BAM: Aerobic Plate Count
January 2001
Bacteriological Analytical Manual
Chapter 3
Aerobic Plate Count
Authors
Chapter Contents
Conventional Plate Count Method
Spiral Plate Method
References
The aerobic plate count (APC) is intended to indicate the level of microorganism in a
product. Detailed procedures for determining the APC of foods have been developed by
the Association of Official Analytical Chemists (AOAC) (3) and the American Public
Health Association (APHA) (1). The conventional plate count method for examining
frozen, chilled, precooked, or prepared foods, outlined below, conforms to AOAC
Official Methods of Analysis, sec. 966.23, with one procedural change (966.23C). The
suitable colony counting range (10) is 25-250. The automated spiral plate count method
for the examination of foods and cosmetics (5), outlined below, conforms to AOAC
Official Methods of Analysis, sec. 977.27. For procedural details of the standard plate
count, see ref. 2.Guidelines for calculating and reporting plate counts have been changed
to conform with the anticipated changes in the 16th edition of Standard Methods for the
Examination of Dairy Products (2) and the International Dairy Federation (IDF)
procedures (6).
Conventional Plate Count Method
Equipment and materials
1. Work area, level table with ample surface in room that is clean, well-
lighted (100 foot-candles at working surface) and well-ventilated, and
reasonably free of dust and drafts. The microbial density of air in working
area, measured in fallout pour plates taken during plating, should not
exceed 15 colonies/plate during 15 min exposure.
2. Storage space, free of dust and insects and adequate for protection of
equipment and supplies
3. Petri dishes, glass or plastic (at least 15 x 90 mm)
4. Pipets with pipet aids (no mouth pipetting) or pipettors, 1, 5, and 10 ml,
graduated in 0.1 ml units
5. Dilution bottles, 6 oz (160 ml), borosilicate-resistant glass, with rubber
stoppers or plastic screw caps
6. Pipet and petri dish containers, adequate for protection
7. Circulating water bath, for tempering agar, thermostatically controlled to
45 ± 1°C
8. Incubator, 35 ± 1°C; milk, 32 ± 1°C
9. Colony counter, dark-field, Quebec, or equivalent, with suitable light
source and grid plate
10. Tally register
11. Dilution blanks, 90 ± 1 ml Butterfield's phosphate-buffered dilution water
(R11); milk, 99 ± 2 ml
12. Plate count agar (standard methods) (M124)
13. Refrigerator, to cool and maintain samples at 0-5°C; milk, 0-4.4°C
14. Freezer, to maintain frozen samples from -15 to -20°C
15. Thermometers (mercury) appropriate range; accuracy checked with a
thermometer certified by the National Institute of Standards and
Technology (NIST)
Procedure for analysis of frozen, chilled, precooked, or prepared foods
Using separate sterile pipets, prepare decimal dilutions of 10-2, 10-3, 10-4, and
others as appropriate, of food homogenate (see Chapter 1 for sample preparation)
by transferring 10 ml of previous dilution to 90 ml of diluent. Avoid sampling
foam. Shake all dilutions 25 times in 30 cm (1 ft) arc within 7 s. Pipet 1 ml of
each dilution into separate, duplicate, appropriately marked petri dishes. Reshake
dilution bottle 25 times in 30 cm arc within 7 s if it stands more than 3 min before
it is pipetted into petri dish. Add 12-15 ml plate count agar (cooled to 45 ± 1°C) to
each plate within 15 min of original dilution. For milk samples, pour an agar
control, pour a dilution water control and pipet water for a pipet control. Add agar
to the latter two for each series of samples. Add agar immediately to petri dishes
when sample diluent contains hygroscopic materials, e.g., flour and starch. Pour
agar and dilution water control plates for each series of samples. Immediately mix
sample dilutions and agar medium thoroughly and uniformly by alternate rotation
and back-and-forth motion of plates on flat level surface. Let agar solidify. Invert
solidified petri dishes, and incubate promptly for 48 ± 2 h at 35°C. Do not stack
plates when pouring agar or when agar is solidifying.
