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2. MATERIAL
Tip, Pippet, Test tube, spectro-20, D.W, 鍮꾩빱, Centrifuge, Fecl3, H2SO4, Ethylacetate ethanol, �궪媛곹뵆由ъ뒪�겕, Tritonx-100, cleanbanch
3. METHOD
1. �삁�븸�쓣 �긽�삩�뿉�꽌 �쓳怨좊맆�븣源뚯�� 湲곕떎由� �썑 �깋�옣怨좎뿉�꽌 �깋媛�
2. �빟 3000rpm 5min媛� �썝�떖 遺꾨━
3. sup留� �뵲濡� 紐⑥���떎.
(1) cholesterol standard curve �옉�꽦
a.
�몴以��슜�븸(1mg/ml)
0.5
1.0
1.5
2.0
2.5
ethylacetate
-etanol(ml)
4.5
4
3.5
3.0
2.5
�뜦�뼱 3000rpm 5min �썝�떖遺꾨━
b. sup. 0.5ml + Fecl3 1ml + H2SO4 1ml�쓣 �꽎�뒗�떎.
c. A550�뿉�꽌 �씉愿묐룄 痢≪젙
d. standard curve �옉�꽦
(2) cholesterol 珥앸웾
a. �삁泥� 0.5ml + ethylacetate ethanol 5ml瑜� �꽎�뒗�떎.
b. 3000rpm, 5遺� �썝�떖遺꾨━
c. sup. 0.5ml + Fecl3 1ml + H2SO4 1ml�쓣 �꽎�뒗�떎.
(3) free cholesterol �뼇 痢≪젙
a. (2) 異붿텧�븸 0.6ml + Tritonx-100 2ml�쓣 �꽎�뒗�떎.
b. 10遺꾧컙 �긽�삩�뿉�꽌 incubation
c. 3000rpm 5min centrifuge
d. pptf,f 1ml acetone 1�쉶 washinl
e. acetone �셿�쟾�엳 �젣嫄고썑 ethylacetate 0.3ml + Fecl3 1ml + H2SO4 1ml�쓣 �꽎�뒗�떎.
f. A550�뿉�꽌 �씉愿묐룄 痢≪젙
g. �뼇 痢≪젙
4���(�깮�왂)
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