LABORATORY CATALOGUE for milk analysis
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CONTENTS Fh[\WY[
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8kjohec[jh_YZ[j[hc_dWj_ede\\WjYedj[djWYYehZ_d]je:h$D$=[hX[hÊic[j^eZ ') A detailed description by graduate chemist Alfred Toepel 8kjohec[jh_YZ[j[hc_dWj_ede\j^[\WjYedj[dje\lWh_ekiZW_hofheZkYji Determination of fat content in cream, ice cream, cheese, butter, milk powder, etc.
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:[j[hc_dWj_ede\\WjYedj[djXkjohec[j[h The butyrometer – an overview of the entire product spectrum
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:[j[hc_dWj_ede\\WjYedj[djWYY[iieh_[i Implements and utensils used in the determination of fat content
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TRADITION PROGRESS CONTINUITY ;jc`Z"9g#C#
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PRODUCTS: J^[YecfWdoZ[l[befi"cWdk\WYjkh[iWdZcWha[ji j^[\ebbem_d][gk_fc[djWdZWYY[iieh_[imehbZm_Z[0 6aaZfj^ebZciVcYVXXZhhdg^Zh[dgi]Z„Gerber method of determining
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Founded: 1904 Managing director: Graduate engineer Konrad Schaefer Authorised signatory: graduate economist Georg Hoernle
Funke-Dr.N.Gerber Labortechnik GmbH Ringstraße 42 12105 Berlin Telephone: (+49-30) 702 006-0 Fax: (+49-30) 702 006-66 E-mail: [email protected] Website: www.funke-gerber.de
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BUTYROMETRIC DETERMINATION OF FAT CONTENT ACCORDING TO GERBER’S METHOD by graduate chemist Alfred Toepel
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PRINCIPLES OF THE METHOD I]Z YZiZgb^cVi^dc d[ [Vi XdciZci VXXdgY^c\ id
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APPLICATION I]^hegdXZhhXVcWZjhZY[dgjcigZViZYVcYeVhiZjg^oZYb^a`l^i]V[ViXdciZcid[%"&+[dgb^a`l]^X]XdciV^chVhj^iVWaZ egZhZgkVi^kZVhlZaaVh[dg]dbd\Zc^oZYb^a`#
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• colourless or only slightly discoloured and free from any substances which might influence the outcome EaZVhZcdiZ/ The required density corresponds to 90 to 91 % by mass. Stronger or weaker concentrations are to be avoided. At 65°C, more highly concentrated sulphuric acid attacks the amyl alcohol, producing olefins through dehydration which influence the result. Weaker concentrations reduce the oxidization effect. Destruction of the fat globule coating is incomplete which can lead to the formation of lumps.
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REQUIRED EQUIPMENT
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PREPARATION OF THE TEST SPECIMEN I]Zb^a`^ci]ZheZX^bZcWdiiaZ^h]ZViZYjeid'%8VcYi]dgdj\]anb^mZYi]gdj\] \ZciaZh]V`^c\#I]^h^hYdcZidWg^c\VWdjiVcZkZcY^hig^Wji^dcd[[ViVcYidegZkZci [gdi]^c\VcYi]Z[dgbVi^dcd[WjiiZg# Milk fat is lighter than water and creams if it is left standing. A fat-rich layer accumulates on the surface. Stirring and careful shaking restore the original distribution. >[i]ZaVnZgd[XgZVbXVccdiWZZkZcanY^hig^WjiZY^ci]^hlVn!i]Zb^a`h]djaYWZhadlan ]ZViZY id (*")%8 VcY \Zcian hl^gaZY VgdjcY jci^a V ]dbd\Zcdjh [Vi Y^hig^Wji^dc ^h VX]^ZkZY#I]Zb^a`^hi]ZcXddaZYid'%8WZ[dgZWZ^c\YgVlc^cidi]Ze^eZiiZ# Foam breaks the fat globule coating. The milk may begin to turn into butter when stirred and uniform distribution of fat is no longer possible. The fat liquefies at 35-40° and the distribution process is speeded up. 6[iZgi]ZiZbeZgVijgZ]VhWZZchZi!i]Zb^a`XVchiVcY[dg(id)b^cjiZhidVaadlVcn edX`Zihd[V^gidY^heZghZ# The volumeters are calibrated at 20°C. Any variations in temperature will influence the volume. Air pockets reduce the density and hence the mass of milk measured.