Guidelines for calculating and reporting APCs in uncommon cases
Official Methods of Analysis (3) does not provide guidelines for counting and
reporting plate counts, whereas Standard Methods for the Examination of Dairy
Products, 16th ed. (2) presents detailed guidelines; for uniformity, therefore, use
APHA guidelines as modified (6,8). Report all aerobic plate counts (2) computed
from duplicate plates. For milk samples, report all aerobic plate (2) counts
computed from duplicate plates containing less than 25 colonies as less than 25
estimated count. Report all aerobic plate counts (2) computed from duplicate
plates containing more than 250 colonies as estimated counts. Counts outside the
normal 25-250 range may give erroneous indications of the actual bacterial
composition of the sample. Dilution factors may exaggerate low counts (less than
25), and crowded plates (greater than 250) may be difficult to count or may
inhibit the growth of some bacteria, resulting in a low count. Report counts less
than 25 or more than 250 colonies as estimated aerobic plate counts (EAPC). Use
the following guide:
16. Normal plates (25-250). Select spreader-free plate(s). Count all colony
forming units (CFU), including those of pinpoint size, on selected plate(s).
Record dilution(s) used and total number of colonies counted.
17. Plates with more than 250 colonies. When number of CFU per plate
exceeds 250, for all dilutions, record the counts as too numerous to count
(TNTC) for all but the plate closest to 250, and count CFU in those
portions of plate that are representative of colony distribution. See ref. 2
for detailed guidelines. Mark calculated APC with EAPC to denote that it
was estimated from counts outside 25-250 per plate range (see D-3).
18. Spreaders. Spreading colonies are usually of 3 distinct types: 1) a chain of
colonies, not too distinctly separated, that appears to be caused by
disintegration of a bacterial clump; 2) one that develops in film of water
between agar and bottom of dish; and 3) one that forms in film of water at
edge or on surface of agar. If plates prepared from sample have excessive
spreader growth so that (a) area covered by spreaders, including total area
of repressed growth, exceeds 50% of plate area, or (b) area of repressed
growth exceeds 25% of plate area, report plates as spreaders. When it is
necessary to count plates containing spreaders not eliminated by (a) or (b)
above, count each of the 3 distinct spreader types as one source. For the
first type, if only one chain exists, count it as a single colony. If one or
more chains appear to originate from separate sources, count each source
as one colony. Do not count each individual growth in such chains as a
separate colony. Types 2 and 3 usually result in distinct colonies and are
counted as such. Combine the spreader count and the colony count to
compute the APC.
19. Plates with no CFU. When plates from all dilutions have no colonies,
report APC as less than 1 times the corresponding lowest dilution used.
Mark calculated APC with asterisk to denote that it was estimated from
counts outside the 25-250 per plate range. When plate(s) from a sample
are known to be contaminated or otherwise unsatisfactory, record the
result(s) as laboratory accident (LA).
Computing and recording counts (see refs 6, 8)
To avoid creating a fictitious impression of precision and accuracy when
computing APC, report only the first two significant digits. Round off to two
significant figures only at the time of conversion to SPC. For milk samples, when
plates for all dilutions have no colonies, report APC as less than 25 colonies
estimated count. Round by raising the second digit to the next highest number
when the third digit is 6, 7, 8, or 9 and use zeros for each successive digit toward
the right from the second digit. Round down when the third digit is 1, 2, 3, or 4.
When the third digit is 5, round up when the second digit is odd and round down
when the second digit is even.
Examples
Calculated Count APC
12,700 13,000
12,400 12,000
15,500 16,000
14,500 14,000
1. Plates with 25-250 CFU.
Formula: N = Σ C / [ (1 * n1) + (0.1 * n2) ] * (d)
a. Calculate the APC as follows:
where N = Number of colonies per ml or g of product
Σ C = Sum of all colonies on all plates counted
n1 = Number of plates in first dilution counted
n2 = Number of plates in second dilution counted
d = Dilution from which the first counts were obtained
Example
1:100 1:1000
232, 244 33, 28
= 537/0.022
= 24,409
= 24,000
b. When counts of duplicate plates fall within and without the 25-250 colony range, use
only those counts that fall within this range.
2. All plates with fewer than 25 CFU. When plates from both dilutions yield fewer than
25 CFU each, record actual plate count but record the count as less than 25 x 1/d when d
is the dilution factor for the dilution from which the first counts were obtained.
