Glycative Stress Research
Online edition : ISSN 2188-3610
Print edition : ISSN 2188-3602
Received : Jan. 26, 2014
Accepted : Mar. 4, 2014
Published online : Mar. 31, 2014

Original Artcle

Evaluation of the anti-glycation effect and the safety of a vinegar beveragecontaining indigestible dextrin and a mixed herbal extract--- A placebo-controlled, double-blind study --Masayuki Yagi, Akihiko Shimoide, Umenoi Hamada, Junko Naito, Masamitsu Ichihashi,
Yoshikazu Yonei
Anti-Aging Research Center/Glycation Stress Research Center, Graduate School of Life and Medical Sciences, Doshisha University

Abstract

Objective: We evaluated the inhibitory effect of the long-term consumption of a vinegar beverage containing indigestible dextrin
and a mixed herbal extract (the test diet) on the production of advanced glycation end products (AGEs) in a placebo-controlled,
randomized, double-blind, parallel-group comparison study design.
Methods: A total of 109 post-menopausal women previously diagnosed with an increased blood glucose or hemoglobin A1c(HbA1c) level, with an abdominal circumference of 90 cm or more, were subjected to an oral rice ingestion test followed bymeasurement of fluorescence intensity of skin AGEs. Among them, 23 women (mean age 57.4 ± 3.9) with high levels of 60-minutepostprandial blood glucose and skin AGE deposition were enrolled and assigned to either the test group (n = 11) or the control
group (n =11). The subjects in the test group took 240 mL of the test diet (corresponding to 840 mg/day of acetic acid, 5 g /day of
indigestible dextrin, and 100 mg/day of mixed herbal extract) daily divided into two equal doses taken before breakfast and dinner
for a period of 8 consecutive weeks. Blood biochemistry parameters and glycation stress markers were measured before and at 8
and 12 weeks after the start of the test diet. The study was approved by an ethical review committee.
Results: A significant decrease in serum aspartate transaminase (AST/GOT) level in the test group compared to control wasobserved after 8 weeks of diet intake ( p < 0.05). With regard to glycation stress markers, no significant differences were observed
between groups in the blood concentrations of fasting glucose, HbA1c, insulin, 3-deoxyglucosone (3DG), N ε- (carboxymethyl)
lysine (CML) or pentosidine. No significant intergroup difference was observed in the fluorescence intensity of skin AGE as
measured with an AGE Reader. In the subclass analysis of the subjects with high postprandial blood glucose ( > 150 mg/dL at 60
minutes), CML content of the skin stratum corneum as determined by the tape stripping method was significantly decreased in
the test group compared to control after 8 weeks of diet intake (p < 0.05). No serious adverse event was observed during the study
period.
Conclusion: The results of the present study suggest that the intake of the test diet causes a reduction in CML content in the skinstratum corneum, a marker of glycation stress, in people with relatively high glycation stress. Combined with the demonstrated
safety of the test diet, this observation indicates a potential for the use of the test diet as a functional food.

KEY WORDS: Glycation stress, vinegar beverage, advanced glycation end products (AGEs), N ε-(carboxymethyl) lysine(CML), tape stripping techniqueIntroduction

Glycation stress is a series of reactions triggered whenreducing sugars, organic acids or aldehydes react with proteinderived amino acids to produce post-translational modification
products, such as carbonyl compounds, succinyl compounds,
racemic compounds, or advanced glycation end products(AGEs). Subsequent accumulation of such compounds causes
stress to cells and tissues and leads to the functional declineof proteins which can bind to receptors for AGEs, inducing or
aggravating inflammation via cytokine production 1-3). Glycationstress is considered a major risk factor for accelerated aging.
Although diabetes, including the prediabetic state, is amajor predisposing factor for glycation stress, glycation canalso occur in the absence of diabetes. This type of glycation is
Glycative Stress Research 2014; 1 (1): 14-24(c) Society for Glycation Stress Research_ 14 _

referred to as normoglycemic glycation or non-hyperglycemic
glycation and is caused most commonly by postprandial
hyperglycemia or, less commonly, by hypertriglyceridemia,
uremic toxin, excessive alcohol intake or excessive fructose
intake. Smoking and a lack of sleep are also known to promote
AGE production 4). These factors should be eliminated toreduce glycation stress.
Recent studies have identified and evaluated various
substances that inhibit the production of AGEs, promote
the degradation of AGEs or antagonize receptors for AGEs.
Previously, we have evaluated the in vitro inhibitory effect ofroman chamomile (Anthemis nobilis), doku-dami (Houttuynia
cordata), hawthorn (Crataegus laevigata (C. oxyacantha)),grape leaf (Vitis vinifera), and a mixture of these herbs 5),
Chrysanthemum morifolium 6,7), Sasa senanensis 8), and extracts
Yoshikazu Yonei, MD, PhD
Anti-Aging Medical Research Center, Graduate School of Life andMedical Sciences, Doshisha University
1-3 Tatara Miyakodani, Kyotanabe City, Kyoto 610-0321, Japan
Tel & Fax: 0774-65-6394 / Email: yyonei@mail.doshisha.ac.jp

Glycative Stress Researchof various types of healthy tea 9) on AGE production. We havealso performed a clinical evaluation of a herbal mixture 10),
Chrysanthemum morifolium 7), and a food product containing
lingonberry (Vaccinium vitis-idaea) and cherry blossom(Prunus lannesiana) as main ingredients 11). The objective of
the present study was to evaluate the inhibitory effect on AGE
production and the safety of a vinegar beverage containing
indigestible dextrin and a mixed herbal extract administeredfor 8 consecutive weeks in a placebo-controlled, randomized,
double-blind design.

even if they were not having a meal. The mean intake rate was
97.1% (88.3%) in the control group, 98.3% (97.3%) in the test
group, and 97.7% (92.8%) in the entire population, with values inparenthesis representing mean rates up to week 8.
The study was conducted between September 2012 andDecember 2012 at TES Holdings Co., Ltd. (Bunkyo-ku, Tokyo,
Japan). After the subjects were given sufficient explanation
regarding purpose and details of the study and participants' rights,
they provided written informed consent. They were also told thatearly withdrawal from the study would not be a detriment.

Test diet

Methods

The compositions of placebo and test diet are shown in Table 1.

