BIOLOGY

1) Which are the fundaments of Biuret s and Benedict s reactions?

2) Which are the limits of both reagents?
3) What is the imortance of a blan!?
1) Biuret reaction
"he formation of biuret that gi#es a #iolet color as a result of the reaction of a
ol$etide of more than three aminoac$l residues %ith &u'O( in strongl$ al!aline
solution) dietides and amino acids *e+cet histidine, serine, and threonine) do not so
react) used for the detection and -uantification of ol$etides, or roteins, in biologic
fluids.
"he Biuret reagent is made of sodium h$dro+ide */aO0) and h$drated coer*II)
sulfate, together %ith otassium sodium tartrate.
1(2
3otassium sodium tartrate
142
is added
to comle+ to stabili5e the curic ions. 3roteins in the al!aline en#ironment reduce
&u
26
to &u
6
, %hich forms a coordination comle+ %ith roteins, leading to a blue to
in! color change. 0istidine is the onl$ amino acid %hich ans%ers this test.
Benedict reaction
In a mild al!aline medium, sodium carbonate con#erts glucose into an edinol form. "his
edinol reduces curic ions to curous roducing curous h$dro+ide. "his is !et in
solution b$ sodium citrate. On boiling, curous h$dro+ide is con#erted into red curous
o+ide %hich is reciitated.
2) Limits Benedict reagent7
It does not react %ith all small sugars, the test is -ualitati#e %hich allo%s us to onl$
dtect the resence of certain molecule, also the test does not allo% us to determine
%hich samle sugar is resent. "his test is sensiti#e, is false ositi#e in the resence of
salic$lic acid, entose sugars and drugs li!e dasone and metrog$l.
Limits of Biuret reagent
"her is a general rogression, onl$ test if roteins are resent *not detailed result of
%hich rotein), -ualitati#e not -uantitati#e.
3) Blan!s are useful %hen there are other substances in the e+erimental tube besides
the substance $ou are tr$ing to measure. 'ince those other substances are not the
chemical that $ou are tr$ing to measure, the$ often interfere %ith the absorbance
reading of the chemical of interest. It gi#es a non secific bac!ground #alue.
Blan! solutions re t$icall$ used to calibrate instruments such as the calorimeter, %hich is
used to measure the absorbance of light %a#elengths b$ a secial solution.
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