografiya maze igatanga igisubizo cya 0.4% bya Azure B. Iya gatatu itatanya imirasire X ku kristalo maze ikavuga imiterere yabwo irimo amazi. Zose uko ari eshatu ziravuga ukuri, ariko nta n'imwe yasubije ibindi bibazo bibiri. Urujijo rutangira iyo igisubizo kimwe gifashwe nk'igisubiza ibyo bibazo uko ari bitatu.

Iyi ngingo itandukanya ibyo bibazo. Kumenya ikinyabutabire hakoreshejwe imirasire ya infraruje byemeza ubwoko bwa molekile habigereranyijwe n'icyitegererezo fatizo. Kromatografiya yemeza ibikigize. Itatanishwa ry'imirasire ryemeza imiterere yacyo mu buryo bukomeye. Guhuza ibisubizo mu kizamini cyo kumenya ikinyabutabire si icyemezo cy'ubuziranenge.

Kumenya ikinyabutabire hakoreshejwe imirasire ya infraruje bisubiza neza ikibazo kimwe

Ikizamini cyo kumenya ubururu bwa metilene giteganywa mu gitabo cyemewe cy'ibipimo by'imiti ni ukugereranya sipekitirumu n'iy'icyitegererezo fatizo, si urutonde rw'impinga zo kugenzura. Mu mbanzirizasoma y'inyandiko ya USP ku bururu bwa metilene, Identification A ivuga isesengura rya sipekitirumu z'imirasire ya infraruje hakoreshejwe ibikoresho by'urumuri bya ATR cyangwa akabumbe ka bromide ya potasiyumu. Sipekitirumu y'icyitegererezo igomba guhura n'iy'icyitegererezo fatizo cya USP, kandi byombi bigategurwa bikanapimwa mu bihe bingana. Nta rutonde rudahinduka rw'imibare y'imiraba iranga ikinyabutabire rugena umwanzuro. Kuwugena ni uguhuza sipekitirumu zombi.

Uko ikizamini cyateguwe bigaragaza uko sipekitirumu za FTIR z'ubururu bwa metilene zitwara. Ibimenyetso bikomeye kurusha ibindi biri mu gice kiranga umwihariko w'ikinyabutabire. Inyandiko za siyansi zihora zishyira umurongo w'ingenzi w'impeta zifite imiyoboro isimburana hafi ya 1590 kugeza kuri 1600 cm-1, aho uburyo bwo kurambuka kw'imiyoboro ya karubone-karubone na karubone-azote yo mu mpeta aromatike bukorana. Indi mirongo iri hafi ya 1490 na 1390 cm-1 igaragaza ihinduka ry'imiterere y'impeta n'imiyoboro ya karubone-azote na karubone-hidrojene. Imirongo iri hafi ya 1355 na 1330 cm-1 ijyanye n'imiyoboro ya karubone-azote yo mu matsinda ya dimetilamino n'ihinduka ry'imiterere y'amatsinda ya metile. Ibimenyetso biri hafi ya 1225 kugeza kuri 1250 cm-1 bijyanye n'urwego rw'impeta zirimo atome zitari karubone, naho imirongo iri hafi ya 1170 kugeza kuri 1130 na 1066 cm-1 ikagaragaza imiyoboro ya karubone-sufure, hamwe no kwihina kwa C-H yo mu mpeta aromatike hafi ya 880 cm-1. Uburyo bwo kurambuka bugaragara ku mibare y'imiraba iri hejuru: C-H yo mu mpeta aromatike hafi ya 3040 cm-1 na C-H ya N-metile hagati ya hafi 2900 na 3000 cm-1. Ubusobanuro bwatangajwe bw'iyo mirongo buratandukana mu makuru arambuye kubera ko urwego rwa phenothiazinium runyeganyega nk'urusobe rukorera hamwe; bityo, imbonerahamwe iyo ari yo yose ihuza imirongo n'imiyoboro uyifate nk'ubusobanuro bwatangajwe, aho kuyifata nk'ubusobanuro bwonyine bushoboka.