Subjects

Eligible subjects were selected according to the following
criteria. A total of 109 post-menopausal women aged
between 50-65 years, previously diagnosed with increased
blood glucose or hemoglobin A1c (HbA1c) levels and an
abdominal circumference of 90 cm or more, were recruited.
After explanation of the study, written informed consent tostudy participation was obtained from all women. An oral
rice ingestion test and skin AGE deposition analysis were
performed as screening tests on all potential subjects. In theoral rice ingestion test, subjects were instructed to eat 200 g ofrice (294 kcal, 67.8 g carbohydrate and 4.2 g protein) with 2.5
g of rice seasoning condiment (11 kcal, 1.0 g carbohydrate and0.6 g protein). Blood glucose levels were measured at 0, 30,
60, and 120 minutes after consumption according to the unified
procedure proposed by the Japanese Association for the Studyof Glycemic Index. A total of 23 women (mean age 57.4 ± 3.9
years) shown to have high levels of 60-minute blood glucose
and skin AGE deposition were included in the full analysis set(FAS) of the study. Except for one subject who was withdrawn
during the study period, no subject met any of the exclusion
criteria, and all of the remaining 22 subjects completed thestudy and complied with the protocol (per protocol set; PPS).
The subject was withdrawn because she experienced acid reflux
when taking the test diet, which was determined to be unrelated
to study diet intake. Subjects taking drugs or supplements that
may affect the blood glucose level, smokers or those with ashort sleep duration (< 5 hours) were excluded. Those meetingthe last two criteria were excluded because these lifestylefactors are known to increase AGE deposition in the skin 4).

Study design

The study was conducted in a two-group (control and test
group), placebo-controlled, randomized, double-blind study
design. Subjects were given either a placebo (control group) ora vinegar beverage containing indigestible dextrin and a mixed
herbal extract as the test diet (test group) for 8 consecutive weeks.
A clinical evaluation consisting of an interview, a physicalexamination, a blood/urine tests, and a skin function test was
performed at 0, 8, and 12 weeks.
Subjects were instructed to take 120 ml of the control or test
diet twice daily, i.e., before breakfast and dinner. The daily intake
amounted to 840 mg of acetic acid, 5 g of indigestible dextrin
and 100mg of mixed herbal extract. Both the placebo and thetest diet were given for 8 weeks, with a follow-up period of 4
weeks. Initially, the placebo/test diet intake period was set at 12
weeks. However, later it was shortened to 8 weeks after foreign
matter contamination was detected in the manufacturing processof the test diet. Subjects were instructed to take the test diet

Table 1. Composition of placebo and test diet.
Ingredient (%)

Placebo diet

Test diet

Acetic acid

0.35

Citric acid

0.1

Indigestible dextrin

2.1

Mixed herbal extract 1)

0.042

Tien-cha extract 2)

0.01

α-G-rutin

0.0125

Sucralose

0.004

0.008

Acesulfame potassium

0.003

0.006

Salt

0.04

Condensed tomato juice

0.05

Other ingredientsf lavoringwater

f lavoringwater1) Mixed herbal extract contains Anthemis nobilis, Houttuynia cordata,
Crataegus laevigata, and Vitis vinifera of leaf.
2) Tien-cha extract contains Rubus suavissimus.

AG herb mixTM, a functional food ingredient developed
and marketed by ARKRAY Inc. (Kyoto, Japan), is a mixtureof powdered hot water extracts of herbs belonging to different
taxonomic groups, including Houttuynia cordata, Crataegus
laevigata (C.oxyacantha), Anthemis nobilis, and Vitis vinifera.
This herbal extract and its formulated product have been shownto inhibit AGE production in vitro, both in a diabetes model inrats and in randomized controlled trials (RCTs) in humans 10-13).
The herbal extract is considered safe as it is composed of raw
materials that have long been consumed as food ingredients,
and because it is produced with the same extraction method as
used for herb tea. Anthemis nobilis has been shown to cause
an allergic reaction in people with multiple allergies, and
Houttuynia cordata, used as a folk medicine, has been reported
to cause photosensitivity and hyponatremia when consumed
in large amounts for long periods of time. However, the daily
intake of the mixed herbal extract contained in the test diet usedin this study was only 100 mg, thus it was not likely to cause
any problems. Moreover, the safety of the mixed herbal extract
has been demonstrated in various studies, including a rec-assay(lethal sensitivity test), a reverse mutation test, an acute oral
toxicity study using male and female rats, and an overdose test
in humans (3,000 mg/day, corresponding to 5 times the regulardose, for 4 weeks).
Fibersol-2H was developed and marketed by Matsutani
Chemical Industry (Hyogo, Itami, Japan). The compound
affects the control of hyperglycemia after a meal and is used as_ 15 _

Anti-glycation effect of vinegar beverage in humans
a functional food for specified health use. Indigestible dextrin
in the test diet has been associated with diarrhea and othergastrointestinal symptoms when consumed in large amounts at
a time. Given that the maximum non-effect level for diarrhea
in women is ≥1.0 g / kg, the daily intake of indigestible dextrin
was set at 5 g in this study.
An inhibitory action on AGE production of tien-cha
extract, containing Rubus suavissimus extract, was confirmed invitro 9). Rutin, Citric acid, sucralose, and potassium acesulfame
are food additives, and their content was within accepted limits.
Acetic acid, salt, and concentrated tomatoes have a long history
as ingredients in various foods and were added to the test dietat concentrations smaller than found in commercially available
food items. These facts assured a sufficient level of safety ofthe test diet. A list of safety studies of the active ingredients
present in the test diet is shown in Table 2.
To ensure product quality, the manufacturing process ofthe vinegar beverage containing indigestible dextrin and mixed
herbal extract is strictly controlled from the reception of raw
materials to the packaging of the end product. Among otheracceptance criteria, the end product has to contain less than
10 3/mL viable cells and no detectable coliform bacteria to passquality control.

Test procedure
Subjective symptoms

Subjective symptoms were divided into physical andmental symptoms and evaluated on a 5-point scale using
the Anti-Aging QOL Common Questionnaire (AAQol), as
described previously 7, 10, 11, 14, 15).

Anthropometry and physical examination

Body height, body weight, blood pressure, and bodycomposition as measured by bioelectrical impedance analysis
with a body composition analyzer (BC-118D Tanita Corp.,
Tokyo, Japan) as described previously 14, 15), were determined.

