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Effect of apple cider Vinegar on blood glucose level in diabetic mice

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Background: In recent years, apple cider vinegar has been singled out as an especially helpful health remedy. It has been widely used in various dosage forms in alternative medicine for several conditions such as diabetes and obesity. In this study, the effect of apple cider vinegar on blood glucose level has been evaluated. Methods: Intraperitoneal (IP) injection of streptozocin 40 mg/kg/day for 4 days was used to induce diabetes in mice. The mice were divided in six groups (n=10). Two concentrations of 0.16% and 1.6% of apple cider vinegar were used in drinking water for 21 days. Normal saline and acetic acid were used as negative controls and glibenclamide by IP injection (5mg/kg) as positive control. For studying any possible combination effects, 0.16% apple cider vinegar and glibenclamide were used together. Also, the effects of apple cider vinegar on glucose tolerance test and amylase serum concentration were evaluated. Results: Our results indicated that apple cider vinegar in both concentrations was not effective after 3 days of the start of its administration. However, on day 7 it reduced blood glucose levels significantly and this was maintained on days 14 and 21. Glucose tolerance test showed that apple cider vinegar was effective in lowering blood glucose level after 60 minutes of glucose administration and this was maintained up to 120 minutes. Also, in both concentrations significantly reduced serum amylase levels 21 days after the start of its administration. Conclusions: Therefore, in this study it has been revealed that apple cider vinegar has considerable reducing effect on blood glucose levels in diabetic mice. The mechanism of this action and its significance remain to be elucidated in future investigations.
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http://journals.tbzmed.ac.ir/PHARM
*Corresponding Author: Seyed Adel Moallem, Department of Pharmacodynamics and Toxicology, School of Pharmacy, Mashhad University of
Medical Sciences, Mashhad, Iran. Tel: +98 51 38823255 , Fax: +98 51 38823251, E
-
mail:
moallem
@mums.ac.ir
Pharmaceutical
Sciences, March 2015, 20, 163-168
Dio:
10.5681/PS.2015.006
Effect of Apple Cider Vinegar on Blood Glucose Level in Diabetic
Mice
Maryam Iman1, Seyed Adel Moallem2,3,4*, Ahmad Barahoyee4
1Chemical Injuries Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran.
2Pharmaceutical Research Center, Mashhad University of Medical Sciences, Mashhad, Iran.
3Medical Toxicology Research Center, Mashhad University of Medical Sciences, Mashhad, Iran.
4Department of Pharmacodynamics and Toxicology, School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran.
Introduction
Apple cider vinegar is an acidic solution produced by
fermenting apples. It contains vitamins, minerals and
many trace elements.1 It contains a potent supply of
potassium. Potassium is essential for soft tissue repair
and the replacement of worn-out tissues within the
body. Cider vinegar improves the health and function
of the vital organs of the body by preventing
excessively alkaline urine. It is a strong detoxifying and
purifying agent. It breaks down fatty, mucous and
phlegm deposits within the body. It also oxidizes and
thins the blood, which is important in preventing high
blood pressure. Cider vinegar has been found to
neutralize any toxic substances that enter the body. It
neutralizes harmful bacteria that may be found in
certain foods, promotes digestion, assimilation and
elimination.2,3 Toxic build-ups with the body can cause
boils, blisters, acne, etc. Cider vinegar detoxifies and
helps with the cleansing and clotting processes of the
blood, by helping along the blood oxidation process.
