low doses aspartame, acesulfame-K, saccharin in mice induce DNA damage in
bone marrow -- "a potential health risk", S Ghoshal et al, U. Kentucky,
Lexington, Drug Chem Toxicol 2008: Rich Murray 2008.10.15
http://rmforall.blogspot.com/2008_10_01_archive.htm
Wednesday, October 15, 2008
http://groups.yahoo.com/group/aspartameNM/message/1567

[ See also for similar results:

http://groups.yahoo.com/group/aspartameNM/message/961
genotoxins, Comet assay in mice: Ace-K, stevia fine; aspartame poor;
sucralose, cyclamate, saccharin bad: Y.F. Sasaki Aug 2002: Murray 2003.01.27
[A detailed look at the data] ]


Drug Chem Toxicol. 2008; 31(4): 447-57.
Genotoxicity testing of low-calorie sweeteners: aspartame, acesulfame-K, and
saccharin.
Bandyopadhyay A,
Ghoshal S,
Mukherjee A.
Centre of Advanced Study, Cell and Chromosome Research, Department of
Botany, University of Calcutta, Kolkata, India.

Authors:
Atrayee Bandyopadhyay a; atrayee.banerjee@...;
Sarbani Ghoshal b;
Anita Mukherjee a
Affiliations:
a Centre of Advanced Study, Cell and Chromosome Research, Department of
Botany, University of Calcutta, Kolkata, India
b Present address: Department of Internal Medicine, University of Kentucky,
Lexington, Kentucky, USA

Low-calorie sweeteners are chemicals that offer the sweetness of sugar
without the calories.

Consumers are increasingly concerned about the quality and safety of many
products present in the diet, in particular, the use of low-calorie
sweeteners, flavorings, colorings, preservatives, and dietary supplements.

In the present study, we evaluated the mutagenicity of the three low-calorie
sweeteners in the Ames/Salmonella/microsome test and their genotoxic
potential by comet assay in the bone marrow cells of mice.

Swiss albino mice, Mus musculus, were orally administered with different
concentrations of
aspartame (ASP; 7, 14, 28, and 35 mg/kg body weight),
acesulfame-K (ASK; 150, 300, and 600 mg/kg body weight),
and saccharin (50, 100, and 200 mg/kg body weight) individually.

Concurrently negative and positive control sets were maintained.

The animals were sacrificed and the bone marrow cells were processed for
comet assay.

The standard plate-incorporation assay was carried with the three sweeteners
in Salmonella typhimurium TA 97a and TA 100 strains both in the absence and
presence of the S9 mix.

The comet parameters of DNA were increased in the bone marrow cells due to
the sweetener-induced DNA strand breaks, as revealed by increased comet-tail
extent and percent DNA in the tail.

ASK and saccharin were found to induce greater DNA damage than ASP.

However, none could act as a potential mutagen in the Ames/Salmonella
/microsome test.

These findings are important, since they represent a potential health risk
associated with the exposure to these agents. PMID: 18850355

Sarbani Ghoshal
Email: sarbanighoshal@...,
Title: Post-Doctoral Scholar
Department: Pharmaceutical Sciences
Address: 561 Charles T Wethington Building 40536-0200
phone 859 323-4993 home phone 859 323-9610

http://www.psgcas.ac.in/downloads/FreePaperpresentation.pdf

POSTER PRESENTATION (11.01.2007) THURSDAY
SESSION IX (4.30 TO 6.00 PM)

PP 18 Atrayee Bandyopadhyay, et.al.,
atrayee.banerjee@...
Center for Advanced Study, Cell & Chromosome Research,
Dept. of Botany, Univ of Calcutta , Kolkata
DNA damage induced by Aspartame a low calorie sweet

PP 10 Ms. Salma Ghosh & Anita Mukherjee,
anitamukherjee28@...
Center of Advanced Study in cell & Chromosome Research,
Dept. of Botany, Univ of Calcutta, Kolkata
Evaluation of DNA damage by ophenylenediamine in Allium assay
___________________________________________________


similar levels of daily formaldehyde and formic acid, causes of
birth defects, come from cigarettes, aspartame, and dark wines and
liquors -- folic acid protects most people: Rich Murray 2008.07.15
http://rmforall.blogspot.com/2008_07_01_archive.htm
Tuesday, July 15, 2008
http://groups.yahoo.com/group/aspartameNM/message/1552

"A smoker who goes through one pack a day will smoke 7,300
cigarettes a year, inhaling the equivalent of nearly 1 gram of
formaldehyde (yikes!)."