Ingaruka ebyiri z'imitegurire zikwiye kwitabwaho kuko zihindura sipekitirumu za IR z'ubururu bwa metilene zidahinduye irangi ubwaryo. Icya mbere, ikinyabutabire kirimo amazi mu miterere yacyo kigaragaza umurongo mugari wa O-H hafi ya 3300 kugeza kuri 3500 cm-1, hamwe no kwihina kwa molekile z'amazi hafi ya 1630 kugeza kuri 1640 cm-1; kandi imiterere itandukanye irimo amazi yimura imirongo yo mu gice kiranga umwihariko ku rugero rushobora gupimwa. Icya kabiri, ibikoresho bya KBr bikozwemo akabumbe bikurura ubuhehere: ubuhehere byanyoye butanga imirongo muri ibyo bice nyine by'amazi, bukaba bwakwitiranywa n'amazi ari mu miterere y'icyitegererezo. Gupima hakoreshejwe ATR birinda gukoresha KBr, ariko ubukana bw'ibimenyetso bya ATR buhinduka hakurikijwe ubujyakuzimu imirasire icengeramo, na bwo buterwa n'umubare w'imiraba. Bityo, sipekitirumu ya ATR n'iy'imirasire inyuzwa muri ya fu imwe ntizishobora guhuzwa mu buryo butaziguye. Itegeko ry'ingirakamaro rikomoka ku gisabwa n'igitabo cyemewe cy'ibipimo by'imiti: gereranya icyitegererezo n'icyitegererezo fatizo byapimwe hakoreshejwe uburyo bumwe, kandi wandike uburyo bwakoreshejwe. Ku bijyanye n'ubutabire bwa katoyoni, umunyu, n'imiterere irimo amazi bituma izi sipekitirumu zibaho, reba inyandiko yerekeye imiterere n'ibiranga ubururu bwa metilene.

Imbonerahamwe yo guhitamo uburyo bw'isesengura

Iyi mbonerahamwe ni igikoresho cyo guhitamo. Soma umurongo wose kugira ngo umenye ikibazo uburyo runaka busubiza, icyo igisubizo cyujuje ibisabwa cyemeza, n'ibitaracyemejwe.

Impamvu guhuza sipekitirumu z'imirasire ya infraruje atari icyemezo cy'ubuziranenge

Guhuza ibisubizo byo kumenya ikinyabutabire bikuraho ko cyaba ari irangi ritari ryo. Ntibitanga urutonde rw'ibiri mu cyitegererezo. Hari imbogamizi eshatu z'ingenzi.

Icya mbere, ibinyabutabire bifitanye isano ya hafi birasa iyo bipimwe n'imirasire ya infraruje. Azure B itandukaniye n'ubururu bwa metilene ku itsinda rimwe gusa rya metile, bityo sipekitirumu yayo ikaba isa cyane n'iy'ubururu bwa metilene. Imvange irimo Azure B ku rugero rw'ijanisha rito iracyahuza neza sipekitirumu yayo n'iy'icyitegererezo fatizo cy'ubururu bwa metilene. Gutahura uwo mwanda ni umurimo wa kromatografiya, usobanurwa mu mfashanyigisho ku myanda y'ubururu bwa metilene n'uburyo bwo kubupima.

Icya kabiri, isesengura rya sipekitirumu z'imirasire ya infraruje ripima icyitegererezo muri rusange kandi rifite ubushobozi buke bwo gutahura ibikigize biri ku rugero ruto. Ingano nto z'amarangi afitanye isano, ibisigazwa by'ibishongesha, cyangwa ibisigazwa by'ibinyabutabire bitari ibya karubone zihishwa n'imirongo ikomeye. Kutabona izindi mpinga ni gihamya idakomeye y'uko nta myanda irimo.

