Vol.

75, 1969] 195

DETERMINATION OF METALS IN BEER AND WINE BY ATOMIC

ABSORPTION SPECTROPHOTOMETRY

By J. P. Weiner, F.R.I.C., and L. Taylor, B.Sc, F.R.I.C.

(Courage, Barclay & Sitnonds Limited, London)

Received 20th August, 1968

Methods in common use for the determination of metals in beer and wine are
often time consuming and difficult to apply for routine quality control. Atomic
absorption spectrophotometry provides a rapid and accurate means for examining
a large number of samples. The application of the technique for estimating the trace
metals Cu, Fe, Zn and Pb in beers and wines is described. A survey of the magnesium
content of beer and brewing materials is also included.

Introduction improving the resistance of beer to light-

The technique of atomic absorption spectro
photometry is finding increasing application
for the determination of metallic elements
in many laboratories since the method is
rapid, specific and sensitive. For most
elements, it is superior to emission measure
ment, being less dependent on flame temper
ature and spectral interference. The degree
of accuracy compares favourably with most
other analytical instrumental methods.
The theory of atomic absorption and the
factors affecting practical performance have
been dealt with in several publications.4'9
Briefly, the element being determined is
passed through a flame to produce an atomic
vapour of the particular element in its
unexcited "ground" state. It is then capable
of absorbing radiation, usually provided by a
hollow-cathode lamp, resulting in excitation
of the atom. The amount of radiation
absorbed is proportional to the concentration
of atoms in the flame.
The importance of trace metals in beer and
wine is now well recognized and is subject
to legislation in the case of lead, copper
and zinc. The Lead in Food Regulations
limit the lead content of beer to 0-5 p.p.m.
and of wine to 1-0 p.p.m., whilst the Food
Standards Committee recommend limits of
7-0 p.p.m. copper and 6-0 p.p.m. zinc for
both beer and wine. The association of
certain metals with beer stability has been
shown in a number of publications,8-8-7'8'10
and the addition of small quantities of metal
salts has been advocated to improve the
quality of beer foam.1'13 The supplementa
tion of zinc-deficient worts with zinc ions,
as a means of stimulating fermentation and
struck odours, has also been recommended.12
Frey, DeWitt & Bellomy have applied
atomic absorption spectrophotometry for
the determination of a number of trace
metals in beer, wort and yeast,8 and they
have also used the technique during the
investigation of the effect of several trace
metals on fermentation.4
In the present paper, the determination of
Cu, Fe, Zn and Pb in beers and wines for
routine quality control examination is de
scribed. Included in the report is the result
of an investigation into the magnesium
content of beer and brewing materials.
Experimental
Apparatus.—Unicam SP 90 Atomic Absorp
tion Spectrophotometer (Unicam Instru
ments Ltd.); Servoscribe Potentiometric Re
corder (Kelvin Electronics Company).
Standard instrumental conditions.—The
operating conditions for the respective metals
are given in Table I.
Reagents.—
(a) Ammonium pyrollidine dithiocarba-
mate solution (APDC). Dissolve 1 g. APDC
in distilled water and dilute to 100 ml.
Prepare fresh daily.
(b) Methyl isobutyl ketone. Saturate
by shaking with distilled water.
(c) Acetic acid (Analytical Grade).
(d) Standard copper solution. Dissolve
3-93 g. of CuSO4.5HaO (Analytical Grade)
and dilute to 1 litre with distilled water.
1 ml. = 1 mg. Cu.
196 WE1KER AND TAYLOR: METALS IN BEER AND WINE [J. Inst. Brew.
TABLE I
Operating Conditions for Determination of Metals

Copper Iron Zinc Lead Magnesium

Sample None Treatment None Treatment None

treatment with APDC* with APDC
& extraction & extraction

Fuel Propane- Acetylene- Propane- Acetylcnc- Acetylene-

air air air air air

Wavelength ni/i 324-8 248-3 213-0 283-3 285-2

Slit width mm. 0-08 0-2 0-4 0-2 0-1

Burner height cm. 1-5 1-3 1-2 1-0 1-0

* APDC = Ammonium pyrollidine dithiocarbamate solution.