Vascular function test

Blood concentrations of endothelin, vascular endothelial
growth factor (VEGF), and nitric oxide (NO) were measured
as parameters for evaluating arteriosclerosis. Fingertip
acceleration pulse wave was also measured with aplethysmometer (SDP-100, Fukuda Denshi, Tokyo, Japan)
to estimate vascular age 16-18). Briefly, the second derivativeof plethysmogram aging index (SDP TGAI) was calculated
from parameters b/a, c/a, d/a, and e/a, and vascular age wascalculated using the following formulas:
Men:
Vascular age = 43.50 × SDPTGAI + 65.90
Women: Vascular age = 41.67 × SDPTGAI + 61.75

Glycation stress markers

Insulin resistance was evaluated by fasting plasma
glucose (FPG), insulin, and HbA1c levels. For analysis of
AGEs and glycation intermediates, serum concentrations of
3-deoxyglucosone (3DG), N ε-(carboxymethyl) lysine (CML),
and pentosidine were measured as described previously 7, 10, 11).
Skin AGE deposition was measured in the medial aspectof the right upper arm (at 10 cm from the olecranon toward
the shoulder) with an AGE Reader (DiagnOptic, Netherland)as described previously 4, 19). CML content in the skin stratum
corneum (in the medial aspect of the right upper arm) was

Table 2. List of safety studies.
Name of ingredient

Indigestible dextrin(as Fibersol-2H)

Reverse mutation test

Negative

A single-dose oral toxicity study in rats (male and female)No sign of toxicity at 10 g/kg

A repeated-dose oral toxicity study in rats

NOAEL: > 5.0 g/kg

A study to find the maximum non-effect level in humans

Maximum non-effect level for diarrhea

Rec-assay ( lethal sensitivity test )Negative

Reverse mutation test

Negative

An acute oral toxicity study in rats (male and female)

LD50 : > 2,000 mg/ kg

An overdose study (in humans)
3,000 mg/day ( 5 times the regular dose) for 4 weeks

No adverse event was observed.

An acute oral toxicity study in mice (male and female)

LD50 : > 5,000 mg/ kg

A 28-day repeated-dose oral toxicity study in rats (male and female)

NOAEL: 600 mg/kg/day

Reverse mutation test

Negative

Micronucleus test

No mutation was detected.

An acute oral toxicity study in mice (male and female)

LD50 : > 42,000 mg/ kg

at solid content of 0.4, 0.5, 0.6, 0.8 and 1.0 g/kg

Mixed herbal extract(as AG herb mix)

Tien-cha extract(Tien-cha Extract M Powder)

Rutin(as glucosyl rutin)

Results

Study details

A 28-day subacute oral toxicity study
50, 200 and 1,000 mg/ kg/day
Abbreviations: NOAEL, no observable adverse effect level; LD50, 50％ lethal dose_ 16 _

Men: 0.8 g/ kg, Women: > 1.0 g/kg

No subacute oral toxicity was observed.

Glycative Stress Researchmeasured as described previously 20). Briefly, an adhesive film
was firmly applied to the skin to collect a sample of the stratum
corneum (tape stripping technique). Three samples werecollected from the same site and CML content was measured.
Samples are typically collected from the left cheek (center
portion between the bottom of the ear lobe and the lip end) orfrom the medial aspect of the right upper arm (at 10 cm fromthe olecranon toward the shoulder).

Oxidation stress markers

Concentrations of 8-hydroxydeoxyguanosine (8-OHdG)
and isoprostane in urine samples collected during the night were
measured as markers for oxidation stress 21-25). These parameters
were measured by Mitsubishi Chemical Medience Corporation(Minato-ku, Tokyo, Japan). In addition, concentrations of
8-OHdG, isoprostane, and creatinine in the first urine collectedin the early morning were measured to calculate creatinineadjusted concentrations of 8-OHdG (8-OHdG/CRE) andisoprostane (isoprostane/CRE).

at 8 and 12 weeks after intake: total cholesterol (TC), LDL
cholesterol (LDL-C), HDL cholesterol (HDL-C), triglyceride(TG), atherogenic index (AI), total bilirubin (TB), aspartate
aminotransferase (AST/GOT), alanine aminotransferase(ALT/GPT), lactate dehydrogenase (LDH), gamma-glutamyltranspeptidase ( γ - GTP), creatine phosphokinase (CPK), uricacid (UA), urea nitrogen (BUN), creatinine (CRE), sodium(Na), potassium (K), chloride (Cl), calcium (Ca), total protein(TP ), albumin (ALB), albumin/globulin ratio (A/G), andiron (Fe ). Urine samples collected during the night were also
analyzed for concentrations of sodium (Na), potassium (K),and calcium (Ca) to detect possible excessive electrolyte intake.
Analysis of glycation stress markers, i.e., 3DG and CML,
was performed by SRL Inc. (Shinjuku-ku, Tokyo, Japan).
Analysis of endothelin, a marker for vascular endothelial
function, and CML content in the skin stratum corneum
was performed at the Life & Medical Science Investigation
Center of A-kit Corporation (Kyotanabe, Kyoto, Japan). The
remaining blood/urine parameters were measured by Mitsubishi
Chemical Medience Corporation.

Immune stress markers

Ethical considerations

Serum and plasma concentrations of high-sensitivity
C-reactive protein (hsCRP) and interleukin-6 (IL-6), bothinflammatory markers, were measured as immune stressmarkers.

Skin function test

The properties and function of the skin were evaluated bymeasuring color difference, moisture content, and melanin/erythema content, as well as by the identification of wrinkles
and spots based on imaging analysis. These tests were
performed in a room with constant temperature and humidity(25°C, 50%) after a 20-minute conditioning period.
For the imaging analysis of facial skin, the VISIA
Evolution System (Canfield Imaging Systems, Fairfield, NJ,
USA) 26) was used to examine pores, spots (visualized aslight spots), melanin (brown spots), hemoglobin (red spots),
wrinkles, color unevenness (texture), porphyrin, and latent
spots (UV spots). This test was performed on the left cheek ofeach subject.
Skin elasticity was evaluated using a cutometer (MPA580;
Courage & Khazaka, Kern, Germany) 29-31). The skin surface
was drawn by negative pressure into the aperture of the probe,
and the length of the skin drawn into the aperture was measured
by the prism. This test was performed on the left cheek (center
portion between the bottom of the ear lobe and the lip end) inthe supine position or on the medial aspect of the right upperarm (at 10 cm from the olecranon toward the shoulder) in asitting position. Results were expressed as skin elasticity index
R2 or R7, respectively.
Skin moisture content was measured in the left cheekwith a moisture meter (Corneometer, CM825; Courage &Khazaka) 32). For skin color analysis, a spectrophotometer(CM-2600d, Konica Minolta Sensing, Osaka, Japan) wasused to measure L *, a *, b *, melanin index (melanin content),
hemoglobin content (Hb index), and blood oxygen saturation(Hb SO 2 index) of the left cheek as described previously 28).

Safety test

For safety evaluation, the following parameters were
measured for the test diet and placebo before ( 0 weeks) andThis study was conducted at a third-party institution incompliance with the ethical principles based on the Declarationof Helsinki, the Private Information Protection Law, and theMinisterial Ordinance on Good Clinical Practice (GCP) for
Drugs (Ministry of Health, Labor and Welfare, Ordinance No.
28 of March 27, 1997). The study protocol was reviewed for
ethical aspects and appropriateness of the study and approvedby the human research ethics committee of the institutional
review board at Tokyo Synergy Clinic (Chuo-ku, Tokyo,
Japan). The study was conducted according to the approved
protocol.
The principal investigator and sub investigators, incooperation with a contract research organization, explained
the details of the study to and obtained written consent fromeach subject based on her free will before initiating the study.