Icya gatatu, icyitegererezo fatizo ni cyo kigena igereranya ryose. Icyitegererezo fatizo cyangiritse, cyahinduriwe sipekitirumu n'amazi ari mu miterere yacyo, cyangwa cyafashwe nabi gihindura ibyo igereranya rishingiraho. Ni na ko bigenda iyo hahujwe uburyo budahuye, nko kugereranya sipekitirumu y'icyitegererezo yapimwe na ATR n'iy'icyitegererezo fatizo yapimwe hakoreshejwe KBr. Kuri buri mwanzuro, andika nimero y'icyiciro cy'icyitegererezo fatizo, uburyo bwakoreshejwe, n'uko cyateguwe.

Imvange zizana indi mburi ya nyuma. Sipekitirumu ihuye ahanini n'iy'icyitegererezo fatizo ariko ifite utubyimba ku mpande z'impinga cyangwa ubukana budasanzwe ntifatwa nk'iyujuje ibisabwa hiyongereyeho ibisobanuro byo ku ruhande. Ni ikimenyetso cy'uko ibigize icyitegererezo bigomba kubanza gutandukanywa mbere yo kubimenya.

Uko uburyo bwunganira bukoreshwa

HPLC y'ubururu bwa metilene itanga igisubizo ku bikigize imirasire ya infraruje idashobora gutanga. Uburyo buriho ubu mu gitabo cyemewe cy'ibipimo by'imiti bukoresha inkingi yo mu bwoko bwa phenyl, hamwe n'imvange y'amazi na acetonitrile ifite ibipimo bihinduka kandi irimo aside trifluoroacetic; gutahura bikorerwa kuri 246 nm, si mu gice cy'imirasire igaragara. Kumenya ikinyabutabire bishingira ku guhuza igihe kimara mu nkingi n'icy'icyitegererezo fatizo; iryo gereranya kandi ritanga igipimo cy'ingano yacyo. Ikizamini cy'imyanda y'ibinyabutabire bya karubone kimenya Azure B hakoreshejwe icyitegererezo fatizo cyayo, ku gihe cyo kugumana kigereranyijwe kiri hafi ya 0.8 ugereranyije n'ubururu bwa metilene kuri 1.0, kandi gisaba ko impinga zombi zitandukanywa neza cyane. Zirikana uburebure bw'umuraba bwatoranyijwe: ibinyabutabire byo mu ruhererekane rwo gukuraho amatsinda ya metile binywa imirasire ku burebure bw'umuraba bugenda bwegereza ibara ritukura, kuva kuri thionine hafi ya 600 nm, bikanyura kuri Azure C, Azure A, na Azure B, bikagera ku bururu bwa metilene hafi ya 670 nm. Bityo, igikoresho gitahura gishyizwe hafi ya 660 nm ntigisubiza kimwe kuri buri kinyabutabire cyo muri urwo ruhererekane, kandi kigaragaza ingano nto kurusha iy'ukuri kuri bimwe mu binyabutabire bifitanye isano. Iyo bikenewe gusesengura urwo ruhererekane rwose, hagomba gukoreshwa uburyo bwa C18 LC-MS/MS hamwe n'ibyitegererezo fatizo byagenzuwe ko bikwiriye kuri buri kinyabutabire gipimwa.