(e) Standard iron solution. Dissolve 0-100
g. of pure iron metal in the minimum quantity
of hydrochloric acid (Analytical Grade) and
make up to 100 ml. with distilled water.
1 ml. = 1 mg. Fe.
(/) Standard zinc solution. Dissolve
4-398 g. of ZnSO4-7H,O (Analytical Grade)
and dilute to 1 litre with distilled water.
1 ml. = 1 mg. Zn.
(g) Standard lead solution. Dissolve 1-600
g. of Pb(NO8)2 (Analytical Grade) in distilled
water, add 10 ml. of nitric add, S.G. 1-42
(Analytical Grade) and dilute to 1 litre.
1 ml. = 1 mg. Pb.
(h) Standard magnesium solution. Dis
solve 0-100 g. of oxide-free magnesium ribbon
in the minimum quantity of hydrochloric
acid (Analytical Grade) and make up to
100 ml. with distilled water. 1 ml. = 1 mg.
MB.
Methods
Copper.—Freshly prepare standards by the
addition of suitably diluted standard copper
solution to decarbonated low copper content
beer to obtain beer standards containing
0, 0-5 and 1-0 p.p.m. copper, respectively.
Using a two-times scale expansion on the
instrument, aspirate the standard beer
solutions followed by the decarbonated beer
samples under test. Plot a calibration
graph of copper concentration against re
corded peak height and read the copper
concentrations in the beer samples.
Iron.—Prepare standards in distilled water
containing 0, 0-5, 1-0, 1-5 and 2-0 p.p.m.
iron. Take 25 ml. of each solution in 50-ml.
stoppered centrifuge tubes, add 2 ml. of
APDC solution, mix and add by pipette
10 ml. methyl isobutyl ketone. Shake
vigorously for several minutes and centrifuge
to separate.
Take 25 ml. of decarbonated beer in a
stoppered centrifuge tube, and treat as above.
Using single scale expansion, aspirate the
solvent layer of the blank and standard
solutions followed by the sample solutions.
Use pure methyl isobutyl ketone solvent as
reference, adjusting while aspirating to
obtain a stable non-luminous or slightly
luminous flame. Plot a calibration graph
of iron concentration against recorded peak
height and read the iron concentrations
in the beer samples.
Zinc.—Freshly prepare standards by add
ing suitably diluted standard zinc solution
to decarbonated low zinc beer to obtain
beer standards containing 0, 0-5, 1-0, 1-5 and
2-0 p.p.m. zinc.
Aspirate the blank and standard beer
solutions followed by the decarbonated
beer samples being examined. Plot a cali
bration graph of zinc concentration against
recorded peak height and read the zinc
concentrations in the beer samples.
Lead.—Prepare standards in distilled water
containing 0, 0-2 and 0-5 p.p.m. lead. Take
50 ml. of each solution, add 5 ml. of acetic
acid followed by 2 ml. of APDC solution,
mix and add by pipette 10 ml. methyl
isobutyl ketone. Shake vigorously for
several minutes and centrifuge to separate.
Take 50 ml. of decarbonated beer; add
5 ml. acetic acid. Bring to boiling and boil
for about 2 min. Cool, add 2 ml. of APDC
Vol. 75, 1969] WEINER AND TAYLOR: METALS IN BEER AND WINE 197