Example
Colonies
1:100 1:1000 EAPC/ml (g)
18 2 <2,500
0 0 <2,500
3. All plates with more than 250 CFU. When plates from both 2 dilutions yield more
than 250 CFU each (but fewer than 100/cm2), estimate the aerobic counts from the plates
(EAPC) nearest 250 and multiply by the dilution.
Example
Colonies
1:100 1:1000 EAPC/ml (g)
TNTC 640 640,000
TNTC, too numerous to count.
EAPC, estimated aerobic plate count.
4. All plates with spreaders and/or laboratory accident. Report respectively as Spreader
(SPR), or Laboratory Accident (LA).
5. All plates with more than an average of 100 CFU per sq cm. Estimate the APC as
greater than 100 times the highest dilution plated, times the area of the plate. The
examples below have an average count of 110 per sq cm.
Example
Colonies/Dilution
1:100 1:1000 EAPC/ml (g)
>6,500,000
TNTC 7,150(a)
EAPC(b)
>5,900,000
TNTC 6,490
EAPC
a
Based on plate area of 65 cm2.
b
EAPC, estimated APC.
c
Based on plate area of 59 cm2.
Spiral Plate Method
The spiral plate count (SPLC) method for microorganisms in milk, foods, and cosmetics
is an official method of the APHA (2) and the AOAC (3). In this method, a mechanical
plater inoculates a rotating agar plate with liquid sample. The sample volume dispensed
decreases as the dispensing stylus moves from the center to the edge of the rotating plate.
The microbial concentration is determined by counting the colonies on a part of the petri
dish where they are easily countable and dividing this count by the appropriate volume.
One inoculation determines microbial densities between 500 and 500,000
microorganisms/ml. Additional dilutions may be made for suspected high microbial
concentrations.
Equipment and materials
1. Spiral plater (Spiral Systems Instruments, Inc., 7830 Old Georgetown
Road, Bethesda, MD 20814)
2. Spiral colony counter (Spiral Systems) with special grid for relating
deposited sample volumes to specific portions of petri dishes
3. Vacuum trap for disposal of liquids (2-4 liter vacuum bottle to act as
vacuum reservoir and vacuum source of 50-60 cm Hg)
4. Disposable micro beakers, 5 ml
5. Petri dishes, plastic or glass, 150 x 15 mm or 100 x 15 mm
6. Plate count agar (standard methods) (M124)
7. Calculator (optional), inexpensive electronic hand calculator is
recommended
8. Polyethylene bags for storing prepared plates
9. Commercial sodium hypochlorite solution, about 5% NaOCl (bleach)
10. Sterile dilution water
11. Syringe, with Luer tip for obstructions in stylus; capacity not critical
12. Work area, storage space, refrigerator, thermometers, tally, incubator, as
described for Conventional Plate Count Method, above.
13. Sodium hypochlorite solution (5.25%). Available commercially.
Preparation of agar plates.
Automatic dispenser with sterile delivery system is recommended to prepare agar
plates. Agar volume dispensed into plates is reproducible and contamination rate
is low compared to hand-pouring of agar in open laboratory. When possible, use
laminar air flow hood along with automated dispenser. Pour same quantity of agar
into all plates so that same height of agar will be presented to spiral plater stylus
tip to maintain contact angle. Agar plates should be level during cooling.
The following method is suggested for prepouring agar plates: Use automatic
dispenser or pour constant amount (about 15 ml/100 mm plate; 50 ml/150 mm
plate) of sterile agar at 60-70°C into each petri dish. Let agar solidify on level
surface with poured plates stacked no higher than 10 dishes. Place solidified agar
plates in polyethylene bags, close with ties or heat-sealer, and store inverted at 0-
4.4°C. Bring prepoured plates to room temperature before inoculation.
Preparation of samples.
As described in Chapter 1, select that part of sample with smallest amount of
connective tissues or fat globules.
Description of spiral plater.
Spiral plater inoculates surface of prepared agar plate to permit enumeration of
microorganisms in solutions containing between 500 and 500,000
microorganisms per ml. Operator with minimum training can inoculate 50 plates
per h. Within range stated, dilution bottles or pipets and other auxiliary equipment
are not required. Required bench space is minimal, and time to check instrument
alignment is less than 2 min. Plater deposits decreasing amount of sample in
Archimedean spiral on surface of prepoured agar plate. Volume of sample on any
portion of plate is known. After incubation, colonies appear along line of spiral. If
colonies on a portion of plate are sufficiently spaced from each other, count them
on special grid which associates a calibrated volume with each area. Estimate
number of microorganisms in sample by dividing number of colonies in a defined
area by volume contained in same area. Studies have shown the method to be
proficient not only with milk (4) but also with other foods (7,10).