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CONDUCTING A TEST = WORK PROCEDURES I]ZhVbZb^a`heZX^bZcbjhiWZiZhiZYil^XZ# EaVXZ ' WjingdbZiZgh ^cid V XaVbe WjingdbZiZg hiVcY# L^i]i]ZV^Yd[VbZVhjg^c\iVe!^cigdYjXZ&%bad[hjae]jg^XVX^Y ^cidi]ZWjingdbZiZgl^i]djilZii^c\i]ZcZX`d[i]ZWjingdbZiZg hZZ;^\#&# '# 8VgZ[jaan ijgc i]Z WdiiaZ l^i] i]Z b^a` heZX^bZc jeh^YZ Ydlc i]gZZ dg [djg i^bZh# I]Zc ^bbZY^ViZan e^eZiiZ &%#,* ba d[ b^a` ^cid i]Z WjingdbZiZg ^c hjX] V lVn i]Vi i]Z WjingdbZiZg cZX` YdZh cdi \Zi lZi VcY i]Z b^a` YdZh cdi b^m l^i] i]Z hjae]jg^X VX^Y# Id Yd hd! aZVc i]Z i^e d[ i]Z e^eZiiZ aViZgVaan Vh YZZean Vhedhh^WaZV\V^chii]ZlVaad[i]ZWjingdbZiZghdi]Vii]Zb^a` [dgbhVaVnZgdcided[i]Zhjae]jg^XVX^YhZZ;^\#'# When the Gerber method was first introduced, 11.0 ml of milk were used. By reducing the quantity of milk 10.75 ml, the determined fat content more closely matches the results of the reference method. If the butyrometer neck is wetted with milk, residues may cling to it. A clear dividing line between the acid and the milk, without a brownishcoloured edge, is the sign of good layering.
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Due to the low density of amyl alcohol, the two liquids do not mix.
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*# I]ZWjingdbZiZg^heaVXZY^ci]ZWjingdbZiZgijWZl^i]i]ZWjaW ed^ci^c\YdlclVgYh#H]V`Zi]ZWjingdbZiZgk^\dgdjhanjci^ai]Z ilda^fj^YhVgZi]dgdj\]anb^mZY#@ZZendjgi]jbWÒgbanegZhhZY Ydlcdci]ZWjingdbZiZghideeZgl]^aZYd^c\hd#Ijgci]ZWjin" gdbZiZgjeVcYYdlchZkZgVai^bZh^cdgYZgidZcVWaZi]Zhjae]j" g^XVX^YgZbV^c^c\^ci]ZWjaWidY^heZghZhZZ;^\#(# When the liquids are mixed, a considerable amount of heat is given off. The gas built up in this way can cause the stopper to shoot out, or the butyrometer to break. The butyrometer tube is used as a safety precaution. Instead of using a butyrometer tube, the butyrometer can be wrapped in a cloth.
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Too lax shaking of the butyrometer or holding it unnecessarily in a slanted position inhibits quick mixing and therefore the rapid oxidation of the entire specimen, thus ruining the careful work done layering the liquid.
