JHSMR
Original Article
Journal of
Health Science
and Medical Research
Variability of Isoflavone Content in Soy Milk Products Commercially
Available in Thailand
Ekkapon Hirattanapun1, Nut Koonrungsesomboon, M.D., Ph.D.1,
Supanimit Teekachunhatean, M.D., Ph.D.1,2
Department of Pharmacology, 2Center of Thai Traditional and Complementary Medicine,
Faculty of Medicine, Chiang Mai University, Mueang, Chiang Mai 50200, Thailand.
1
Received 13 November 2017 l Accepted 19 December 2017 l Published online 24 May 2018
Abstract:
Objective: To determine the isoflavone content in soy milk products commercially available in Thailand.
Material and Methods: Two flavors of each of 4 brands of soy milk were obtained from retail outlets in Chiang Mai,
Thailand. The isoflavone content in soy milk was determined using high-performance liquid chromatography.
Results: Genistin and daidzin, both β-glycoside conjugates, were found to be the main isoflavone components in soy
milk. The total amount of isoflavones varied considerably among different products, ranging from 25.5 to 63.5 mg per
serving (250 mL) of soy milk. Marked variation in total isoflavones was also found even among different lots of the same
product.
Conclusion: The isoflavone content in soy milk products available in Thailand varies substantially among different
products as well as among different lots of the same product. This suggests that standardizing or, at least, declaring the
isoflavone content in soy milk products is needed to facilitate the optimal consumption of soy milk for health benefits.
Keywords: isoflavones, soy milk, Thailand
Contact: Assoc. Prof. Supanimit Teekachunhatean, M.D., Ph.D.1,2
1
Department of Pharmacology, 2Center of Thai Traditional and
Complementary Medicine, Faculty of Medicine, Chiang Mai University,
Mueang, Chiang Mai 50200, Thailand.
E-mail: supanimit.t@cmu.ac.th
JHSMR 2018;36(2):117-126
www.jhsmr.org
117
Isoflavone in Soy Milk in Thailand
Hirattanapun E, et al.
Introduction
Material and Methods
Soy milk product sampling
Two flavors of each of 4 brands of soy milk (coded
as A1, A2, B1, B2, C1, C2, D1, D2) were obtained from
retail outlets in Mueang Chiang Mai district, Chiang Mai
province, Thailand. For each product, 3 different lots (3
samples per lot) were randomly selected based on the
expiry date displayed on the package: Lot 1 expired in
August 2015, Lot 2 expired in October or November 2015,
and Lot 3 expired in March 2016. None of the 8 soy milk
product packages provided information on isoflavone
content.
Soy isoflavones represent the most common group
of phytoestrogens and are structurally similar to 17βestradiol.1 They exhibit 3 aglycone structures, daidzein,
genistein, and glycitein, each of which has a corresponding
β-glycoside conjugate, i.e., daidzin, genistin, and glycitin,
respectively, as well as acetyl and malonyl glycoside
conjugates.2 Interest in isoflavones has increased owing
to evidence that these phytoestrogens have a wide range
of pharmacological activities.3 Promising data from clinical
studies suggest beneficial effects of isoflavones for various
health conditions, primarily those related to women’s health.4
For example, systematic reviews and meta-analyses have
demonstrated a significant reduction in the frequency and
severity of menopausal hot flashes with regular consumption
of isoflavones.5-8 In addition, isoflavones have exhibited
potential health benefits in the prevention of osteoporosis
and cardiovascular disease in menopausal women.9-11
Soybeans (Glycine max L. Merr.) and soy-based
products are one of the richest food sources of isoflavones.12
Soy is found in traditional foods in many Asian countries,
especially Japan, while in Western countries soy consumption
has continuously increased over the last decade.13 At
present, soy milk and other soy-based beverages are widely
consumed worldwide and interest has grown because
of the prospect of their beneficial properties in maintaining
health and preventing diseases.14 However, in spite of the
wide array of soy milk products commercially available
on the market, there is only limited information available
regarding soy isoflavone content in Thailand. This lack
of information hinders the determination of the optimal
consumption of soy milk as a healthful beverage.
The objective of the present study was to determine
the isoflavone content of soy milk products commercially
available in Thailand.
