Kuzindikira methylene blue mu labotale: masipekitiramu a infrared ndi njira zina zothandizira

Tiyerekeze kuti malabotale atatu alandira ufa womwewo wabuluu. Yoyamba imalemba sipekitiramu ya infrared n’kunena kuti ufawo ndi methylene blue. Yachiwiri imachita chromatography n’kunena kuti muli 0.4% Azure B. Yachitatu imapatutsa ma X-ray pogwiritsa ntchito makhiristo n’kutchula mawonekedwe ake okhala madzi. Zonse zitatu ndi zolondola, koma palibe imene yayankha mafunso ena awiriwo. Chisokonezo chimayamba pamene chotsatira chimodzi chimaonedwa ngati chikuyankha zonse zitatu.

Nkhaniyi imasiyanitsa mafunso amenewo. Kuzindikira ndi infrared kumatsimikizira mtundu wa molekyulu poyerekeza ndi muyezo wolozera. Chromatography imatsimikizira zinthu zimene zili mu chitsanzocho ndi kuchuluka kwake. Kupatuka kwa kuwala kumatsimikizira mawonekedwe a chinthu cholimba. Kupambana pa kuyerekezera kozindikira chinthu si satifiketi ya kuyera kwake.

Kuzindikira ndi infrared kumayankha bwino funso limodzi

Mayeso ozindikira methylene blue a m’buku lovomerezeka la miyezo ya mankhwala amayerekezera sipekitiramu ndi muyezo, osati kungoyang’ana mndandanda wa nsonga. Mu chithunzithunzi cha monograph ya USP ya methylene blue, Identification A imatchula infrared spectroscopy pogwiritsa ntchito zipangizo za ATR kapena piritsi lopanikizidwa la potassium bromide. Sipekitiramu ya chitsanzo iyenera kufanana ndi sipekitiramu ya muyezo wolozera wa USP, ndipo chitsanzo ndi muyezowo ziyenera kukonzedwa ndi kuyezedwa m’mikhalidwe yofanana. Palibe mndandanda wokhazikika wa manambala a mafunde ozindikirira umene umapereka chigamulo. Kuyika masipekitiramu pamwamba pa wina ndi mnzake n’kumene kumachipereka.

Kusankha njira imeneyi kumasonyeza mmene masipekitiramu a FTIR a methylene blue amakhalira. Zizindikiro zamphamvu kwambiri zimakhala m’chigawo cha zizindikiro zapadera zozindikirira chinthucho. Mabuku ofufuza nthawi zambiri amaika chizindikiro chachikulu cha mphete zokhala ndi maubale osinthana pafupi ndi 1590 mpaka 1600 cm-1, kumene kutambasuka kwa maubale a carbon-carbon ndi carbon-nitrogen a mphete za aromatic kumachitika mogwirizana. Zizindikiro zina pafupi ndi 1490 ndi 1390 cm-1 zimakhudzana ndi kusintha kwa mawonekedwe a mphete, carbon-nitrogen, ndi carbon-hydrogen. Zizindikiro pafupi ndi 1355 ndi 1330 cm-1 zimakhudza maubale a carbon-nitrogen a dimethylamino ndi kusintha kwa mawonekedwe a methyl. Zizindikiro pafupi ndi 1225 mpaka 1250 cm-1 zimachokera ku chimango cha mphete zokhala ndi maatomu osiyanasiyana, pamene zizindikiro pafupi ndi 1170 mpaka 1130 ndi 1066 cm-1 zimakhudzana ndi carbon-sulfur, ndipo kupindika kwa aromatic C-H kumapezeka pafupi ndi 880 cm-1. Zizindikiro za kutambasuka zimapezeka pamwamba kwambiri: aromatic C-H pafupi ndi 3040 cm-1 ndi N-methyl C-H pakati pa pafupifupi 2900 ndi 3000 cm-1. Mafotokozedwe ofalitsidwa amasiyana pa mfundo zina chifukwa chimango cha phenothiazinium chimanjenjemera ngati dongosolo limodzi logwirizana, choncho onani tebulo lililonse lofotokoza zizindikiro ngati zimene ofufuzawo ananena, osati ngati kufotokoza kokha kotheka.

