Improvement in Anthrone Method for Determination of Carbohydrates
u3 :I qualitative ( 2 ) and quantitative h plot oi the optical deiisitj. a g a i i ~ tweiglit of glucuse in the
tietermination of' carbo1iydr;iteh has beer1 siiiiple gives a a~ruiglit-liiierelationship froni 0 to 80 micrograms.
considerably iinpet1t.d by the darkening n-it11time oi the anthro~ie 'The acetic acid arid ethyl alcohol produced by the hydrolysis oi
reagent, iv1iic.h is usually 0.1 to 0.2% anthrone in 'JEi70sulfuric the ethyl acetate do riot interfere v i t h the reaction in any way.
acid (21. The tIar:;ening of the reagent has been shown to be iii- 'The ethyl acetate solutiori of anthrone will keep several R-eeks
versely related 10 the purity of the sulfuric acid employed. when stored in :in umber glass-stoppered bottle. Thus, the I
\Vheti thcs reqyiit j, used quantitatively, this darkening phe- troublesonie developinent of color in the sulfuric wid-nnthronc
iionienoii creates :L iwed either fur careful staridarization with w- rragent eniployed in ~ a r l i e rprocedures is avoided.
spect to the age oi the reagent used or for preparation of a i l e \ ~ Using the values reported by Morris (4)as conversion factors,
s t a n d u d curve iur td:ich set of carbohydrate deterniinations. :I single standard curve prepared from glucose uill suffice for the
.I Iiioililicatioii of lforris' method (4) has bee11 devised which quantitative determination of a11 carbolij.drates listed. 13). deter-
:ivuids tlie necessity of dissolving the anthrone in sulfuric acid iiiining the conversion factors, inuny other hexoses and hesose-
prior to the determination, thus eliininating color variations yielding conipounds, including those listed by Dreywood ( 1 ),
:irking ironi this sourcc~and :illowing a single standardieatioii Viles and Silverman ( 5 ) , and Ko~valdand r\IcC:orinack (3),cn11
curve Lo suficc . be quantitatively estiniated in terms of glucose.
To 2 nil. of tlie carboliydwte solutiori (containing ail amount of
carbohydr:tte that will give a color intensity in the rauge given by
0 and 80 microgranis of glucose) in a 19 X 150 111111. tcst tube is
added 0.5 nil. of a solution of 2% anthrone (recryst:illized from
benzene nnd light, petroleum et'her) in reagent grade or e.?. ethyl
:tcetate. Then 5 nil. of concentrated sulfuric acid are carefully
layered into the tube. The tube is gently swirled until the ethyl
scetate has hydrolyzed, as indicated by the floc of anthrone
ivhich appe:trs. More rapid swirling then thorou hly mises the
contents of the tube and dissolves the anthrone. t h e developed
color may be read after 10 minutes a t 620 me in a spectrophotom-
eter against distilled water and corrected for the absorption of a
blank containing onlv the reagents and water.
Errors in Volumetric Analysis Arising from Adsorption
yOS\VAY ( 1 ) in his study of errors in voluinelric andysis niorc, r:idioclieniists have long liceii aivare of the fact that C U I I -
(J made no inention of adsorption as a possible source of crro1'. siderable losses may occur hen very dilute solutio~isare stored
However, the import:ince of adsorption in one nnalytical method i n glass contaiuers. .Igeneral study of the adsorption of certaiil
WLS strikingly denioustrated by Schoonover's (6) studies of :I ions onto soft gl:iss, borosilicate gl , and iiietallic surfaces,
colorimetric niethotl Cor the determination of silver. Further- carried out by Hensley, Long, and \Villard (5), iiidicatcd that it
\vas necessary, for ivork IJeing done in this laboratory, to deter-
niine the mignitude of the losses to be expected froni adsorption
iii pipeting very dilute solutions of silver perchlorate in 0.1 .Y
c perchloric acid.
In view of the sniall aiiiount of silver involved, and because
radioactivity offered a sensitive and convenient method of meas-
urement, radioactive Xg111 obtained by neutron irradiation of
palladiuni in the Oak Ridge pile was used for this study. Carrier-
i'ree Ag111 of high specific activity was recovered froni the palla-
dium by electroseparatioii froni cyanide solution according to the
inethotl of Griess and Rogers ( d ) , following which the silver ac-
tivitj. \vas dissolved b>, electrolysis into a few niilliliters of 0.1 -11
percliloric acid for use in tlie adsorption studies. The concentra-
tion of silver in thc resulting solution could be calculated lis less
than 1 0 - b J I (C), but as especially purified reagents were not
used, the concentration was probably greater. Standard solu-
tions containing higher concentratioiis of silver were prepared by
adding portions of this trace1 solutiori to kno\\n aniounts of 2.00 Jf
p(Ag*i silver perchlorate o r dilutions thereof and bringing to volunit:
I I I ! I I I \vith 0.1 III perchloric acid. Solutions \rere .stord in glass-
0 1 2 3 4 5 6 7 8 9
stoppered bottles coated with ceresine \vas i n orclei, to niininiize
Figure 1. Adsorption on Kirk Pipet changes in concentratioiis \\-it11tirne.
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