Nā ʻenehana Chromatography Layer Thin ma ka lāʻau lapaʻau
Pendahuluan
ʻO ka Thin Layer Chromatography (TLC) kahi ʻenehana loiloi maʻamau i hoʻohana ʻia i ka ʻoihana lāʻau lapaʻau. Ua ʻike ʻia kēia ʻano hana no kona maʻalahi, wikiwiki, a me ke kumukūʻai kūpono, a me kona hiki ke hāʻawi i ka ʻike qualitative koʻikoʻi e pili ana i ka haku mele ʻana o kahi laʻana. I loko o nā keʻena hoʻokolohua lāʻau lapaʻau, hoʻohana pinepine ʻia ʻo TLC i ka noiʻi a me ka hoʻomohala ʻana i nā lāʻau lapaʻau, ka hoʻāʻo ʻana i nā mea maka, a me ka kaohi maikaʻi o ka huahana i hoʻopau ʻia. ʻOiai ke hoʻohana nui ʻia nei nā mea hana loiloi hou e like me HPLC a me LC-MS, pili mau ʻo TLC ma muli o kona maʻalahi o ka hana a me kona hiki ke lilo i ʻano loiloi mua.
Nā Kumumanaʻo Kumu o ka Chromatography Papa Lahilahi
Hoʻokumu ʻia ke kumumanaʻo o TLC ma ka hoʻokaʻawale ʻana o nā ʻāpana o kahi hui ʻana ma muli o ko lākou ʻokoʻa i ka pilina no ʻelua mau pae: kahi pae paʻa a me kahi pae neʻe. ʻO ka pae paʻa he papa lahilahi ia o ka adsorbent e like me ka silica gel, alumina, a i ʻole cellulose i uhi ʻia ma luna o ke aniani, alumini, a i ʻole ka pā palakiko. ʻO ka pae neʻe, ʻoiai, he hui solvent a solvent paha e neʻe ana ma o ka pae paʻa ma muli o ka hana capillary.
Ke ʻike ʻia kahi hāpana ma lalo o kahi pā a waiho ʻia ka pā i loko o kahi ipu e ulu ana me kahi pae neʻe, e hele ka mea hoʻoheheʻe i luna. ʻO nā ʻāpana i ʻoi aku ka hoʻoheheʻe ʻia i loko o ka pae neʻe a launa pū iki me ka pae paʻa e hele mamao aku, ʻoiai ʻo nā ʻāpana e launa pū ikaika me ka pae paʻa e paʻa a hele wikiwiki iki. ʻO ka hopena, he kaʻawale ma ke ʻano o nā kiko ma nā wahi like ʻole.
Nā ʻĀpana Nui ma TLC
I ka hana lāʻau lapaʻau, ʻo ka holomua o TLC e hilinaʻi nui ʻia i ke koho ʻana a me ka maikaʻi o kāna mau ʻāpana nui:
1. Pā TLC
ʻO ka maʻamau, ua uhi ʻia nā papa me ka polar silica gel (SiO₂). He kūpono ka silica gel no ka hoʻokaʻawale ʻana i nā ʻano hui lāʻau lapaʻau he nui, ʻoiai nā mea me nā polarities like ʻole. Loaʻa pū nā papa i hoʻohuli ʻia, e like me C18, no nā hui nonpolar a i ʻole kekahi mau hui i paʻakikī ke hoʻokaʻawale i ka pae maʻamau.
2. Pae lawe lima (eluent)
Hiki i ka eluent ke lilo i hoʻokahi mea hoʻoheheʻe a i ʻole he hui. Aia ke koho i nā waiwai kemika o ka analyte, ʻoiai kona polarity. ʻO nā hui maʻamau e komo pū me ka chloroform-methanol, ethyl acetate-hexane, a i ʻole toluene-acetate. Hoʻonohonoho pinepine ʻia ka lakio o ka mea hoʻoheheʻe e hoʻokō i nā hoʻokaʻawale ʻoi.
3. Moku hoʻomohala
Pono e pani paʻa ʻia ka ipu i mea e hoʻopiha ʻia ai ke keʻena me ka mahu hoʻoheheʻe. Kōkua kēia saturation i ka hoʻoulu ʻana i ka ulu mau a me nā kiko maikaʻi aʻe.
