ʻO UV-Vis spectrophotometers nā mea hana maʻalahi i hoʻohana nui ʻia i nā ʻano ʻepekema like ʻole. ʻOiai ko lākou koʻikoʻi, nui ka poʻe ʻaʻole maopopo i ke ʻano o kēia mau mea hana, kā lākou noi, a pehea lākou e hana ai. Ke manaʻo nei kēia ʻatikala e hāʻawi i ka wehewehe hohonu o nā spectrophotometers UV-Vis, e uhi ana i kā lākou mau kumu, hoʻohana, a me nā kūlana i hoʻohana ʻia ai lākou.
He aha ka UV-Vis Spectrophotometer?
ʻO ka spectrophotometer UV-Vis kahi mea analytical i hoʻohana ʻia e ana i ka ikaika o ka mālamalama i nā ʻāpana ultraviolet (UV) a ʻike ʻia (Vis) o ka spectrum electromagnetic. He mea koʻikoʻi kēia mau mea hana no ka nānā ʻana i nā waiwai optical o nā mea, ka hoʻoholo ʻana i ko lākou ʻike ʻana, a me ka hoʻomaopopo ʻana i kā lākou ʻano ma lalo o nā kūlana māmā.
Pehea e hana ai ka UV-Vis Spectrophotometer?
ʻO ka hana o kahi spectrophotometer UV-Vis e pili ana i kekahi mau mea nui a me nā ʻanuʻu:
Puna Māmā:
Loaʻa i ka spectrophotometer kahi kumu kukui, maʻamau ka hui pū ʻana o ke kukui deuterium (no ke kukui UV) a me ke kukui tungsten (no ke kukui ʻike ʻia). Hoʻopuka kēia mau kukui i ka mālamalama ma waena o ka UV a ʻike ʻia.
Monochromator:
ʻO ke kukui i hoʻopuka ʻia e ke kumu e hele i loko o kahi monochromator, e hoʻokaʻawale iā ia i nā lōʻihi nalu pākahi. Hana ʻia kēia me ka hoʻohana ʻana i ka prism a i ʻole ka diffraction grating.
Laʻana Paʻa:
Hoʻonohonoho ʻia ke kukui monochromatic ma o ka mea paʻa hāpana, kahi i waiho ʻia ai ka hāpana hāpana i loko o kahi cuvette, kahi pahu liʻiliʻi i hana ʻia i ke aniani a i ʻole quartz.
ʻIke:
Ma hope o ka hele ʻana i ka hāpana, hiki ke kukui i kahi mea ʻike. ʻIke ka mea ʻike i ka ikaika o ke kukui i hoʻouna ʻia a hoʻololi iā ia i hōʻailona uila.
ʻIkepili ʻikepili:
Hoʻohana ʻia ka hōʻailona uila e ke kamepiula a i ʻole microprocessor, kahi e hoʻopuka ai i kahi kikoʻī e hōʻike ana i ka hoʻopalekana a i ʻole ka lawe ʻana o ka hāpana ma nā lōʻihi nalu like ʻole.
Nā Kumu o ka UV-Vis Spectrophotometry
ʻO ke kumu kumu ma hope o UV-Vis spectrophotometry ʻo ia ka Beer-Lambert Law, e pili ana i ka hoʻomoʻa ʻana o ka mālamalama i nā waiwai o ka mea e hele ai ka mālamalama. Hōʻike ʻia ke kānāwai penei:
=⋅⋅
ma hea:
ʻO A ka absorbance (ʻaʻohe ʻāpana, ʻoiai he ratio).
ʻo ia ka helu hoʻopalekana molar (L/mol·cm), he mea mau e hōʻike ana i ka ikaika o ka hoʻopaʻa ʻana o ka mea i ka mālamalama i kekahi lōʻihi nalu.
ʻo ia ka ʻike ʻana o nā ʻano mea hoʻomoe i ka hāpana (mol/L).
ʻo ia ka lōʻihi o ke ala e hele ai ka mālamalama i ka hāpana (cm).
Hoʻohālikelike pololei ka Absorbance i ka noʻonoʻo a me ka lōʻihi o ke ala, e hana ana i ka spectrophotometry UV-Vis i mea hana ikaika no ka loiloi quantitative.
Nā noi o nā UV-Vis Spectrophotometers
Loaʻa i nā spectrophotometers UV-Vis ka laulā o nā noi ma nā kahua like ʻole:
1. Kemika
Ka hoʻoholo ʻana o ka manaʻo:
Hoʻohana mau ʻia nā spectrophotometers UV-Vis no ka hoʻoholo ʻana i ka neʻe ʻana o nā solutes i loko o kahi hopena. No ka laʻana, hiki ke ana ʻia ka neʻe ʻana o ka DNA, nā protein, a i ʻole nā biomolecules ʻē aʻe e ko lākou komo ʻana i nā lōʻihi nalu.