Journal of Health Science and Medical Research
Sample preparation
Sample preparation was completed using a modified
version of a method described in the Association of
Official Agricultural Chemists (AOAC) Official Methods of
Analysis. 15 Using that method, 1 mL of soy milk was
mixed with 9 mL of 80.0% methanol in water. Then, 350 µL
of 2 M NaOH was added and the mixture was sonicated
for 30 minutesafter which 250 µL of 100.0% acetic acid
was added. One mL of the mixture was centrifuged. Ten µL
of clear supernatant was then mixed with 30 µL of mobile
phase and spiked with 10 µL of internal standard (IS, 10,000
ng/mL of chloramphenicol). Five µL of the mixture was
injected into a high-performance liquid chromatography
(HPLC) system. Each sample was analyzed in triplicate.
High-performance liquid chromatographic
condition
The assay used to quantify the isoflavones was
modified from the isocratic reversed-phase HPLC method
developed by Cesar Ida, et al. 16 The chromatographic
system consisted of a C18 reverse phase column (Inertsil®,
250 mm x 4.6 I.D., 5 µm, GL Science, Tokyo, Japan) equipped
118
JHSMR 2018;36(2):117-126
Hirattanapun E, et al.
Isoflavone in Soy Milk in Thailand
with a guard column of the same material (Inertsil® ODS-3,
10 mm x 4.0 I.D., 5 µm, GL Science, Tokyo, Japan). The
chromatographic analysis included a mobile phase A
(0.1% acetic acid in water) and a mobile phase B (100.0%
methanol). Separation was performed at 40 °C. Gradient
elutions of 85.0% A with 15.0% B for 20 min, 30.0% A
with 70.0% B for 5 min, and 85.0% A with 15.0% B for 7 min
were accomplished. The mobile phase was maintained
at a flow rate of 1 mL/min, and the results were quantified
by UV absorption at 259 nm.
was 100 ng/mL, which is the lowest concentration of an
analyte that can be reliably measured. Of the 5 samples
spiked at 100 ng/mL each, the average coefficients of
variation (% CV) at the LLOQ concentration (calculated using
Equation (1) for daidzin, genistin, glycitin, daidzein, genistein,
and glycitein were 2.0%, 1.2%, 2.6%, 0.2%, 0.5%, and
2.4%, respectively. The average % deviation at LLOQ
concentration (calculated using Equation 2) for those 6
isoflavones were 0.7%, 7.5%, 5.6%, 8.1%, 7.9%, and 5.3%,
respectively.
Quantification of isoflavones and assay validation
Standard β-glycosides (daidzin, genistin, and
glycitin) and aglycones (daidzein, genistein, and glycitein)
were spiked in serial dilution (in 100.0% methanol) to
obtain a standard calibration curve. That curve ranged
between 100 and 6,400 ng/mL for daidzin and genistin,
and between 100 and 2,000 ng/mL for daidzein, genistein,
glycitin, and glycitein. The HPLC chromatogram of standard
isoflavones and their retention times are shown in
Figure 1A. All peaks were clearly separated and no interference from other substances was observed. The regression equations for testing the linearity of the standard
calibration curves are as follows:
y=0.2706x-1.1487 (r2=1.0000), for daidzin
y=0.4055x+2.0608 (r2=0.9999), for genistin
y=0.2252x-0.0279 (r2=0.9999), for glycitin
y=0.4117x-0.8728 (r2=0.9996), for daidzein
y=0.5420x-0.2867 (r2=0.9996), for genistein
y=0.2808x+1.2273 (r2=0.9998), for glycitein
The isoflavone content of the soy milk samples were
determined using a calibration curve of the peak height
ratios of isoflavones and using the linear regression of an
internal standard versus respective isoflavone concentrations.
The lower limit of quantification (LLOQ) of the 6
isoflavones under the HPLC conditions described above
Journal of Health Science and Medical Research
Standard deviation
% CV=
×100
Mean
(Equation 1)
(Equation 2)
(measured
concentration-spiked
concentration)
% Deviation=
×100
Spiked concentration
Intraday and interday validation procedures were
carried out to assess the precision of the technique.
For intraday validation, 5 samples from each of 3 quality
control (QC) samples (300, 2,500, and 5,000 ng/mL of
daidzin and genistin; 300, 900, and 1,800 ng/mL of daidzein,
genistein, glycitin, and glycitein) were evaluated with a
single calibration curve. For interday validation, 5 sets of
the 3 different concentrations of QC samples were studied
concurrently on 5 different days with 5 standard calibration
curves. The % CV (calculated using Equation 1) and %
deviation (calculated using Equation 2) of intraday and
interday assay validation for 6 isoflavonesare shown in
Table 1. None of the % CV values were above 6.2%, and
none of the % deviation values were greater than 9.4%
or less than -7.8%.