That's about 2.5 mg daily formaldehyde intake for 20 cigarettes,
over the 2 mg USA FDA alarm level for formaldehyde in average
2 liters daily drinking water, while a single 12 oz can of diet soda
also results in about 2 mg formaldehyde toxic products in the body,
including formic acid, a notorious cause of birth defects.

Dark wines and liquors usually supply even more methanol, which
the body always turns into formaldehyde and formic acid -- the
major cause of "morning after" hangovers.

High levels of folic acid, a safe, affordable vitamin in fruits and
vegetables, largely prevents formaldehyde and formic acid toxicity
in most people.

It is certain that high levels of aspartame use,
above 2 liters daily for months and years,
must lead to chronic formaldehyde-formic acid toxicity.

Fully 11 % of aspartame is methanol -- 1,120 mg aspartame
in 2 liters diet soda, almost six 12-oz cans, gives 123 mg methanol
(wood alcohol). The methanol is immediately released
into the body after drinking .
Within hours, the liver turns much of the methanol into formaldehyde,
and then much of that into formic acid, both of which in time
are partially eliminated as carbon dioxide and water.

However, about 30 % of the methanol remains in the body
as cumulative durable toxic metabolites of formaldehyde
and formic acid -- 37 mg daily,
a gram every month, accumulating in and affecting every tissue.

If only 10 % of the methanol is retained daily as formaldehyde,
that would give 12 mg daily formaldehyde accumulation -- about
60 times more than the 0.2 mg from 10 % retention
of the 2 mg EPA daily limit for formaldehyde in drinking water.

Bear in mind that the EPA limit for formaldehyde in drinking water is
1 ppm, or 2 mg daily for a typical daily consumption of 2 liters of water.


formaldehyde and formic acid in FEMA trailers and other sources
(aspartame, dark wines and liquors, tobacco smoke):
Murray 2008.01.30
http://rmforall.blogspot.com/2008_01_01_archive.htm
Wednesday, January 30, 2008
http://groups.yahoo.com/group/aspartameNM/message/1508

The FEMA trailers give about the same amount of formaldehyde
and formic acid daily as from a quart of dark wine or liquor,
or two quarts (6 12-oz cans) of aspartame diet soda,
from their over 1 tenth gram methanol impurity
(one part in 10,000), which the body quickly makes into
formaldehyde and then formic acid -- enough to be the major cause
of "morning after" alcohol hangovers.

Methanol and formaldehyde and formic acid also result from
many fruits and vegetables, tobacco and wood smoke, heater
and vehicle exhaust, household chemicals and cleaners, cosmetics,
and new cars, drapes, carpets, furniture, particleboard,
mobile homes, buildings, leather... so all these sources add up
and interact with many other toxic chemicals.


methanol impurity in alcohol drinks [ and aspartame ] is turned into
neurotoxic formic acid, prevented by folic acid, re Fetal Alcohol
Syndrome, BM Kapur, DC Lehotay, PL Carlen at U. Toronto,
Alc Clin Exp Res 2007 Dec. plain text: detailed biochemistry,
CL Nie et al. 2007.07.18: Murray 2008.02.24
http://rmforall.blogspot.com/2008_02_01_archive.htm
Sunday, February 24, 2008
http://groups.yahoo.com/group/aspartameNM/message/1524


http://groups.yahoo.com/group/aspartameNM/message/1469
highly toxic formaldehyde, the cause of alcohol hangovers, is
made by the body from 100 mg doses of methanol from
dark wines and liquors, dimethyl dicarbonate, and aspartame:
Murray 2007.08.31

http://groups.yahoo.com/group/aspartameNM/message/1286
methanol products (formaldehyde and formic acid) are main cause
of alcohol hangover symptoms [same as from similar amounts of
methanol, the 11% part of aspartame]: YS Woo et al, 2005 Dec:
Murray 2006.01.20

Addict Biol. 2005 Dec;10(4): 351-5.
Concentration changes of methanol in blood samples during
an experimentally induced alcohol hangover state.
Woo YS, Yoon SJ, Lee HK, Lee CU, Chae JH, Lee CT, Kim DJ.
Chuncheon National Hospital, Department of Psychiatry,
The Catholic University of Korea, Seoul, Korea.
http://www.cuk.ac.kr/eng/ sysop@...
Songsin Campus: 02-740-9714 Songsim Campus: 02-2164-4116
Songeui Campus: 02-2164-4114
http://www.cuk.ac.kr/eng/sub055.htm eight hospitals

[ Han-Kyu Lee ]

A hangover is characterized by the unpleasant physical and mental
symptoms that occur between 8 and 16 hours after drinking alcohol.