Statistical analysis

All results were expressed as mean ± standard deviation.
Dunnett’s test was used to compare data obtained before andat 8 and 12 weeks after diet intake, while unpaired MannWhitney U-test was used for comparisons between two groups.
In addition, a subclass analysis was performed in the group of
subjects with a 60-minute postprandial blood glucose level of
150 mg/dL or more (7 subjects from the control group and 10
from the test group), as determined by screening examination.
All analyses were performed using IBM SPSS Statistics
20 software (IBM Japan, Tokyo, Japan), with a two-sided
significance level of 5%. Safety evaluation was based on blood
parameter assessment and the occurrence of individual adverse
events.

Results
Subjective and objective symptoms

No significant changes or differences were observed in thescores for physical and mental symptoms, as assessed by the
AAQol questionnaire, within or between the control and testarms over the study period (data not shown).

_ 17 _

Anti-glycation effect of vinegar beverage in humans
insulin level in the test group compared to control was observedat week 12, but was within the range of physiological variation.
No significant intergroup differences were observed in
the serum concentrations of 3DG, CML, or pentosidine or in
the CML content in the skin stratum corneum during the study
period. The fluorescence intensity of skin AGEs was similar
between the two groups at week 8, but significantly higher inthe test group compared to the control group at week 12, i.e.,
after discontinuation of the test diet.
A subclass analysis was performed in 10 subjects fromthe test group and 7 from the control group with a 60-minutepostprandial blood glucose level of 150 mg/dL or more. CML
content in the skin stratum corneum in the test group was
significantly decreased compared to control at week 8 (p < 0.05)
and remained at a low level even after discontinuation of thetest diet, although no significant difference was observed at 12
weeks ( Fig. 1 ).

Physical examination

No significant changes or differences were observed inbody height, body weight, body composition, basal metabolic
rate, blood pressure or pulse rate within or between the control
and test arms over the study period ( Table 3 ).

Blood chemistry

The results of blood chemistry are summarized in Table 4.
The AST (GOT ) level in the test arm was significantly decreased
compared to control at week 8, but returned to baseline level
after discontinuation of the test diet. No significant intergroupdifference was observed in any other parameter.

Vascular function test

No significant intergroup difference was observed inestimated vascular age as determined by fingertip acceleration
pulse wave analysis or by serum concentrations of endothelin,
VEGF, and NOX ( Table 5 ).

Oxidation stress parameters

In the test group, a significant increase in urine 8-OHdG
concentration was observed at weeks 8 and 12 as compared
to before diet intake (p = 0.009 and p = 0.009, respectively)( Table 7 ). Meanwhile, no significant difference was foundin this parameter between the test and control group. No
significant changes or differences were observed in urine
isoprostane concentrations within or between groups.

Glycation stress parameters

Glycation stress parameters are summarized in Table 6.
No significant intergroup difference was observed in FPG or
HbA1c levels during the study period. A significantly increased

Table 3. Anthropometry and physical examination.
Parameter

Unit Reference
range

Age

years

ー

Body heightcmーBody weightkgーBody fat%ー

Fat masskgーFat-free masskgーMuscle mass

kg

ー

BMI

kg/m2

ー

Basal metabolic
rate

kcal

ー

Diet

n

Control diet 11
Test diet

11

Control diet 11
Test diet

11

Control diet 11
Test diet

11

Control diet 11
Test diet

11

Control diet 11
Test diet

11

Control diet 11
Test diet

11

Control diet 11
Test diet

11

Control diet 11
Test diet

11

Control diet 110W

8W

Dunnettmean ±

SD

57.45 ±

4.44

ー

±

ー

ー

ー

±

ー

ー

±

ー

3.72

ー

ー

ー

±

ー

4.00

ー

±

ー

ー

ー

ー

±

ー

ー

6.49

ー

±

ー

±

ー

ー

5.45

0.989

8.01

1.000

3.89

0.943

5.11

0.990

3.97

0.969

5.24

0.996

2.21

0.995

3.75

0.983

2.02

0.994

3.43

0.984

1.99

0.985

2.47

0.997

152.43 ±153.18 ±59.16 ±
60.05 ±
35.05 ±
35.80 ±
20.87 ±
21.80 ±
38.28 ±
38.23 ±
36.09 ±
36.05 ±
25.45 ±
25.48 ±

5.20
8.79
3.75
5.15
3.87
5.61
2.10
4.04
1.92
3.68
1.89
2.57

1128.36 ± 70.40

ーー

59.18 ±

5.20

1.000

58.88 ±

60.10 ±

8.57

1.000

60.12 ±

35.06 ±

3.67

1.000

34.59 ±

35.84 ±

5.13

1.000

35.55 ±

20.88 ±

3.79

1.000

20.52 ±

21.83 ±

5.56

1.000

21.63 ±

38.30 ±

2.12

1.000

38.35 ±

38.27 ±

3.94

1.000

38.49 ±

36.11 ±

1.93

0.999

36.16 ±

36.08 ±

3.59

1.000

36.27 ±

25.46 ±

1.95

1.000

25.34 ±

25.52 ±

2.56

0.999

25.55 ±

1129.00 ± 71.21

1.000 1128.64 ± 74.24

111133.55 ± 130.05

1134.55 ± 126.05

Systolic blood mmHg 100-139 Control diet 11
pressure
Test diet 11

118.82 ± 12.68

114.41 ± 12.42

0.667

71.27 ± 10.83

0.997

76.05 ±

71.91 ±

8.03

0.461

67.50 ±

7.79

0.335

65.73 ±

7.66

0.181

Diastolic blood
mmHg 50-80
pressure
Pulse rate

bpm

40-80

Test diet

114.73 ± 14.80

Control diet 1171.55 ± 10.32

11

8.51

Test dietControl diet 11
Test diet11

71.64 ±

7.13

71.91 ± 10.00

mean ± SD

Dunnett's Independent samples t-testSD

57.27 ±

vs. 0W

12W

mean ±

113.36 ± 16.23

Abbreviations: BMI; body mass index_ 18 _

1.000 1139.18 ± 118.86
0.969

113.18 ± 15.27
118.55 ± 15.37
71.64 ± 11.09
73.73 ± 10.28
68.18 ±
67.00 ±

vs. 0W

1.000
0.992
0.961
0.998
1.000
0.772

7.38

0.455

8.10

0.321

At 8W

At 12W

ー

ー

ー

ー

0.907

0.568

0.949

0.683

0.959

0.727

0.881

0.404

0.923

0.442

0.837

0.460

0.934

0.470

0.509

0.750

0.216

0.375

0.497

0.598

Glycative Stress Research

Table 4. Serum parameters.
Parameter

Unit

Reference
range

Total protein

g/dL

6.7 - 8.3

Albuming/dL

3.8 - 5.3

A /G

1.1 - 2.0

AST(GOT)IU/L/37℃

10 - 4 0

ALT(GPT)