Plating procedure
Check stylus tip angle daily and adjust if necessary. (Use vacuum to hold
microscope cover slip against face of stylus tip; if cover slip plane is parallel at
about l mm from surface of platform, tip is properly oriented). Liquids are moved
through system by vacuum. Clean stylus tip by rinsing for 1 s with sodium
hypochlorite solution followed by sterile dilution water for 1 s before sample
introduction. This rinse procedure between processing of each sample minimizes
cross-contamination. After rinsing, draw sample into tip of Teflon tubing by
vacuum applied to 2-way valve. When tubing and syringe are filled with sample,
close valve attached to syringe. Place agar plate on platform, place stylus tip on
agar surface, and start motor. During inoculation, label petri plate lid. After agar
has been inoculated, stylus lifts from agar surface and spiral plater automatically
stops. Remove inoculated plate from platform and cover it. Move stylus back to
starting position. Vacuum-rinse system with hypochlorite and water, and then
introduce new sample. Invert plates and promptly place them in incubator for 48 ±
3 h at 35 ± 1°C.
Sterility controls
Check sterility of spiral plater for each series of samples by plating sterile dilution
water. CAUTION: Prepoured plates should not be contaminated by a surface
colony or be below room temperature (water can well-up from agar). They should
not be excessively dry, as indicated by large wrinkles or glazed appearance. They
should not have water droplets on surface of agar or differences greater than 2
mm in agar depth, and they should not be stored at 0-4.4°C for longer than l
month. Reduced flow rate through tubing indicates obstructions or material in
system. To clear obstructions, remove valve from syringe, insert hand-held
syringe with Luer fitting containing water, and apply pressure. Use alcohol rinse
to remove residual material adhering to walls of system. Dissolve accumulated
residue with chromic acid. Rinse well after cleaning.
Counting grid
14. Description. Use same counting grid for both 100 and 150 mm petri
dishes. A mask is supplied for use with 100 mm dishes. Counting grid is
divided into 8 equal wedges; each wedge is divided by 4 arcs labeled l, 2,
3, and 4 from outside grid edge. Other lines within these arcs are added for
ease of counting. A segment is the area between 2 arc lines within a
wedge. Number of areas counted (e.g., 3) means number of segments
counted within a wedge. Spiral plater deposits sample on agar plate in the
same way each time. The grid relates colonies on spiral plate to the
volume in which they were contained. When colonies are counted with
grid, sample volume becomes greater as counting starts at outside edge of
plate and proceeds toward center of plate.
15. Calibration. The volume of sample represented by various parts of the
counting grid is shown in operator's manual that accompanies spiral plater.
Grid area constants have been checked by the manufacturer and are
accurate. To verify these values, prepare 11 bacterial concentrations in
range of 106-103 cells/ml by making 1:1 dilutions of bacterial suspension
(use a nonspreader). Plate all Incubate both sets of plates for 48 ± 3 h at 35
± 1°C. Calculate concentrations for each dilution. Count spiral plates over
grid surface, using counting rule of 20 (described in H, below), and record
number of colonies counted and grid area over which they were counted.
Each spiral colony count for a particular grid area, divided by aerobic
count/ml for corresponding spirally plated bacterial concentrations,
indicates volume deposited on that particular grid area. Use the following
formula:
To check total volume dispensed by spiral plater, weigh amount dispensed from
stylus tip. Collect in tared 5 ml plastic beaker and weigh on analytical balance (±
0.2 mg).
Figure 1. 10 cm plate, area (3b)
Examination and reporting of spiral plate counts.
Counting rule of 20. After incubation, center spiral plate over grid by adjusting
holding arms on viewer. Choose any wedge and begin counting colonies from
outer edge of first segment toward center until 20 colonies have been counted.
Complete by counting remaining colonies in segment where 20th colony occurs.
