I-SPE isineminyaka ikhona, futhi ngesizathu esihle. Uma ososayensi befuna ukususa izingxenye ezingemuva kumasampula abo, babhekana nenselele yokwenza lokho ngaphandle kokunciphisa ikhono labo lokunquma ngokunembile nangokunembile ubukhona nenani lenhlanganisela yabo abayithakaselayo. I-SPE iyindlela eyodwa ososayensi abavame ukuyisebenzisa ukuze basize ukulungiselela amasampula abo okokusebenza okubucayi okusetshenziselwa ukuhlaziya inani. I-SPE iqinile, isebenzela uxhaxha olubanzi lwezinhlobo zamasampula, futhi imikhiqizo emisha ye-SPE nezindlela ziyaqhubeka nokuthuthukiswa. Umnyombo wokuthuthukisa lezo zindlela ukwazisa ukuthi noma igama elithi “chromatography” lingaveli egameni lenqubo, i-SPE nokho iwuhlobo lokuhlukaniswa kwe-chromatographic.

I-SPE: I-Chromatography Ethule

Kunesisho sakudala esithi “uma isihlahla siwela ehlathini, kungekho muntu osizwayo, siyawukhipha umsindo na?” Leso sisho sisikhumbuza i-SPE. Lokho kungase kubonakale kuyinqaba ukukusho, kodwa uma sicabanga nge-SPE, umbuzo uba “uma ukuhlukana kwenzeka futhi kungekho mtshina lapho ongawuqopha, ingabe i-chromatography yenzeka ngempela?” Endabeni ye-SPE, impendulo ingu-"yebo!" Lapho usungula noma uxazulula inkinga indlela ye-SPE, kungasiza kakhulu ukukhumbula ukuthi i-SPE imane iyichromatography ngaphandle kwechromatogram. Uma ucabanga ngakho, wayengeyena yini uMikhail Tsvet, owaziwa ngokuthi “uyise we-chromatography,” wenza lokho esingakubiza ngokuthi “SPE” namuhla? Lapho ehlukanisa izingxube zakhe zombala wezitshalo ngokuvumela amandla adonsela phansi ukuba awathwale, ancibilike entweni encibilikayo, ngoshoki ophansi, ingabe kwakuhluke kakhulu kunendlela yesimanje ye-SPE?

Ukuqonda Isampula Yakho

Njengoba i-SPE isekelwe ezimisweni ze-chromatographic, enhliziyweni yazo zonke izindlela ezinhle ze-SPE ubuhlobo phakathi kwabahlaziyi, i-matrix, isigaba esimile (i-SPE sorbent), nesigaba seselula (izincibilikisi ezisetshenziselwa ukugeza noma ukukhipha isampula) .

Ukuqonda imvelo yesampula yakho ngangokunokwenzeka kuyindawo engcono kakhulu yokuqala uma kufanele uthuthukise noma uxazulule indlela ye-SPE. Ukuze ugweme ukuzama okungadingekile kanye nephutha ngesikhathi sokuthuthukiswa kwendlela, izincazelo zezakhiwo zomzimba nezamakhemikhali zabo bobabili abahlaziyi bakho kanye ne-matrix ziwusizo kakhulu. Uma usuwazi ngesampula yakho, uzoba sesimweni esingcono sokufanisa leyo sampuli nomkhiqizo ofanelekile we-SPE. Isibonelo, ukwazi i-polarity ehlobene yabahlaziyi uma kuqhathaniswa nomunye nomunye kanye ne-matrix kungakusiza unqume ukuthi ukusebenzisa i-polarity ukuhlukanisa abahlaziyi ku-matrix kuyindlela efanele yini. Ukwazi ukuthi ingabe abahlaziyi bakho abathathi hlangothi noma bangaba khona ezifundeni ezikhokhiswayo kungasiza futhi ukukuqondisa emikhiqizweni ye-SPE egxile ekubambeni noma ekuvezeni okungathathi hlangothi, okunezinhlamvu ezinhle, noma izinhlobo ezinecala elibi. Le mibono yomibili imelela izici ezimbili ze-analyte ezivame ukusetshenziswa kakhulu ukuze kuthuthukiswe lapho kwakhiwa izindlela ze-SPE nokukhetha imikhiqizo ye-SPE. Uma ungakwazi ukuchaza abahlaziyi bakho kanye nezingxenye ezivelele ze-matrix kule migomo, usendleleni eya ekukhetheni isiqondiso esihle sokuthuthukiswa kwendlela yakho ye-SPE.

Ukwehlukana Ngokuzwana

Izimiso ezichaza ukuhlukana okwenzeka ngaphakathi kwekholomu ye-LC, ngokwesibonelo, ziyasebenza ekuhlukaniseni kwe-SPE. Isisekelo sanoma yikuphi ukuhlukaniswa kwe-chromatographic sisungula isistimu enamazinga ahlukahlukene okusebenzelana phakathi kwezingxenye zesampula nezigaba ezimbili ezikhona kukholomu noma i-cartridge ye-SPE, isigaba seselula kanye nesigaba esimile.

Esinye sezinyathelo zokuqala zokuzizwa ukhululekile ngokuthuthukiswa kwendlela ye-SPE ukujwayelana nezinhlobo ezimbili ezivame ukuhlangatshezwana nazo ezisetshenziswa ekuhlukaniseni kwe-SPE: i-polarity kanye/noma isimo sokushaja.