solution, mix and add by pipette 10 ml. 120

methyl isobutyl ketone. Shake vigorously

for several minutes and centrifuge to separate. 100
Using a two-times scale expansion, aspirate
the solvent layer of the blank and standard
solutions followed by the sample solutions.
Use pure methyl isobutyl ketone solvent as
reference, adjusting while aspirating to obtain
a stable non-luminous or slightly luminous
flame. Plot a calibration graph of lead
concentrations against recorded peak heights
and read the lead concentrations in the beer
samples.
Magnesium.—Freshly prepare standards
in distilled water containing 0, 0*5, 1-0, 1-5
and 2-0 p.p.m. magnesium. 05 1-0 IS 20
Copper (mg/litre)
Aspirate the blank and standard solutions
followed by the beer samples suitably diluted Fig. 1.—Enhancement of response for copper due to
with distilled water. Normally 1 in 50 or alcohol strength of beer. The figures along
1 in 100 dilutions of beer are suitable. Plot side the lines indicate amounts of added
alcohol (%).
a calibration graph and read magnesium
concentrations in the diluted beer samples.
standards and for this reason copper standards
are best prepared by addition to beer. The
Results and Discussions effect of dissolved sugar solids is negligible,
Recoveries obtained for various metals at least up to a concentration of 2%. The
added to beer are given in Table 11. use of propane as fuel gives slightly better
The calibration curve for low levels of sensitivity than the use of acetylene. The
copper is a straight line. Direct aspiration sensitivity (1% absorption) for the deter
of beer shows enhancement of response mination of copper is about 0-05 p.p.m.
for copper compared to aqueous standards. If a greater sensitivity is required, an
The extent of this enhancement is roughly extraction procedure similar to that described
proportional to the alcoholic strength of the for iron may be employed. It has been
beer (Fig. 1). It has been found that this observed that slightly low and more erratic
effect may yield results 10-25% high if results are obtained with unfiltered beers
the instrument is calibrated to aqueous containing yeast in suspension.
The sensitivity of the method for iron is
lower than for copper and direct aspiration
TABLE II
of beer is only suitable for use as a sorting
Determination by Atomic Absorption Spec-
trophotometry of metals added to beer test for levels above 0-5 p.p.m. Using the
extraction procedure given, a greatly im
proved sensitivity can be obtained, not only
Added
metal 0/ because of the concentration involved but
(p.p.m.) Detected also owing to enhancement caused by the
use of an organic solvent. The acetylene-air
Copper 05 100
flame is preferred because of the greater
10 05
Iron 0-5 100
1-0 00 TABLE III
Zinc 0-5 110 Zinc Contents of Bottled and Canned Beers
1-0 110 by Atomic Absorption Spectrophotometry
Lead 0-2 05
0-4 100
00 03 Zinc (p.p.m.)
0-8 00 Range Average
1-0 01
Magnesium 50 00 Bottled beers 0-1-1-6 0-33
100 05 Canned beers 0-1-4-4 0-87
198 WEINER AND TAYLOR! METALS IN BEER AND WINE []. Inst. Brew.

usually originates from the end scaling

0.5 p.p.m. standard "y
r time-consuming and laborious operation using
1.0 p.p.m. standard
I Beer control
I Added 0.2 p.p.m.
2^jAddcd 0.4 p.p.m.
r to break down possible lead complexes in beer,
Added 0.6 p.p.m.
c small amounts of lead. The extracted lead
Added 0.8 p.p.m.
Added 1.0 p.p.m.
Fig. 2.—Recovery of added amounts of lead from
beer.
stability of the flame. The calibration
curve is a straight line for low levels of iron.
The atomic absorption method for zinc
is very sensitive and is successfully carried
out by direct aspiration of beer. The
calibration curve obtained is slightly curved
towards the concentration axis. A survey
of zinc levels in bottled and canned beers
gave the results shown in Table III. These
results confirm the findings of Frey et al.6
that canned beers contain relatively higher
levels of zinc than bottled beers. Our
experience is that when zinc is present it
TABLE IV
Magnesium Contents of Mashing Liquors,
Worts and Beers by Atomic Absorption
Spectrophotometry
compound used.
Determination of lead in trace amounts is a
the usual colorimetric methods, which in
volve wet or dry ashing and complicated
preparation procedures to isolate the lead
in solution. The atomic absorption tech
nique offers a rapid and sensitive alternative.
Our investigations indicate that a simple
boiling procedure with acetic acid is sufficient
resulting in the complete recovery of very
complex is not very stable and it is therefore
necessary to complete the determination with
the minimum delay after extraction. Fig. 2
shows recorder traces demonstrating the
recovery of added amounts of lead from beer.
If it is required to determine iron in the
beer sample, the same extract as was used
for lead may be aspirated and compared to
suitable standards as described under the
method for iron.
Of all the metals magnesium is the most
sensitive to detection by atomic absorption
and considerable dilution of beer is necessary.
The calibration curve obtained is somewhat
curved. Using an acetylene-air flame, no
interferences were noticed. The method
has been used in an investigation of
magnesium levels in beer and brewing
materials. Table IV gives the figures ob
tained for mashing liquors, worts and beers.
The changes occurring in the magnesium
content of wort during fermentation were
investigated with the results shown in Table
V. Determination of the magnesium content
of yeast after wet oxidation of the organic
matter with nitric and sulphuric acids gave

TABLE V
Magnesium Changes Occurring in the Magnesium Content
Material analysed (p.p.m.) of Wort During Fermentation by Different
Yeast strains
Mashing liquor C-4-7-8
Malt—soluble hot extract (calculated
to original malt) 620-850 Magineshim (p.p.m.)