In this counting procedure, numbers such as 3b, 4c (Fig. l) refer to area segments
from outer edge of wedge to designated arc line. Any count irregularities in
sample composition are controlled by counting the same segments in the opposite
wedge and recording results. Example of spirally inoculated plate (Fig. l)
demonstrates method for determining microbial count. Two segments of each
wedge were counted on opposite sides of plate with 31 and 30 colonies,
respectively. The sample volume contained in the darkened segments is 0.0015
ml. To estimate number of microorganisms, divide count by volume contained in
all segments counted. See example under Fig. l.
If 20 CFU are not within the 4 segments of the wedge, count CFU on entire plate.
If the number of colonies exceeds 75 in second, third, or fourth segment, which
also contains the 20th colony, the estimated number of microorganisms will
generally be low because of coincidence error associated with crowding of
colonies. In this case, count each circumferentially adjacent segment in all 8
wedges, counting at least 50 colonies, e.g., if the first 2 segments of a wedge
contain 19 colonies and the third segment contains the 20th and 76th (or more),
count colonies in all circumferentially adjacent first and second segments in all 8
wedges. Calculate contained volume in counted segments of wedges and divide
into number of colonies.
When fewer than 20 colonies are counted on the total plate, report results as "less
than 500 estimated SPLC per ml." If colony count exceeds 75 in first segment of
wedge, report results as "greater than 500,000 estimated SPLC per ml." Do not
count spiral plates with irregular distribution of colonies caused by dispensing
errors. Report results of such plates as laboratory accident (LA). If spreader
covers entire plate, discard plate. If spreader covers half of plate area, count only
those colonies that are well distributed in spreader-free areas.
Compute SPLC unless restricted by detection of inhibitory substances in sample,
excessive spreader growth, or laboratory accidents. Round off counts as described
in I-D, above. Report counts as SPLC or estimated SPLC per ml.
References
1. American Public Health Association. 1984. Compendium of Methods for the
Microbiological Examination of Foods, 2nd ed. APHA, Washington, DC
2. American Public Health Association. 1993. Standard Methods for the
Examination of Dairy Products, 16th ed. APHA, Washington, DC.
3. Association of Official Analytical Chemists. 1990. Official Methods of Analysis,
15th ed. AOAC, Arlington, VA.
4. Donnelly, C.B., J.E. Gilchrist, J.T. Peeler, and J.E. Campbell. 1976. Spiral plate
count method for the examination of raw and pasteurized milk. Appl. Environ.
Microbiol. 32:21-27.
5. Gilchrist, J.E., C.B. Donnelly, J.T. Peeler, and J.E. Campbell. 1977.
Collaborative study comparing the spiral plate and aerobic plate count methods. J.
Assoc. Off. Anal. Chem. 60:807-812.
6. International Dairy Federation. 1987. Milk and Milk Products: Enumeration of
Microorganisms--Colony Count at 3°C. Provisional IDF Standard 100A. IDF,
Brussels, Belgium.
7. Jarvis, B., V.H. Lach, and J.M. Wood. 1977. Evaluation of the spiral plate maker
for the enumeration of microorganisms in foods. J. Appl. Bacteriol. 43:149-157.
8. Niemela, S. 1983. Statistical evaluation of Results from Quantitative
Microbiological Examinations. Report No. 1, 2nd ed. Nordic Committee in Food
Analysis, Uppsala, Sweden.
9. Tomasiewicz, D.M., D.K. Hotchkiss, G.W. Reinbold, R.B. Read, Jr., and P.A.
Hartman. 1980. The most suitable number of colonies on plates for counting. J.
Food Prot. 43:282-286.
10. Zipkes, M.R., J.E. Gilchrist, and J.T. Peeler. 1981. Comparison of yeast and
mold counts by spiral, pour, and streak plate methods. J. Assoc. Off. Anal. Chem.
64:1465-1469.
Hypertext Source: Bacteriological Analytical Manual, Edition 8, Revision A, 1998.
Chapter 3.
*Authors: Larry Maturin and James T. Peeler
Compendium
3.1 See Appendix A of Volume 2.
3.2 Commercial Sterility: the condition achieved by the application of heat, alone or in
combination with other treatments, to render a food free from viable forms of
microorganisms, including spores, capable of growing in the food at temperature s at
which the food is designed normally to be held during distribution and storage. If the
normal temperature for storage or handling of the product is higher than 40°C then
analyse for Thermophiles. Otherwise, analyse for Mesophiles only.