4. Nā laʻana a me nā kūlana hoʻohālikelike
I ka loiloi lāʻau lapaʻau, hoʻohālikelike pinepine ʻia nā laʻana me nā kūlana e hōʻoia i ka ʻike a i ʻole ka maʻemaʻe. ʻO ka ʻike ʻana i ke kūlana a me ka laʻana ma ka pā hoʻokahi e hoʻonui ai i ka hilinaʻi o ka wehewehe ʻana.
5. Ka ʻike ʻana i nā kiko
Hiki ke ʻike maka ʻia nā kiko inā he kala ko lākou, a i ʻole e hoʻohana ana i ke kukui UV (254 nm a i ʻole 366 nm). Ma kekahi ʻaoʻao, hiki ke hoʻohana ʻia nā reagents pīpī e like me ka ninhydrin (no nā waikawa amino), Dragendorff (no nā alkaloids), anisaldehyde sulfate (no nā metabolites like ʻole), a i ʻole iodine.
Nā Pae o ka Hoʻokō ʻana o KLT
ʻO ke kaʻina hana TLC ma ka hale kūʻai lāʻau lapaʻau e pili ana i nā ʻanuʻu koʻikoʻi penei:
1. Hoʻomākaukau ʻana i ka pā
Inā e hoʻohana ana i nā papa mākaukau e hoʻohana, he maʻalahi kēia hana. Eia nō naʻe, i kekahi mau hihia, pono paha e hoʻāla ʻia nā papa ma ka hoʻomehana ʻana (e laʻa, a i 100–120°C) e wehe i ka makū e hiki ke hoʻopilikia i ka hoʻokaʻawale ʻana.
2. Ka ʻike ʻana i nā laʻana
Hoʻohana ʻia ke kaulahao no ke kaha kiʻi ʻana i ka laʻana me ka hoʻohana ʻana i kahi capillary a i ʻole micropipette ma ka laina hoʻomaka, me kahi mamao kikoʻī ma waena o nā kiko. Pono e liʻiliʻi a mānoanoa nā kiko e pale aku ai i ka laha ʻana o ke kiko.
3. Hoʻomohala ʻana i ka Chromatogram
Hoʻokomo ʻia ka pā i loko o ke keʻena hoʻomohala me ka pae solvent ma lalo o ka laina kiko. ʻAe ʻia ka solvent e piʻi i ka pae i makemake ʻia, a laila wehe ʻia ka pā a hoʻomaloʻo ʻia.
4. Ka nānā ʻana a me ka ʻike ʻana i nā kiko
Ma hope o ka maloʻo ʻana, ʻike ʻia nā kiko me nā reagents UV a i ʻole nā mea pīpī. A laila ana ʻia nā kūlana kiko e helu ai i ka waiwai Rf (Retardation factor):
\[
R_f = \frac{\text{mamao huakaʻi wahi}}{\text{mamao huakaʻi mua o ka mea hoʻoheheʻe}}
\]
Hiki ke hoʻohana ʻia ka waiwai Rf ma ke ʻano he palena ʻike, ʻoiai ke hoʻohālikelike ʻia me nā kūlana a lawe ʻia ma lalo o nā kūlana loiloi like.
Nā noi TLC ma ka hale kūʻai lāʻau lapaʻau
He nui nā noi koʻikoʻi o TLC i ka honua lāʻau lapaʻau, e like me:
1. Ka ʻike ʻana i nā mea maka no nā lāʻau lapaʻau
Hoʻohana pinepine ʻia ka TLC e hōʻoia i ka hoʻokō ʻana o nā mea maka i loaʻa i nā kikoʻī. No ka laʻana, hiki ke hana ʻia ka ʻike ʻana i kahi hui hana kikoʻī ma ka hoʻohālikelike ʻana i ka Rf o ka laʻana i kahi kūlana kūhelu.
2. Hoʻāʻo maʻemaʻe a me ka ʻike ʻana i ka haumia
I ke kaohi ʻana i ka maikaʻi, hiki i ka TLC ke hōʻike i ke alo o nā haumia a i ʻole nā huahana hōʻino. ʻO nā kiko keu ma mua o ke kiko mua e hōʻike ana i nā haumia hiki.
