凝膠層析法分離酪蛋白與核黃素課件

2.GelChromatographySeparateCaseinandRiboflavinbySephadexG-75

Purposebasicoperationofgelchromatography(凝膠層析)

principleofseparatingriboflavin(核黃素)andcaseinbySephadexG-75welcometousethesePowerPointtemplates,NewContentdesign,10yearsexperienceprinciple1.ConceptionGelchromatography,

in

analyticalchemistry,techniqueforseparatingchemicalsubstancesbyexploitingthedifferencesintheratesatwhichtheypassthroughabedofaporous,semisolidsubstance(gel).Themethodisespeciallyusefulforseparatingenzymes,proteins,peptides,andaminoacidsfromeachotherandfromsubstancesoflow

molecularweight.

2.固定相（凝膠）三維空間網(wǎng)狀結(jié)構(gòu)

交聯(lián)劑越多交聯(lián)度越大網(wǎng)孔結(jié)構(gòu)越緊密交聯(lián)劑越少交聯(lián)度越小網(wǎng)孔結(jié)構(gòu)越疏松Sephadex?

Beadsarepreparedbycross-linkingdextran

(葡聚糖)withepichlorohydrin

(環(huán)氧氯丙烷).Widelyusedinindustrialprocesses.Alsousefulindesalting(脫鹽)andbufferexchange.Sephadex的分級范圍

G10<700G15<1,500G251,000~5,000G-501,500~30,000G-753,000~70,000G1004,000~150,000G1505,000~400,000G2005,000~800,000吸水性*G類的型號—每克干凝膠吸水量（ml）x10*如G-75—每克干凝膠吸水量為7.5mlVe-VoKd=————Vi分配系數(shù)Kd

：特點(diǎn)：（1）與被分離物質(zhì)分子的大小和凝膠顆粒孔隙的大小有關(guān)（2）與柱的長短粗細(xì)無關(guān)

每個溶質(zhì)分子在流動相和固定相之間有一個特定的分配系數(shù)(Kd)應(yīng)用脫鹽測分子量分離純化VeAlbuminNaClVeVe優(yōu)點(diǎn)缺點(diǎn)凝膠層析的優(yōu)缺點(diǎn)凝膠是一種不帶電荷的惰性載體，結(jié)構(gòu)穩(wěn)定要求樣品和洗脫液的粘度很低操作條