UV-Vis spectrophotometers ke lisebelisoa tse tsoetseng pele tse sebelisoang haholo makaleng a fapaneng a mahlale.Ho sa tsotellehe bohlokoa ba tsona, batho ba bangata ha ba utloisise ka botlalo hore na lisebelisoa tsena ke eng, lisebelisoa tsa tsona le hore na li sebetsa joang.Sengoliloeng sena se ikemiselitse ho fana ka tlhaloso e tebileng ea li-spectrophotometer tsa UV-Vis, tse koahelang melao-motheo ea tsona, ts'ebeliso le maemo ao li hiriloeng tlas'a tsona.
UV-Vis Spectrophotometer ke eng?
UV-Vis spectrophotometer ke sesebelisoa sa tlhahlobo se sebelisetsoang ho lekanya matla a khanya ho ultraviolet (UV) le libakeng tse bonahalang (Vis) tsa mahlaseli a motlakase.Lisebelisoa tsena ke tsa bohlokoa bakeng sa ho sekaseka thepa ea optical ea lintho, ho tseba ho tsepamisa mohopolo oa tsona, le ho utloisisa boitšoaro ba tsona tlas'a maemo a fapaneng a leseli.
UV-Vis Spectrophotometer e sebetsa Joang?
Ts'ebetso ea UV-Vis spectrophotometer e kenyelletsa likarolo le mehato e 'maloa ea bohlokoa:
Mohloli oa Leseli:
spectrophotometer e na le mohloli oa leseli, hangata motsoako oa lebone la deuterium (bakeng sa lebone la UV) le lebone la tungsten (bakeng sa leseli le bonahalang).Mabone ana a ntša khanya ho pholletsa le UV le ponahalo e bonahalang.
Monochromator:
Leseli le hlahisoang ke mohloli le feta ka monochromator, e leng e arolang ka bolelele ba maqhubu ka bomong.Hangata sena se etsoa ho sebelisoa prism kapa grating ea diffraction.
Mohlala oa Mohlokomeli:
Leseli la monochromatic le lebisitsoe ka sets'oants'o sa mohlala, moo tharollo ea mohlala e kenngoa ka cuvette, setshelo se senyenyane se entsoeng ka khalase kapa quartz.
Mohloekisi:
Ka mor'a ho feta ka har'a sampuli, leseli le fihla ho detector.Mohloli o lekanya matla a lebone le fetisoang ebe o le fetola lets'oao la motlakase.
Tlhahlobo ea Lintlha:
Joale lets'oao la motlakase le sebetsoa ke komporo kapa microprocessor, e hlahisang spectrum e bonts'ang ho monya kapa ho fetisoa ha sampuli ka bolelele bo fapaneng ba maqhubu.
Melao-motheo ea UV-Vis Spectrophotometry
Molao-motheo oa motheo oa UV-Vis spectrophotometry ke Molao oa Beer-Lambert, o amanang le ho monya ha leseli le litšobotsi tsa thepa eo leseli le tsamaeang ka eona.Molao o hlalosoa tjena:
=⋅⋅
moo:
A ke ho monya (ha ho na li-unit, kaha ke karo-karolelano).
ke molar absorptivity coefficient (L/mol·cm), e sa fetoheng e bontšang hore na ntho e monya khanya ka matla hakae boleleleng bo itseng ba maqhubu.
ke bongata ba mefuta e monyang ka har'a sampole (mol/L).
ke bolelele ba tsela eo leseli le tsamaeang ka eona sampuling (cm).
Absorbance e lekana ka kotloloho le mahloriso le bolelele ba tsela, e etsa hore UV-Vis spectrophotometry e be sesebelisoa se matla sa tlhahlobo ea bongata.
Lisebelisoa tsa UV-Vis Spectrophotometers
UV-Vis spectrophotometers e na le mefuta e mengata ea lits'ebetso ho pholletsa le masimo a fapaneng:
1. K'hemistri
Boikemisetso ba ho tsepamisa maikutlo:
Li-spectrophotometers tsa UV-Vis li sebelisoa khafetsa ho fumana hore na solute e kae ka tharollo.Ka mohlala, bongata ba DNA, liprotheine kapa li-biomolecules tse ling li ka lekanyetsoa ka ho monya ha tsona ka bolelele bo itseng ba maqhubu.