This definition does not apply to low acid foods which are to be kept under refrigeration
or frozen and labelled as such or to tomato products which have a pH of 4.7 or less after
heat processing.
3.3 Hermetically Sealed Containers: containers designed and intended to be secure
against the entry of microorganisms, including spores. These containers include metal
cans, glass jars, flexible packages such as retort pouches and Tetrapaks, etc.
3.4 Low Acid Food: a food, other than alcoholic beverages, where any component of the
food has a pH greater than 4.6 and a water activity greater than 0.85.
3.5 Acid and Acidified Food: a food with a pH equal to or lower than 4.6.
3.6 Water Activity: the ratio of the vapour pressure of a food to the vapour pressure of
pure water, at the same temperature and pressure.
3.7 Refrigeration: exposure to a temperature of 4°C or less but not frozen.
3.8 Semi-Liquid Products: products that may pour with difficulty, but can be mixed by
shaking the container.
Prior to opening containers containing liquid products, the contents should be mixed by
shaking the unopened container.
3.9 Semi-Solid Products: products that have a high viscosity; they pour with great
difficulty, and cannot be mixed by shaking the unopened container.
Sodium benzoate
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Sodium benzoate
IUPAC name[hide]
Sodium benzoate
Other names[hide]
E211, benzoate of soda
Identifiers
CAS number 532-32-1
PubChem 517055
RTECS number DH6650000
SMILES [show]
Properties
Molecular formula NaC6H5CO2
Molar mass 144.11 g/mol
Density 1.497 g/cm3
Melting point 300 °C
Solubility in water soluble
Solubility soluble in ethanol
Acidity (pKa) 8.0
Hazards
Flash point 100 °C
Autoignition
500 °C
temperature
LD50 4100 mg/kg (oral, rat)
(what is this?) (verify)
Except where noted otherwise, data are given for materials in
their standard state (at 25 °C, 100 kPa)
Infobox references
Sodium benzoate (E211) has the chemical formula NaC6H5CO2. It is the sodium salt of
benzoic acid and exists in this form when dissolved in water. It can be produced by
reacting sodium hydroxide with benzoic acid.
Contents
[hide]
• 1 Uses
• 2 Mechanism of food preservation
• 3 Safety and health
o 3.1 Hyperactivity
• 4 Compendial status
• 5 See also
• 6 References
• 7 External links
[edit] Uses
Sodium benzoate is a preservative. It is bacteriostatic and fungistatic under acidic
conditions. It is used most prevalently in acidic foods such as salad dressings (vinegar),
carbonated drinks (carbonic acid), jams and fruit juices (citric acid), pickles (vinegar),
and condiments. It is also found in alcohol-based mouthwash and silver polish.[citation needed]
It can also be found in cough syrups like Robitussin.[1] Sodium benzoate is declared on a
product label as 'sodium benzoate' or E211.
It is also used in fireworks as a fuel in whistle mix, a powder that emits a whistling noise
when compressed into a tube and ignited. The fuel is also one of the fastest burning
rocket fuels and provides a lot of thrust and smoke. It does have its downsides, there is a
high danger of explosion when the motor is pressed because of the fuel's sensitivity to
impact. That is why only professional pyrotechnicians should make it.
Sodium benzoate is produced by the neutralization of benzoic acid with sodium
hydroxide.[2] Benzoic acid is detectable at low levels in cranberries, prunes, greengage
plums, cinnamon, ripe cloves, and apples. Though benzoic acid is a more effective
preservative, sodium benzoate is more commonly used as a food additive because
benzoic acid does not dissolve well in water.[2] Concentration as a preservative is limited
by the FDA in the U.S. to 0.1% by weight. The International Programme on Chemical
Safety found no adverse effects in humans at doses of 647–825 mg/kg of body weight per
day.[3][4]
Cats have a significantly lower tolerance against benzoic acid and its salts than rats and
mice.[5] Sodium benzoate is, however, allowed as an animal food additive at up to 0.1%,
according to AFCO's official publication.[6]
[edit] Mechanism of food preservation
The mechanism starts with the absorption of benzoic acid into the cell. If the intracellular
pH changes to 5 or lower, the anaerobic fermentation of glucose through
phosphofructokinase is decreased by 95%.[7]
[edit] Safety and health
Main article: benzene in soft drinks
In combination with ascorbic acid (vitamin C, E300), sodium benzoate and potassium
benzoate does form benzene, a known carcinogen; however the levels are below those
considered dangerous for consumption [8]. Heat, light and shelf life can affect the rate at
which benzene is formed.
