Chlorophyll yog cov xim ntsuab pom hauv cov nroj tsuag siab dua thiab tag nrho lwm cov kab mob photosynthetic. Chlorophyll A thiab chlorophyll B yog soluble nyob rau hauv ethanol, ether thiab acetone thiab lwm yam kuab tshuaj, insoluble nyob rau hauv dej thiab roj av ether, yog li ntawd, siv tau polar solvents xws li acetone, methanol, ethanol, ethyl acetate thiab thiaj li nyob rau extract chlorophyll.
Chlorophyll extraction txheej txheem:
Chlorophyll extraction yog npaj nyob rau hauv ib nrab tsaus chav tsev ntawm 25 degree C. Cov kauj ruam extraction yog raws li nram no:
(1) Siv 1000 grams ntawm cov nplooj ntsuab tshiab thiab tsoo lawv hauv Wechsler.
(2) Muab cov crushed 1000 grams ntawm nplooj ntsuab rau hauv acetone nrog ib qho me me ntawm calcium carbonate (kub 20 degree) rau kev rho tawm, kom txog thaum cov nplooj tawg tsis muaj xim tom qab lim thiab ntxuav.
(3) Muab cov lim dej acetone tso rau hauv lub qhov dej uas muaj 1 litre roj av ether thiab 100mL ntawm acetone, thiab maj mam tig, thaum ntxiv dej distilled kom txog thaum stratified. Feem ntau ntawm cov acetone thiab dej-soluble impurities nyob rau hauv cov dej txheej yog muab pov tseg, tsuas yog cov roj av ether tov.
(4) Tom qab cov kua roj av ether tau ua kom huv dua nrog dej distilled, nws raug ntxuav ntau dua 5 zaug nrog 200ml80 feem pua methanol tov uas muaj roj av ether thiab 0.01g oxalic acid, thiab thaum kawg daj-ntsuab ncua kev kawm ntawv tau txais .
(5) Qhuav lub suspension nrog anhydrous sodium sulfate, thiab infiltrate rau hauv ib kem ua los ntawm 3cm tuab sucrose hmoov. Tom qab ntawd ntxuav cov pigments precipitated kom tshem tawm carotenoids thiab ua rau lawv tsuas muaj chlorophyll ntuj.
(6) Sucrose kem uas muaj chlorophyll ntuj muab faib ua ob txheej, txheej ntsuab muaj 4-10 hli chlorophyll B txheej, lwm txheej xiav yog 2-6 hli chlorophyll A txheej.
(7) Qhov chaw nyob hauv nruab nrab ntawm cov txheej xiav (li ib nrab ntawm cov txheej xiav) yog muab tso rau hauv ether, qhov raug tshem tawm yog lim thiab muab rho tawm, ntxuav nrog dej distilled, qhuav nrog sodium sulfate, thiab lim nrog cov tais diav kom tau chlorophyll. A.
(8) Tshem tawm qhov nruab nrab ntawm cov txheej ntsuab hauv (6), thiab muab tso rau hauv ether sai sai rau lim thiab ntxuav kom chlorophyll B ether tov.
Yuav ua li cas cais thaum lub sij hawm rho tawm?
Chromatography yog ib txoj hauv kev tseem ceeb rau kev sib cais, purification thiab txheeb xyuas cov organic tebchaw, tshwj xeeb tshaj yog nyob rau hauv microanalysis yog dav siv. Pigments nyob rau hauv txiv hmab txiv ntoo thiab zaub tsuas yog muaj roj-soluble carotene, lutein, chlorophyll thiab dej-soluble anthocyanin. Hauv kev sim extraction, peb tuaj yeem siv lub hauv paus ntsiab lus ntawm cov dej-soluble anthocyanin lim dej zoo sib xws, uas tuaj yeem siv cov txheej nyias nyias chromatography, kem chromatography, kev ua haujlwm siab ua kua chromatography (HPLC) rau kev sib cais ntawm beta-carotene, lutein. thiab chlorophyll, tsis muaj zog vim qhov polarity ntawm peb hom xim nyob rau hauv lem, tuaj yeem tsim nyog xaiv ib qho organic hnyav lossis sib txawv ntawm cov kuab tshuaj sib xyaw ua tus neeg sawv cev nthuav dav thiab elution tus neeg sawv cev, qhov pom kev sib cais tau txiav txim siab.