I-polarity

Uma uzosebenzisa i-polarity ukuhlanza isampula yakho, enye yezinketho zokuqala okufanele uzenze ukunquma ukuthi iyiphi "imodi" engcono kakhulu. Kungcono kakhulu ukusebenza nge-SPE ephakathi nendawo emaphakathi kanye nesigaba seselula esingeyona i-polar (okungukuthi imodi evamile) noma okuphambene, indawo ye-SPE engeyona ingxenye ehlangene ehlanganiswe nesigaba seselula esinendawo epholile (okungukuthi imodi ehlehlisiwe, eqanjwe kanjalo nje ngoba iphambene. "yemodi evamile" esungulwe ekuqaleni).

Njengoba uhlola imikhiqizo ye-SPE, uzothola ukuthi izigaba ze-SPE zikhona ezinhlobonhlobo ze-polarity. Ngaphezu kwalokho, ukukhetha kwe-solvent yesigaba seselula nakho kunikeza izinhlobonhlobo ze-polarity, evame ukufundeka kakhulu ngokusebenzisa izixube zezincibilikisi, amabhafa, noma ezinye izithasiselo. Kunezinga elikhulu le-finesse elingenzeka uma usebenzisa umehluko we-polarity njengesici esiyinhloko ongasisebenzisa ukuze uhlukanise abahlaziyi bakho kusukela ekuphazamisekeni kwe-matrix (noma kusuka komunye nomunye).

Vele ukhumbule isisho samakhemikhali sakudala esithi “njengokuncibilika njengokufana” lapho ubheka i-polarity njengomshayeli wokuhlukana. Uma inhlanganisela ifana kakhulu ne-polarity yesigaba esihambayo noma esimile, maningi amathuba okuba ihlanganyele ngokuqinile. Ukusebenzisana okuqinile nesigaba esimile kuholela ekugcinweni okude endaweni ye-SPE. Ukusebenzisana okuqinile nesigaba seselula kuholela ekugcinweni okuncane kanye nokubonakala kwangaphambi kwesikhathi.

Isimo sokushaja

Uma abahlaziyi bezithakazelo behlala bekhona endaweni ekhokhisiwe noma bekwazi ukubekwa esimweni sokukhokhiswa yizimo zesixazululo abachithwa ngaso (isb. i-pH), bese kuba enye indlela enamandla yokuwahlukanisa ne-matrix (noma ngayinye). enye) ngokusebenzisa imidiya ye-SPE engabaheha ngezindleko zabo.

Kulokhu, imithetho yakudala yokukhanga i-electrostatic iyasebenza. Ngokungafani nokuhlukaniswa okuncike ezicini ze-polarity kanye nemodeli "efana nencibilika njenge" yokusebenzisana, ukusebenzisana kwezwe okukhokhisiwe kusebenza emthethweni wokuthi "okuphikisayo kuheha." Isibonelo, ungaba nendawo ye-SPE enenkokhiso eqondile endaweni yayo. Ukuze ulinganisele leyo ndawo ene-positive charge, ngokuvamile kukhona uhlobo olushajwa kabi (i-anion) oluboshelwe kuyo ekuqaleni. Uma i-analyte yakho eshajwa kabi yethulwa ohlelweni, inamandla okukhipha i-anion eboshwe ekuqaleni futhi ixhumane nendawo ye-SPE eshajwe kahle. Lokhu kubangela ukugcinwa komhlaziyi esigabeni se-SPE. Lokhu kushintshwa kwama-anion kubizwa nge-“Anion Exchange” futhi kuyisibonelo esisodwa nje sesigaba esibanzi semikhiqizo ye-SPE ye-“Ion Exchange”. Kulesi sibonelo, izinhlobo zezilwane ezikhokhiswa kahle zizoba nesisusa esinamandla sokuhlala esigabeni samaselula futhi zingasebenzisani nendawo ye-SPE eshajwa kahle, ngakho ngeke zigcinwe. Futhi, ngaphandle uma indawo ye-SPE inezinye izici ngaphezu kwezakhiwo zayo zokushintshisana nge-ion, izinhlobo ezingathathi hlangothi nazo zizogcinwa kancane (yize, imikhiqizo enjalo ye-SPE ehlanganisiwe ikhona, ekuvumela ukuthi usebenzise ukushintshaniswa kwe-ion kanye nezindlela zokugcina isigaba esihlehlisiwe endaweni efanayo ye-SPE. ).

Umehluko obalulekile okufanele ukhunjulwe lapho kusetshenziswa izindlela zokushintshanisa i-ion uhlobo lwesimo sokushaja somhlaziyi. Uma i-analyte ihlale ikhokhiswa, kungakhathaliseki ukuthi i-pH yesisombululo ikuyo, ibhekwa njengezinhlobo "eziqinile". Uma i-analyte ikhokhiswa kuphela ngaphansi kwezimo ezithile ze-pH, kubhekwa njengezinhlobo "ezibuthakathaka". Leso isici esibalulekile okufanele usiqonde mayelana nabahlaziyi bakho ngoba sizonquma ukuthi yiluphi uhlobo lwemidiya ye-SPE okufanele ulusebenzise. Ngokuvamile, ukucabanga ngezinto eziphikisanayo ezihamba ndawonye kuzosiza lapha. Kutuswa ukuhlanganisa i-ion exchange SPE sorbent ebuthakathaka nezinhlobo "eziqinile" kanye ne-sorbent yokushintshanisa i-ion eqinile ne-analyte "ebuthakathaka".