After inducing experimental hangover in normal individuals,
we measured the methanol concentration prior to
and after alcohol consumption
and we assessed the association between the hangover condition
and the blood methanol level.

A total of 18 normal adult males participated in this study.

They did not have any previous histories of psychiatric
or medical disorders.

The blood ethanol concentration prior to the alcohol intake
(2.26+/-2.08) was not significantly different from that
13 hours after the alcohol consumption (3.12+/-2.38).

However, the difference of methanol concentration
between the day of experiment (prior to the alcohol intake)
and the next day (13 hours after the alcohol intake)
was significant (2.62+/-1.33/l vs. 3.88+/-2.10/l, respectively).

A significant positive correlation was observed
between the changes of blood methanol concentration
and hangover subjective scale score increment when covarying
for the changes of blood ethanol level (r=0.498, p<0.05).

This result suggests the possible correlation of methanol
as well as its toxic metabolite to hangover. PMID: 16318957

[ The toxic metabolite of methanol is formaldehyde, which in turn
partially becomes formic acid -- both potent cumulative toxins
that are the actual cause of the toxicity of methanol.]

This study by Jones AW (1987) found next-morning hangover
from red wine with 100 to 150 mg methanol
(9.5 % w/v ethanol, 100 mg/l methanol, 0.01 %).
Fully 11% of aspartame is methanol --
1,120 mg aspartame in 2 L diet soda,
almost six 12-oz cans, gives 123 mg methanol (wood alcohol).

Pharmacol Toxicol. 1987 Mar; 60(3): 217-20.
Elimination half-life of methanol during hangover.
Jones AW. wayne.jones@...;
Department of Forensic Toxicology,
University Hospital, SE-581 85 Linkoping, Sweden.

This paper reports the elimination half-life of methanol in human
volunteers.
Experiments were made during the morning after the subjects had
consumed 1000-1500 ml red wine
(9.5 % w/v ethanol, 100 mg/l methanol)
the previous evening. [ 100 to 150 mg methanol ]
The washout of methanol from the body
coincided with the onset of hangover.
The concentrations of ethanol and methanol in blood were
determined indirectly by analysis of end-expired alveolar air.
In the morning when blood-ethanol dropped
below the Km of liver alcohol dehydrogenase (ADH)
of about 100 mg/l (2.2 mM),
the disappearance half-life of ethanol was 21, 22, 18 and 15 min.
in 4 test subjects respectively.
The corresponding elimination half-lives of methanol
were 213, 110, 133 and 142 min. in these same individuals.
The experimental design outlined in this paper can be used
to obtain useful data on elimination kinetics of methanol
in human volunteers without undue ethical limitations.
Circumstantial evidence is presented to link methanol
or its toxic metabolic products, formaldehyde and formic acid,
with the pathogenesis of hangover. PMID: 3588516 ]


http://groups.yahoo.com/group/aspartameNM/message/1143
methanol (formaldehyde, formic acid) disposition, Bouchard M et al, full
plain text, 2001 -- substantial sources are degradation of fruit pectins,
liquors, aspartame, smoke: Murray 2005.01.05


http://groups.yahoo.com/group/aspartameNM/message/925
aspartame puts formaldehyde adducts into tissues, Part 1 of 2 full text,
Trocho & Alemany 1998.06.26: Murray 2002.12.22


opportunities re BA Magnuson, GA Burdock et al., Aspartame Safety Evaluation
2007 Sept., Critical Reviews in Toxicology: Rich Murray 2008.07.11
http://rmforall.blogspot.com/2008_07_01_archive.htm
Friday, July 11, 2008 full text
http://groups.yahoo.com/group/aspartameNM/message/1550 partial text

Thrasher JD, Kilburn KH. toxicologist@...;
Embryo toxicity and teratogenicity of formaldehyde. [100 references]
Arch Environ Health 2001 Jul-Aug; 56(4): 300-11.
Sam-1 Trust, Alto, New Mexico, USA. (702) 987-4590 (505) 937-1150
http://www.drthrasher.org/formaldehyde_embryo_toxicity.html full text