When a mixture of cider vinegar and water is taken
before a meal (particularly food served in restaurants or
at picnics where the preparation or duration of food left
uncovered and not refrigerated is questionable), it
seems to prevent diarrhea or digestive upsets.4,5 Cider
vinegar can be taken alone or used in cooking. The best
method of using apple cider vinegar is in its natural
liquid form. Cider vinegar is thought to be beneficial in
the treatment of arthritis, asthma, nose bleeds,
osteoporosis, cancer, Candida, high cholesterol, colds,
constipation, muscle cramps, colitis, diabetes, diarrhea,
depression, dizziness, ear discharge, eczema, fatigue,
gallstones, kidney stones, hay fever, headaches,
heartburn, hiccups, indigestion, insomnia, kidney and
bladder problems, metabolism, nasal congestion, sore
throats, stiff joints, ulcers and weight loss.6 Some
reports showed that vinegar effect the glucose and
insulin responses to a sucrose or starch load. It is near
25 years that several in vivo and in vitro studies have
analysed the effect of vinegar on glucose metabolism in
Background:
In recent years, apple cider vinegar has been singled out as an especially
helpful health remedy. It has been widely used in various dosage forms in alternative
medicine for several conditions such as diabetes and obesity. In this study, the effect
of apple cider vinegar on blood glucose level has been evaluated. Methods:
Intraperitoneal (IP) injection of streptozocin 40 mg/kg/day for 4 days was used to
induce diabetes in mice. The mice were divided in six groups (n=10). Two
concentrations of 0.16% and 1.6% of apple cider vinegar were used in drinking water
for 21 days. Normal saline and acetic acid were used as negative controls and
glibenclamide by IP injection (5mg/kg) as positive control. For studying any possible
combination effects, 0.16% apple cider vinegar and glibenclamide were used together.
Also, the effects of apple cider vinegar on glucose tolerance test and amylase serum
concentration were evaluated. Results: Our results indicated that apple cider vinegar
in both concentrations was not effective after 3 days of the start of its administration.
However, on day 7 it reduced blood glucose levels significantly and this was
maintained on days 14 and 21. Glucose tolerance test showed that apple cider vinegar
was effective in lowering blood glucose level after 60 minutes of glucose
administration and this was maintained up to 120 minutes. Also, in both
concentrations significantly reduced serum amylase levels 21 days after the start of its
administration. Conclusions: Therefore, in this study it has been revealed that apple
cider vinegar has considerable reducing effect on blood glucose levels in diabetic
mice. The mechanism of this action and its significance remain to be elucidated in
future investigations.
A R T I C L E I N F O
A B S T R A C T
Article Type:
Original Research
Article History:
Received: 22 Dece mber 2014
Accepted: 4 February 2015
Keywords:
Apple Cider Vinegar
Glucose
Streptozocin
Alpha Amylase
Mice
Diabetes
[164]
Apple cider vi negar in diabetes
healthy subjects and in subjects with diabetes
mellitus.7,8
One of the major worldwide health problems is
diabetes. Diabetes appears to be increasing in most
countries, due to increasing population growth, aging,
urbanization, and increasing prevalence of obesity and
physical inactivity. Diabetes is a metabolic disease
which affects not only the glucose metabolism but also
lipid and protein metabolism. Diabetes can lead to
increased cardiovascular mortality, nephropathy,
neuropathy and retinopathy.9
There are mainly two types of diabetes–Type 1 and
Type 2. In Type 1 diabetes, the hormone insulin is not
produced while Type 2 diabetes mellitus (T2DM) is
characterized by a relative decreased sensitivity of
target tissues to the action of this hormone and
progressive impairment of insulin secretion.
Approximately one T2DM patients are treated with oral
hypoglycemic agents to stimulate insulin secretion.10,11
T2DM is managed through a program that consists of
lifestyle modifications including appropriate diet and
exercise programs and addition of oral
antihyperglycemic agents.12,13Although oral
hypoglycemic agents (insulin) are the mainstay of
treatment of diabetes and are effective in controlling
hyperglycemia, they have prominent side effects and
fail to significantly alter the course of diabetic
complications.
The common side effects associated with the main
classes of drugs used for the treatment of T2DM are
hypoglycemia, weight gain, gastrointestinal disorders,
peripheral edema and liver disease.14 The present study
investigates the hypoglycemic effects of apple cider
vinegar and its combination effects with
antihyperglycemic agents. It is thought to be beneficial
for decreasing uses of antihyperglycemic agents and
their side effects.
Methods and Materials
Apple cider vinegar with 16% concentration was
obtained from Septiko Co. (Mashhad, Iran) and
streptozosin (Zanosar®) from Pharmacia & Upjohn
(Mich, USA). Amylase kit was obtained from Zist
Chimi Co. (Iran). Glibenclamide (Daroupakhsh Co,
Iran) was used. Accu-chek Glucometer (Roche,
Canada) was used to determine blood glucose levels.
