Inqaku
Ukuchonga imethylene blue elabhoratri: iispektra ze-infrared neendlela ezixhasanayo
Khawucinge ngeelabhoratri ezintathu ezifumana umgubo oluhlaza okwesibhakabhaka ofanayo. Eyokuqala irekhoda ispektram ye-infrared ize ithi yimethylene blue. Eyesibini yenza ikromatografi ize inike ingxelo ye-0.4% Azure B. Eyesithathu isebenzisa ukwahluka kwemitha ye-X kwiikristale ize ichaze uhlobo lwehydrate. Zontathu zichanile, kodwa akukho nanye ephendule eminye imibuzo emibini. Ukudideka kuqala xa isiphumo esinye sitolikwa ngokungathi siphendula yomithathu.
Eli nqaku liyahlula loo mibuzo. Ukuchonga nge-infrared kuqinisekisa ukuba yeyiphi imolekyuli ekhoyo ngokuyithelekisa nomgangatho wesalathiso. Ikromatografi iqinisekisa ukwakheka komxube. Ukwahluka kwemitha kuqinisekisa uhlobo lwemo eqinileyo. Ukungqinelana okuphumeleleyo ekuchongeni akusosiqinisekiso sobunyulu.
Ukuchonga nge-infrared kuphendula umbuzo omnye kakuhle
Uvavanyo lokuchonga imethylene blue oluchazwe kwincwadi esemgangathweni yamayeza luthelekisa ispektram nomgangatho wesalathiso; aluloluhlu lokuphawula iincopho. Ngokwe-mboniso wangaphambili wemonografi ye-USP yemethylene blue, i-Identification A ibhekisa kwispektroskopi se-infrared sisebenzisa izixhobo zamehlo ze-ATR okanye ipellet yepotassium bromide. Ispektram yesampulu kufuneka ingqinelane nespektram yomgangatho wesalathiso we-USP, isampulu nomgangatho zilungiswe zaza zarekhodwa phantsi kweemeko ezilinganayo. Akukho luhlu lumiselweyo lwamanani amaza okuchonga olugqibayo ngesiphumo. Kukubekwa kweespektra phezu kwenye okuqinisekisayo.
Olu khetho lwendlela lubonisa indlela iispektra ze-FTIR zemethylene blue eziziphatha ngayo. Iimpawu ezinamandla kakhulu zikummandla womnwe wespektram. Uncwadi lwezenzululwazi luhlala lunxulumanisa ibhendi ephambili yeeringi ezidityanisiweyo nommandla okufuphi ne-1590 ukuya ku-1600 cm-1, apho iindlela zokungcangcazela zokolulwa kweebhondi zecarbon-carbon nezecarbon-nitrogen kwiiringi ezinevumba elimnandi zidibana khona. Ezinye iibhendi ezikufuphi ne-1490 ne-1390 cm-1 zineempawu zokuguquka kwemilo yeringi, kwecarbon-nitrogen, nakwecarbon-hydrogen. Iibhendi ezikufuphi ne-1355 ne-1330 cm-1 zibandakanya iibhondi zecarbon-nitrogen zedimethylamino nokuguquka kwemilo yemethyl. Iimpawu ezikufuphi ne-1225 ukuya ku-1250 cm-1 zezesakhelo seheterocyclic, ngoxa iibhendi ezikufuphi ne-1170 ukuya ku-1130 nakwi-1066 cm-1 zineempawu zecarbon-sulfur, kunye nokugoba kwe-C-H yeeringi ezinevumba elimnandi kufuphi ne-880 cm-1. Iindlela zokungcangcazela zokolulwa zivela kumanani amaza aphezulu: i-C-H yeeringi ezinevumba elimnandi kufuphi ne-3040 cm-1, ne-N-methyl C-H phakathi kwe-2900 ne-3000 cm-1 malunga noko. Ukunxulunyaniswa kweebhendi okupapashiweyo kuyahluka kwiinkcukacha kuba isakhelo sephenothiazinium singcangcazela njengenkqubo edibeneyo, ngoko nayiphi na itheyibhile enye yokunxulumanisa mayithathwe njengengxelo, hayi njengokuphela kwengcaciso enokwenzeka.