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RESULT AND DEGREE OF ACCURACY -# 6[iZggZbdk^c\i]ZWjingdbZiZg[gdbi]ZlViZgWVi]!^ih]djaYWZ]ZaYkZgi^XVaan ViV]Z^\]il]ZgZi]ZbZc^hXjhd[i]ZXdajbcd[[Vi^hViZnZaZkZa#L^i]i]Z]Zae d[i]ZhideeZg!bVg`i]ZYZbVgXVi^dca^cZWZilZZci]ZgZh^YjVab^mijgZVcY[Vi dcVl]daZhjWY^k^h^dcd[i]ZWjingdbZiZghXVaZVcYgZVYd[[i]Z]Z^\]id[i]Z [ViXdajbcVii]ZadlZhied^cid[i]ZbZc^hXjh#>[i]ZgZVY^c\iV`Zhiddadc\!i]Z WjingdbZiZgbjhiWZeaVXZYWVX`^cidi]ZlViZgWVi]hZZ;^\h#+VcY,# >[i]ZbZc^hXjhd[i]Z[ViXdajbc^hcdiViZnZaZkZa!VeVgVaaVmZggdggZhjaih#
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BUTYROMETRIC DETERMINATION OF THE FAT CONTENT OF VARIOUS DAIRY PRODUCTS FOREWORD:
The butyrometric determination of the fat content in milk has been and is being replaced at a progressive rate by other routine tests (with appliances such as LactoStar, for instance). However, milk products such as cheese, ice cream, etc. either cannot be tested with such appliances or can only be tested after elaborate specimen preparation. For these products, butyrometric methods are a good alternative for routine analysis.
1.0 FIELD OF APPLICATION 9ZiZgb^cVi^dcd[[ViXdciZci^cb^a`VcYkVg^djhYV^gnegdYjXih#
2.0 VOLUMES JcaZhhhiViZYdi]Zgl^hZ!i]Z[daadl^c\fjVci^i^Zhd[X]Zb^XVahVcYiZhiheZX^bZch VgZjhZY/ Hjae]jg^XVX^Y/ 6bnaVaXd]da/ B^a`dgYV^gnegdYjXi/
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3.0 BRIEF DESCRIPTION OF THE BUTYROMETRIC DETERMINATION OF FAT CONTENT: 3.1 ... IN MILK (ACCORDING TO GERBER’S METHOD): EZg[ZXian XaZVcZY b^a` WjingdbZiZgh! bdhi ^bedgiVcian [gZZ d[ [Vi gZh^YjZh! VgZ ÒaaZY ^ci]Z[daadl^c\dgYZg/&%bad[hjae]jg^XVX^YYZch^in/-&- $"%#%%(\$ba!&%#,*ba d[b^a`!VcY&bad[VbnaVaXd]da#I]Zb^a`VcYVbnaVaXd]daVgZidWZÒaaZY^caVnZgh hdi]Vii]ZnYdcdib^mWZ[dgZh]V`^c\#6[iZghZVa^c\i]ZWjingdbZiZg!i]ZXdciZcihVgZ i]dgdj\]anb^mZYWnh]V`^c\VcYijgc^c\i]ZWjingdbZiZgjeh^YZYdlchZkZgVai^bZh# I]gdj\]XVgZ[jaVY_jhibZcid[i]ZhZVa^c\hideeZg!i]ZXdciZcihd[i]ZWjingdbZiZgVgZ gZ\jaViZY^chjX]VlVni]Vii]ZhXVaZ^h[jaaWjicda^fj^YZciZghi]ZWjaW#I]ZWjingd" bZiZg^hXZcig^[j\ZY^ci]Z]ZViZYXZcig^[j\Z!VcYi]ZciZbeZgZY^cV+*8lViZgWVi][dg *b^cjiZh#I]ZY^k^Y^c\a^cZWZilZZci]Zhjae]jg^XVX^Yb^mijgZVcYi]Z[ViXdajbc^hhZi ViV[jaahXVaZa^cZVcYi]ZjeeZgZcYd[i]Z[ViXdajbc^hgZVYd[[Vii]ZadlZgbZc^hXjh#
3.2 ... IN HOMOGENISED MILK As above, except that the specimens are centrifuged three times, for 5 minutes each time. In between centrifugings, the butyrometers are heated in a 65°C water bath for 5 minutes (for more detail, see page 19).