IU/L/37℃

5 - 45LD(LDH)

IU/L/37℃ 120 - 240

γ - GT

IU/L/37℃≤ 30

Total bilirubin

mg /dL

0.2 - 1.2

CPK

IU/L/37℃

4 0 - 150

Creatinine

mg /dL0.47 - 0.79

Urea nitrogen

mg /dL

8.0 - 20.0

Uric acidmg /dL

2.5 - 7.0

Total cholesterol

mg /dL120 - 219

HDL cholesterol

mg /dL40 - 95

LDL cholesterol

mg /dL65 - 139

TG (triglyceride)

mg /dL30 - 149

SodiummgEq/L137 - 147

Potassium

mgEq/L

3.5 - 5.0

Chloride

mgEq/L98 - 108

Calcium

mg/dL

8.4 - 10.4

Serum iron

μg/dL

40 - 180AI(atherogenic index)

<4

0W

8W

Dunnett's

12W

Dunnett's

Independent
samples t-test

mean ± SD

mean ± SD

vs. 0Wmean ± SD

vs. 0W

At 8W At 12W

11

7.35 ± 0.33

7.57 ± 0.32

0.292

7.54 ± 0.44

0.413

Test diet11

7.34 ± 0.37

7.50 ± 0.39

0.525

7.60 ± 0.38

0.202

Control diet11

4.36 ± 0.14

4.46 ± 0.17

0.355

4.43 ± 0.23

0.639

Test diet11

4.37 ± 0.15

4.43 ± 0.22

0.698

4.47 ± 0.18

0.346

Control diet11

1.49 ± 0.18

1.45 ± 0.18

0.770

1.45 ± 0.17

0.844

Test diet11

1.49 ± 0.15

1.45 ± 0.21

0.811

1.45 ± 0.16

0.764

Control diet11

19.91 ± 2.95

21.18 ± 4.83

0.604

21.27 ± 2.10

0.564

Test diet11

24.36 ± 6.09

20.80 ± 4.39

0.307

23.45 ± 7.03

0.913

Control diet11

16.82 ± 4.77

16.91 ± 5.11

0.999

17.64 ± 4.70

0.893

Test diet11

21.73 ± 7.51

19.30 ± 5.58

0.678

22.00 ± 8.67

0.995

Control diet11

204.55 ± 53.95

196.55 ± 37.52

0.871

198.45 ± 34.25

0.922

Test diet11

216.36 ± 19.73

205.70 ± 18.32

0.380

206.82 ± 21.59

0.438

Control diet11

26.82 ± 17.55

23.64 ± 9.05

0.782

21.73 ± 9.63

0.548

Test diet1140.55 ± 34.52

28.40 ± 19.57

0.561

38.55 ± 32.98

0.982

Control diet

110.75 ± 0.25

0.79 ± 0.19

0.879

0.76 ± 0.16

0.992

Test diet

110.80 ± 0.29

0.89 ± 0.28

0.773

0.88 ± 0.43

0.799

Control diet11

109.64 ± 53.84

115.18 ± 65.40

0.953

94.18 ± 23.19

0.699

Test diet11

84.27 ± 32.42

84.60 ± 26.60

1.000

94.36 ± 36.21

0.687

Control diet

11

0.67 ± 0.09

0.63 ± 0.09

0.479

0.66 ± 0.08

0.971

Test diet

110.64 ± 0.07

0.62 ± 0.07

0.876

0.64 ± 0.07

0.984

Control diet1112.84 ± 3.34

13.57 ± 3.44

0.826

13.34 ± 3.26

0.915

Test diet11

12.34 ± 2.18

12.64 ± 2.22

0.938

12.57 ± 2.58

0.960

Control diet11

5.12 ± 1.50

4.83 ± 1.21

0.855

4.95 ± 1.64

0.951

Test diet

11

4.74 ± 1.25

4.76 ± 1.03

0.998

4.49 ± 1.10

0.833

Control diet11234.36 ± 29.69

238.27 ± 38.58

0.950

235.45 ± 34.37

0.996

Test diet11

238.09 ± 31.48

245.70 ± 30.84

0.829

243.09 ± 38.51

0.917

Control diet11

63.91 ± 14.58

71.73 ± 20.07

0.459

68.45 ± 15.73

0.756

Test diet11

62.36 ± 10.59

66.10 ± 11.09

0.645

65.27 ± 10.47

0.752

Control diet11148.09 ± 26.63

147.36 ± 36.81

0.998

148.00 ± 34.92

1.000

Test diet11

154.45 ± 29.54

156.10 ± 29.17

0.989

155.91 ± 34.48

0.991

Control diet11

102.82 ± 52.17

84.55 ± 37.62

0.525

88.64 ± 40.40

0.670

Test diet

11102.00 ± 24.08

104.60 ± 39.88

0.984

111.00 ± 51.40

0.821

Control diet11142.00 ± 1.26

140.55 ± 1.21

0.016

141.18 ± 1.17

0.215

Test diet11

142.18 ± 1.33

141.70 ± 1.57

0.619

141.64 ± 1.03

0.530

Control diet11

4.30 ± 0.24

4.37 ± 0.27

0.722

4.40 ± 0.24

0.551

Test diet

11

4.32 ± 0.27

4.44 ± 0.39

0.562

4.29 ± 0.24

0.968

Control diet11

104.64 ± 1.57

103.45 ± 1.04

0.079

103.55 ± 1.29

0.109

Test diet11

104.91 ± 1.45

104.90 ± 2.13

1.000

103.91 ± 1.51

0.303

Control diet11

9.77 ± 0.30

9.81 ± 0.30

0.946

9.76 ± 0.33

0.997

Test diet119.83 ± 0.25

9.84 ± 0.27

0.991

9.82 ± 0.27

0.995

Control diet11

98.09 ± 34.67

99.91 ± 35.80

0.988

94.64 ± 28.39

0.957

Test diet11

94.00 ± 27.04

96.70 ± 21.80

0.961

105.73 ± 29.13

0.483

Control diet11

2.82 ± 0.97

2.50 ± 0.91

0.633

2.59 ± 0.87

0.787

Test diet11

2.89 ± 0.65

2.81 ± 0.81

0.957

2.78 ± 0.79

0.919

Diet

n

Control diet

0.407

0.549

0.419

0.650

0.726

0.821

0.029

0.117

0.333

0.830

0.729

0.767

0.537

0.634

0.723

0.451

0.891

0.153

0.288

0.597

0.660

0.734

0.271

0.735

0.910

0.731

0.146

0.555

0.931

0.880

0.145

0.172

0.116

0.697

0.716

0.307

0.180

0.897

0.854

1.000

0.812

0.119

0.167

0.584

Abbreviations: AST/GOT, aspartate aminotransferase; ALT/GPT, alanine aminotransferase; LD/LDH, lactate dehydrogenase; y-GT, gamma-glutamyl transpeptidase; CPK,
creatine phosphokinase_ 19 _

