体外消化实验文档格式.doc

体外消化实验文档格式.doc

《体外消化实验文档格式.doc》由会员分享，可在线阅读，更多相关《体外消化实验文档格式.doc（4页珍藏版）》请在冰点文库上搜索。

Constituentsandconcentrationsofthevarioussyntheticjuicesoftheinvitrodigestionmodelrepresentingfedconditions

Saliva唾液

Gastricjuice胃液

Duodenaljuice十二指肠液

Bilejuice胆汁

Inorganicsolution

10mlKCl89.6g/l

15.7mlNaCl175.3g/l

40mlNaCl175.3g/l

30mlNaCl175.3g/l

10mlKSCN20g/l

3.0mlNaH2PO488.8g/l

40mlNaHCO384.7g/l

68.3mlNaHCO384.7g/l

10mlNaH2PO488.8g/l

9.2mlKCl89.6g/l

10mlKH2PO48g/l

4.2mlKCl89.6g/l

10mlNaSO457g/l

18mlCaCl2·

2H2O22.2g/l

6.3mlKCl89.6g/l

150μlHCl37%g/g

1.7mlNaCl175.3g/l

10mlNH4Cl30.6g/l

10mlMgCl25g/l

20mlNaHCO384.7g/l

6.5mlHCl37%g/g

180μlHCl37%g/g

Organicsolution

8mlurea25g/l

10mlglucose65g/l

4mlurea25g/l

10mlurea25g/l

10mlglucuronicacid（葡萄糖醛酸）2g/l

3.4mlurea25g/l

10mlglucosaminehydrochloride（盐酸氨基葡萄糖）33g/l

Addtomixtureorganic

+inorganicsolution

290mgα-amylase

1gBSA

9mlCaCl2·

10mlCaCl2·

15mguricacid

2.5gpepsin

1.8gBSA

25mgmucin

3gmucin

9gpancreatin

30gbile

1.5glipase

pH

6.8±

0.2

1.30±

0.02

8.1±

8.2±

Theinorganicandorganicsolutionsareaugmentedto500mlwithdistilledwater.Aftermixingoftheinorganicandorganicsolutions,somefurtherconstituentsareaddedanddissolved.Ifnecessary,thepHofthejuicesisadjustedtotheappropriateinterval.

另外，

抗性淀粉含量测定：

1、参考Li等（2008）方法（Characterizationofmaizeamylose-extender（ae）mutantstarches.PartI:

Relationshipbetweenresistantstarchcontentsandmolecularstructures）

2.3.Resistantstarch（RS）content

RScontentsoftheae-mutantstarchsamplesweredeterminedusingtheAOACMethod991.43fortotaldietaryfiber（AOAC,2003）andEnglyst’smethod（1992）forcomparison.

FortheAOAC991.43method,starch（1.0g,dry-starchbasis,dsb）wassuspendedinaMes-trisbuffersolution（0.05M,40ml）andhydrolyzedwith500uofa-amylasefromBacilluslicheniformis（SigmaChemical,Cat.No.A3403）inaboilingwater-bathfor30minwithstir.ThesamplewasthendigestedwithproteasefromBacilluslicheniformis（5mg,SigmaChemical,Cat.No.P3910）at60º

Cinashakerwaterbathfor30min.ThesampledispersionwasadjustedtopH4.4–4.6byaddingHClandthenhydrolyzedwithamyloglucosidase（300U,Sigmachemical,Cat.No.A9913）at60º

Cinashakerwater-bathfor30min.Thedigestedsamplewasfilteredthroughacelitelayerinacrucibleandwashedtwicewith15mlof78%ethanol,twicewith15mlof100%ethanolandrinsedwith15mlofacetone.Theremainingsamplewasdriedinanovenat100º

Covernight.Theresistantstarchcontentwascalculatedusingtheequation:

（%）RScontent=Remainingsampleweight（g,dsb）/initialsampleweight（g,dsb）×

100%

ForRSdeterminedusingEnglyst’smethod（1992）,Starch（1.000g,db）in20mLofsodiumacetatebuffer（0.1M,pH5.2）wascookedinaboilingwater-bathfor30min.Thestarchdispersionwascooleddownto37º

C,mixedwithanenzymesolution（5mL）consistingofpancreatinextractandamyloglucosidase,andincubatedinawater-bathat37º

C.Thepancreatinextractwaspreparedasfollows;

3.0gofpancreatin（Sigma,Cat.No.P7545）wassuspendedin20mLdeionizedwater,stirredfor10minatroomtemperature,andcentrifugedat1500gfor10min.Theenzymesolutionwaspreparedbymixing13.5mlsupernatantofpancreatinextract,210Uamyloglucosidase（Sigma,Cat.No.A7095）,and1.0mLdeionizedwater.Therapiddigestiblestarch（RDS）wasdefinedasthetotalstarchdigestedwithinthefirst20min,andtheslowlydigestiblestarch（SDS）wasthestarchdigestedbetween20and120min（Englystetal.,1992）.Theresistantstarchcontentwascalculatedasfollows:

（%）RScontent=100%×

（totalstarch–RDS-SDS）（g,dsb）/totalstarch（g,dsb）