Data analysis
Descriptive analysis was performed; the quantities
of isoflavone content are presented as mean (±standard
119
JHSMR 2018;36(2):117-126
Isoflavone in Soy Milk in Thailand
Hirattanapun E, et al.
deviation; S.D.) per one serving (250 mL) of soy milk. The
values of isoflavones in the form of β-glycoside conjugates
used in the following text refer to the aglycone equivalent
weights.
Figure 1 HPLC chromatograms of (A) standard isoflavones (3,200 ng/mL of daidzin and genistin; 1,600 ng/mL of daidzein,
genistein, glycitin, and glycitein) and (B) isoflavone content in soy milk (a sample from D1).
Journal of Health Science and Medical Research
120
JHSMR 2018;36(2):117-126
Hirattanapun E, et al.
Isoflavone in Soy Milk in Thailand
Table 1 Intraday and interday assay validation of isoflavone content measurements
Concentration of QC
sample (ng/mL)
Precision (% CV)
Deviation (%)
Precision (% CV)
Deviation (%)
Daidzin
300
2,500
5,000
1.5
0.2
0.5
3.1
2.1
1.0
4.2
2.9
4.7
3.9
2.8
0.8
Genistin
300
2,500
5,000
0.7
0.3
0.6
8.8
9.4
8.9
1.2
0.7
3.3
7.3
8.9
7.2
Glycitin
300
900
1,800
0.9
1.5
2.7
-3.6
-1.6
-0.1
1.5
3.0
4.3
-5.2
-5.5
-4.0
Daidzein
300
900
1,800
0.9
0.6
0.4
-5.8
-1.6
1.9
2.2
3.1
5.6
-5.6
-4.1
-4.2
Genistein
300
900
1,800
1.0
1.3
1.8
-4.4
-2.4
1.7
2.0
3.7
6.2
-4.6
-5.0
-4.4
Glycitein
300
900
1,800
0.9
1.3
0.5
-1.6
-6.4
5.4
1.7
1.0
5.2
-2.3
-7.8
0.0
Isoflavones
Intraday
Interday
CV=the coefficient of variation, QC=quality control
Results
content among the same lot of the same product did
not vary to a great extent for any of the soy milk products
(% CV=0.2–5.5) (Figure 2).
The majority of isoflavones in soy milk were in the
form of β-glycoside conjugates, accounting for approximately 84.0–97.0% of the total isoflavones (Table 2).
Genistein, daidzein, and glycitein and their β-glycoside
forms accounted for 55.5% (S.D.=5.7), 40.1% (S.D.=3.5),
and 4.4% (S.D.=2.5), respectively, of the total isoflavone
content (Table 2 and Figure 3). The ratios of genistein and
its β-glycoside conjugateto total isoflavones varied
The HPLC chromatogram of isoflavone content
in soymilk is shown in Figure 1B. The mean quantities of
total isoflavones in soy milk products varied widely,
from 25.5 mg (in B1) to 63.5 mg (in D2) per 250 mL of
product (Table 2). Marked variation in total isoflavones
among different lots of the same product was found
across all brands, with the calculated % CV ranging from
17.7 (in B1) to 43.3 (in A1) (Figure 2). In all the brands
except B, the amount of total isoflavones in Lot 2 was
lower than Lot 1 and Lot 3. However, total isoflavone
Journal of Health Science and Medical Research
121
JHSMR 2018;36(2):117-126
Isoflavone in Soy Milk in Thailand
Hirattanapun E, et al.
from 0.5 to 0.6 while those of daidzein and its β-glycoside
conjugate were between 0.4 and 0.5 (Figure 3). The
amount of genistein and its β-glycoside form was higher
than daidzein and its β-glycoside form in all products.
