I-Chlorophyll i-pigment eluhlaza etholakala ezitshalweni eziphakeme kanye nazo zonke ezinye izinto eziphilayo ze-photosynthetic. I-Chlorophyll A ne-chlorophyll B iyancibilika ku-ethanol, i-ether kanye ne-acetone nezinye izincibilikisi, ezingancibiliki emanzini nase-petroleum ether, ngakho-ke, zingasebenzisa izincibilikisi ze-polar njenge-acetone, i-methanol, i-ethanol, i-ethyl acetate nokunye ukukhipha i-chlorophyll.
Inqubo yokukhipha i-chlorophyll:
Ukukhishwa kwe-Chlorophyll kulungiselelwa endaweni-emnyama engu-25℃C. Izinyathelo zokuyikhipha zimi kanje:
(1) Thatha amagremu ayi-1000 wamaqabunga aluhlaza bese uwachoboza ku-Wechsler.
(2) Faka amagremu angu-1000 amaqabunga aluhlaza okotshanisiwe ku-acetone enenani elincane le-calcium carbonate (izinga lokushisa elingu-20℃) ukuze likhishwe, kuze kube yilapho izingcezu zamaqabunga zingenambala ngemva kokuhlunga nokuhlanza.
(3) Faka ukukhishwa kwe-acetone ehlungiwe kufaneli equkethe ilitha elingu-1 le-petroleum ether kanye no-100mL we-acetone, bese uphendukisa ngobumnene, kuyilapho ungeza amanzi ahlutshiwe kuze kube yi-stratified. Iningi le-acetone namanzi{4}}okungcola okuncibilikayo kungqimba lwamanzi kuyalahlwa, kusale isisombululo se-petroleum ether kuphela.
(4) Ngemva kokuthi ikhambi le-petroleum ether lihlanjululwe futhi ngamanzi acwecwe, lihlanzwa izikhathi ezingaphezu kwezi-5 ngesisombululo se-methanol esingu-200ml80% esiqukethe i-petroleum ether kanye ne-0.01g oxalic acid, futhi ekugcineni kutholwe ukumiswa okuphuzi{5}okuluhlaza.
(5) Yomisa ukumiswa nge-anhydrous sodium sulfate, bese uyishutheka kukholomu eyenziwe ngempushana ye-sucrose engu-3cm. Bese uhlanza ama-pigments anciphile ukuze ususe ama-carotenoid futhi uwenze aqukethe i-chlorophyll yemvelo kuphela.
(6) Ikholomu ye-Sucrose equkethe i-chlorophyll yemvelo ihlukaniswe izingqimba ezimbili, ungqimba oluluhlaza lunongqimba lwe-chlorophyll B olungu-4-10mm, olunye ungqimba oluluhlaza okwesibhakabhaka luyi-2-6mm chlorophyll A.
(7) Ingxenye etholakala phakathi nongqimba oluluhlaza okwesibhakabhaka (cishe ingxenye yongqimba oluluhlaza okwesibhakabhaka) ifakwa ku-ether, ukumiswa kuyahlungwa futhi kukhishwe, kuhlanzwe ngamanzi acwecwe, omiswe nge-sodium sulfate, bese kuhlungwa ngezitsha ukuze kutholwe i-chlorophyll A.
(8) Khipha ingxenye emaphakathi yongqimba oluluhlaza ku-(6), bese uyifaka ngokushesha ku-ether ukuze ihlungwe futhi iwashwe ukwenza isixazululo se-chlorophyll B ether.
Indlela yokuhlukanisa ngesikhathi sokukhipha?
I-Chromatography iyindlela ebalulekile yokuhlukanisa, ukuhlanzwa kanye nokuhlonza izinhlanganisela eziphilayo, ikakhulukazi ku-microanalysis isetshenziswa kabanzi. Izingulube zezithelo nemifino ikakhulukazi zifaka amafutha-i-soluble carotene, i-lutein, i-chlorophyll namanzi-i-anthocyanin encibilikayo. Ezivivinyweni zokukhipha, singasebenzisa isimiso sokuhlanganisa amanzi afanayo{4}}okuhlunga i-anthocyanin encibilikayo, yona engase isebenzise-ungqimba lwe-chromatography, ikholomu chromatography, ukusebenza okuphezulu kwe-liquid chromatography (HPLC) ekuhlukaniseni i-beta{6}}izinhlobo ze-carotene, i-lutein to the chlorgrity ebuthakathaka ngenxa yezinhlobo ezintathu ze-chlorgrity. futhi, kungaba ngokufanelekile ukukhetha isinyibilikisi esisodwa se-organic noma isilinganiso esihlukile se-solvent exubile njenge-ejenti yokunweba kanye ne-ejenti ye-elution, Izimo ezilungile zokuhlukanisa zanqunywa.