Reaction Kinetics:
Lisebelisoa tsena li thusa ho ithuta litekanyetso tsa karabelo ea lik'hemik'hale ka ho beha leihlo phetoho ea ho monya ha li-reactants kapa lihlahisoa ha nako e ntse e ea.
Tlhahlobo ea Lik'hemik'hale:
Li sebelisetsoa tlhahlobo ea boleng le bongata ba lik'hemik'hale tsa lik'hemik'hale, ho thusa ho tseba lintho tse ipapisitseng le pono ea tsona ea ho monya.
2. Biochemistry le Molecular Biology
Protheine le Nucleic Acid Quantification:
UV-Vis spectrophotometry e bohlokoa ho biochemistry bakeng sa ho lekanya mahloriso le bohloeki ba nucleic acids (DNA le RNA) le liprotheine.
Ts'ebetso ea enzyme:
Mosebetsi oa li-enzyme o ka ithutoa ka ho lekanya ho monya ha li-substrates kapa lihlahisoa tse amehang karabelong ea enzymatic.
3. Saense ea Tikoloho
Teko ea Boleng ba Metsi:
UV-Vis spectrophotometers e sebelisoa ho bona le ho lekanya lintho tse silafatsang metsing, joalo ka li-nitrate, phosphates, le litšepe tse boima.
Tlhokomelo ea Boleng ba Moea:
Li thusa ho beha leihlo lintho tse silafatsang moea ka ho lekanya ho monya ha likhase tse kang ozone le nitrogen dioxide.
4. Tlhahlobo ea Tleliniki le Meriana
Teko le Nts'etsopele ea Lithethefatsi:
Indastering ea meriana, UV-Vis spectrophotometers e sebelisoa ho hlahloba mahloriso le bohloeki ba lithethefatsi le ho ithuta botsitso le ho senyeha ha metsoako ea meriana.
Litlhahlobo tsa Kliniki:
Lisebelisoa tsena li thusa tlhahlobo ea bongaka ka ho metha hore na ke lintho life tse fapaneng marong a 'mele, joalo ka tsoekere, cholesterol le bilirubin.
5. Indasteri ea Lijo le Lino
Taolo ea Boleng:
UV-Vis spectrophotometry e sebelisoa ho netefatsa boleng le polokeho ea lijo le lino ka ho lekanya bongata ba li-additives, preservatives le litšila.
Tlhahlobo ea Phepo:
Tekanyetso ea livithamini, liminerale le limatlafatsi tse ling lihlahisoa tsa lijo li ka khethoa ho sebelisoa mokhoa ona.
Mefuta ea UV-Vis Spectrophotometers
Lisebelisoa tsa UV-Vis spectrophotometers
'na ka litlhophiso tse fapaneng, e' ngoe le e 'ngoe e loketse lits'ebetso tse ikhethileng:
Single-Beam Spectrophotometers:
Tsena li na le tsela e le 'ngoe ea khanya, ho bolelang hore litšupiso le litekanyo tsa sampole li nkuoa ka tatellano.Li bonolo ebile ha li na litšenyehelo empa li ka fosahetse ka lebaka la ho feto-fetoha ho ka bang teng ho matla a mohloli o khanyang.
Double-Beam Spectrophotometers:
Lisebelisoa tsena li arola leseli ka litsela tse peli, e 'ngoe e feta sampoleng' me e 'ngoe e feta ka litšupiso.Seta sena se lumella ho metha ka nako e le 'ngoe, ho lefella ho fetoha ha khanya le ho fana ka liphetho tse nepahetseng haholoanyane.
Babali ba Microplate:
E etselitsoe ho hlahloba boemo bo phahameng, babali ba li-microplate ba ka lekanya lisampole tse ngata ka nako e le 'ngoe ba sebelisa li-microplates tse nang le liliba tse ngata, tse sebelisoang hangata ho biotechnology le indasteri ea meriana.