All reagents were of analytical grade.
Animals
Mice at 6-8 weeks of age, weighing 25-30 g were
obtained from Avicenna Research Institute (Mashhad,
Iran). The mice were housed in a standard environment
at a constant temperature of 25˚C under a 12-h
light/dark cycle with free access to food and drinking
water.
Diabetic-induced mice
Intraperitoneal (IP) injection of streptozocin 40
mg/kg/day for 4 days was used to induce diabetes in
mice. Two weeks after streptozosin injection, blood
glucose levels were measured and those with fasting
glucose levels above 11.1 mmol/l were included in the
study.15,16
Treatment regimens with apple cider vinegar
Mice could not tolerate gavage of 8% and 16% of apple
cider vinegar, probably due to the high acidity of these
concentrations. Various experiments showed that
concentrations of 0.16% and 1.6% of apple cider
vinegar in drinking water up to 21 days are well
tolerated.
Experimental Protocols
Streptozocin-diabetic mice were divided randomly into
6 groups (n=10). The method of treatment was
“drinking water”. Two concentrations of 0.16% and
1.6% of apple cider vinegar were used in drinking
water for 21 days.17 Normal saline and acetic acid were
used as negative controls and glibenclamide by IP
injection (5mg/kg) as positive control. For studying any
possible combination effects, 1.6% apple cider vinegar
and glibenclamide were also used together. Also, the
effects of apple cider vinegar on glucose tolerance test
and amylase serum concentration were evaluated. The
glucose levels in the plasma were determined in groups
after 3, 7, 14, 21 days from drug administration. All of
samples were obtained after 4h of fasting. The plasma
glucose levels were expressed in mmol/l.
Blood collection for measurement of blood glucose
levels
Stressful methods of blood collection can affect blood
parameters such as glucose levels. To decrease stress,
the mice were acclimated to the restraining device and
were warmed and placed in the restraining tube. After 4
hour of fasting, a 2 mm distal section of the mouse
sterilized tail is scratched using a syringe needle and
gently squeezed to obtain two drops of blood, the first
of which is discarded. The second drop is applied
directly to the test strip to obtain blood glucose
measurement. In our experience, one reading per time
point was sufficient to obtain an accurate reading,
however additional readings per time point were
performed randomly for results validation.
Glucose tolerance test
The test is usually used to diagnose prediabetes and
diabetes, insulin resistance, and sometimes reactive
hypoglycaemia. The glucose is most often given orally
to determine how quickly it is cleared from the blood.
The test may be performed as part of a panel of tests,
such as the comprehensive metabolic panel in medical
practice.18
In the last day of treatment, after the fasting plasma
glucose was tested, all groups received 5 g/kg of
glucose by gavage. The glucose levels in the plasma
were determined in groups after 30, 60 and 120 minutes
of glucose administration.
[165]
Iman et al.
Measurement of blood α-amylase
Amylase is an enzyme that its catalytic function is t o
hydrolyze sugar and starch. It digests polysaccharides
into smaller disaccharide units, eventually converting
them into monosaccharaides such as glucose. α-
Amylase is the major form of amylase found in humans
and other mammals. It is produced by the pancreas t o
help digestion. An amylase test measures the amount of
this enzyme in a sample of blood. In the last day of
treatment, mice were sacrificed and 1.5-2 ml blood
samples were obtained by cardiac Puncture. Amylase
activity was determined by using a diagnostic kit. The
substrate was ethylidene-p-nitrophenyl maltoheptaoside
(EPS-G7). Absorbance, which is directly related to α-
amylase activity, was measured at 405 nm and 37 ºC
using an auto analyzer (Alcyon 300® Plus, Molecular
Devices Corporation, Sunnyvale, CA). Before
application, the auto analyzer was calibrated with the
control sera N and P (TrueLab N and TrueLab P®,
respectively; Zist Chimi., Iran) and a calibrator solution
(TrueCal U®, Zist Chimi Co., Iran). After calibration,
the auto analyzer mixes 6 μl of enzyme sample with
300 μl of substrate solution, automatically and
calculates the enzyme activity (IU/L).