3. Ke nānā ʻana i ke kaʻina hana synthesis
I ka noiʻi a me ka hoʻomohala ʻana i nā lāʻau lapaʻau, hoʻohana ʻia ʻo TLC e nānā i nā hopena kemika: inā paha e loaʻa ana nā reactants, inā paha i hoʻokumu ʻia nā huahana, a i ka wā e hiki ai ka hopena i kona wahi kūpono loa.
4. Ka loiloi phytopharmaceutical a me nā mea kanu
Hoʻohana nui ʻia ka TLC i ka nānā ʻana i nā mea kanu lāʻau lapaʻau. Hoʻopaʻa pū kekahi mau pharmacopoeias i nā ʻano TLC no ka ʻike ʻana i nā pūhui māka i nā huahana lāʻau.
5. Hoʻāʻo kūpaʻa maʻalahi
I nā haʻawina paʻa mua, hiki i ka TLC ke kōkua i ka ʻike ʻana i ka hoʻokumu ʻia ʻana o nā huahana palaho ma muli o ka wela, ka mālamalama, a i ʻole ka makū.
Nā Pōmaikaʻi a me nā Palena o TLC
Hanohano
- Wikiwiki a hoʻokele waiwai hoʻi i ka hoʻohālikelike ʻia me nā ʻano hana hana paʻakikī.
- He maʻalahi ke hana a ʻaʻole pono nā lako pipiʻi.
– Hiki ke loli, no ka mea, hiki iā ia ke hoʻokaʻawale i nā ʻano hui he nui me nā ʻano like ʻole o nā eluents a me nā pae paʻa.
– Ka nānā pono ʻana, kūpono no nā pae mua o ka nānā ʻana a i ʻole ka noiʻi ʻana.
Nā Palena
– I kekahi manawa, ʻaʻole maikaʻi ka hana hou ʻana inā ʻaʻole i kāohi pono ʻia nā kūlana (ka saturation o ka moku, ka makū, ka maikaʻi o ka solvent).
- Ua kaupalena ʻia ka helu ʻana, ʻoiai hiki ke hoʻomaikaʻi ʻia me ka densitometry (TLC Scanner).
– ʻOi aku ka haʻahaʻa o ka ʻike ma mua o ka HPLC a i ʻole LC-MS.
Nā Hoʻomohala Hou: HPTLC
Ma ke ʻano he hoʻomohala ʻana o TLC, ua ʻike ʻia ʻo High-Performance Thin-Layer Chromatography (HPTLC). Hoʻohana ʻo HPTLC i nā papa me nā ʻāpana ʻoi aku ka maikaʻi a me kahi papa like, e hopena ana i nā hoʻokaʻawale ʻoi aku ka ʻoi a ʻoi aku ka naʻau i hiki ke kālailai ʻia me ka hoʻohana ʻana i nā scanner densitometric. I ka ʻoihana lāʻau lapaʻau, hāʻawi ʻo HPTLC i kahi kaulike ma waena o ka maʻalahi o TLC a me ka pololei o nā ʻano hana hana hou.
Ka hopena
ʻO ka Thin Layer Chromatography (TLC) ke ʻano hana loiloi koʻikoʻi i ka ʻoihana lāʻau lapaʻau ma muli o kona maʻalahi, wikiwiki, a me ka maʻalahi. He mea pono ka TLC no ka ʻike ʻana, ka hoʻāʻo ʻana i ka maʻemaʻe, ka nānā ʻana i ka synthesis, a me ka nānā ʻana i nā huahana kūlohelohe. ʻOiai nā palena i ka helu ʻana a me ka ʻike, ʻo TLC ke koho mua no ka nānā ʻana a me ka hoʻāʻo ʻana. Me ka hoʻomohala ʻana o HPTLC, ua hoʻonui ʻia nā hiki o TLC, e hiki ai iā ia ke hoʻokō i nā pono o ka loiloi lāʻau lapaʻau ʻoi aku ka paʻakikī a me ke kūlana maʻamau.
Inā makemake ʻoe, hiki iaʻu ke hoʻohui i nā laʻana hihia haʻawina (e laʻa, ka ʻike ʻana i ka paracetamol, caffeine, a i ʻole nā mea hōʻailona i loko o ka turmeric) me nā ʻōlelo paipai no nā pae mobile a me nā ʻano hana ʻike maka i hoʻohana pinepine ʻia ma ka hale hana.