3.3 ... IN SKIM MILK AND WHEY Use of skim milk butyrometers with narrowed scales according to Sichler’s method. 7jingdbZiZghVgZXZcig^[j\ZYil^XZVcYeaVXZY^cV+*8lViZgWVi][dg*b^cjiZh^c WZilZZcXZcig^[j\^c\h#
3.4 ... IN CONDENSED MILK (UNSWEETENED) I]ZXdcYZchZYb^a`^h]ZViZYid*%8VcYVaadlZYidXddaV\V^c#I]Zc^i^hb^mZYl^i] lViZg^cV&/&gVi^d#I]^hY^aji^dc^hi]ZciZhiZYa^`Zb^a`VXXdgY^c\id
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3.5 ... IN BUTTERMILK (MOHR AND BAUR’S MODIFICATION) I]Z WjingdbZiZg ^h ÒaaZY l^i] &% ba d[ hjae]jg^X VX^Y YZch^in/ -(% $" %#%%( \$ba# >chiZVYd[&%#,*ba!&%bad[WjiiZgb^a`VcY'#%bad[VbnaVaXd]daVgZe^eZiiZY^c#6[iZg hZVa^c\! i]Z WjingdbZiZg ^h h]V`Zc VcY ^bbZY^ViZan XZcig^[j\ZY [dg &% b^cjiZh# I]^h egZkZcihVccdn^c\WadX`V\Zh#I]ZgZVY^c\^hiV`ZcV[iZgiZbeZg^c\i]ZheZX^bZcVi+*8 $"'8#;ViXdciZci2gZVY"d[[kVajZm%,*#
3.6 ... IN POWDERED MILK ACCORDING TO TEICHERT’S METHOD Use of powdered milk butyrometer according to Teichert’s method. I]ZWjingdbZiZg^hÒaaZYl^i]&%bad[hjae]jg^XVX^YYZch^in/-&- $"%#%%(\$ba#,#*ba d[lViZgVcY&bad[VbnaVaXd]daVgZaVnZgZYdcid^i#'#*\d[edlYZgZYb^a`VgZlZ^\]ZY^cV lZ^\]^c\WdViVcYigVch[ZggZY^cidi]ZWjingdbZiZgi]gdj\]V[jccZa!jh^c\V]V^gWgjh]#6[" iZghZVa^c\!i]ZWjingdbZiZg^hi]dgdj\]anh]V`Zc!VcY^ciZgb^iiZcianeaVXZY^cV+*8lViZg WVi]hZkZgVai^bZh#>i^hXZcig^[j\ZYil^XZ!*b^cjiZhZVX]i^bZ!^ci]Z]ZViZYXZcig^[j\ZVcY i]ZkVajZ^hgZVYd[[V[iZgi]ZWjingdbZiZg^heaVXZY^ci]ZlViZgWVi][dg*b^cjiZh#
3.7 ... IN CREAM ACC. TO ROEDER’S METH. (WEIGHING METH.) Use of cream butyrometer according to Roeder’s method. *\d[XgZVbVgZlZ^\]ZYdji^cidi]Z\aVhhWZV`Zgh^ijViZY^ci]ZhideeZgVcYigVch[ZggZY ^cidi]ZWjingdbZiZg#Hjae]jg^XVX^YYZch^in/*'' $"%#%%*\$ba^hedjgZYi]gdj\]i]Z jeeZgdeZc^c\d[i]ZWjingdbZiZgjci^a^igZVX]Zh_jhidkZgi]ZjeeZgZY\Zd[i]Z\aVhh WZV`Zg#6[iZghZVa^c\!i]ZWjingdbZiZg^heaVXZY^cV,%8lViZgWVi]VcYh]V`ZcgZeZVi" ZYanjci^ai]ZegdiZ^c^hXdbeaZiZanY^hhdakZY#Hjae]jg^XVX^Y^hedjgZY^cjci^a^igZVX]Zh i]ZWZ\^cc^c\d[i]ZhXVaZ!