I-UV-Vis spectrophotometers zizixhobo ezinobunkunkqele ezisetyenziswa ngokubanzi kwiinkalo ezahlukeneyo zenzululwazi. Ngaphandle kokubaluleka kwazo, abantu abaninzi abaqondi ngokupheleleyo ukuba zeziphi ezi zixhobo, izicelo zabo, kunye nendlela ezisebenza ngayo. Eli nqaku lijolise ekunikeni inkcazo enzulu ye-UV-Vis spectrophotometers, equka imigaqo-siseko, ukusetyenziswa, kunye neemeko abaqeshwe phantsi kwazo.
Yintoni i-UV-Vis Spectrophotometer?
I-UV-Vis spectrophotometer sisixhobo sohlalutyo esisetyenziselwa ukulinganisa ubunzulu bokukhanya kwi-ultraviolet (UV) kunye nemimandla ebonakalayo (Vis) ye-electromagnetic spectrum. Ezi zixhobo zibalulekile ekuhlalutyeni iimpawu ze-optical yezinto, ukugqiba ukugxila kwazo, kunye nokuqonda ukuziphatha kwazo phantsi kweemeko ezahlukeneyo zokukhanya.
Isebenza njani i-UV-Vis Spectrophotometer?
Ukusebenza kwe-UV-Vis spectrophotometer kubandakanya amacandelo aphambili kunye namanyathelo:
Umthombo wokukhanya:
I-spectrophotometer iqulethe umthombo wokukhanya, ngokuqhelekileyo ukudibanisa isibane se-deuterium (ukukhanya kwe-UV) kunye nesibane se-tungsten (sokukhanya okubonakalayo). Ezi zibane zikhupha ukukhanya kwi-UV kunye ne-spectra ebonakalayo.
I-Monochromator:
Ukukhanya okukhutshwa ngumthombo kudlula kwi-monochromator, eyahlula ibe ngamaza obude obubodwa. Oku kuqhele ukwenziwa kusetyenziswa iprism okanye i-diffraction grating.
Isibambi Isampulu:
Ukukhanya kwe-monochromatic kuqondiswe kumnini wesampuli, apho isisombululo sesampuli sifakwe kwi-cuvette, isitya esincinci esenziwe ngeglasi okanye i-quartz.
Isikhangeli:
Emva kokudlula kwisampuli, ukukhanya kufikelela kwi-detector. I-detector ilinganisa ubunzulu bokukhanya okugqithisiweyo kwaye iguqule ibe ngumqondiso wombane.
Uhlalutyo lwedatha:
Umqondiso wombane ucutshungulwa yikhompyuter okanye i-microprocessor, eyenza i-spectrum ebonisa ukufunxa okanye ukuhanjiswa kwesampuli kubude bamaza ahlukeneyo.
Imigaqo-siseko ye-UV-Vis Spectrophotometry
Umgaqo osisiseko emva kwe-UV-Vis spectrophotometry nguMthetho weBeer-Lambert, onxibelelanisa ukufunxwa kokukhanya kwiimpawu zemathiriyeli ehamba ngayo ukukhanya. Umthetho uchazwa ngolu hlobo:
=⋅⋅
apho:
A yi-absorbeance (akukho yunithi, njengoko kumlinganiselo).
yi-molar absorptivity coefficient (L/mol·cm), into engaguqukiyo ebonisa ukuba into ifunxa ngamandla kangakanani na ukukhanya kubude obuthile.
kukuxinana kweentlobo ezifunxayo kwisampulu (mol/L).
bubude bendlela apho ukukhanya kuhamba khona kwisampulu (cm).
I-Absorbance ihambelana ngokuthe ngqo kugxininiso kunye nobude bendlela, okwenza i-UV-Vis spectrophotometry ibe sisixhobo esinamandla sohlalutyo lobungakanani.
Ukusetyenziswa kwe-UV-Vis Spectrophotometers
I-UV-Vis spectrophotometers inoluhlu olubanzi lwezicelo kwiindawo ezahlukeneyo:
1. IKhemistri
Ukumiselwa koQwalaselo:
I-UV-Vis spectrophotometers isetyenziswa rhoqo ukumisela ukuxinwa kwe-solutes kwisisombululo. Ngokomzekelo, i-concentration ye-DNA, iiprotheni, okanye ezinye ii-biomolecules zinokulinganiswa ngokufunxa kwazo kumaza athile.