Anti-glycation effect of vinegar beverage in humans

Table 5. Vascular endothelial function parameters.
Parameter

Unit

Estimatedvascular age

Years

SDPTGAI

−

b/a

−

c /a

−

d /a

−

e /a

−

b -a

ms

c -a

ms

d -a

ms

e -a

ms

a-a

ms

PTGAI

−

Fingertip acceleration pulse wave

n

Diet

Control diet 11
Test diet

11

Control diet 11
Test diet

11

Control diet 11
Test diet

11

Control diet 11
Test diet

11

Control diet 11
Test diet

11

Control diet 11
Test diet

11

Control diet 11
Test diet

11

Control diet 11
Test diet

11

Control diet 11
Test diet

11

Control diet 11
Test diet

11

Control diet 11
Test diet

11

Control diet 11
Test diet

Endothelin

pg/mL(vascular endothelial growth factor)

VEGF

pg/mL

NO

μmol/L

11

Control diet 11
Test diet

11

Control diet 11
Test diet

11

Control diet 11
Test diet11

mean ± SD

mean ± SD

8W

Dunnett's
vs. 0Wmean ± SD

60.50 ± 11.33

58.32 ± 8.58

0.831

62.95 ± 10.12

0W

65.27 ± 8.13
-0.01 ± 0.27
0.11 ± 0.23

-0.48 ± 0.07
-0.42 ± 0.12
-0.20 ± 0.11
-0.21 ± 0.05
-0.37 ± 0.11
-0.42 ± 0.11
0.11 ± 0.04
0.10 ± 0.06

95.45 ± 19.06
93.64 ± 9.71

173.27 ± 18.38
164.91 ± 14.60
222.55 ± 14.89
226.73 ± 21.60
304.36 ± 21.03
311.09 ± 28.29
914.36 ± 99.64
896.18 ± 94.47
1.14 ± 0.12
1.24 ± 0.21
1.25 ± 0.23
1.27 ± 0.38

19.68 ± 6.25

27.11 ± 13.12
32.29 ± 24.71
26.65 ± 13.29

60.77 ± 12.01-0.07 ± 0.20
-0.01 ± 0.30
-0.51 ± 0.08
-0.49 ± 0.13
-0.15 ± 0.08
-0.18 ± 0.08
-0.38 ± 0.11
-0.38 ± 0.11
0.09 ± 0.03
0.08 ± 0.06

86.73 ± 12.34
87.09 ± 12.44

168.00 ± 11.70
161.45 ± 14.67

225.09 ± 15.06
222.36 ± 6.12

303.64 ± 16.90
304.73 ± 9.60

918.55 ± 87.08
926.18 ± 86.91
1.20 ± 0.12
1.22 ± 0.20
1.39 ± 0.29
1.27 ± 0.38

27.48 ± 12.47
63.53 ± 59.29
48.41 ± 26.44
42.70 ± 19.37

0.414

12W

61.68 ± 6.23
0.04 ± 0.24

0.812

0.02 ± 0.17

0.420

-0.45 ± 0.10

0.647

-0.44 ± 0.07

0.272

-0.17 ± 0.09

0.333

-0.17 ± 0.09

0.600

-0.40 ± 0.11

0.984

-0.36 ± 0.10

0.539

0.08 ± 0.04

0.634
0.566
0.322
0.356
0.647
0.824
0.878
0.736
0.993
0.668
0.991
0.676

Dunnett's Independent samples t-test

0.07 ± 0.03

88.18 ± 14.01
88.73 ± 13.92

163.27 ± 15.98
167.09 ± 17.24
224.18 ± 12.02
222.91 ± 14.73
303.09 ± 15.83
306.00 ± 16.10
928.18 ± 72.64
906.00 ± 99.70
1.25 ± 0.16

0.449

vs. 0W
0.793
0.559
0.849
0.594
0.647
0.878
0.669
0.490
0.699
0.295
0.261
0.397
0.444
0.544
0.244
0.924
0.947
0.789
0.980
0.769
0.905
0.957
0.109

0.956

1.22 ± 0.16

0.251

−

−

−

0.985

−

−

−

0.179
0.089
0.229
0.039

29.42 ± 12.69
58.94 ± 38.02
31.14 ± 22.28
22.05 ± 12.00

0.979

0.080
0.135
0.991
0.699

At 8W

At 12W

0.530

0.110

0.519

0.156

0.392

0.262

0.481

0.943

0.257

0.032

0.749

0.874

0.655

0.625

0.731

0.067

0.431

0.539

0.542

0.696

0.403

0.924

0.308

0.052

0.357

−

0.109

0.091

0.917

0.622

Abbreviations: SDPTGAI, second derivative of plethysmogram aging index; PTGAI, plethysmogram aging index; NO, nitric oxide

Table 6. Glycation stress-related parameters.
Parameter

Unit

Reference
range

Glucosemg/dL

70 - 109

Insulin

μU/mL

1.7 - 10.4

HbA1c/NGSP%

4.6 - 6.2

HbA1c/JDS%

4.3 - 5.8

3-deoxyglucosone

ng/mL 3.76 - 18.14

CM L

μg/mL

−

Pentosidine

pmol/mL−

Skin AGE deposition(AF level)−−

CML content instratum corneum

μg/mg
protein

−

Diet

n

0W
mean ± S D

8Wmean ± S D

Control diet 1187.91 ±

5.74

88.73 ± 6.59

Control diet 113.80 ±

2.65

3.48 ± 1.61

Test diet

11

Test diet

11

Test diet

11

Test diet

11

Test diet

11

Test diet

11

Test diet

11

Test diet

11

Test diet

11

Control diet 11
Control diet 1190.45 ±
3.15 ±
5.53 ±
5.65 ±
5.15 ±
5.26 ±

Control diet 11

16.82 ±

Control diet 114.45 ±

16.73 ±
4.40 ±

9.11
0.98
0.34
0.35
0.31
0.34

92.60 ± 11.42
4.04 ± 1.25
5.54 ± 0.27
5.59 ± 0.37
5.15 ± 0.26
5.22 ± 0.34

3.57

24.81 ± 6.89

0.64

4.78 ± 1.04

4.13
0.77

25.89 ± 5.13
4.39 ± 0.64

Control diet 11

94.52 ± 15.60

73.43 ± 27.35

Control diet 112.30 ±

2.09 ± 0.25

Control diet 11

89.13 ± 26.33
2.24 ±

0.25
0.22

103.31 ± 24.99
112.12 ± 24.13

66.29 ± 15.30
2.18 ± 0.23

100.71 ± 18.17
93.85 ± 27.17

Dunnett's
12W
vs. 0W mean ± S D
0.934
0.814
0.919
0.245
0.996
0.903
1.000
0.948
0.001
0.000
0.616
1.000
0.038
0.028
0.129
0.755
0.933
0.182

