The Blue Bottle Experiment Revisited

How Blue? How Sweet?

A. Gilbert Cook, Randi M. ~ o l l i v e rand
, ~ ~Janelle
~
E. ~illiarns'*~
Valparaiso University, Valparaiso, IN 46383
The "Blue Bottle" experiment was first popularized by
Campbell with his article in this Journal in 1963 (1)and
later in his book (2).The reaction consists of a flask about
half full of a colorless liquid that, when shaken, turns blue.
When left to stand the liquid turns colorless again. The
cyclic process--being shaken, turning blue, being leR to
stand, and turning colorless+an be repeated many times.
By making a series of guided observations with this simple but colorful system, the student can determine a rate
law and a plausible mechanism for the reaction without
ever knowing the contents of the flask. The activation energy of the reaction can also be measured by making observations a t various tem~eratures.This ~ o ~ u lex~eriment
ar
has been carried out b; general c h e m i i t j studenis at Valparaiso University since 1963 with p e s t success.
The Reaction
The contents of the flask are an aqueous solution of sodium hydroxide, glucose, and methylene blue. The relative
amounts of each of these needed to make the solution is
approximately 20 g of sodium hydroxide, 20 g of gluwse,
and 1mL of 1% alcoholic methylene blue solution per liter
of water.
The rate law for the reaction is

Table 1. Dye Structures

Thfazines

R4

y&*

R1

R3

R i = (CH3)2N,R2 = (CH3)zN'
RI = NHz, R2 = (CH3)zNi
RI = (CHdNH,Rz = (CH3)Nf
RI = (CH3)zN.R2 = (CH3)N.R6 = CH3
RI = CzHsNH. R2 = CzH5NHf. R4 = R5 = CH3
Ri = HzN, Rz = HzNt
Rx = HzN, Rz = H2Nt, Rs = CH3
R i = (CH3)2N,R2 = (CH3)2N', R3 = NO2

Ri = (CH3)zN.R2 = 0
Oxazines

rate = k[D,I[CHI[OHI
where Do, is the oxidized (blue) form of methylene blue,
and CH is the carbohydrate, glucose.
The following is a simple mechanism for this reaction.

fast D,
O,+D d

R3
Ri=NaO,Rz=O,R7=0
Ri = (CzH5)2N,R2 = HzN,' Rs = CH3
Ri = H2N, R2 = HzNt, Rs and Re = fused benzene
Ri =OH, R2 = (CzH5)2N,R3 = OH, R4 = HzNCO
Azines

where D is the reduced (wlorless) form of the dye methylene blue, and X- represents the oxidation products from
the glucose (arabinoic, formic, oxalic, and erythronic acids
(3,411.The enthalpy of the reaction has been reported to be
5.5 kcal/mol(5).
How Blue?
Are other colors beside blue possible for this reaction?
Methylene blue is a thiazine dye (structure 1 shown in
Table 1).There are manv other dves in the thiazine dve
family that might be usckd, as wcil as dyes in the related
oxazine and azme dye families. Wc havc found that many
Preliminary report presented at the 106th Annual Meeting of the
Indiana Academy of Science, 1990.
'PRISMS Program (Precollege Research In the Sciences for Minority Students)participants during summers of 1990and 1991. Program cosponsored by GTE and Valparaiso University.
'Theodore Roosevelt High School. Gary, IN 46407.
3West Side High School, Gary, IN 46406.

160

Journal of Chemical Education

- . R = H except where noted.

of these other dyes also catalyze the oxidation of an alkaline aqueous solution of glucose when mixed with oxygen.
However, the rates of reaction and sometimes the wlors
of the oxidized dyes vary from that observed with methylene blue. The color of the oxidized form of the dye in aqueous base and the relative rates of oxidation of glucose are
shown in Table 2. Two of the oxazine dyes (cresyl violet
acetate and resazuriu) fluoresce red in their oxidized
states when excited by a 365-11111 U V lamp, whereas fluo-