I-Rection Kinetics:
Ezi zixhobo zinceda ekufundeni amaxabiso eempendulo zeekhemikhali ngokubeka iliso kwinguqu yokufunxa i-reactants okanye iimveliso ngokuhamba kwexesha.
Uhlalutyo lweMichiza:
Zisetyenziselwa ukuhlalutya okusemgangathweni kunye nobungakanani bemichiza yeekhemikhali, kunceda ukuchonga izinto ezisekelwe kwi-spectra yabo yokuthatha.
2. I-Biochemistry kunye ne-Molecular Biology
Iprotheyini kunye neNucleic Acid Ubungakanani:
I-UV-Vis spectrophotometry ibalulekile kwi-biochemistry yokulinganisa ukugxila kunye nokucoceka kwe-nucleic acids (i-DNA kunye ne-RNA) kunye neeprotheni.
Umsebenzi we-Enzyme:
Umsebenzi wee-enzyme unokuphononongwa ngokulinganisa ukufunxa kwee-substrates okanye iimveliso ezibandakanyekayo kwi-enzymatic reactions.
3. INzululwazi yezokusiNgqongileyo
Uvavanyo loMgangatho waManzi:
I-UV-Vis spectrophotometers isetyenziselwa ukubona kunye nokulinganisa ukungcola emanzini, njenge-nitrate, i-phosphates, kunye neentsimbi ezinzima.
UkuJonga uMgangatho woMoya:
Zinceda ekubekeni iliso kwizinto ezingcolisa umoya ngokulinganisa ukufunxwa kweegesi ezinjengeozone kunye nenitrogen dioxide.
4. Uhlalutyo lweKlinikhi kunye noLwamachiza
Uvavanyo kunye noPhuhliso lweziyobisi:
Kwishishini lamachiza, ii-spectrophotometers ze-UV-Vis zisetyenziselwa ukuhlalutya ugxininiso kunye nokucoceka kwamachiza kunye nokufunda ukuzinza kunye nokuthotywa kwe-pharmaceutical compounds.
Uxilongo lweKlinikhi:
Ezi zixhobo zinceda kuxilongo lweklinikhi ngokulinganisa ingxinano yezinto ezahlukeneyo kulwelo lomzimba, njengeglucose, cholesterol, kunye nebilirubin.
5. Ishishini lokutya kunye neziselo
Ulawulo lwemeko:
I-UV-Vis spectrophotometry isetyenziselwa ukuqinisekisa umgangatho kunye nokhuseleko lokutya kunye neziselo ngokulinganisa i-concentration ye-additives, i-preservatives, kunye ne-contaminants.
Uhlalutyo lwesondlo:
Ukuxinwa kweevithamini, iiminerali, kunye nezinye izondlo kwiimveliso zokutya kunokumiselwa ngokusebenzisa obu buchule.
Iindidi ze-UV-Vis Spectrophotometers
UV-Vis spectrophotometers co
kum kuqwalaselo olwahlukeneyo, nganye ilungele usetyenziso oluthile:
I-Spectrophotometers yeBeam enye:
Ezi zinendlela enye yokukhanya, oku kuthetha ukuba ireferensi kunye nemilinganiselo yesampulu ithathwa ngokulandelelana. Zilula kwaye azibizi kakhulu kodwa zinokuchaneka kancinci ngenxa yokuguquguquka okunokwenzeka kumthombo wokukhanya.
IiSpectrophotometers eziphindwe kabini:
Ezi zixhobo zahlula ukukhanya kwiindlela ezimbini, enye idlula kwisampuli kwaye enye idlula kwireferensi. Olu seto luvumela umlinganiselo ngaxeshanye, ubuyekeze ukuguquguquka kokuqina kokukhanya kunye nokubonelela ngeziphumo ezichanekileyo.
Abafundi beMicroplate:
Eyilelwe ukukhangela okuphezulu, abafundi be-microplate banokulinganisa iisampuli ezininzi ngaxeshanye besebenzisa ii-microplates ezinamaqula amaninzi, asetyenziswa ngokuqhelekileyo kwi-biotechnology kunye namashishini amayeza.