&bad[VbnaVaXd]da^hVYYZYVcYi]ZWjingdbZiZg^hhZVaZY! h]V`Zc!VcYeaVXZY^ci]Z,%8lViZgWVi][dgVcdi]Zg*b^cjiZh#I]Zc!^i^hXZcig^[j\ZY [dg*b^cjiZhVcYiZbeZgZY^cV+*8lViZgWVi]#I]ZgZVY^c\^hiV`ZcVi+*8!i]Z[Vi XdajbcVY_jhiZYidi]ZoZgded^ci!i]ZkVajZgZVYd[[Vii]ZadlZgbZc^hXjh#
3.8 ... IN CREAM ACC. TO SCHULZ-KLEY’S METH. (WEIGHING METH.) Use of cream butyrometer according to Schulz-Kley’s method. &%bad[hjae]jg^XVX^YYZch^in/-&- $"%#%%(\$ba!*bad[lViZg!XV#*\d[XgZVb bZVhjgZYWnY^[[ZgZci^ValZ^\]^c\jh^c\Vhng^c\ZdglZ^\]^c\e^eZiiZViiVX]ZYidi]Z hXVaZ!VcY&bad[VbnaVaXd]daVgZhjXXZhh^kZan^cigdYjXZY^cidi]ZWjingdbZiZg#6[iZg hZVa^c\!i]ZXdciZcihVgZb^mZYWnh]V`^c\VcYijgc^c\i]ZWjingdbZiZgjeh^YZYdlc VcYXZcig^[j\ZY[dg*b^cjiZh^ci]Z]ZViZYXZcig^[j\Z#I]ZkVajZ^hgZVYd[[V[iZg* b^cjiZhd[iZbeZg^c\^cV+*8lViZgWVi]#CdiZ/YdcdiVaadlbdgZi]Vc&*b^cjiZh ideVhhWZilZZcVYY^c\i]ZlViZgVcYh]V`^c\i]ZheZX^bZcYjZidi]Zedhh^W^a^ind[ VYZXgZVhZ^ci]Z]ZVid[gZVXi^dcVhVgZhjaid[i]ZVYY^i^dcd[lViZg#I]ZY^hhdaji^dc egdXZhhbjhiWZXdbeaZiZYl^i]^c+%hZXdcYh# ;ViXdciZci2gZVY"d[[kVajZm*$XgZVbcZilZ^\]i#
3.9 ... IN CREAM ACC. TO KOEHLER’S METH. (MEASURING METH.) Use of cream butyrometer according to Koehler’s method. &%bad[hjae]jg^XVX^YYZch^in/-&- $"%#%%(\$ba!*#%*bad[XgZVb!*bad[lViZg! VcY&bad[VbnaVaXd]daVgZedjgZY^cijgc^cidi]ZXgZVbWjingdbZiZg#L]Zcjh^c\V XgZVbhng^c\Z!^ibjhiWZg^chZYhZkZgVai^bZhl^i]lViZgWZ[dgZi]Z*bad[lViZgVgZ ^cigdYjXZY#I]ZWjingdbZiZg^hhZVaZY!h]V`Zc!XZcig^[j\ZY[dg*b^cjiZh!VcYi]Zci]Z kVajZ^hgZVYd[[V[iZgiZbeZg^c\i]ZheZX^bZc^cV+*8lViZgWVi]#I]ZkVajZ^hgZVY d[[[gdbi]ZoZgded^ci#
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3.10 ... IN CHEESE ACC. TO VAN GULIK’S METH. (WEIGHING METH.) (see ISO 3433) Use of the cheese butyrometer according to Van Gulik’s method. 6[iZgedjg^c\&*bad[hjae]jg^XVX^YYZch^in/*'' $"%#%%*\$ba^cidi]ZKVc
3.11 ... IN ICE CREAM ACC. TO KOEHLER’S METH. (MEASURING METH.)