Table 2 Isoflavone content of the 8 soy milk products investigated in this study
Soy milk
product
A1
A2
B1
B2
C1
C2
D1
D2
Isoflavone content (mg/250 mL of soy milk)
β-glycosides
Aglycones
Total
Daidzin*
Genistin*
Glycitin*
Daidzein
Genistein
Glycitein
13.5 (±5.6)
12.2 (±3.4)
8.3 (±2.9)
9.7 (±3.5)
23.2 (±1.9)
21.4 (±11.6)
25.9 (±13.4)
26.9 (±13.5)
24.8 (±10.6)
22.5 (±4.3)
11.6 (±1.5)
14.1 (±3.2)
34.4 (±12.4)
28.2 (±10.4)
29.0 (±10.0)
30.1 (±9.8)
0.2 (±0.3)
0.1 (±0.2)
1.5 (±0.2)
1.8 (±0.5)
2.8 (±0.5)
3.0 (±0.7)
3.3 (±0.7)
3.3 (±0.6)
1.5 (±0.8)
1.4 (±0.3)
1.9 (±1.1)
2.0 (±1.3)
0.7 (±0.2)
1.5 (±0.9)
1.6 (±0.9)
1.6 (±0.9)
1.8 (±1.5)
1.6 (±0.8)
2.2 (±1.3)
2.1 (±1.3)
0.6 (±0.1)
1.4 (±0.9)
1.2 (±0.6)
1.3 (±0.6)
0.0 (±0.0)
0.1 (±0.1)
0.0 (±0.0)
0.0 (±0.0)
0.3 (±0.2)
0.4 (±0.2)
0.4 (±0.1)
0.3 (±0.2)
41.8 (±18.1)
37.9 (±8.1)
25.5 (±4.5)
29.7 (±8.5)
61.9 (±14.0)
56.0 (±24.0)
61.3 (±25.6)
63.5 (±25.4)
32.6
66.9
26.4
33.7
97.8
32.5
Inter-product
42.7
variation (% CV)
Data are presented as mean (±standard deviation) of isoflavone content (for each type, n=9: 3 lots, 3 samples per lot).
*Data represent content in aglycone equivalent weight.
Figure 2 Total isoflavones in the 8 soy milk products (3 samples per lot) investigated in this study
Journal of Health Science and Medical Research
122
JHSMR 2018;36(2):117-126
Hirattanapun E, et al.
Isoflavone in Soy Milk in Thailand
Figure 3 Proportion of each aglycone and its corresponding β-glycoside conjugate to total isoflavones in the 8 soy
milk products (n=9: 3 lots, 3 samples per lot) investigated in this study
Discussion
to extensive variation in isoflavone content in soy milk
include the characteristics of the raw material (i.e., seed
variety and quality) rather than the largely standardized
manufacturing methods and storage conditions. This
finding indicates that producing soy milk products with
standardized isoflavone content would be a challenging
task for manufacturers.
The majority of the isoflavones found in soymilk
were β-glycosides (i.e., genistin and daidzin), while small
amounts of the aglycone forms were detected in all the
products. This observation is compatible with the current
knowledge of soy isoflavones: isoflavones in nonfermented
soy-based products appear predominantly in the form
of glycoside conjugates.24,25 Although the isoflavone content
investigated in this study was limited to the β-glycoside
conjugates and aglycones, this should have only a negligible effect on the total isoflavone content in soy milk,
as published studies suggest that isoflavones in the form
of malonyl- and acetyl-glycoside conjugates are generally
converted into β-glycoside forms during soy milk prepa-
The present study found substantial variation in
total and individual isoflavone content among the 8 soy
milk products commercially available in Thailand. The value
of total isoflavones in 1 serving (250 mL) of soy milk in
this study is comparable to the range of values (5 to 96
mg per 250 mL) for soy-based beverages previously
reported.17-19 The variation in the quantity of soy isoflavones
commonly observed might be attributable to several factors.
Differences in the characteristics of soybeans used (e.g.,
soy variety, seed quality, planting location, crop year, and
even planting dates within the same crop year), manufacturing methods (e.g., soaking, grinding, and heating
processing), storage conditions, as well as the quality of
beans used per 250 mL serving could contribute to the
considerable variation in isoflavone content in soymilk.20-23
The present study found a large difference in isoflavone
content among lots of the same product, while the content
within lots did not vary greatly. Based on this observation,
it is reasonable to postulate that the key factors contributing
Journal of Health Science and Medical Research
123
JHSMR 2018;36(2):117-126
Isoflavone in Soy Milk in Thailand
Hirattanapun E, et al.