Statistical Analysis
One-way ANOVA and t-student statistical tests were
used to assess the significance of the differences. In
case of significant F value, multiple comparison Tukey-
Kramer tests were used to compare the means of
different treatment groups. Results with p<0.05 were
considered to be statistically significant.
Results and discussion
Measurement of blood glucose
The effect of apple cider vinegar on glucose levels in
the plasma was determined in mice after 3, 7, 14, 21
days of drug administration. Glucose blood levels in
diabetic mice treated with normal saline and acetic acid
were increased during 21 days (Fig. 1). Apple cider
vinegar in both concentrations was not effective after 3
days of the start of its administration. However, on day
7 it reduced blood glucose levels significantly and this
was maintained on days 14 and 21 (p < 0.05) (Fig. 1).
Comparisons of treated groups showed no significant
difference in their glucose levels at days 7, 14 and 21 (p
> 0.05), indicating that apple cider vinegar at both
doses has antihyperglycemic effects comparable to
glibenclamide.
Figure 1. Glucose blood l evels during 21 days.
Glucose tolerance test
In the last day of treatment after determining the fasting
blood glucose, all groups received 5 g/kg of glucose by
gavage. The glucose levels in the plasma were
determined in groups after 30, 60 and 120 minutes. The
results showed that apple cider vinegar in both
concentrations significantly reduced blood glucose
level after 60 minutes of glucose administration in
comparison to its level after 30 minutes (p < 0.05) and
this was maintained up to 120 minutes after the start of
its consumption by diabetic mice (p < 0.05) (Fig. 2).
This was also apparent in the positive control group.
There was no dose-dependent effect as no significant
difference was observed between the two apple cider
vinegar groups (p > 0.05).
As the glucose level did not rise significantly after
glucose administration after 30 minutes in the
combination treated group (apple cider vinegar +
glibenclamide), there was no significant reduction in
the glucose levels after 60 and 120 minutes as
compared t o glucose level at 30 minutes (p > 0.05).
However, the glucose levels in this group were the
lowest among all treated groups (Fig. 2).
[166]
Apple cider vi negar in diabetes
Figure 2. Glucose blood levels after administrations of 5g/kg glucose in diabetic-mice at day 21.
Measurement of blood α- amylase
In the last day of treatment, α-amylase was determined.
Apple cider vinegar in both concentrations significantly
reduced α-amylase levels in serum after 21 days (p <
0.05) (Fig. 3). The higher concentration of apple cider
vinegar has more effect on reducing α-amylase serum
levels compared t o the lower concentration of apple
cider vinegar (p < 0.05). Interestingly, glibenclamide
administration did not affect α-amylase serum levels
alone or in combination to apple cider vinegar,
compared to normal saline and apple cider vinegar,
respectively (Fig. 3).
Figure 3. Amylase blood l evels after 21 d ays of treatment.
In previous studies, it was demonstrated that vinegar
affect glucose metabolism in healthy or in patients with
diabetes mellitus, improves insulin sensitivity in
healthy or in patients with insulin-resistant and could
further delay gastric emptying, so do postprandial
hypoglycaemia. But some studies showing no
beneficial effect on glucose metabolism that show
several factors affect vinegar effects.19-21 There is much
interest in identifying diet patterns that could possibly
reduce hyperglycemia. The aim of this study was to
investigate the effect of apple cider vinegar on blood
glucose level. Our results indicated that apple cider
vinegar on days 7 and 14 reduced blood glucose levels
significantly (p<0.05) and was effective in lowering
blood glucose level after 60 minutes of glucose
administration and this was maintained up to 120
minutes. Also, apple cider vinegar in both
concentrations significantly (p<0.05) reduced serum
amylase levels 21 days after the start of its consumption
by diabetic mice. So these results showed the favorable
effects of apple cider vinegar on glucose level and it
can be used in patient with diabetes type 2.
Conclusions
Our results indicated that apple cider vinegar in both
concentrations was not effective after 3 days of the start
[167]
Iman et al.
of its administration in diabetic mice. However, on day
7 it reduced blood glucose levels significantly (p<0.05)
and this was maintained on days 14. A slight rise in
blood glucose levels was observed on day 21 that was
not significant (p < 0.05). The findings of the present
investigation showed that blood glucose levels in the
acetic acid group were comparable to the normal saline
group at days 3, 7 and 14. However, they were higher
than the normal saline group at day 21 suggesting that
continuous consumption of acetic acid probably due to
its acidic property might induce stress in treated mice.