90.36 ± 6.41
91.18 ± 6.31
3.95 ± 2.17
4.65 ± 1.73
5.40 ± 0.24
5.45 ± 0.45
5.03 ± 0.21
5.07 ± 0.42

19.05 ± 3.660
19.94 ± 4.289
5.78 ± 1.045
5.61 ± 0.824

87.26 ± 15.221
93.98 ± 16.634
2.06 ± 0.268
2.25 ± 0.276

94.08 ± 14.458
89.52 ± 25.502

Dunnett's Independent samples t-test
vs. 0W
At 8W At 12W
0.562
0.975
0.982
0.028
0.487
0.366
0.483
0.382
0.474
0.185
0.004
0.001
0.613
0.798
0.071
0.990
0.450
0.085

0.951

0.468

0.087

0.026

0.329

0.328

0.516

0.339

0.741

0.599

0.613

0.820

0.824

0.081

0.077

0.017

0.176

0.246

Abbreviations: HbA1c/NGSP, hemoglobin A1c/National Glycohemoglobin Standardization Program; HbA1c/JDS, hemoglobin A1c/ Japan Diabetes Society; CML,
carboxymethyl lysine; AGE, advanced glycation end products; AF, autofluorescence_ 20 _Glycative Stress Research

1.20

Inflammation-related parameters

No significant changes or differences were observed in
hsCRP or IL-6 levels within or between groups over the study
period ( Table 8 ).

Placebo diet (n=7)
Test diet (n=10)

CML content (change ratio)

1.10

1.00

Urine electrolytes＊

Concentrations of sodium, potassium, and calcium present
in urine samples collected during the night are summarized inTable 9. No significant changes or differences were observed inurine electrolyte concentrations within or between groups overthe study period.

0.90
0.80
0.70
0.60

diet intake

0W

Skin function test

follow up

8W

No significant changes or differences were observed in
the results of the imaging analysis with the VISIA Evolution
system, moisture content analysis, or skin elasticity test withinor between groups over the study period ( Table 10 ).
In the color difference test, the Hb SO 2 index (blood
oxygen saturation) in the test group (measured in the leftcheek) was significantly higher than that in the control groupat week 8, and it remained at a similar level until week 12, i.e.,
after discontinuation of the test diet, although no significant
intergroup difference was observed. No significant intergroupdifference was observed in other parameters.

12W

Test period (week)

Fig. 1. Change in carboxymethyl lysine (CML) content in
the stratum corneum.

Subclass analysis of the subjects with high postprandial blood glucose(>150 mg/dL at 60 minutes).
CML content in the stratum corneum was measured by the tape
stripping method. The change ratio was calculated as follows:* p < 0.05, Mann-Whitney test.

Table 7. Oxidation stress-related parameters.
Parameter

Unit

8OHdG

pg/mg・Cr

Urine
isoprostane

pg/mg・Cr

DietnControl diet

0W

8W

mean ±

SD

mean ± SD

117.21 ±

2.06

Test diet11

6.98 ±

Control diet
Test dietDunnett's

12W

Dunnett's

vs. 0Wmean ± SD

9.52 ± 2.23

0.063

10.21 ± 2.95

0.014

2.38

10.03 ± 2.06

0.009

10.05 ± 2.58

0.009

11

267.09 ± 102.38

291.55 ± 97.15

0.754

287.36 ± 69.30

0.822

11

308.36 ± 111.31

346.73 ± 93.29

0.508

323.09 ± 55.08

0.898

8W

Dunnett's

12W

Dunnett's

vs. 0W

Independent samples t-test
At 8W

At 12W

0.504

0.956

0.634

0.898

Abbreviations: 8OHdG, 8-hydroxydeoxyguanosine

Table 8. Inflammation-related parameters
Parameter

Unit

High-sensitivity
CRP

mg/dL

Interleukin-6

pg/mL

0W

Diet

n

mean ± SD

mean ± SD

Control diet11

0.06 ± 0.05

0.05 ± 0.02

0.522

Test diet11

0.08 ± 0.07

0.07 ± 0.07

Control diet110.93 ± 0.62

Test diet110.99 ± 0.47

mean ±

SDvs. 0W

−

±

−

−

0.856

−

±

−

−

0.84 ± 0.51

0.712

−

±

−

−

0.88 ± 0.39

0.550

−

±

−

−

8W

Dunnett's

vs. 0W

Independent samples t-test
At 8W

At 12W

0.530

−

0.529

−

Abbreviations: CRP, C-reactive protein

Table 9. Results of urine electrolyte analysis.
Parameter

Unit

Urine sodium

mEq/L

Urine potassium mEq/L
Urine calcium

mg/dL

0W

12W

Dunnett's

DietnControl diet11

94.45 ± 45.96

93.18 ± 51.38

0.998

97.73 ± 56.44

0.984

Test diet11

77.91 ± 42.80

103.55 ± 41.20

0.328

110.36 ± 52.09

0.183

Control diet11

20.20 ± 12.26

27.75 ± 21.92

0.532

27.67 ± 19.39

0.538

Test diet1117.85 ± 15.50

25.45 ± 14.36

0.439

28.65 ± 17.94

0.213

Control diet11

7.65 ±

3.93

12.18 ± 7.61

0.123

8.79 ±

4.67

0.851

Test diet11

12.55 ±

9.51

16.53 ± 13.86

0.589

14.38 ±

7.51

0.888

mean ± SD

mean ± SD_ 21 _

vs. 0Wmean ± SD

vs. 0W

Independent samples t-test
At 8W

At 12W

0.168

0.168

0.992

0.992

0.894

0.894

Anti-glycation effect of vinegar beverage in humans

Table 10. Skin function test.