Li-Spectrophotometers tse nkehang tsa UV-Vis:
Lisebelisoa tsena tse kopanetsoeng, tse tšoaroang ka letsoho li sebelisetsoa mosebetsi oa masimo le tlhahlobo ea sebaka sa marang-rang, tse fanang ka boiketlo le bonolo bakeng sa ho lekola tikoloho le taolo ea boleng.
Mekhoa e tsoetseng pele le mefuta e fapaneng
UV-Vis spectrophotometry e fetohile ho kenyelletsa mekhoa e tsoetseng pele le mefuta e fapaneng:
1. Derivative Spectrophotometry
Mokhoa ona o kenyelletsa ho bala ho tsoa ho sekhahla sa ho monya, ho ntlafatsa tharollo ea litlhoro tse fetang le ho ntlafatsa ho nepahala ha litekanyo tsa mahloriso metsoakong e rarahaneng.
2. Spectrophotometry ea Phallo e Emeng
E sebelisetsoa ho ithuta kinetics ea karabelo e potlakileng, spectrophotometry e emisitsoeng e kopanya li-reactants ka potlako 'me e lekanya liphetoho tsa ho monya ka nako ea sebele, e fana ka leseli mabapi le mekhoa e potlakileng ea lik'hemik'hale le lik'hemik'hale.
3. Photoacoustic Spectroscopy
Mokhoa ona o lekanya maqhubu a molumo a hlahisoang ke ho monya ha leseli le fetotsoeng, a fanang ka kutlo e phahameng bakeng sa ho ithuta lisampole tse tiileng le tse opaque moo spectrophotometry ea setso ea UV-Vis e kanna ea se sebetse hantle.
Melemo le Mefokolo
Melemo
E sa Senyeheng:
UV-Vis spectrophotometry ka kakaretso ha e senyehe, e boloka sampole bakeng sa tlhahlobo e tsoelang pele.
Kutloelo-bohloko e Phahameng le ho Nepaha:
Mokhoa ona o fana ka kutlo e phahameng le ho nepahala, ho etsa hore e lokele ho bona le ho lekanya likhakanyo tse tlase tsa bahlahlobisisi.
Mefuta-futa:
E ka sekaseka mefuta e mengata ea lintho, ho kenyeletsoa metsoako ea lintho tse phelang le e sa tsitsang, libakeng tse fapaneng (tse tiileng, tse metsi le tsa khase).
E potlakile ebile e bonolo:
Litekanyo hangata li potlakile ebile li hlakile, li lumella tlhahlobo e sebetsang le e tloaelehileng.
Mefokolo
Litšitiso:
Ho ba teng ha lintho tse kena-kenanang tse monyang ka maqhubu a tšoanang ho ka thatafatsa tlhahlobo.
Tokiso ea Mohlala:
Mehlala e meng e ka hloka ho lokisoa haholo kapa ho hlapolloa, ho ka hlahisa liphoso.
Litaba tse Lekanyelitsoeng:
UV-Vis spectrophotometry e fana haholo ka leseli mabapi le ho tsepamisa mohopolo le ho monya ha metsoako empa e haelloa ke lintlha tse qaqileng tsa sebopeho, tse hlokang mekhoa e tlatselletsang joalo ka mass spectrometry kapa NMR.
UV-Vis spectrophotometers ke lisebelisoa tsa bohlokoa tsa mahlale a sejoale-joale, tse fanang ka mokhoa o feto-fetohang le o matla oa ho sekaseka lintho tse ngata.Likopo tsa bona li akaretsa likarolo tse fapaneng, ho kenyelletsa k'hemistri, biochemistry, saense ea tikoloho, tlhahlobo ea bongaka le indasteri ea lijo.Ho utloisisa melao-motheo, ts'ebetso le ts'ebeliso ea UV-Vis spectrophotometry e lumella bo-ramahlale le litsebi ho sebelisa bokhoni ba bona ba ho etsa lipatlisiso le nts'etsopele, taolo ea boleng le merero ea tlhahlobo.Leha e na le mefokolo, UV-Vis spectrophotometer e ntse e le lejoe la motheo la lilaboratori tsa tlhahlobo, e tlatsetsang haholo tsoelopeleng ea mahlale le thekenoloji.