Mean
vsiIiiaq
Yeast strain W (») («)
Bottled pale ale (O.G. 1032) 70 Initial wort
Draught pale ale (O.G. 1040) 86 (O.G. 1040).. .. 82 82 82 82
Bottled pale ale (O.G. 1050) 105 After 1 day .. .. 70 72 76 74
Bottled pale ale (O.G. 1068) 158 After 2 days .. .. 69 70 68 65
Strong ale (O.G. 108S) 190 After 3 days .. .. 66 68 73 71
Pilsner lager (O.G. 1032) .. 69 After 4 days .. .. 67 69 75 70
Pilsner lager (O.G. 1045) .. 89 After 7 days .. .. 72 75 78 75
Vol. 75, 1989] WE1NER AND TAYLOR: METALS IN BEER AND WINE 199

TABLE VI account of the much higher levels normally

Determination op Several Trace Metals in present.
Table Wines by Atomic Absorption Spectro- Standards may be prepared by the addi
photometry
tion of the metal solutions to the wine sample,
but for routine control purposes the use of
White Rose Red a 10% alcohol solution for the preparation
Metal (p.p.m.) wines wines wines
of the standards is found convenient and
Copper satisfactory.
range 0-2-1-7 0-4-1-1 0-25-1-4 Some typical trace metal figures obtained
average 0-71 0-73 0-74 on a selection of wines are given in Table
iron

range 2-8-9-4 3-5-10-0 0-6-150

VI.
average S-26 6-87 016
Zinc Acknowledgements.—The authors thank the
range 0-4-2-2 0-6-1-5 0-8-4-5 Directors of Courage, Barclay & Simonds
average 10 00 21
Limited for permission to publish this
Lead
range 0-10-0-28 0-05-0-08 paper.
average 0-16 007
References
1. Brenner, M. W., U.S. Patent No. 3,266, 902,
0-35 to 0-41% magnesium on the dry weight Aug. 16th, 1000.
2. Chapon, L., this Journal, 1065, 290.
of the yeast.
3. Clark, A. G., this Journal, 1960, 318.
The results in Table IV indicate that most 4. Elwell, W. T., & Gidley, J. A. F., Atomic
of the magnesium present in wort is derived Absorption Speclrophotometry, 2nd Ed.,
from malt and that the contribution from Pcrgamon Press Ltd., 1066.
5. Frey, S. W., DeWitt, W. G., & Bellomy, B. R.,
other sources is negligible. The results Proc. A. M. Amer. Soc. Brew. Chent., 1966,
suggest that there is ample magnesium 172.
in wort to satisfy requirements of yeast 6. Frey, S. W., DeWitt, W. G., & Bellomy, B. R.,
growth and metabolism and that magnesium Proc. A. M. Amer. Soc. Brew. Chew., 1967.
109.
deficiency is unlikely to occur.
7. Hudson, J. R.. this Journal, 1955, 127.
8. Hudson, J. R., this Journal, 1958, 157.
Examination of wines 0. Kahn, H. L., /. Chem. Educ, 1966,43, A7 and
A103.
The application of atomic absorption 10. Michel, G., Gagnairc, B., & Lebrcton, P.,
spectrophotometry to the analysis of table Bull. Soc. Chem. Biol., 1056, 931.
wines for trace metals has been very success 11. Nassan, A., Amer. Brewer, Feb. 1957, 49.
ful. The methods used are similar to those 12. Stone, I., U.S. Patent, 3, 164, 472, 6th Jan.,
1065.
detailed for beer, except that direct aspiration 13. Stone, I., U.S. Patent, 3, 223, 529. 14th Dec,
of the sample is satisfactory for iron on 1066.