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PYTOESTROGENS
INTRODUCTION:
Phytoestrogens are plant-derived xenoestrogens functioning as the primary female sexhormone
estrogen not generated within the endocrine system but consumed by eating phytoestrogonic
plants. Also called "dietary estrogens", are a diverse group of naturally occurring nonsteroidal
plant compounds that, because of their structural similarity with estradiol (17-β-estradiol), have
the ability to cause estrogenic or/and antiestrogenic effects.
ESTROGEN:
Estrogen is a hormone that comprises a group of compounds, including estrone, estradiol and
estriol. It is the main sex hormone in women and is essential to the menstrual cycle. Estrone is
produced during menopause, estradiol is the predominant form in non-pregnant females and
estriol is the primary estrogen of pregnancy. Although estrogen exists in men as well as women,
it is found in higher amounts in women, especially those capable of reproducing. Estrogen
contributes to the development of secondary sex characteristics, which are the defining
differences between men and women that don’t relate to the reproductive system. In women,
these characteristics include breasts, a widened pelvis, and increased amounts of body fat in the
buttock, thigh and hip region. Estrogen also contributes to the fact that women have less facial
hair and smoother skin then men. Estrogen is an essential part of a woman’s reproductive
process. It regulates the menstrual cycle and prepares the uterus for pregnancy by enriching and
thickening the endometrium. Two hormones, the luteinizing hormone (LH) and the follicle
stimulating hormone (FSH), help to control how the body produces estrogen in women who
ovulate. Estrogen is manufactured mostly in the ovaries, by developing egg follicles. In addition,
estrogen is produced by the corpus luteum in the ovary, as well as by the placenta. The liver,
breasts and adrenal glands may also contribute to estrogen production, although in smaller
quantities. Estrogen can be broken down into three distinct compounds: estrone, estradiol and
estriol. During a woman’s reproductive life, which starts with the onset of menstruation and
continues until menopause, the main type of estrogen produced is estradiol. Enzymatic actions
produce estradiol from androgens. Testosterone contributes to the production of estradiol, while
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the estrogen estrone is made from andostenedione. Estrogen is important to a woman’s health,
not only as it relates to her reproductive cycle. Although it can cause women to retain fluid, and
early exposure through early menses can increase a woman's risk of developing breast cancer,
estrogen has its benefits. It can contribute to increasing high density lipoprotein (HDL), which is
considered the “good” cholesterol, and lower the low density lipoprotein (LDL), which is the
“bad” cholesterol. After menopause, women experience a reduction in estrogen. This can lead to
vaginal dryness, memory problems, hot flashes, fatigue, irritability and possibly one of the most
devastating problems, a decrease in bone density.
BIOSYNTHESIS:
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In females, synthesis of estrogens starts in theca interna cells in the ovary, by the synthesis of
androstenedione from cholesterol. Androstenedione is a substance of moderate androgenic
activity. This compound crosses the basal membrane into the surrounding granulosa cells, where
it is converted to oestrone or oestradiol, either immediately or through testosterone. The
conversion of testosterone to oestradiol, and of androstenedione to oestrone, is catalyzed by the
enzyme aromatase.
Oestradiol levels vary through the menstrual cycle, with levels highest just before ovulation.
MECHANISM OF ACTION:
Estrogens readily diffuse across the cell membrane. Once inside the cell, they bind to and
activate estrogen receptors which in turn modulate the expression of many genes. Additionally,
estrogens have been shown to activate a G protein-coupled receptor, GPR30.
ESTROGEN RECEPTORS:
There are two different forms of estrogen receptors ER alpha(ER-α ) and beta (ER-β).
LOCATION:
ER-α
Uterus, Testis, Pituitary, Ovary, Epidydimis, and adrenal gland.
ER-β
Brain, Kidney, Prostate, Ovary, lung, Bladder, Intestine, and epididymis.
