Maitiro eChromatography yeChikamu Chitete muFarmasi
Pendauluan
Thin Layer Chromatography (TLC) inzira inoshandiswa zvakanyanya mukuongorora mishonga. Nzira iyi inozivikanwa nekureruka kwayo, kumhanya kwayo, uye kukwanisika kwayo kutenga, pamwe nekukwanisa kwayo kupa ruzivo rwakakosha nezvekugadzirwa kwesampuli. Mumarabhoritari emishonga, TLC inowanzo shandiswa mukutsvagisa nekugadzira mishonga, kuyedza zvinhu zvisina kugadzirwa, uye kudzora mhando yechigadzirwa. Kunyangwe zvishandiso zvemazuva ano zvekuongorora zvakaita seHPLC neLC-MS zviri kushandiswa zvakanyanya, TLC ichiri kukosha nekuda kwekureruka kwayo kushanda uye kugona kwayo senzira yekutanga yekuongorora.
Nheyo dzeKuongorora Kromatography yeChikamu Chitete
Nheyo yeTLC yakavakirwa pakupatsanurwa kwezvikamu zvemusanganiswa zvichibva pakusiyana kwazvo muhukama hwezvikamu zviviri: chikamu chisingachinji uye chikamu chinofamba. Chikamu chisingachinji chinowanzova chikamu chakatete che adsorbent chakadai se silica gel, alumina, kana cellulose yakafukidzwa pagirazi, aruminiyamu, kana purasitiki. Chikamu chinofamba, panguva imwe chete, musanganiswa we solvent kana solvent unofamba nechikamu chisingachinji nekuda kwe capillary action.
Kana sampuro ikaonekwa pasi peplate uye plate yacho ikaiswa mumudziyo uri kukura une chikamu chinofamba, chinhu chinonyungudutsa chinofamba chichienda kumusoro. Zvikamu zvinonyunguduka zvakanyanya muchikamu chinofamba uye zvisingashande zvakanyanya nechikamu chinomira zvichafamba kure, nepo zvikamu zvinobatana zvakasimba nechikamu chinomira zvichachengetwa uye hazvifambi nekukurumidza. Mugumisiro wacho kuparadzaniswa muchimiro chemavara panzvimbo dzakasiyana.
Zvikamu Zvikuru muTLC
Mukurapa, kubudirira kweTLC kunoenderana zvakanyanya nekusarudzwa uye kunaka kwezvikamu zvayo zvikuru:
1. TLC plate
Maplate acho anowanzo putirwa ne polar silica gel (SiO₂). Silica gel yakakodzera kupatsanura mhando dzakawanda dzemishonga, kunyanya idzo dzine polarities dzakasiyana. Maplate e reversed-phase, akadai seC18, anowanikwawo kune mamwe macompounds asiri polar kana mamwe macompounds akaoma kupatsanura muchikamu chenguva dzose.
2. Chikamu chekufambisa (eluent)
Iyo eluent inogona kuva solvent imwe chete kana musanganiswa. Sarudzo yacho inoenderana nehunhu hwemakemikari hweanalyte, kunyanya polarity yayo. Misanganiswa yakajairika inosanganisira chloroform–methanol, ethyl acetate–hexane, kana toluene–acetate. Chiyero chesolvent chinowanzo gadziriswa kuti chiwane kupatsanurwa kwakapinza.
3. Ngarava yekuvandudza
Mudziyo unofanirwa kuvharwa zvakasimba kuti mukamuri muve nehutsi hunonyungudutsa. Kuzara uku kunobatsira kugadzira kukura kwakasimba uye nzvimbo dziri nani.
4. Mienzaniso uye mwero wekuenzanisa
Mukuongorora mishonga, sampuli dzinowanzo enzaniswa nezvinodiwa kuti dzisimbise hunhu kana kuchena. Kuona chiyero nemuenzaniso pandiro imwe chete kunowedzera kuvimbika kwedudziro.
5. Kuonekwa kwenzvimbo
Mavara anogona kuonekwa nemaziso kana akapendwa, kana kushandisa chiedza cheUV (254 nm kana 366 nm). Neimwe nzira, ma reagents ekupfapfaidza akadai se ninhydrin (yeamino acids), Dragendorff (yealkaloids), anisaldehyde sulfate (yea metabolites akasiyana-siyana), kana iodine inogona kushandiswa.
Matanho eKushandiswa kweKLT
Maitiro eTLC mu pharmacy anowanzo sanganisira matanho akakosha anotevera:
1. Kugadzirira ndiro
Kana uchishandisa maplate akagadzirira kushandiswa, danho iri riri nyore. Zvisinei, mune dzimwe nguva, maplate acho angangoda kuvhurwa nekudziisa (semuenzaniso, kusvika ku100–120°C) kubvisa hunyoro hunogona kukanganisa kupatsanurwa.
2. Kuonekwa kwemuenzaniso
Muenzaniso unoonekwa uchishandisa capillary kana micropipette pamutsetse wekutanga, paine daro rakati pakati pemavanga. Mavanga acho anofanira kunge madiki uye akakora kudzivirira kuti nzvimbo yacho isapararire.