ration.26,27 Additionally, available evidence clearly shows
that basic hydrolysis, which was applied during sample
preparation, would break ester bonds and convert the
malonyl- and acetyl-glycoside isoflavone forms to their
respective β-glucosides.28 The ratio of genistein and its
β-glycoside conjugate to daidzein and its β-glycoside
conjugate observed in the present study was around
1:1 to 2:1, which is comparable to Thai soybean varieties
reported in previous research.29
Genistein and its β-glycoside conjugate were the
main isoflavones in soy milk, accounting for 49.0–64.0% of
the total isoflavones. Variations in the proportion (or the
amount) of genistein and its glycoside conjugate in different
soy milk products may be of clinical relevance, since lines
of evidence suggest that genistein appears to be the
most biologically active component, responsible for the
beneficial effects of isoflavones in soy-based products.30,31
At least 15 mg/day (in aglycone equivalents) of genistein
consistently contributed to a significant reduction in the
symptoms of hot flashes in several clinical trials.32,33
Furthermore, the administration of genistein (54 mg/day)
for a year was found to exhibit significant positive effects
on bone mineral density in osteopenic menopausal and
postmenopausal women.34,35 Although isoflavones in soy
milk appear mainly in the β-glycoside form (genistin)
rather than the aglycone form (genistein), this seems
to have only modest effects on health benefits because
genistein wouldbe absorbed in equal amounts irrespective
of whether the glycoside form or the aglycone form had
been initially consumed.36-38 The presence of the sugar
moiety in the β-glycoside form may only delay the
absorption rate of genistein while having no significant
effect on the total absorption of isoflavones.36
Based on the results of this study, one glass of soy
milk (250 mL) would result in an intake of around 25–60
Journal of Health Science and Medical Research
mg of total isoflavones, which is consistent with the results
of a recent evaluation of the epidemiologic literature.39
That value is at least equivalent to the Japanese daily
intake of isoflavones from soybeans and soy-based foods
(an estimated 28 mg/day).40 The literature suggests that
the daily dose of total isoflavones that would benefit
menopausal health by reducing the frequency and severity
of hot flashes,5,6 as well as preventing osteoporosis10 and
cardiovascular disease,11 is about 40–100 mg (in aglycone
equivalent weight). Thus, the daily consumption of 1–3
servings of soy milk (250 mL/serving) would be required
to obtain the potential health benefits from soy isoflavones,
if soy milk were the sole isoflavone source in the diet.
Given the growing interest in and increasing
consumption of soy-based beverages, it would be useful
to consumers if nutritional details regarding isoflavone
content were provided on the packaging of soy milk
products. Accurate labelling of isoflavone content would
provide a valuable reference for the rational consumption
of soy isoflavones and could have an impact on consumers’ dietary choices. The information would also help
researchers more accurately estimate isoflavone
consumptionand facilitate epidemiological studies ofthe
beneficial health effects of soy isoflavones.
Conclusion
The isoflavone content in soy milk products
commercially available in Thailand varies substantially
among different products, as well as among different
lots of the same product. This suggests a need to
standardize isoflavone content in soy milk, or at least to
declare the isoflavone content on the package, in order
to help consumers and nutritionists determine the intake
of soy milk sufficient for obtaining health benefits from soy
isoflavones.
124
JHSMR 2018;36(2):117-126
Hirattanapun E, et al.
Isoflavone in Soy Milk in Thailand
Acknowledgment
The authors would like to thank Mr. Boonyium
Kumsornand Ms. Sujitra Techatoei for their supervision in
analysis and assay validation of isoflavone content. The
authors are grateful to Dr. G. Lamar Robert for his
assistance in editing the manuscript.
This work received financial support from the
Faculty of Medicine, Chiang Mai University, Thailand.
9.
10.
Disclosures statement
The authors declare that there are no conflicts of
interest regarding the publication of this article. None of
the authors have a direct financial relationship with any
of the commercial entities mentioned in this paper.
11.
References
12.
1. Setchell KD. Phytoestrogens: the biochemistry, physiology,
and implications for human health of soy isoflavones. Am J
Clin Nutr 1998;68(6 Suppl):S1333-46.
2. Wang H, Murphy PA. Isoflavone content in commercial
soybean foods. J Agric Food Chem 1994;42:1666-73.
3. Zaheer K, Humayoun Akhtar M. An updated review of dietary
isoflavones: nutrition, processing, bioavailability and impacts
on human health. Crit Rev Food Sci Nutr 2017;57:1280-93.