"The major difference is that the Japanese demonstrated the incorporation of
FA and its metabolites into the placenta and fetus. The quantity of
radioactivity remaining in maternal and fetal tissues
at 48 hours was 26.9 % of the administered dose." [ Ref. 14-16 ] The DNA
fraction contained 20 % and 50% of total incorporated radioactivity in the
maternal and fetal liver at 6 and 24 hours when compared to the acid
insoluble fraction (Fig. 1). Of primary interest is that the incorporated
radioactivity persisted longer in the fetal liver and brain when compared to
the mothers."


The ASE review missed the actual details of this provocative study by an
experienced team that continues to use the sensitive, fast, low cost
automated Comet assay to measure DNA damage.

The team tested 39 food additives, including 7 sweeteners, using groups of
just 4 mice at a uniform oral dose of 2000 mg/kg bw, testing 8 organs at 3
or 24 hours later.

However, each test group had an assigned control group, somehow selected
from a total of 21 4-rat control groups.

Although the aspartame control levels happened to be much higher than the
averages for all 21 control groups, the aspartame values were about twice
the control values, close to statistical significance, 3 hours after the
oral dose, for stomach, colon, liver, bladder, and lung.

Results for DNA damage for Ace-K and stevia were nil, while sucralose,
cyclamate, and saccharin were significant for many tissues.

This result, once again, cries out for thorough replication, with much
larger groups of mice, using a variety of dose levels and test times, and
testing many more tissues.

A separate report confirmed that stevia was not genotoxic.

http://groups.yahoo.com/group/aspartameNM/message/935
Comet assay finds DNA damage from sucralose, cyclamate, saccharin in mice:
Sasaki YF & Tsuda S Aug 2002: Murray 2003.01.01
[ Also borderline evidence, in this pilot study of 39 food additives, using
test groups of 4 mice, for DNA damage from for stomach, colon, liver,
bladder, and lung 3 hr after oral dose of 2000 mg/kg aspartame -- a very
high dose. Methanol is the only component of aspartame that can lead to DNA
damage. ]

http://groups.yahoo.com/group/aspartameNM/message/961
genotoxins, Comet assay in mice: Ace-K, stevia fine; aspartame poor;
sucralose, cyclamate, saccharin bad: Y.F. Sasaki Aug 2002: Murray 2003.01.27
[A detailed look at the data] ]

"Comparing the mean control values [average of all 21 control groups] to the
values for the other 7 sweeteners:

Best is acesulfame K, with no significant or high values.

Good is glycyrrhizin (derived from licorice), two 1.4 ratios for Stomach and
Brain.

Next is stevia, with one high value [above ratio 1.4], 9.48+-1.99 for
Bladder, 2000 mg 3 hr, ratio 1.8 .

Aspartame has high values for 2000 mg 3 hr for Stomach, Colon, Liver,
Bladder, Lung.

Sucralose has 3 significant values and 13 high values, for Stomach, Colon,
Kidney, Bladder, Lung, Brain.

Sodium cyclamate has 4 significant values and 10 high values for Stomach,
Colon, Liver, Kidney, Bladder, Lung, Brain, Bone.

Saccharin has 3 highly significant values for Colon, and 13 high values for
Stomach, Colon, Kidney, Lung, Brain, Bone.

Sodium saccharin has 5 highly significant values for Stomach and Colon, and
14 high values for Stomach, Liver, Kidney, Bladder, Lung, Brain, Bone."


"For Liver, 5 of the 21 control groups, with values 1.67, 1.63, 1.29, 1.06,
1.65 would make some 3 hr aspartame values approach or reach significance.

Ratios about 2 for different tissues with aspartame that would be close to
significant would exist for many of the 21 control groups: Stomach 1 Colon 5
Liver 5 Bladder 11 Lung 5 .