3. Kugadzirwa kweChromatogram
Ndiro inoiswa mukamuri riri kukura rine mwero we solvent uri pasi pemutsetse wenzvimbo. Solvent inobvumirwa kukwira kusvika padanho raunoda, ipapo ndiro inobviswa yoomeswa.
4. Kuona nekuona mavara
Mushure mekuoma, mavara acho anoonekwa neUV kana maspray reagents. Nzvimbo dzemapundu dzinozoyerwa kuti paverengwe kukosha kweRf (Retardation factor):
\[
R_f = \frac{\text{spot travel distance}}{\text{solvent front travel distance}}
\]
Kukosha kweRf kunogona kushandiswa sechiratidzo chekuzivikanwa, kunyanya kana kuchienzaniswa nemazinga uye kuchiitwa pasi pemamiriro akafanana ekuongorora.
Zvikumbiro zveTLC muFarmacy
TLC ine mabasa akasiyana-siyana akakosha munyika yemishonga, anosanganisira:
1. Kuzivikanwa kwezvinhu zvekushandisa mumishonga
TLC inowanzo shandiswa kuona kuti zvinhu zvakagadzirwa zvinosangana nezvinodiwa. Semuenzaniso, kuzivikanwa kwechinhu chinoshanda kunogona kuitwa nekuenzanisa Rf yemuenzaniso nechiyero chepamutemo.
2. Kuyedzwa kwekuchena uye kuonekwa kwekuchena
Mukutonga kwemhando, TLC inogona kuratidza kuvapo kwetsvina kana zvigadzirwa zvekuora. Mamwe madonhwe ari kunze kwenzvimbo huru anoratidza kusviba kunogona kuitika.
3. Kutarisa maitiro ekugadzira
Mukutsvaga nekugadzira mishonga, TLC inoshandiswa kutarisa mashandiro emakemikari: kana ma "reactants" achiripo, kana zvigadzirwa zvakaumbwa, uye kana "reaction" yacho yasvika padanho rayo rakanaka.
4. Kuongorora mishonga yemishonga uye mishonga yemiti
TLC inoshandiswa zvakanyanya mukuongorora zvinotorwa mumiti yemishonga. Mamwe ma pharmacopoeias anonyorawo nzira dzeTLC dzekuziva makemikari ezviratidzo mumiti.
5. Bvunzo iri nyore yekugadzikana
Muzvidzidzo zvekutanga zvekugadzikana, TLC inogona kubatsira kuona kuumbwa kwezvigadzirwa zvekuora nekuda kwekupisa, chiedza, kana hunyoro.
Zvakanakira uye Zvisina Kunaka zveTLC
Keunggulan
- Inokurumidza uye isingadhuri zvichienzaniswa nenzira dzakaoma dzekushandisa.
- Zviri nyore kuita uye hazvidi michina inodhura.
- Inochinjika, nekuti inogona kuparadzanisa mhando dzakawanda dzemakemikari ane kusiyana muzvikamu zve eluents uye zvikamu zvisingachinji.
- Kuongorora kunobudirira, kwakakodzera matanho ekutanga ekuongorora kana kutsvagisa.
Miganhu
– Kuberekazve dzimwe nguva kwakashata kana mamiriro ekunze asina kudzorwa zvakanyanya (kuwanda kwemidziyo, hunyoro, kunaka kwezvinonyungudutsa).
- Kuyera huwandu hwemasero (quantification) kwakaganhurirwa, kunyangwe kuchigona kuvandudzwa nekushandisa densitometry (TLC Scanner).
– Kunzwa kunowanzova kwakaderera pane HPLC kana LC-MS.
Kufambira Mberi Kwemazuva Ano: HPTLC
Sekuvandudzwa kweTLC, High-Performance Thin-Layer Chromatography (HPTLC) inozivikanwa. HPTLC inoshandisa maplate ane zvidimbu zvakapfava uye layer yakafanana, zvichikonzera kupatsanurwa kwakapinza uye kwakaoma kunogona kuongororwa nehuwandu uchishandisa densitometric scanners. Muindasitiri yemishonga, HPTLC inopa chiyero pakati pekureruka kweTLC uye kunyatsojeka kwenzira dzemazuva ano dzekushandisa.
Mhedziso
Thin Layer Chromatography (TLC) ichiri nzira yakakosha yekuongorora muindasitiri yemishonga nekuda kwekureruka kwayo, kumhanya kwayo, uye kuchinjika kwayo. TLC inobatsira pakuziva, kuyedza kuchena, kutarisa kugadzirwa kwezvinhu, uye kuongorora zvigadzirwa zvechisikigo. Pasinei nemiganhu muhuwandu uye kunzwisisika, TLC inoramba iri sarudzo huru yekuongorora nekuyedza nguva dzose. Nekuvandudzwa kweHPTLC, kugona kweTLC kwakawedzera, zvichiita kuti ikwanise kusangana nezvinodiwa zvekuongorora mishonga kwakasimba uye kwakajairika.
Kana muchida, ndinogona kuwedzera mienzaniso yezvidzidzo (semuenzaniso, kuziva paracetamol, caffeine, kana mamarker compounds mu turmeric) pamwe chete nemazano ekufamba-famba uye nzira dzekuona zvinhu dzinowanzoshandiswa murabhoritari.