Use of the ice cream butyrometer according to Koehler’s method. 6cn^X^c\dggdj\]eVgi^XaZhZ#\#[gj^i!ZiX#bjhiWZgZbdkZY#I]Z^XZXgZVb^h i]dgdj\]anb^mZYV[iZg^i]VhgZVX]ZYgddbiZbeZgVijgZ0edhh^WaZV^gedX`ZihXVc WZVabdhiXdbeaZiZangZbdkZYWnZkVXjVi^dc# &% b d[ hjae]jg^X VX^Y YZch^in/ #-&- $" %#%%( \$ba! * ba d[ ^XZ XgZVb! * ba d[ lViZg!VcY&bad[VbnaVaXd]daVgZ^cigdYjXZY^cijgc^cidi]Z^XZXgZVbWjingdbZiZg# L]Zc jh^c\ V hng^c\Z! ^i bjhi WZ g^chZY hZkZgVa i^bZh l^i] lViZg WZ[dgZ i]Z * ba d[lViZgVgZ^cigdYjXZY#H]djaYi]ZWjingdbZiZgegdkZidcdiWZhj[ÒX^Zcian[jaa!je id ' ba d[ lViZg XVc WZ VYYZY# I]Z WjingdbZiZg ^h hZVaZY! h]V`Zc! XZcig^[j\ZY [dg *b^cjiZh!VcY!V[iZgiZbeZg^c\^cV+*8lViZgWVi][dg*b^cjiZh!i]ZkVajZ^hiV`Zc#
3.12 ... IN ICE CREAM ACC. TO ROEDER’S METH. (WEIGHING METH.) Use of ice cream butyrometer according to Roeder’s method. *\d[lZaa"b^mZY^XZXgZVbVgZlZ^\]ZYdji^cidi]Z\aVhhWZV`Zgh^ijViZY^ci]Z hideeZg VcY igVch[ZggZY ^cid i]Z WjingdbZiZg# Hjae]jg^X VX^Y YZch^in/ *'' $" %#%%*\$ba^hedjgZYi]gdj\]i]ZjeeZgdeZc^c\d[i]ZWjingdbZiZgjci^a^igZVX]Zh _jhi dkZg i]Z jeeZg ZY\Z d[ i]Z \aVhh WZV`Zg# 6[iZg hZVa^c\! i]Z WjingdbZiZg ^h eaVXZY^cV,%8lViZgWVi]VcYh]V`ZcgZeZViZYanjci^ai]ZegdiZ^c^hXdbeaZiZan Y^hhdakZY#&bad[VbnaVaXd]da^hVYYZY!VcYi]Zchjae]jg^XVX^Y^hedjgZY^cjci^a ^i gZVX]Zh i]Z &% bVg` dc i]Z hXVaZ# I]Z WjingdbZiZg ^h hZVaZY! h]V`Zc! VcY eaVXZY^ci]Z,%8lViZgWVi][dgVcVYY^i^dcVa&%b^cjiZh#9jg^c\i]^hi^bZ!i]Z WjingdbZiZg^hh]V`ZcVigZ\jaVg^ciZgkVah#I]^hegdXZhh^h[daadlZYWn,b^cjiZhd[ XZcig^[j\^c\VcYiZbeZg^c\^cV+*8lViZgWVi]#I]ZkVajZ^hiV`ZcVi+*8!i]Z[Vi XdajbcVY_jhiZYidi]ZoZgded^ci#I]ZkVajZ^hgZVYd[[Vii]ZadlZgbZc^hXjh#
3.13 ... IN BUTTER ACC. TO ROEDER’S METH. (WEIGHING METH.) Use of butter butyrometer according to Roeder’s method. *\d[WjiiZgVgZlZ^\]ZYdji^cidi]Z\aVhhWZV`Zgh^ijViZY^ci]ZhideeZgVcYigVch[ZggZY ^cidi]ZWjingdbZiZg#Hjae]jg^XVX^YYZch^in/*'' $"%#%%*\$ba^hedjgZYi]gdj\]i]Z jeeZgdeZc^c\d[i]ZWjingdbZiZgjci^a^igZVX]Zh_jhidkZgi]ZjeeZgZY\Zd[i]Z\aVhh WZV`Zg#6[iZghZVa^c\!i]ZWjingdbZiZg^heaVXZY^cV,%8lViZgWVi]VcYh]V`ZcgZeZVi" ZYanjci^ai]ZegdiZ^c^hXdbeaZiZanY^hhdakZY#Hjae]jg^XVX^Y^hedjgZY^cjci^a^igZVX]Zh i]ZWZ\^cc^c\d[i]ZhXVaZ![daadlZYWn&bad[VbnaVaXd]da#I]ZWjingdbZiZg^hhZVaZY! h]V`Zc!VcYeaVXZY^ci]Z,%8lViZgWVi][dgVcVYY^i^dcVa&%b^cjiZh#I]^hegdXZhh^h [daadlZYWn*b^cjiZhd[XZcig^[j\^c\VcYVeegdm^bViZan*b^cjiZhd[iZbeZg^c\^cV+*8 lViZgWVi]#I]ZkVajZ^hgZVYd[[Vi+*8Vii]ZadlZgbZch^Xjh#