Iziphumo ezimbini zokulungiselela zifuna ingqalelo kuba ziguqula iispektra ze-IR zemethylene blue ngaphandle kokuguqula idayi. Okokuqala, izinto ezinamanzi kwimo yehydrate zibonisa ibhendi ebanzi ye-O-H malunga ne-3300 ukuya ku-3500 cm-1 kunye nokugoba kwamanzi kufuphi ne-1630 ukuya ku-1640 cm-1, kwaye iintlobo ezahlukeneyo zehydrate zitshintsha indawo yeebhendi zomnwe wespektram ngendlela enokulinganiswa. Okwesibini, imatrix yepellet ye-KBr itsala ukufuma: ukufuma okufunxiweyo kuvelisa iibhendi kanye kuloo mimandla yamanzi yaye kunokubonakala ngokungathi ngamanzi ehydrate aphuma kwisampulu. Ukulinganisa nge-ATR kuyayiphepha imatrix ye-KBr, kodwa amandla eebhendi ze-ATR ayatshintsha ngokobunzulu bokungena kwemitha obuxhomekeke kwinani lamaza, ngoko ispektram ye-ATR nespektram yokudluliselwa kwemitha kumgubo ofanayo azinakubekwa ngqo phezu kwenye ukuze zithelekiswe. Umgaqo osebenzayo ulandela imfuneko yencwadi esemgangathweni: thelekisa isampulu nesalathiso ezirekhodwe ngendlela efanayo, uze ubhale ukuba yeyiphi esetyenzisiweyo. Ngekhemistri esisiseko yekatiyoni, yetyuwa, neyeehydrate ezibangela ezi spektra, jonga isalathiso sesakhiwo neempawu zemethylene blue.
Itheyibhile yokukhetha indlela yohlalutyo
Le theyibhile sisixhobo sokukhetha. Funda kumqolo wonke ukuze ufumanise ukuba indlela iphendula wuphi umbuzo, isiphumo esiphumeleleyo siqinisekisa ntoni, nokuba yintoni engekangqinwa.
| Indlela | Umbuzo ewuphendulayo | Oko kuqinisekiswa sisiphumo esiphumeleleyo | Oko ingenakuqinisekisa |
|---|---|---|---|
| I-FTIR xa ithelekiswa nomgangatho wesalathiso | Ingaba le nto yimethylene blue? | Ukuchongwa kwemolekyuli okungqinelana nespektram yesalathiso | Ubunyulu, iprofayili yokungcola, uhlobo lwehydrate ngokwayo, okanye uxinaniso |
| Uvavanyo lomlinganiselo nge-HPLC-UV nokungcola kwezinto eziphilayo (ikholamu yephenyl, 246 nm) | Ingakanani imethylene blue kwaye ingakanani i-Azure B? | Ukuchongwa ngekromatografi ngokungqinelana kokugcinwa; umlinganiselo wesithako kunye nezixa zokungcola ezichaziweyo | Uhlobo lwemo eqinileyo; ukuchongwa kweencopho ezingalindelekanga ngaphandle kwemigangatho yesalathiso |
| I-LC-MS/MS enemigangatho yesalathiso eqinisekisiweyo | Zeziphi iidayi ezisuswe amaqela emethyl ezikhoyo? | Ukwahlulwa nokuchongwa kuthotho lwe-MB, Azure A, Azure B, Azure C, thionine | Ukuthobela incwadi esemgangathweni; olu lulwandiso lwenqanaba lophando, hayi uvavanyo lwesiqhelo lwemonografi |
| I-XRD yomgubo | Yeyiphi ihydrate yekristale ekhoyo? | Isigaba semo eqinileyo, imixube yezigaba engaphezu komda wokubhaqwa, nobukristale | Ukuchongwa kwemolekyuli zokungcola okungekho kwimo yekristale; uxinaniso lwesinyibiliko |
| Ukufunxwa kwe-UV-Vis | Ingaba isinyibiliko sifunxa njengoko kulindelekile? | Iincopho zokufunxa ezineempawu ezithile nokuziphatha okuxhomekeke kuxinaniso | Ukuchongwa okanye ubunyulu ngokwayo |
| Ukuchongwa kwechloride nokulahleka kobunzima ekomisweni (ezixhasayo) | Ingaba iayoni echasene nekatiyoni ikhona; bungakanani ubunzima obunokuphuma bungumphunga? | Uhlobo lwetyuwa nobunzima obulahleka ekomisweni phantsi kweemeko ezichaziweyo | Yeyiphi imolekyuli enika umbala |
Kutheni ukungqinelana kwe-infrared kungesosiqinisekiso sobunyulu
Ukungqinelana ekuchongeni kukhupha amathuba okuba yidayi engalunganga. Akudwelisi zonke izinto ezikwisampulu. Imida emithathu ibaluleke kakhulu.