Skin viscoelasticity

Moisture content (%)

Color difference analysis

Imaging analysis by VISIA (score)

Parameter
Brown spot

× 100

Pore

× 100

Porphyrin

× 100

Red spot

× 100

Spot

× 100

Texture× 100

UV spot

× 100

Wrinkle

× 100

L*

−

a*

−

b*

−

MelaninIndex−

Hb Index−

Hb SO 2
Index−

Moisturecontent

Apex of left cheek

R2

Dunnett's Independent samples t-test

0W

8W

Dunnett's

12W

mean ± SD

mean ± SD

vs. 0Wmean ± SD

vs. 0W11

7.52 ± 1.93

7.69 ± 2.27

0.976

7.76 ± 2.39

0.953

Test diet11

6.94 ± 2.58

7.46 ± 2.08

0.808

7.45 ± 2.01

0.816

Control diet11

1.26 ± 0.52

1.50 ± 0.54

0.542

1.52 ± 0.71

0.498

Test diet11

1.59 ± 1.25

1.84 ± 1.42

0.871

1.77 ± 1.36

0.924

Control diet

110.30 ± 0.30

0.25 ± 0.29

0.867

0.26 ± 0.23

0.918

Test diet11

0.15 ± 0.21

0.15 ± 0.14

0.999

0.13 ± 0.15

0.947

Control diet11

1.26 ± 0.45

1.37 ± 0.32

0.751

1.31 ± 0.51

0.934

Test diet11

1.40 ± 0.64

1.51 ± 0.85

0.930

1.52 ± 0.76

0.913

Control diet11

2.38 ± 1.00

2.59 ± 1.16

0.856

2.48 ± 1.02

0.970

Test diet11

2.11 ± 0.79

2.27 ± 0.91

0.879

2.32 ± 0.86

0.789

Control diet11

1.14 ± 0.43

1.22 ± 0.51

0.909

1.28 ± 0.62

0.765

Test diet11

1.63 ± 1.57

1.81 ± 1.67

0.953

1.80 ± 1.71

0.959

Control diet11

2.51 ± 1.64

2.64 ± 1.81

0.985

3.25 ± 2.71

0.630

Test diet11

2.32 ± 1.52

2.56 ± 1.66

0.912

2.66 ± 1.59

0.834

Control diet11

2.79 ± 2.28

2.86 ± 2.75

0.996

2.77 ± 1.98

1.000

Test diet11

3.54 ± 3.02

3.55 ± 2.66

1.000

3.09 ± 2.62

0.900

Control diet11

61.77 ± 2.34

62.39 ± 2.56

0.774

62.86 ± 2.39

0.474

Test diet11

61.88 ± 1.89

62.44 ± 2.15

0.714

62.76 ± 1.63

0.457

Control diet11

7.01 ± 1.21

6.75 ± 1.30

0.856

6.74 ± 1.40

0.849

Test diet11

7.18 ± 1.02

7.32 ± 1.57

0.957

7.09 ± 1.30

0.979

Control diet1117.82 ± 1.60

17.34 ± 1.93

0.778

16.92 ± 2.14

0.444

Test diet11

18.01 ± 1.96

17.75 ± 2.26

0.941

17.69 ± 2.13

0.912

Control diet11

1.04 ± 0.17

0.99 ± 0.21

0.772

0.94 ± 0.21

0.409

Test diet11

1.05 ± 0.15

1.01 ± 0.18

0.780

0.99 ± 0.16

0.624

Control diet110.92 ± 0.15

0.92 ± 0.17

1.000

0.95 ± 0.18

0.904

Test diet110.94 ± 0.19

0.98 ± 0.24

0.855

0.94 ± 0.20

0.998

Control diet11

56.45 ± 6.11

57.02 ± 7.57

0.968

58.82 ± 5.20

0.593

Test diet11

56.49 ± 5.67

60.23 ± 3.95

0.169

61.25 ± 5.56

0.068

Control diet11

63.57 ± 6.77

60.72 ± 8.61

0.545

59.05 ± 5.54

0.247

Diet

n

Control diet

Test diet11

59.71 ± 3.73

54.81 ± 6.89

0.093

57.87 ± 5.93

0.671

Medial aspect ofright upper arm

Control diet

110.81 ± 0.03

0.80 ± 0.06

0.640

0.80 ± 0.04

0.678

Test diet

110.81 ± 0.05

0.80 ± 0.05

0.761

0.81 ± 0.04

0.967

R7

Medial aspect ofright upper arm

Control diet

110.60 ± 0.05

0.56 ± 0.06

0.250

0.57 ± 0.06

0.534

Test diet

110.59 ± 0.06

0.56 ± 0.08

0.426

0.59 ± 0.05

0.944

R2

Left cheek

Control diet

110.69 ± 0.07

0.71 ± 0.07

0.830

0.75 ± 0.06

0.129

Test diet

110.73 ± 0.07

0.72 ± 0.05

0.995

0.72 ± 0.06

0.986

R7

Left cheek

Control diet

110.33 ± 0.06

0.35 ± 0.04

0.570

0.38 ± 0.06

0.098

Test diet

110.33 ± 0.06

0.35 ± 0.04

0.453

0.34 ± 0.05

0.714

Abbreviations: 8OHdG, 8-hydroxydeoxyguanosine_ 22 _At 8W

At 12W

0.443

0.572

0.922

0.583

0.263

0.712

0.894

0.715

0.645

0.327

0.538

0.872

0.524

0.516

0.946

0.387

0.911

0.668

0.287

0.551

0.603

0.226

0.749

0.248

0.396

0.626

0.036

0.152

0.534

0.344

0.842

0.465

0.849

0.488

0.563

0.089

0.926

0.212

Glycative Stress Research

Safety test

No serious adverse events related to the test diet were observed.
One adverse event was assessed as probably related to the test
diet, but it was mild and did not require study discontinuation.
The event occurred in early October and was resolved by
November 22. The study was continued, and no aggravationof symptoms was observed. In brief, a 51-year-old woman
reported a tendency for gas retention, possibly due to atransient disturbance of the intestinal bacterial flora caused byindigestible dextrin. With no serious adverse event observed
during the study period, the test diet was considered safe.

Discussion

In this study we investigated the long-term effects of theintake of a vinegar beverage containing indigestible dextrin
and a mixed herbal extract on a panel of glycation stressmarkers. A main result of the study showed that intake of thetest diet caused a reduction in CML content in the skin stratum
corneum, a maker of glycation stress, in people with a relatively
high level of glycation stress, indicating the diet has an antiglycation effect. As a second result of the study we found thetest diet to be safe for long-term use as no serious adverse
events could be observed during the study period. Indigestible
dextrin acts by suppressing hyperglycemia after a meal.
The tien-cha extract containing Rubus suavissimus has beenshown to inhibit AGE production in vitro 9). AG herb mixTM
has been shown to inhibit AGE production in in vitro studies,
experimentally induced diabetic rats, and RCTs in humans 5, 10) .
While no significant intergroup difference was observedin any of the glycation stress markers evaluated in the entire
population, a subclass analysis involving 17 subjects with a
60-minute postprandial blood glucose level of 150 mg/dL or
more showed a significant improvement in CML content in
the stratum corneum after intake of the test diet ( p < 0.05 ).
The pentosidine level was unchanged in the control arm anddecreased in the test group. Although no significant difference
was observed between groups, this observation also supports
the inhibitory effect of the test diet on AGE production. The
tape stripping technique 20), used in this study for determining
CML content in the stratum corneum, is promising as a noninvasive method for evaluating glycation stress in the skin.
No significant changes were observed in the blood

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Conflict of interest

The authors have no conflict of interest in this study.

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