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The addition of potassium sulphate serves to increase the boiling temperature of the sulphuric acid and the addition of copper sulphate serves as an oxidation catalyst. They are also available as Kjeldahl tabs (art. no. 4230/4231). If the process is executed with tabs, then 5± 0.1 g of milk are mixed with 20 ml of sulphuric acid and 2 Kjeldahl tabs and left to sit for 5 min. Then the temperature program can be carried out. During the heating of the sample, foam is not allowed to climb higher then 4 - 5 cm below the flask opening. The sulphuric acid is added in such a way that any copper sulphate solution, potassium sulphate or milk which may have adhered to the flask neck is flushed down. If the flask is sealed airtight, it can also be stored for later digestion. To determine the specific digestion time, it is advisable to execute preliminary tests with samples high in protein and fat. Substantial crystallisation is a sign of too little sulphuric acid and can lead to low protein values. It is therefore advisable to reduce the loss of sulphuric acid by minimising the amount sucked. Before the hot digestion flasks can be taken from the digestion block, it must be ensured that no condensed fluid has collected in the extraction apparatus. If it has, the suction volume must be increased and the condensed liquid removed. The undiluted digestion should not be stored in the flask for a long period of time (overnight) for any reason. There is a risk that the sample will solidify and it is difficult to bring it into solution form again. After the sample is cooled down and dilluted with 70 ml of water, there is no problem to keep it overnight.
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KJELDAHL’S NITROGEN DETERMINATION METHOD
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The water distiller must be set up on a stable laboratory bench with an even, horizontal support which is located near a cold water supply and a drain. The water pressure must be at least 0.5 bar. Before starting the operation, all hoses must be connected and the coolant engaged. The storage tank must be correctly positioned and the fluid level checked. The water vapour discharge hose must be introduced into the digestion flask. The water distiller is equipped with a safety gate. During the first distillation, the water vapour comes into contact with cold pipes and glass parts. This leads to increased build-up of condensation which can in turn lead to excessive sample dilution and liquid volume in the digestion vessel. A trial run is therefore essential. The discharge of water vapour with a temperature of approx. 106°C creates loud noises. This is no cause for alarm. The distillation is conducted until a distillation volume of 150 ml is obtained. About 2 minutes before the end of the distillation, the Erlenmeyer flask is lowered in such a way that the end of the discharge pipe is no longer submerged in the acid solution. The pipe must be flushed with water. This water is collected in the Erlenmeyer flask.