Ephathwayo UV-Vis Spectrophotometers:
Ezi zixhobo zihlangeneyo, ziphathwa ngesandla zisetyenziselwa umsebenzi kunye nohlalutyo lwendawo, zibonelela ngokulula kunye nokuguquguquka kokuhlolwa kokusingqongileyo kunye nolawulo lomgangatho.
Ubuchwephesha obuphezulu kunye nokwahluka
I-UV-Vis spectrophotometry iye yavela ukubandakanya ubuchule obuphambili kunye nokwahluka:
1. I-Spectrophotometry ephuma kuyo
Obu buchwephesha bubandakanya ukubala i-derivative ye-absorpct spectrum, ukuphucula ukusonjululwa kweencopho ezidlulanayo kunye nokuphucula ukuchaneka kwemilinganiselo yoxinaniso kwimixube enzima.
2. I-Spectrophotometry emisiweyo-Ukuhamba
Isetyenziselwa ukufunda i-kinetics yokuphendula ngokukhawuleza, i-spectrophotometry yokuyeka i-spectrophotometry ixuba i-reactants ngokukhawuleza kwaye ilinganisa utshintsho lwe-absorption ngexesha langempela, inika ulwazi kwiinkqubo ze-biochemical kunye neekhemikhali ezikhawulezayo.
3. I-Photoacoustic Spectroscopy
Le ndlela ilinganisa amaza esandi aveliswa kukufunxwa kokukhanya okumodareyithiweyo, enika ubuntununtunu obuphezulu bokufunda iisampulu eziqinileyo kunye ne-opaque apho i-UV-Vis spectrophotometry yemveli isenokungasebenzi.
Izinto eziluncedo kunye neMida
Iingenelo
Ayingonakalisi:
I-UV-Vis spectrophotometry ngokuqhelekileyo ayinakonakalisa, igcina isampuli ukuze ihlalutye ngakumbi.
Uvakalelo oluPhezulu kunye nokuchaneka:
Ubuchwephesha bubonelela ngobuntununtunu obuphezulu kunye nokuchaneka, buyenza ilungele ukufumanisa kunye nokulinganisa ukugxila okuphantsi kwabahlalutyi.
Ukuguquguquka:
Iyakwazi ukuhlalutya uluhlu olubanzi lwezinto, kubandakanywa iikhompawundi eziphilayo kunye ne-inorganic, kwiindawo ezahlukeneyo (eziqinileyo, ezimanzi, kunye negesi).
Ngokukhawuleza kwaye kulula:
Imilinganiselo idla ngokukhawuleza kwaye ithe ngqo, ivumela uhlalutyo olusebenzayo noluqhelekileyo.
Ukulinganiselwa
Iziphazamiso:
Ubukho bezinto eziphazamisayo ezifunxa kubude obufanayo bunokwenza kube nzima uhlalutyo.
Ukulungiselela Isampulu:
Ezinye iisampulu zinokufuna ukulungiswa okubanzi okanye ukuhlanjululwa, okunokuthi kungenise iimpazamo.
Ulwazi olunyiniweyo:
I-UV-Vis spectrophotometry ibonelela ikakhulu ngolwazi kugxininiso kunye nokufunxa iikhompawundi kodwa iswele ulwazi oluneenkcukacha lolwakhiwo, olufuna iindlela ezincedisanayo ezifana ne-mass spectrometry okanye i-NMR.
I-UV-Vis spectrophotometers zizixhobo eziyimfuneko kwisayensi yanamhlanje, enika indlela eguquguqukayo nenamandla yokuhlalutya uluhlu olubanzi lwezinto. Izicelo zabo zithatha indawo kwiinkalo ezahlukeneyo, kubandakanya ikhemistri, i-biochemistry, isayensi yokusingqongileyo, uxilongo lwezonyango, kunye neshishini lokutya. Ukuqonda imigaqo, ukusebenza, kunye nokusetyenziswa kwe-UV-Vis spectrophotometry ivumela izazinzulu kunye neengcali ukuba zisebenzise amandla azo apheleleyo kuphando nophuhliso, ulawulo lomgangatho, kunye neenjongo zohlalutyo. Ngaphandle kwemida yayo, i-UV-Vis spectrophotometer ihlala ililitye lembombo leelabhoratri zohlalutyo, inegalelo elikhulu kwinkqubela phambili kwisayensi nakwitekhnoloji.