4. Messina M. Soy foods, isoflavones, and the health of postmenopausal women. Am J Clin Nutr 2014;100(Suppl 1):S423-30.
5. Taku K, Melby MK, Kronenberg F, Kurzer MS, Messina M.
Extracted or synthesized soybean isoflavones reduce menopausal hot flash frequency and severity: systematic review and
meta-analysis of randomized controlled trials. Menopause
2012;19:776-90.
6. Howes LG, Howes JB, Knight DC. Isoflavone therapy for
menopausal flushes: a systematic review and meta-analysis.
Maturitas 2006;55:203-11.
7. Franco OH, Chowdhury R, Troup J, Voortman T, Kunutsor S,
Kavousi M, et al. Use of plant-based therapies and menopausal
symptoms: asystematic review and meta-analysis. JAMA
2016;315:2554-63.
8. Sarri G, Pedder H, Dias S, Guo Y, Lumsden MA. Vasomotor
Journal of Health Science and Medical Research
13.
14.
15.
16.
17.
18.
125
symptoms resulting from natural menopause: a systematic
review and network meta-analysis of treatment effects from
the National Institute for Health and Care Excellence guideline
on menopause. BJOG 2017;124:1514-23.
Ma DF, Qin LQ, Wang PY, Katoh R. Soy isoflavone intake
increases bone mineral density in the spine of menopausal
women: meta-analysis of randomized controlled trials. Clin
Nutr 2008;27:57-64.
Taku K, Melby MK, Takebayashi J, Mizuno S, Ishimi Y, Omori T,
et al. Effect of soy isoflavone extract supplements on bone
mineral density in menopausal women: meta-analysis of
randomized controlled trials. Asia Pac J Clin Nutr 2010;19:
33-42.
Hodis HN, Mack WJ, Kono N, Azen SP, Shoupe D, HwangLevine J, et al. Isoflavone soy protein supplementation and
atherosclerosis progression in healthy postmenopausal
women: a randomized controlled trial. Stroke 2011;42:316875.
Gacek M. Soy and legume seeds as sources of isoflavones:
selected individual determinants of their consumption in a
group of perimenopausal women. Prz Menopauzalny 2014;13:
27-31.
Barrett JR. The science of soy: what do we really know?
Environ Health Perspect 2006;114:A352-8.
Sethi S, Tyagi SK, Anurag RK. Plant-based milk alternatives
an emerging segment of functional beverages: a review.
J Food Sci Technol 2016;53:3408-23.
Klump SP, Allred MC, MacDonald JL, Ballam JM. Determination of isoflavones in soy and selected foods containing
soy by extraction, saponification, and liquid chromatography:
collaborative study. J AOAC Int 2001;84:1865-83.
Cesar Ida C, Braga FC, Soares CD, Nunan Ede A, Pianetti GA,
Condessa FA, et al. Development and validation of a RPHPLC method for quantification of isoflavone aglycones in
hydrolyzed soy dry extracts. J Chromatogr B Analyt Technol
Biomed Life Sci 2006;836:74-8.
Genovese MI, Lajolo FM. Isoflavones in soy-based foods
consumed in Brazil: levels, distribution, and estimated intake.
J Agric Food Chem 2002;50:5987-93.
Teekachunhatean S, Rojanasthien N, Sangdee C. Isoflavone
contents in UHT and fresh soymilk available in amphur
Muang, Chiang Mai. Chiang Mai Med Bull 2004;43:151-61.
JHSMR 2018;36(2):117-126
Isoflavone in Soy Milk in Thailand
Hirattanapun E, et al.
30. McCarty MF. Isoflavones made simple - genistein’s agonist
activity for the beta-type estrogen receptor mediates their
health benefits. Med Hypotheses 2006;66:1093-114.
31. Marini H, Bitto A, Altavilla D, Burnett BP, Polito F, Di Stefano V,
et al. Breast safety and efficacy of genistein aglycone for
postmenopausal bone loss: a follow-up study. J Clin Endocrinol
Metab 2008;93:4787-96.
32. Lethaby A, Marjoribanks J, Kronenberg F, Roberts H, Eden J,
Brown J. Phytoestrogens for menopausal vasomotor symptoms.
Cochrane Database Syst Rev 2013:CD001395.
