PLAN 01 THi INVESTIGATION. 31 



The -oluhle protein- were determined ly the following method. 

 hv .]. S. Oiaml>erlain : 



An amount <>f air-dried barley or malt equivalent to l!n trains of dry material 

 was extracted with water of su.-h a volume that the total resulting mixture 

 ainoiintc.l to exactly inn ,-,-. in order to know the \olume of liquid in such 

 an extraction it w.i ry to determine the \ohinie occupied by the residue 



from LMI -rams .,f the dry barley after extraetiou. which was found to be 

 in.77 re. In calculating this volume, the figure obtained by II. T. P.rown " 

 tor the specific gra\it\ of tin- dry residue f extracted barley was used, namely. 

 l.r.T. Subtracting the volume occupied by the dry residue of extracted barley 

 from the inn CC gives tin- \olume of liquid actually present. An aliquot of this 

 \olume was taken after tiltration and the nitrogen determined therein. In 

 practiee. howexer. an amount of bar:. iken such that in the proportion 



of _' gnuna Of d- MCO to lOn ,,- ,,r the resultin- extraction mixture 



there will be pfesi-nl. after allowing for the \oluine occupied by the extracted 

 barley, exactly Inn , of liquid. In this way aliqiiots .>f In cr. _'.", CC, ", 

 ild be easily obtained: tha 1 H -rums of dry barley in the 



proportion of I'll; ]nn \ v ill require a volume of extraction liquid equaling 

 1 rj.nr, CC, ;nid _'_'.! -rim- of dry Narley will lea \ e after extraction a dry 

 residue equaling' U.IM; ,-. Therefore the \ohime of liquid present equals 

 1 1 LM ir. I _ 



The amount of air-dr\ barley to b, H then easily calculated in each 



iroin the perceiita-e of moisture in the sjtmple. and this weight of air dry 

 material was added and extracted under the conditions just des.-ribed. l-'or 

 the extractioii. distilletl \\ a i . in feni|N'rature \va> DMd, .md the bottles 



in which the extraction took place \\.-n- shaken in a revolving shaker for six 

 hours. The mixture \\ . then filtered as rapidly as |mssihle thnxiirh folded 

 tiller papers, the iir>t IHUMIOII of filtrate, \\hen doiidv . beimr poured back upon 

 the tiller paper until a dear filtrate was obtained. In an aliquot of this clear 

 filtrate the amount of nitrogen \\as determined, which, multiplinl by (>. !'.">, gave 

 the protein, representing the soluble protein. 



The soluble noncoairulable protein was determined by boiling i'n cc of tin 

 abo\e filtrate o\er a small flame until the volume was reduced to about in ec. 

 After diluting to the original volume, the liquid was filtered, washed, and the 

 noneoairulahle nitro-en deterndned b\ usint: the whole of Hie filtrate. 



The soluble eoau'iihiliie proteins wen- determined by subtracting the soluble 

 noncoairulable protein from the total protein. 



The determinations made l>y Walil. \\liieh re(jnire >perial mention. 

 \\ere a- follow- : 



!! the soluble protein determination rn grams of tinely -round barley were 

 extracted with l'.~>n -rams of water for six hours at is (\ ztl, stirring well 

 every fifteen minutes. The loss on eva|Nration < approximately n.l L'ram) was 

 made up by adding water until the total weight equaled 300 grams. The extract 

 Mltered dear, maintaining approximately the same temperature. The total 

 soluble nitrogen was determined in an aliquot of the filtrate according to Kjel- 

 dahl's method. 



The coagnlable nitrogen was determined by boiling 100 cc of the above fil- 

 trate for thirty minutes, keeping the volume constant, filtering, and estimating 

 the nitrogen in the precipitate. The factor fi.LT. was used in all determinations 

 in changing the percentage of nitrogen into protein. 



cit. 

 L'-lf, Hull ILM n'.. :; ' 