This increased stress can lead to a rise of glucose blood
levels. Although the cider vinegar has acidic property,
probably its “therapeutic” effect in lowering glucose
levels was able to overcome this property.
Glucose tolerance test showed that apple cider vinegar
was effective in lowering blood glucose level after 60
minutes of glucose administration and this was
maintained up to 120 minutes. Also, apple cider
vinegar in both concentrations significantly (p<0.05)
reduced serum amylase levels 21 days after the start of
its consumption by diabetic mice.
In this study it has been revealed that apple cider
vinegar has considerable reducing effect on blood
glucose levels in diabetic mice suggesting a useful
outcome in reducing the risk diabetes due to its
antihyperglycemic effect in diabetic mice.
Although the full mechanism of this effect is unclear,
one probable mechanism could be the effect of apple
cider vinegar α-amylase. Reduction of α-amylase in
liver cells can suppress the conversion of carbohydrates
(polysaccharides) into smaller saccharide units such as
glucose leading to a reduction in blood glucose levels.
Whether apple cider vinegar has any effect on insulin
action in peripheral tissues, such as skeletal muscles
and adipocytes any other probable mechanisms are
unclear that can be further studied. More work is
needed to determine the exact nature of the active
ingredients.
Authors' contributions
MI conceived of the study and helped to draft the
manuscript, SAM is head of study and participated in
its design and coordination and helped to draft the
manuscript. AB participated in the taking result and
performed the statistical analysis.
Acknowledgements
We appreciate the technical assistance support of
Mashhad University of Medical Science.
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... Starch hydrolysis process to produce glucose monomer compounds which can be absorbed from gastrointestinal tract involves various enzymes ranging from ptyalin in the oral cavity to hydrolytic enzymes such as alpha-amylase and alphaglucosidase. 16 These enzymes that contributed to hydrolyzing amylum to be oligo and monosaccharide form are called amylolytic enzymes. 17 Monosaccharides absorbed will be transported through the portal vessels to the liver. ...
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... Many animal studies and human intervention studies focused on the hypoglycemic action of FAJ (Tables 4 and 5). For example, Streptozocin-diabetic mice took the concentrations of 0.6% and 1.6% ACV orally for 7 days, and their blood sugar levels were significantly reduced (Iman, Moallem et al., 2015). At the same time, both doses significantly reduced serum amylase levels 21 days after the start of its administration. ...
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... Various types of organic acids, such as acetic acid, butyric acid, citric acid, lactic acid, malic acid, sorbic acid, propionic acid, as well as their salts, are used to improve the health of fish (Hossain et al. 2007;Ng et al., 2009;Safari et al., 2017) and shrimp (Pourmozaffar et al. 2017). Apple cider vinegar is an acidic solution, which contains organic acids such as acetic acid and malic acid, vitamin B and C, and minerals (Iman et al., 2015). Previous study demonstrated that using organic acid in diet resulted in improve blood indexes and healthy in rainbow trout fingerling (Taheri Mirghaed et al., 2019). ...
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... Experimental research supports the role of polyphenols in the prevention of cardiovascular diseases, cancers, neurodegenerative diseases like Alzheimer's disease, osteoporosis, and diabetes [23]. ACV has been shown to have various health and therapeutic effects such as antimicrobial, antioxidant, antidiabetic, antitumor, antiobesity, and neuroprotective effects [23,24]. Up till now, studies on the possible effect of ACV on LPS-induced behavioral deficits including anxiety, depression, and motor impairment, are lacking in literature. ...
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Recent changes in lifestyle, including physical inactivity and unhealthy diets, are likely to have played an important role in the global epidemic of obesity, type 2 diabetes, and metabolic syndrome. Implementation of a healthier lifestyle, with an increase in physical activity and a reduction of body weight, based on the regulation of calories and fat intake, are the basis for the prevention and treatment of both type 2 diabetes and metabolic syndrome. Intervention studies based on changes in lifestyle in individuals at risk found that diabetes incidence was reduced by 42% to 63%. Similarly, intensive lifestyle changes in patients with the metabolic syndrome have been shown to reduce the prevalence of the syndrome by 20% to 48%. Reduction of body weight, improvement of the quality of diet, and promotion of physical activity are the main approaches to prevent and treat patients with type 2 diabetes or metabolic syndrome.