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Benefits and Functions of Estrogen:
Helps maintain memory
Increases sexual interest and joy
Improves mood and happiness
Improves sleep quality
Relieves hot flashes
Increases vaginal secretions
Increases concentration and cognitive abilities
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Enhances energy
Increases reasoning and ideation -- richer language
Prevents joint pains in the fingers, hands and large joints associated with aging
Increases eye (lacrimal) secretions that otherwise lead to dry, itchy eye syndrome
Decreases wrinkles
Stops those tell-tale aging lines of the upper lip
Maintains the amount of collagen and elasticity in the skin
Maintains bone density along with progesterone and testosterone
Increases the water content of skin and is responsible for its thickness and softness
Reduces the overall risk of heart disease by 40 to 50%
Helps with fine motor skills
Helps maintain muscle
Decreases blood pressure -- natural vasodilator
Increases metabolic rate
Reduces risk of cataracts
Protects against macular degeneration
Increases HDL by 10 to 15%
Decreases LDL and prevents its oxidation
Decreases lipoprotein(a) -- Lp(a) -- a highly atherogenic substance
Improves insulin sensitivity -- thereby preventing type II diabetes
Decreases the accumulation of plaque on arteries
Regulates body temperature
Stimulates the production of cholineacetyltransferase, an enzyme which prevents
Alzheimer's Disease
Improves serotonin formation, a anti-depressant brain neurotransmitter, which decreases
depression, irritability, anxiety, and pain sensitivity
Helps maintain the elasticity of arteries
Dilates small arteries
Inhibits platelet adhesiveness -- preventing heart attacks and strokes
Enhances magnesium uptake and utilization
Enhances the production of nerve-growth factor
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Acts as a natural calcium channel blocker to keep arteries open
Reduces homocysteine
Decreases risk of colon cancer
Helps prevent tooth loss
MENOPAUSE:
Menopause is the transition period in a woman’s life when her ovaries stop producing eggs, body
produces less estrogen and progesterone. Menopause is a natural event that normally occurs
between ages of 45 and 55.
PREMATURE MENOPAUSE:
Menopause is more likely to occur at a slight earlier age in women who smoke, have never been
pregnant or live at high altitudes.
PERIMENOPAUSE:
The hormonal changes associated with menopause actually begin prior to the last menstrual
period, during a 3-5 year period called the perimenopause.
SURGICAL MENOPAUSE:
Surgical menopause is menopause induced by the removal of the ovaries. Women who have had
surgical menopause often have a sudden and more severe onset of symptoms of menopause.
As estrogen levels decrease during menopause the following symptoms occur:
Hot flashes
Sleep disturbances
Urogenital atrophy
Osteoporosis
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Skin dryness
Aging
CLINICAL USES OF ESTROGENS:
Oral contraceptives:
Since estrogen circulating in the blood can negatively feed-back to reduce circulating levels of
FSH and LH, most oral contraceptives contain a synthetic estrogen, along with a synthetic
progestin. Even in men, the major hormone involved in LH feedback is estradiol, not
testosterone.
Hormone replacement therapy:
Estrogen and other hormones are given to postmenopausal women in order to prevent
osteoporosis as well as treat the symptoms of menopause such as hot flushes, vaginal dryness,
urinary stress incontinence, chilly sensations, dizziness, fatigue, irritability, and sweating.
Fractures of the spine, wrist, and hips decrease by 50-70% and spinal bone density increases by
~5% in those women treated with estrogen within 3 years of the onset of menopause and for 5–
10 years thereafter.
Before the specific dangers of conjugated equine estrogens were well understood, standard
therapy was 0.625 mg/day of conjugated equine estrogens (such as Premarin). There are,
however, risks associated with conjugated equine estrogen therapy. Among the older
postmenopausal women studied as part of the Women's Health Initiative (WHI), an orally
administered conjugated equine estrogen supplement was found to be associated with an
increased risk of dangerous blood clotting. The WHI studies used one type of estrogen
supplement, a high oral dose of conjugated equine estrogens (Premarin alone and with
medroxyprogesterone acetate as PremPro).
In a study by the NIH, esterified estrogens were not proven to pose the same risks to health as
conjugated equine estrogens. Hormone replacement therapy has favorable effects on serum
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cholesterol levels, and when initiated immediately upon menopause may reduce the incidence of
cardiovascular disease, although this hypothesis has yet to be tested in randomized trials.
Estrogen appears to have a protector affect on atherosclerosis it lowers LDL and triglycerides, it
raises HDL levels and has endothelial vasodilatation properties plus an anti-inflammatory
component.