Okokuqala, izinto ezisondeleleneyo ngokwesakhiwo zikhangeleka zifana kwi-infrared. I-Azure B yahluka kwimethylene blue ngeqela elinye lemethyl, ngoko ispektram yayo ifana kakhulu neyemethylene blue. Umxube onepesenti ezimbalwa ze-Azure B usangqinelana kakuhle xa ispektram yawo ibekwe phezu kweyesalathiso semethylene blue. Ukubhaqa olo kungcola ngumsebenzi wekromatografi, ochazwe kwisikhokelo sokungcola kwemethylene blue neendlela zovavanyo.
Okwesibini, ispektroskopi se-infrared yindlela ehlalutya isampulu iyonke, enovakalelo olulinganiselweyo kwizithako ezikwizixa ezincinane. Izixa ezincinane zeedayi ezinxulumeneyo, izinyibilikisi ezishiyekileyo, okanye iintsalela ezingaphiliyo zifihleka phantsi kweebhendi eziphambili. Ukungabikho kweencopho ezongezelelekileyo bubungqina obubuthathaka bokungabikho kokungcola.
Okwesithathu, umgangatho wesalathiso ulawula lonke uthelekiso. Umgangatho owonakeleyo, oneespektra ezitshintshe ngenxa yamanzi ehydrate, okanye ophathwe ngendlela engafanelekanga utshintsha isiseko sovavanyo. Kuyafana naxa kuthelekiswa iindlela ezingahambelaniyo, njengokugweba ispektram yesampulu ye-ATR ngokuyithelekisa nesalathiso se-KBr. Rekhoda ibhetshi yomgangatho wesalathiso, indlela esetyenzisiweyo, nendlela yokulungiselela kunye nesigqibo ngasinye.
Imixube yongeza isilumkiso sokugqibela. Ispektram engqinelana ubukhulu becala, kodwa enamagxa kwiincopho okanye amandla eebhendi angaqhelekanga, ayisosiphumo esiphumeleleyo esinamanqaku asezantsi. Luphawu lokuba kufuneka kwahlulwe izithako zesampulu ngaphambi kokuyichonga.
Iindlela ezixhasanayo xa zisetyenziswa
I-HPLC yemethylene blue inika impendulo ngokwakheka komxube engakwaziyo ukuyinika i-infrared. Indlela yangoku yencwadi esemgangathweni isebenzisa ikholamu yohlobo lwephenyl enomxube wamanzi neacetonitrile otshintsha ngokuthe ngcembe, oqulethe itrifluoroacetic acid, ize ibhaqe kwi-246 nm, hayi kwibhendi yokukhanya okubonakalayo. Ukuchongwa kuvela ekungqinelaneni kwexesha lokugcinwa kunye nomgangatho wesalathiso; olo thelekiso lukwanika umlinganiselo wesithako. Uvavanyo lokungcola kwezinto eziphilayo luchonga i-Azure B ngesalathiso sayo, kugcino olunxulumeneyo olukufuphi ne-0.8 xa luthelekiswa nemethylene blue kwi-1.0, kwaye lufuna ukwahlulwa okuphezulu phakathi kweencopho ezimbini. Qaphela ukhetho lobude bamaza: uthotho lwezinto ezisuswe amaqela emethyl lufunxa ngokuthe ngcembe ngakumbi kwicala elibomvu, ukusuka kwithionine kufuphi ne-600 nm, ludlule kwi-Azure C, Azure A, ne-Azure B, luye kwimethylene blue kufuphi ne-670 nm. Ngoko isixhobo sokubhaqa esimiselwe kufuphi ne-660 nm siphendula ngokungalinganiyo kulo lonke uthotho yaye sibonisa izixa eziphantsi kuneziyinyani kwezinye iihomologue. Xa kufuneka iprofayili epheleleyo yalo lonke uthotho, kufanele kusetyenziswe indlela ye-C18 LC-MS/MS enemigangatho yesalathiso eqinisekisiweyo yento nganye ehlalutywayo.