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To investigate the influence of sodium acetate and acetic acid from vinegar on blood glucose and acetate response to a mixed meal in healthy subjects. Five healthy subjects consumed in random order six test meals consisting of 100 g of sliced lettuce dressed with olive oil (Blank), olive oil plus 1 g acetic acid in the form of vinegar (AcOH), or olive oil plus sodium acetate in the form of vinegar neutralized to pH 6.0 with sodium bicarbonate (AcNa). On three occasions test meals were followed by a challenge consisting of 50 g carbohydrate portions of white bread (Bread). Glucose and acetate concentrations were measured in arterialized capillary blood before and until 95 min after the meals. Ultrasonography was performed in four other subjects to measure gastric emptying times after AcOH + Bread and AcNa + Bread. Blood acetate response over 95 min was markedly reduced after AcOH and AcOH+Bread meals compared to AcNa and AcNa + Bread. Similarly, the glucose response was depressed by 31.4% (P = 0.0228) after AcOH+Bread with respect to AcNa + Bread and Blank + Bread. No difference was observed between gastric emptying times after AcOH + Bread and AcNa + Bread. The results suggest that oral acetic acid and acetate might have a different effect on acetataemia and that a limited dose of vinegar, in the form of salad dressing, is sufficient to influence significantly the glycaemic response to a mixed meal in normal subjects by a mechanism related to acidity but not to gastric emptying.
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A study was done to determine survival and growth characteristics of acid-adapted, acid-shocked, and control cells of Escherichia coli O157:H7 inoculated into tryptic soy broth (TSB) acidified with organic acids and three commercial brands of apple cider and orange juice. The three types of cells behaved similarly in TSB acidified with acetic acid; however, in TSB (pH 3.9) acidified with lactic acid, acid-adapted cells were more tolerant than acid-shocked cells which, in turn, were more tolerant than control cells. The ability of the three types of cells to grow after inoculation into acidified TSB, then plated on tryptic soy agar containing sodium chloride was determined. Tolerance of acid-adapted cells and, less markedly, acid-shocked cells to sodium chloride was diminished, compared to control cells. The pathogen showed extraordinary tolerance to the low pH of apple cider and orange juice held at 5 or 25 degrees C for up to 42 days. Growth occurred in one brand of apple cider (pH 3.98) incubated at 25 degrees C. Regardless of test parameters, there was no indication that cell types differed in tolerance to the acidic environment in apple cider or orange juice. Survival of control, acid-adapted, and acid-shocked cells heated in apple cider and orange juice was studied. Within each apple cider or orange juice, D(52 degrees C)-values of acid-adapted cells were considerably higher than those of acid-shocked or control cells, which indicates that heat tolerance can be substantially enhanced by acid adaptation compared to acid shock.
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Diluted solutions of various household sanitizers (apple cider vinegar, white vinegar, bleach, and a reconstituted lemon juice product) were tested for their effectiveness in reducing counts of inoculated Escherichia coli and naturally present aerobic, mesophilic bacteria on lettuce. Sanitization treatments were carried out at 4 degrees C and at room temperature (ca. 21 degrees C) with and without agitation and at different exposure times (0, 1, 5, and 10 min). Of the sanitizers tested, 35% white vinegar (1.9% acetic acid) was the most effective in reducing E. coli levels (with a 5-log10 reduction after 5 min with agitation and after 10 min without agitation) and in reducing aerobic plate counts (with a >2-log10 reduction after 10 min with agitation). Lettuce samples treated with diluted household sanitizers were analyzed for consumer acceptability by sensory evaluation using a 9-point hedonic scale. The sanitized samples did not differ in acceptability (P > 0.05), except for samples treated with white vinegar. Samples treated with the white vinegar for 10 min were noticeably sour and slightly wilted in appearance. Consumer acceptability was maintained with all sanitization treatments, including those involving 35% white vinegar.