Itatanishwa ry'imirasire ku ifu risubiza ikibazo cy'imiterere mu buryo bukomeye, amakuru uburyo bwombi twavuze haruguru bwangiza cyangwa butitaho: gushongesha ifu kugira ngo ikorerwe kromatografiya bikuraho amakuru yerekeye kristalo, naho kunyeganyega kwa molekile kugaragaza imiyoboro aho kugaragaza uko molekile zitondetse ku ntera ndende. Ishusho y'itatanishwa ry'imirasire itandukanya ibyiciro by'imiterere irimo amazi n'urwego rwo kugira imiterere ya kristalo, kandi ubururu bwa metilene bukeneye iryo genzura kurusha ibindi binyabutabire byinshi. Ubushakashatsi bwa Rager na bagenzi be ku miterere mu buryo bukomeye bwagaragaje imiterere itanu itandukanye irimo amazi: imwe ifite molekile eshanu z'amazi, ebyiri zifite molekile ebyiri z'amazi, imwe ifite molekile imwe y'amazi, n'indi ifite ingano y'amazi iri hagati; kandi nta miterere nyayo ifite molekile eshatu z'amazi babonye mu yo basesenguye. Uko ifatwa muri laboratwari ni ingenzi hano: ubuhehere n'ubushyuhe bihindura ihamye ry'imiterere irimo amazi, kandi gusya cyane bishobora guhindura icyiciro cy'imiterere cyangwa kwangiza imiterere ya kristalo. Bityo, genzura kandi wandike ubuhehere bugereranyije n'ubushyuhe, kandi ugabanye gusya uko bishoboka. Ibirango by'imiterere irimo amazi n'ingaruka zayo ku buremere bupimwa bisobanurwa birambuye mu miterere y'ubururu bwa metilene irimo amazi n'ibirango byayo.

Kunywa imirasire ya UV-Vis bifite uruhare rwunganira. Ahantu hasanzwe hazwi ho kunywa imirasire ku rugero rwo hejuru hemeza imyitwarire yitezwe y'igisubizo kandi hagafasha gupima ingano iri muri cyo. Ariko igisubizo cy'ubururu kinywa imirasire ku burebure bw'umuraba bwitezwe kiba gishobora kuba ubururu bwa metilene, ntikibyemeza, kuko amarangi afitanye isano na cyo anywa imirasire ku burebure bw'umuraba buri hafi.

Ibyo kubaza mbere yo kugura

Hindura iri tandukaniro mo ibibazo byo kubaza igihe ugura. Baza icyiciro cyapimwe, uburyo bwakoreshejwe mu kumenya ikinyabutabire, niba ikizamini cy'imyanda cya HPLC cyarakoresheje icyitegererezo fatizo cya Azure B, n'iba imiterere irimo amazi yarasesenguwe cyangwa yarafashwe nk'ihari gusa. Ikiruta byose, baza niba icyemezo gikubiyemo ibisabwa byose cyangwa ibizamini byatoranyijwe gusa. Abacuruzi benshi bavuga ko ibicuruzwa byabo byujuje ibipimo by'igitabo cyemewe cy'imiti bapima ibyuma biremereye gusa, bigatuma ibibazo byo kumenya ikinyabutabire, ingano yacyo, imyanda y'ibinyabutabire bya karubone, ibishongesha, n'udukoko tutagaragara n'amaso bisigara bidasubijwe.

Blupreme ipima ibisabwa byose bya USP, birimo kumenya ikinyabutabire, ubuziranenge, imyanda y'ibinyabutabire bya karubone, ibisigazwa by'ibishongesha, imyanda y'ibintu shingiro by'ubutabire, ibisigazwa nyuma yo gutwika, ingano ntarengwa z'udukoko tutagaragara n'amaso, na endotoxine za bagiteri, ku bufatanye n'uruganda rukora ubururu bwa metilene bwo ku rwego rw'imiti. Icyemezo rusange cy'isesengura cya buri cyiciro gitangazwa ku rubuga rwa Blupreme rwo gupima ubuziranenge, kugira ngo ishingiro ry'ibipimo by'ingano n'ibisubizo by'imyanda bisomwe mu buryo butaziguye aho kubitekereza. Ibisubizo biri hafi ya 100% na byo bigomba kugaragaza ishingiro ry'imibare yabyo, nk'uko imfashanyigisho ku bipimo by'ingano birenze 100 ku ijana ibisobanura. Guhuza ibisubizo byo kumenya ikinyabutabire bitangiza igenzura. Inyandiko yuzuye y'ibisubizo ni yo irisoza.