Professor Peter James Piper of the University of Sheffield claims that sodium benzoate
by itself can damage and inactivate vital parts of DNA in a cell's mitochondria.
Mitochondria consume oxygen to generate ATP, the body's energy currency. If they are
damaged due to disease, the cell malfunctions and may enter apoptosis. There are many
illnesses now tied to DNA damage, including Parkinson's and other neurodegenerative
diseases, but above all, the aging process in general.[9][10][11][12][13]
[edit] Hyperactivity
Research published in 2007 for the UK's Food Standards Agency (FSA) suggests that
certain artificial colours, when paired with sodium benzoate (E211) may be linked to
hyperactive behaviour. The results were inconsistent regarding sodium benzoate, so the
FSA recommended further study.[14][15][16]
Professor Jim Stevenson from Southampton University, and author of the report, said:
"This has been a major study investigating an important area of research. The results
suggest that consumption of certain mixtures of artificial food colours and sodium
benzoate preservative are associated with increases in hyperactive behaviour in children.
However, parents should not think that simply taking these additives out of food will
prevent hyperactive disorders. We know that many other influences are at work but this
at least is one a child can avoid."[16]
Two mixtures of additives were tested in the research:[16]
Mix A:
• Sunset yellow (E110)
• Tartrazine (E102)
• Carmoisine (E122)
• Ponceau 4R (E124)
• Sodium benzoate (E211)
Mix B:
• Sunset yellow (E110)
• Quinoline yellow (E104)
• Carmoisine (E122)
• Allura red (E129)
• Sodium benzoate (E211)
Sodium benzoate was included in both mixes, but the effects observed were not
consistent. The Food Standards Agency therefore considers that, if real, the observed
increases in hyperactive behaviour were more likely to be linked to one or more of the
specific colours tested.
On 10 April 2008, the Foods Standard Agency called for a voluntary removal of the
colours (but not sodium benzoate) by 2009.[17] In addition, it recommended that there
should be action to phase them out in food and drink in the European Union (EU) over a
specified period.[18]
In response to consumer insistence on a more natural product and E211's links to DNA
damage and ADHD, the Coca Cola Company is in the process of phasing Sodium
Benzoate out of Diet Coke. The company has stated that it plans to remove E211 from its
other products — including Sprite, Fanta, and Oasis — as soon as a satisfactory
alternative is discovered.[19]
[edit] Compendial status
• British Pharmacopoeia[20][21][22]
• Food Chemical Codex[20][clarification needed]
• European Pharmacopoeia[20][clarification needed]
• Japanese Pharmacopoeia [23][clarification needed]
• United States Pharmacopeia 29[24]
[edit] See also
• Acceptable daily intake
• Food-induced purpura
[edit] References
1. ^ Sodium benzoate in Robitussin cough
2. ^ a b INTERNATIONAL PROGRAMME ON CHEMICAL SAFETY
3. ^ Concise International Chemical Assessment Document 26: Benzoic acid and
sodium benzoate
4. ^ Cosmetic Ingredient Review Expert Panel Bindu Nair (2001). "Final Report on
the Safety Assessment of Benzyl Alcohol, Benzoic Acid, and Sodium Benzoate".
Int J Tox 20 (Suppl 3): 23–50. doi:10.1080/10915810152630729.
5. ^ Bedford PG, Clarke EG (January 1972). "Experimental benzoic acid poisoning
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14. ^ Food Standards Agency issues revised advice on certain artificial colours 6
September 2007
15. ^ Food Colorings and Hyperactivity "Myomancy" 7 September 2007
16. ^ a b c Agency revises advice on certain artificial colours Food Standards Agency
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17. ^ BBC Europe-wide food colour ban call 10 April 2008
18. ^ FSA Board discusses colours advice 10 April 2008
19. ^ The Daily Mail DNA Damage Fear 24 May 2008
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http://en.wikipedia.org/wiki/Sodium_benzoate