Kinetics pane:
Kōkua kēia mau mea hana i ke aʻo ʻana i ka nui o nā hopena kemika ma o ka nānā ʻana i ka hoʻololi ʻana o ka hoʻopaʻa ʻana o nā reactants a i ʻole nā huahana i ka manawa.
Nānā Kemika:
Hoʻohana ʻia lākou no ka loiloi qualitative a me quantitative o nā pūhui kemika, e kōkua ana i ka ʻike ʻana i nā mea ma muli o kā lākou absorbance spectra.
2. Biochemistry and Molecular Biology
Ka helu ʻana o ka protein a me ka nucleic acid:
Pono ka spectrophotometry o UV-Vis i ka biochemistry no ke ana ʻana i ka ʻike a me ka maʻemaʻe o nā waikawa nucleic (DNA a me RNA) a me nā protein.
Hana Enzyme:
Hiki ke aʻo ʻia ka hana o nā enzymes ma ke ana ʻana i ka absorbance o nā substrates a i ʻole nā huahana i komo i nā hopena enzymatic.
3. ʻEpekema Kaiapuni
ʻO ka hoʻāʻo ʻana i ka maikaʻi o ka wai:
Hoʻohana ʻia nā spectrophotometers UV-Vis e ʻike a helu i nā mea haumia i loko o ka wai, e like me nā nitrates, phosphates, a me nā metala kaumaha.
Nānā maikaʻi o ka ea:
Kōkua lākou i ka nānā ʻana i nā mea haumia ea ma ke ana ʻana i ka hoʻopaʻa ʻana o nā kinoea e like me ka ozone a me ka nitrogen dioxide.
4. Ka Hoʻonaʻauao Kauka a me ka lāʻau lapaʻau
ʻO ka hoʻāʻo a me ka hoʻomohala ʻana i ka lāʻau lapaʻau:
I ka ʻoihana lāʻau lapaʻau, hoʻohana ʻia nā spectrophotometers UV-Vis e nānā i ka neʻe ʻana a me ka maʻemaʻe o nā lāʻau lapaʻau a e aʻo i ka paʻa a me ka hoʻohaʻahaʻa ʻana o nā pūhui lāʻau.
Nā hōʻailona hōʻailona:
Kōkua kēia mau mea hana i nā diagnostics clinical ma ke ana ʻana i ka neʻe ʻana o nā mea like ʻole i loko o nā wai o ke kino, e like me ka glucose, cholesterol, a me bilirubin.
5. ʻOihana meaʻai a me nā mea inu
Mana maikaʻi:
Hoʻohana ʻia ka spectrophotometry UV-Vis e hōʻoia i ka maikaʻi a me ka palekana o nā meaʻai a me nā mea inu ma ke ana ʻana i ka ʻike o nā additives, preservatives, a me nā mea haumia.
Nānā Meaʻai:
Hiki ke hoʻoholo ʻia ke ʻano o nā huaora, nā minela, a me nā meaʻai ʻē aʻe i nā huahana meaʻai me ka hoʻohana ʻana i kēia ʻenehana.
Nā ʻano o nā UV-Vis Spectrophotometers
ʻO UV-Vis spectrophotometers co
iaʻu i nā hoʻonohonoho like ʻole, kūpono kēlā me kēia no nā noi kikoʻī:
Nā Kiʻi Kiʻi Kiʻi Hoʻokahi-Beam:
Loaʻa i kēia mau ala māmā hoʻokahi, ʻo ia hoʻi, lawe ʻia ka kuhikuhi a me nā ana laʻana i ka sequentially. ʻOi aku ka maʻalahi a ʻoi aku ka maikaʻi o ke kumukūʻai akā hiki ke liʻiliʻi i ka pololei ma muli o ka hiki ke loli i ka ikaika o ke kumu māmā.
Nā kiʻi kiʻi kiʻi kiʻi pālua:
Hoʻokaʻawale kēia mau mea kani i ke kukui i ʻelua ala, hele kekahi ma ka hāpana a ʻo kekahi ma kahi kuhikuhi. Hāʻawi kēia hoʻonohonoho i ke ana like ʻana, e uku ana i ka fluctuations o ka māmā māmā a hāʻawi i nā hopena pololei.
Mea Heluhelu Microplate:
Hoʻolālā ʻia no ka kānana kiʻekiʻe, hiki i ka poʻe heluhelu microplate ke ana i nā laʻana he nui i ka manawa like me ka hoʻohana ʻana i nā microplates me nā pūnāwai he nui, i hoʻohana mau ʻia i ka biotechnology a me nā ʻoihana lāʻau.