Ukwahluka kwemitha kumgubo kuphendula umbuzo wemo eqinileyo othi iindlela ezimbini ezingasentla ziwutshabalalise okanye zingawuhoyi: ukunyibilikisa umgubo ukwenza ikromatografi kususa ulwazi lwekristale, kwaye ukungcangcazela kweemolekyuli kunika ulwazi ngeebhondi kunokunika ulwazi ngolungelelwaniso olude lwesakhiwo. Iipateni zokwahluka kwemitha zahlula izigaba zehydrate nobukristale, kwaye imethylene blue ifuna olo hlolo ngaphezu kwezinto ezininzi. Uphando lwemo eqinileyo lukaRager noogxa bakhe lwachonga izakhiwo ezintlanu ezahlukeneyo zehydrate, kuquka ipentahydrate, iidihydrate ezimbini, imonohydrate, nehydrate ephakathi, lwaza alwafumana trihydrate yokwenene phakathi kweentlobo ezichongiweyo. Indlela yokuphatha elabhoratri ibalulekile apha: ukufuma nobushushu kutshintsha uzinzo lwehydrate, yaye ukugaya ngamandla kunokuguqula isigaba okanye kunciphise ubukristale, ngoko lawula uze urekhode ukufuma okunxulumeneyo nobushushu, kwaye unciphise ukugaya kangangoko. Ukubhalwa kohlobo lwehydrate kwileyibhile nempembelelo yako kubunzima obulinganisiweyo kuxoxwa ngakumbi kwiintlobo zehydrate zemethylene blue nokubhalwa kweeleyibhile.
Ukufunxwa kwe-UV-Vis kudlala indima yokuxhasa. Iincopho eziqhelekileyo ziqinisekisa ukuziphatha okulindelekileyo kwesinyibiliko zize zenze kube nokwenzeka ukusebenza ngoxinaniso, kodwa isinyibiliko esiluhlaza okwesibhakabhaka esifunxa kubude bamaza obulindelekileyo siyahambelana nemethylene blue ngaphandle kokungqina ukuba yiyo, kuba iidayi ezinxulumeneyo zifunxa kufuphi nalapho.
Oko kufuneka ukubuze ngaphambi kokuthenga
Guqula lo mahluko ube yimibuzo yokuthenga. Buza ukuba yeyiphi ibhetshi evavanyiweyo, yeyiphi indlela esetyenzisiweyo ukuchonga, ukuba uvavanyo lokungcola lwe-HPLC lusebenzise isalathiso se-Azure B, nokuba uhlobo lwehydrate luchongiwe okanye lucingelwe nje. Ngaphezu kwako konke, buza ukuba isiqinisekiso sibandakanya zonke iimfuno ezichaziweyo okanye iimvavanyo ezikhethiweyo kuphela. Abathengisi abaninzi abathi bayahambelana nencwadi esemgangathweni bavavanya kuphela iintsimbi ezinzima, nto leyo eshiya imibuzo yokuchonga, yomlinganiselo wesithako, yokungcola kwezinto eziphilayo, yezinyibilikisi, neyeentsholongwane ingaphendulwanga.
I-Blupreme ivavanya zonke iimfuno ze-USP, ezibandakanya ukuchongwa, ubunyulu, ukungcola kwezinto eziphilayo, izinyibilikisi ezishiyekileyo, ukungcola kweziqalelo, iintsalela emva kokutshiswa, imida yeentsholongwane, nee-endotoxin zeebhaktheriya, isebenzisana nomvelisi wemethylene blue yamayeza. Isiqinisekiso sohlalutyo sebhetshi nganye esifumaneka kuluntu sipapashwa kuvavanyo lomgangatho lwe-Blupreme, ukuze isiseko somlinganiselo wesithako neziphumo zokungcola zifundwe ngqo kunokuba kuqikelelwe. Iziphumo ezikufuphi ne-100% zisafuna ukuba kuchazwe isiseko sokubalwa kwazo, njengoko kuchaza isikhokelo somlinganiselo wesithako ongaphezu kwe-100 pesenti. Ukungqinelana ekuchongeni kuvula uphando. Ingxelo epheleleyo iyaluvala.