3
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The mixture of methyl red and bromcresol green (see 2.5, 2.6) serves as the indicator. The indicator is responsible for the colour change and signals the end of the titration. The neutral background improves the accuracy and reproducibility of the results. This means that the titrations are always carried out under optical conditions that are as similar to each other as possible.
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The blind trial is important for the calculation of the nitrogen content of the sample.
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Kjeldahl’s digestion method is not specific to amino acids and proteins and includes all organically bonded nitrogen. Other non-protein compounds are also are digested and collected (NPN: non-protein nitrogen). However, the proportion of these compounds is very small and is disregarded in the calculation. If the non-protein containing nitrogen should also be established, then method must be executed in accordance with DIN EN ISO 8968-4. If only the protein nitrogen should be determined, then the milk proteins must first be separated. 5±0.1 ml of milk diluted with 5±0.1 ml of water is washed in stages with in total 60 ml of 15 % (w/v) trichloroacetic acid in accordance with DIN EN ISO 8968-5, the proteins are precipitated and finally filtered out into a hard paper filter. The filtrate contains the components of the non-protein nitrogen and the filtered-out precipitate contains the protein nitrogen. The filter with the precipitate is put into a digestion vessel and Kjeldahl’s nitrogen determination method is carried out as described above. The protein content is calculated by multiplying by a factor of 6.38. The value 6.38 is specific to milk and dairy products and was established because milk proteins have a nitrogen content of 15.65 % (100:15.65 = 6.38).
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Caution! The addition of hydrogen peroxide causes an intense reaction. The blind trial is executed with 2 ml of water and 0.25 g of sucrose. The effectiveness of the decomposition is tested with 0.08 g of tryptophan or 0.06 g of lysine hydrochloride.
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In order to monitor the effectiveness of the decomposition, 0.18 g of tryptophan or 0.16 g or lysine hydrochloride and 0.67 g of sucrose are used. 98 % of the nitrogen content must be recovered. If that is not the case, either the decomposition temperature or time is insufficient or the sample is charred.
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4200
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4210
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4220
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pH VALUE MEASUREMENT Anna Politis, graduate engineer of nutrition technology and of biotechnology, reports
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FREEZING POINT DETERMINATION DcZd[i]ZbV^c[dXjhZhd[;jc`Z"9g#C#
HISTORY The German chemist Beckmann, known for the thermometer named after him, began using the freezing point of milk in as early as 1895 to detect if it had been adulterated with water. The American Hortvet worked intensively with this method in 1920 and improved some of its essential features. The first thermistor cyroscopes were brought to the market in the 1960s. However, they had to be operated entirely by hand. At the beginning of the 1970s, the first automatic thermistor cyroscopes became available. With this development it was possible to determine the freezing point automatically at the push of a button. A decisive step in the improvement of thermistor cryoscopy was displayed at the “FoodTec” tradeshow in 1984: Funke-Gerber introduced the first device with automatic calibration. This successful and intensive development work reached a new peak at the “FoodTec” in 1988, where Funke-Gerber presented a fully automatic freezing point determination mechanism with a capacity of 220 samples per hour. With the introduction of an indirect freezing point measurement device (e.g. LactoStar) for routine analysis, interest was focused primarily on reference devices which are able to determine freezing points in accordance with the applicable standards and regulations. These devices must satisfy the strictest requirements with regard to measuring accuracy. For this reason Funke-Gerber developed a programmable cryoscope with a resolution of 0.1 m °C. This instrument has proven its accuracy and reliability in countless laboratories all over the world. The product range has been expanded with a multi-sample device (CryoStarautomatic). Since January 2007, these instruments have been equipped with a graphic colour display. This makes it possible to show the entire freezing curve, in particular the process of the plateau search, with a patented screen presentation.
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