33. Williamson-Hughes PS, Flickinger BD, Messina MJ, Empie
MW. Isoflavone supplements containing predominantly
genistein reduce hot flash symptoms: a critical review of
published studies. Menopause 2006;13:831-9.
34. Marini H, Minutoli L, Polito F, Bitto A, Altavilla D, Atteritano M,
et al. Effects of the phytoestrogen genistein on bone metabolism in osteopenic postmenopausal women: a randomized
trial. Ann Intern Med 2007;146:839-47.
35. Morabito N, Crisafulli A, Vergara C, Gaudio A, Lasco A, Frisina
N, et al. Effects of genistein and hormone-replacement therapy
on bone loss in early postmenopausal women: a randomized
double-blind placebo-controlled study. J Bone Miner Res
2002;17:1904-12.
36. Kano M, Takayanagi T, Harada K, Sawada S, Ishikawa F.
Bioavailability of isoflavones after ingestion of soy beverages
in healthy adults. J Nutr 2006;136:2291-6.
37. Xu X, Wang HJ, Murphy PA, Hendrich S. Neither background
diet nor type of soy food affects short-term isoflavone
bioavailability in women. J Nutr 2000;130:798-801.
38. Zubik L, Meydani M. Bioavailability of soybean isoflavones
from aglycone and glucoside forms in American women. Am
J Clin Nutr 2003;77:1459-65.
39. Messina M. Soy and health update: evaluation of the clinical
and epidemiologic literature. Nutrients 2016;8:E754.
40. Nakamura Y, Tsuji S, Tonogai Y. Determination of the levels of
isoflavonoids in soybeans and soy-derived foods and estimation
of isoflavonoids in the Japanese daily intake. J AOAC Int 2000;
83:635-50.
19. Techatoei S, Sangdee C, Rojanasthien N, Manorot M, Teekachunhatean S. Isoflavone contents in Thai and imported soybased beverages commercially available in Thailand.
Chiang Mai Med Bull 2011;50:59-72.
20. Lee SJ, Ahn JK, Kim SH, Kim JT, Han SJ, Jung MY, et al.
Variation in isoflavone of soybean cultivars with location and
storage duration. J Agric Food Chem 2003;51:3382-9.
21. Eisen B, Ungar Y, Shimoni E. Stability of isoflavones in soy
milk stored at elevated and ambient temperatures. J Agric
Food Chem 2003;51:2212-5.
22. Xu Z, Wu Q, Godber JS. Stabilities of daidzin, glycitin, genistin,
and generation of derivatives during heating. J Agric Food
Chem 2002;50:7402-6.
23. Zhang J, Ge Y, Han F, Li B, Yan S, Sun J, et al. Isoflavone
content of soybean cultivars from maturity group 0 to VI
grown in northern and southern China. J Am Oil Chem Soc
2014;91:1019-28.
24. Wiseman H, Casey K, Clarke DB, Barnes KA, Bowey E.
Isoflavone aglycon and glucoconjugate content of high- and
low-soy U.K. foods used in nutritional studies. J Agric Food
Chem 2002;50:1404-10.
25. Nguyen HT, Pourian M, Bystrom B, Dahlin I, Duc PT, Nguyen
TV, et al. Low aglycone content in commercial soy drink
products. Asia Pac J Clin Nutr 2012;21:52-6.
26. Jackson CJC, Dini JP, Lavandier C, Rupasinghe HPV,
Faulkner H, Poysa V, et al. Effects of processing on the
content and composition of isoflavones during manufacturing
of soy beverage and tofu. Process Biochem 2002;37:1117-23.
27. Barnes S, Kirk M, Coward L. Isoflavones and their conjugates in soy foods: extraction conditions and analysis by
HPLC-mass spectrometry. J Agric Food Chem 1994;42:2466-74.
28. Delmonte P, Perry J, Rader JI. Determination of isoflavones in
dietary supplements containing soy, Red Clover and kudzu:
extraction followed by basic or acid hydrolysis. J Chromatogr
A 2006;1107:59-69.
29. Teekachunhatean S, Hanprasertpong N, Teekachunhatean T.
Factors affecting isoflavone content in soybean seeds grown
in Thailand. Int J Agronomy 2013;2013: Article ID 163573, 11 pages.
Journal of Health Science and Medical Research
126
JHSMR 2018;36(2):117-126