The aspartame values at 3 hr are compared with the mean values for the 21
control groups:

Stomach ---- Colon ---- Liver ---- Kidney ---- Bladder ---- Lung

DNA Migration at 3 hr from 2000 mg/kg dose
8.49+-0.48, 9.18+-0.56, 3.26+-0.16, 1.91+-0.26, 10.7+-2.77, 4.13+-1.26

mean of 21 control groups
6.31,------ 5.81,------ 2.15,------ 2.25, ------ 5.40, ----- 2.61,

range of values for 21 control groups
4.3-8.6 --- 4.0-8.1 --- 1.1-3.6 --- 1.2-2.9 ---- 3.6-7.1 --- 1.6-4.7

ratio = DNA Migration/control mean
1.4, ------ 1.6, ------ 1.5, ------ 0.9, ------- 2.0, ------ 1.6

Brain ---------- Bone [marrow]

0.37+-0.70, ---- 1.01+-0.59 DNA Migration at 3 hr from 2000 mg/kg dose


1.48, ---------- 1.12, mean of 21 control groups


0.8-2.6 -------- 0.6-1.9 range of values for 21 control groups


0.3 ----------------- 0.9, ratio = DNA Migration/control mean

Wouldn't the average of all the 21 control groups be the best control values
to use?

What would then be the appropriate statistical test?

How many mice would it take to reach significance for the 5 tissues with
ratios over 1.4: Stomach, Colon, Liver, Bladder, Lung?

Aspartame at 24 hours had levels too low to reach significance with any of
the 21 control groups."

[ ASE reference 306: ]
Mutat Res 2002 Aug 26; 519(1-2): 103-19
The comet assay with 8 mouse organs: results with 39 currently used food
additives.
Sasaki YF, Kawaguchi S, Kamaya A, Ohshita M, Kabasawa K, Iwama K, Taniguchi
K, Tsuda S.
Laboratory of Genotoxicity, Faculty of Chemical and Biological Engineering,
Hachinohe National College of Technology, Tamonoki Uwanotai 16-1, Aomori
039-1192, Japan.
yfsasaki-c@... ; s.tsuda@...

We determined the genotoxicity of 39 chemicals currently in use as food
additives. They fell into six categories-dyes, color fixatives and
preservatives, preservatives, antioxidants, fungicides, and sweeteners.

We tested groups of four male ddY mice once orally with each additive at up
to 0.5xLD(50) or the limit dose (2000 mg/kg) and performed the comet assay
on the glandular stomach, colon, liver, kidney, urinary bladder, lung,
brain, and bone marrow 3 and 24 h after treatment.

Of all the additives, dyes were the most genotoxic.
Amaranth, Allura Red, New Coccine, Tartrazine, Erythrosine, Phloxine, and
Rose Bengal induced dose-related DNA damage in the glandular stomach, colon,
and/or urinary bladder.
All seven dyes induced DNA damage in the gastrointestinal organs at a low
dose (10 or 100 mg/kg).

Among them, Amaranth, Allura Red, New Coccine, and Tartrazine induced DNA
damage in the colon at close to the acceptable daily intakes (ADIs).

Two antioxidants (butylated hydroxyanisole (BHA) and butylated
hydroxytoluene (BHT)), three fungicides (biphenyl, sodium o-phenylphenol,
and thiabendazole), and four sweeteners (sodium cyclamate, saccharin, sodium
saccharin, and sucralose) also induced DNA damage in gastrointestinal
organs.

Based on these results, we believe that more extensive assessment of food
additives in current use is warranted. PMID: 12160896


details on 6 epidemiological studies since 2004 on diet soda (mainly
aspartame) correlations, as well as 14 other mainstream studies on
aspartame toxicity since summer 2005: Murray 2007.11.18
http://rmforall.blogspot.com/2007_11_01_archive.htm
Wednesday, November 14, 2007
http://groups.yahoo.com/group/aspartameNM/message/1490


http://groups.yahoo.com/group/aspartameNM/message/1340
aspartame groups and books: updated research review of 2004.07.16:
Murray 2006.05.11


European Food Safety Authority EFSA wants aspartame (methanol, formaldehyde,
formic acid) safety data by 2008.10.31 for 'Meeting of National Experts on
Aspartame' in December: Rich Murray 2008.10.10
http://rmforall.blogspot.com/2008_10_01_archive.htm
Friday, October 10, 2008
http://groups.yahoo.com/group/aspartameNM/message/1566
___________________________________________________


Rich Murray, MA Room For All rmforall@...
505-501-2298 1943 Otowi Road, Santa Fe, New Mexico 87505

"Of course, everyone chooses, as a natural priority, to enjoy peace,
joy, and love by helping to find, quickly share, and positively act
upon evidence about healthy and safe food, drink, and environment."

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