Zotsatira ziwiri za kukonzekera chitsanzo ziyenera kuyang’aniridwa chifukwa zimasintha masipekitiramu a IR a methylene blue popanda kusintha utotowo. Choyamba, chinthu chokhala ndi madzi mu makhiristo chimasonyeza chizindikiro chotambalala cha O-H pafupifupi 3300 mpaka 3500 cm-1 komanso kupindika kwa madzi pafupi ndi 1630 mpaka 1640 cm-1, ndipo mawonekedwe osiyanasiyana okhala madzi amasuntha zizindikiro zapadera ndi kusiyana komwe kungayesedwe. Chachiwiri, zinthu za m’piritsi la KBr zimakoka chinyezi: chinyezi cholowamo chimapanga zizindikiro m’zigawo zomwezo za madzi ndipo chingawoneke ngati madzi a mu makhiristo a chitsanzo. Kuyeza ndi ATR kumapewa kugwiritsa ntchito KBr, koma mphamvu za zizindikiro za ATR zimasintha malinga ndi kuya komwe kuwala kumalowa, komwe kumadalira nambala ya mafunde. Choncho sipekitiramu ya ATR ndi sipekitiramu ya kuwala kodutsa mu ufa womwewo sizingayikidwe mwachindunji pamwamba pa wina ndi mnzake kuti ziyerekezedwe. Lamulo logwira ntchito limatsatira zofunika za buku la miyezo: yerekezerani chitsanzo ndi muyezo wolozera zoyezedwa ndi njira yomweyo, ndipo lembani njira imene inagwiritsidwa ntchito. Kuti mudziwe umagwirira wa ayoni wokhala ndi chaji yowonjezera, mchere, ndi mawonekedwe okhala madzi amene amayambitsa masipekitiramuwa, onani zofotokozera kapangidwe ndi katundu wa methylene blue.

Tebulo losankhira njira yowunikira

Tebuloli ndi chida chosankhira njira. Werengani mzere uliwonse kuti mudziwe funso limene njirayo imayankha, zimene zotsatira zopambana zimatsimikizira, ndi zimene zimakhalabe zosatsimikizika.

Chifukwa chake kufanana kwa infrared si satifiketi ya kuyera

Kufanana kozindikirira kumachotsa kuthekera kwakuti ndi utoto wolakwika. Sikulemba zonse zimene zili mu chitsanzocho. Malire atatu ndi ofunika kwambiri.

Choyamba, zinthu zofanana kwambiri zimaoneka zofanana mu infrared. Azure B imasiyana ndi methylene blue ndi gulu limodzi lokha la methyl, choncho sipekitiramu yake imafanana kwambiri ndi ya methylene blue. Chosakaniza chokhala ndi Azure B ya maperesenti ochepa chimafananabe bwino chikayikidwa pamwamba pa sipekitiramu yolozera ya methylene blue. Kuzindikira chonyansa chimenecho ndi ntchito ya chromatography, yofotokozedwa mu malangizo a zonyansa za methylene blue ndi njira zoyesera.

Chachiwiri, infrared spectroscopy imayesa chitsanzo chonse pamodzi ndipo imakhala ndi mphamvu zochepa zozindikira zinthu zomwe zilimo pang’ono. Utoto wofanana nawo, zosungunulira zotsala, kapena zotsalira zosakhala za organic zikakhala zochepa zimabisika pansi pa zizindikiro zazikulu. Kusapezeka kwa nsonga zina ndi umboni wofooka wakuti palibe zonyansa.

Chachitatu, muyezo wolozera ndiwo umalamulira kuyerekezera konse. Muyezo wowonongeka, wosintha chifukwa cha madzi mu makhiristo, kapena wosamalidwa molakwika umasintha maziko a chigamulo. Momwemonso zimachitika ngati njira zosiyanasiyana zikuyerekezedwa, monga kuweruza sipekitiramu ya chitsanzo cha ATR pogwiritsa ntchito muyezo wa KBr. Lembani gulu lopangira muyezowo, njira yoyezera, ndi kukonzekera kwake pamodzi ndi chigamulo chilichonse.

Zosakaniza zimabweretsa chenjezo lomaliza. Sipekitiramu yomwe imafanana kwambiri koma ili ndi mapewa a nsonga kapena mphamvu za zizindikiro zosazolowereka si kupambana kongofunika mawu owonjezera. Ndi chizindikiro chakuti chitsanzocho chiyenera kulekanitsidwa musanachizindikire.

Kugwiritsa ntchito njira zothandizira

HPLC ya methylene blue imapereka yankho la zinthu zimene zilimo ndi kuchuluka kwake, lomwe infrared silingapereke. Njira ya panopa ya buku la miyezo imagwiritsa ntchito kolamu ya mtundu wa phenyl ndi chosakaniza cha madzi ndi acetonitrile chomwe kuchuluka kwa zigawo zake kumasinthidwa pang’onopang’ono, chokhala ndi trifluoroacetic acid, ndipo imazindikira pa 246 nm, osati m’chigawo cha kuwala kooneka. Kuzindikira kumachokera ku kufanana kwa nthawi yosungidwa mu kolamu ndi ya muyezo wolozera; kuyerekezera komweko kumaperekanso kuyeza kuchuluka kwa chinthu. Mayeso a zonyansa za organic amazindikira Azure B ndi chinthu chake cholozera, pa nthawi yosungidwa yoyerekeza ya pafupi ndi 0.8 poyerekezera ndi methylene blue pa 1.0, ndipo amafuna kulekanitsidwa kwabwino kwambiri pakati pa nsonga ziwirizo. Dziwani chifukwa chosankhira utali wa mafunde: mitundu yochotsedwa magulu a methyl imayamba kuyamwa kuwala kumene kumayandikira kwambiri mbali yofiira, kuchokera ku thionine pafupi ndi 600 nm kudzera ku Azure C, Azure A, ndi Azure B mpaka ku methylene blue pafupi ndi 670 nm. Choncho chozindikira chokhazikika pafupi ndi 660 nm chimayankha mosiyana pa mitundu imeneyi ndipo chimasonyeza kuchuluka kochepa kuposa chenicheni kwa ina mwa mitundu yofananayo. Kufufuza kwathunthu kwa mitundu yonseyi, kukafunika, kuyenera kuchitika ndi njira ya C18 LC-MS/MS yokhala ndi miyezo yotsimikiziridwa kuti ndi yoyenera pa chinthu chilichonse choyezedwa.