ʻO nā ʻano kiʻi kiʻi kiʻi UV-Vis paʻa:
Hoʻohana ʻia kēia mau mea paʻa, paʻa lima no ka hana kahua a me ka nānā ʻana ma ka pūnaewele, e hāʻawi ana i ka maʻalahi a me ka maʻalahi no ka nānā ʻana i ke kaiapuni a me ka hoʻokele maikaʻi.
Nā ʻenehana kiʻekiʻe a me nā ʻano like ʻole
Ua ulu aʻe ka spectrophotometry UV-Vis e hoʻokomo i nā ʻenehana holomua a me nā ʻano like ʻole:
1. Hoʻohālikelike ʻano kiʻi kiʻi
Hoʻopili kēia ʻenehana i ka helu ʻana i ka derivative o ka spectrum absorbance, hoʻomaikaʻi i ka hoʻonā ʻana o ka overlapping peaks a me ka hoʻomaikaʻi ʻana i ka pololei o nā ana hoʻopaʻa ʻana i nā huila paʻakikī.
2. Kāohi-Kahe Spectrophotometry
Hoʻohana ʻia no ke aʻo ʻana i nā kinetics pane wikiwiki, hoʻohui ʻia ka spectrophotometry stop-flow i nā reactants a ana i nā loli o ka absorbance i ka manawa maoli, e hāʻawi ana i nā ʻike i nā kaʻina biochemical wikiwiki a me nā kaʻina hana kemika.
3. Photoacoustic Spectroscopy
ʻO kēia ala e ana i nā hawewe kani i hana ʻia e ka absorption o ka māmā modulated, e hāʻawi ana i ka ʻike kiʻekiʻe no ke aʻo ʻana i nā laʻana paʻa a me ka opaque kahi i kūpono ʻole ai ka spectrophotometry UV-Vis.
Pōmaikaʻi a me nā palena
Nā pono
Hoʻopau ʻole:
ʻO UV-Vis spectrophotometry maʻamau ʻaʻole luku, mālama i ka hāpana no ka nānā hou ʻana.
ʻO ka ʻike kiʻekiʻe a me ka pololei:
Hāʻawi ka ʻenehana i ka ʻike kiʻekiʻe a me ka pololei, kūpono ia no ka ʻike ʻana a me ka helu ʻana i nā haʻahaʻa haʻahaʻa o nā analytes.
ʻAno kūpono:
Hiki iā ia ke kālailai i nā ʻano mea like ʻole, me nā pūhui organik a me nā mea ʻole, ma nā mokuʻāina like ʻole (paʻa, wai, a me ke kinoea).
wikiwiki a maʻalahi:
ʻO ka maʻamau ka wikiwiki a me ka pololei o nā ana, e ʻae ana i ka loiloi kūpono a maʻamau.
Nā palena
Hoʻopilikia:
ʻO ka loaʻa ʻana o nā mea keakea e hoʻomoʻa i nā lōʻihi nalu like hiki ke paʻakikī i ka nānā ʻana.
Hoʻomākaukau laʻana:
Pono paha kekahi mau laʻana i ka hoʻomākaukau nui ʻana a i ʻole ka hoʻoheheʻe ʻana, hiki ke hoʻokomo i nā hewa.
ʻIke palena ʻole:
Hāʻawi mua ʻo UV-Vis spectrophotometry i ka ʻike e pili ana i ka noʻonoʻo a me ke komo ʻana o nā pūhui akā ʻaʻohe ʻike kikoʻī kikoʻī, e koi ana i nā ʻenehana hoʻohui e like me mass spectrometry a i ʻole NMR.
ʻO nā spectrophotometers UV-Vis he mea pono loa ia i ka ʻepekema hou, e hāʻawi ana i kahi ala maʻalahi a ikaika hoʻi no ka nānā ʻana i nā ʻano mea like ʻole. Hoʻopili ʻia kā lākou mau noi ma nā ʻano like ʻole, me ka chemistry, biochemistry, environmental science, clinical diagnostics, a me ka ʻoihana meaʻai. ʻO ka hoʻomaopopo ʻana i nā loina, ka hana, a me ka hoʻohana ʻana o UV-Vis spectrophotometry e hiki ai i nā ʻepekema a me nā ʻoihana ke hoʻohana i kona hiki piha no ka noiʻi a me ka hoʻomohala ʻana, ka mana maikaʻi, a me nā kumu analytical. ʻOiai kona mau palena, ʻo ka UV-Vis spectrophotometer ke ʻano he pōhaku kihi o nā laboratories analytical, e hāʻawi nui ana i ka holomua o ka ʻepekema a me ka ʻenehana.