Kupatuka kwa kuwala pogwiritsa ntchito ufa kumayankha funso la mawonekedwe a chinthu cholimba limene njira ziwiri zapamwambazi zimawononga kapena kunyalanyaza: kusungunula ufa kuti uchitidwe chromatography kumachotsa chidziwitso cha makhiristo, ndipo kunjenjemera kwa mamolekyulu kumasonyeza maubale osati mmene mamolekyulu akonzedwera m’malo otambalala. Zithunzi za kupatuka kwa kuwala zimasiyanitsa mawonekedwe okhala madzi ndi kuchuluka kwa makhiristo, ndipo methylene blue imafunika kufufuzidwa motere kuposa zinthu zambiri. Kafukufuku wa zinthu zolimba wa Rager ndi anzake unazindikira makhiristo okhala madzi a mitundu isanu yosiyana, kuphatikizapo pentahydrate, mitundu iwiri ya dihydrate, monohydrate, ndi mtundu wokhala ndi madzi apakati, ndipo sunapeze trihydrate yeniyeni pakati pa mitundu yomwe inafotokozedwa. Kasamalidwe ka mu labotale ndi kofunika apa: chinyezi ndi kutentha zimasintha kukhazikika kwa mawonekedwe okhala madzi, ndipo kuphwanya mwamphamvu kungasinthe mawonekedwewo kapena kuchepetsa kapangidwe ka makhiristo. Choncho sungani ndi kulemba chinyezi choyerekeza ndi kutentha, ndipo chepetsani kuphwanya momwe mungathere. Kulemba mawonekedwe okhala madzi pa chizindikiro ndi mmene zimakhudzira kulemera koyezedwa zikufotokozedwanso mu mawonekedwe a methylene blue okhala madzi ndi kulemba zizindikiro.

Kuyamwa kuwala kwa UV-Vis kumagwira ntchito yothandizira. Malo odziwika oyamwa kuwala kwambiri amatsimikizira khalidwe loyembekezeredwa la yankho ndipo amathandiza kuyeza kuchuluka kwa chinthu mmenemo. Koma yankho labuluu loyamwa kuwala pa utali wa mafunde woyembekezeredwa limagwirizana ndi methylene blue popanda kutsimikizira kuti ndi imeneyo, chifukwa mitundu ya utoto yofanana nayo imayamwa kuwala pafupi ndi pamenepo.

Zimene muyenera kufunsa musanagule

Sinthani kusiyana kumeneku kukhala mafunso ofunsa pogula. Funsani kuti ndi gulu liti lopangidwa limene linayesedwa, ndi njira iti imene inatsimikizira mtundu wa chinthu, ngati mayeso a zonyansa a HPLC anagwiritsa ntchito chinthu cholozera cha Azure B, komanso ngati mawonekedwe okhala madzi anafufuzidwa kapena anangoganiziridwa. Koposa zonse, funsani ngati satifiketiyo ikukhudza miyezo yonse kapena mayeso ena osankhidwa okha. Ogulitsa ambiri amene amanena kuti zinthu zawo zikukwaniritsa buku lovomerezeka la miyezo amangoyesa zitsulo zolemera zokha, zomwe zimasiya mafunso a kuzindikira, kuchuluka kwa chinthu, zonyansa za organic, zosungunulira, ndi tizilombo tating’onoting’ono osayankhidwa.

Blupreme imayesa zofunika zonse za USP, kuphatikizapo kuzindikira, kuyera, zonyansa za organic, zosungunulira zotsala, zonyansa za maelementi, zotsalira pambuyo powotcha, malire a tizilombo tating’onoting’ono, ndi ma endotoxin a mabakiteriya, mogwirizana ndi wopanga methylene blue wa m’gawo la mankhwala. Satifiketi yowunikira ya gulu lililonse lopangidwa imasindikizidwa kuti anthu onse aione pa mayeso a khalidwe a Blupreme, kotero maziko oyeza kuchuluka kwa chinthu ndi zotsatira za zonyansa zitha kuwerengedwa mwachindunji m’malo mongoyerekezeredwa. Zotsatira zapafupi ndi 100% zimafunikabe kutchula maziko owerengera, monga momwe malangizo a kuchuluka koyezedwa kopitirira 100 peresenti amafotokozera. Kufanana kozindikirira kumayambitsa kufufuza. Zolemba zonse za mayeso zimakumaliza.