Research is underway to determine if risks of estrogen supplement use are the same for all
methods of delivery. In particular, estrogen applied topically may have a different spectrum of
side-effects than when administered orally, and transdermal estrogens do not affect clotting as
they are absorbed directly into the systemic circulation, avoiding first-pass metabolism in the
liver. This route of administration is thus preferred in women with a history of thrombo-embolic
disease.
Estrogen is also used in the therapy of vaginal atrophy, hypoestrogenism (as a result of
hypogonadism, castration, or primary ovarian failure), amenorrhea, dysmenorrhea, and
oligomenorrhea. Estrogens can also be used to suppress lactation after child birth.
Breast cancer
About 80% of breast cancers, once established, rely on supplies of the hormone estrogen to
grow: they are known as hormone-sensitive or hormone-receptor-positive cancers. Suppression
of production of estrogen in the body is a treatment for these cancers.
Hormone-receptor-positive breast cancers are treated with drugs which suppress production of
estrogen in the body. This technique, in the context of treatment of breast cancer, is known
variously as hormonal therapy, hormone therapy, or anti-estrogen therapy (not to be confused
with hormone replacement therapy). Certain foods such as soy may also suppress the
proliferative effects of estrogen and are used as an alternative to hormone therapy.
Prostate cancer
Under certain circumstances, estrogen may also be used in males for treatment of prostate
cancer.
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Miscellaneous
In humans and mice, estrogen promotes wound healing.
At one time, estrogen was used to induce growth attenuation in tall girls. Recently, estrogen-
induced growth attenuation was used as part of the controversial Ashley Treatment to keep a
developmentally disabled girl from growing to adult size.
Most recently, estrogen has been used in experimental research as a way to treat patients
suffering from bulimia nervosa, in addition to Cognitive Behavioral Therapy, which is the
established standard for treatment in bulimia cases. The estrogen research hypothesizes that the
disease may be linked to a hormonal imbalance in the brain.
Estrogen has also been used in studies which indicate that it may be an effective drug for use in
the treatment of traumatic liver injury.
ESTROGEN PREPARATIONS:
1. Ethinyl Estradiol
1. Hormone Replacement: 0.005 to 0.02 mg/day
2. Oral Contraceptive: 0.03 to 0.10 mg/day
2. Micronized estradiol (1-2 mg/day)
3. Estradiol Cypionate (2-5 mg every 3-4 weeks)
4. Estradiol valerate (2-20 mg every other week)
5. Estropipate (1.25-2.5 mg/day)
6. Conjugated Estrogen (e.g. Premarin)
1. Oral (0.3-1.25 mg/day)
2. Injectable (0.2-2 mg/day)
3. Transdermal
7. Diethylstilbestrol (0.1-0.5 mg/day)
8. Quinestrol (0.1-0.2 mg/week)
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9. Chlorotrianisene (12-25 mg/day)
10. Methallenestril (3-9 mg/day)
11. Esterified Estrogen (Menest): 0.3 to 1.25 mg/day
Estrogen replacement therapy has been widely used by women in menopause. It provides relief
from acute menopausal symptoms and in the later stage, it prevents complications like
osteoporosis. But long term use of estrogens may produce the following side effects:
Women can experience side effects during hormone therapy; these can be divided into more
minor side effects, and more serious side effects. The more minor side effects are more common
than the serious side effects, and are generally perceived by women as "annoying." These
symptoms include:
Nausea, vomiting
Pain, cramps or tenderness in the abdomen
Yellowing of the skin and/or whites of the eyes
Breast tenderness or enlargement
Enlargement of benign (fibroid) tumors
Irregular bleeding or spotting
Change in amount of cervical secretion
Vaginal yeast infections
Retention of excess fluid. (This may make some conditions worsen, such as asthma,
epilepsy, migraine, heart disease, or kidney disease)
A spotty darkening of the skin, particularly on the face
Reddening of the skin
Skin rashes
Worsening of porphyria
Headache, migraines
Dizziness, faintness
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Changes in vision (including intolerance to contact lenses)
Mental depression
Involuntary muscle spasms
Hair loss or abnormal hairiness
Changes in weight. (Most women experience weight gain instead of weight loss)
Changes in sex drive. (Most women experience decreased libido)
Possible changes in blood sugar
Shortness of breath
Coughing up blood
Severe stomach pain or swelling
Acne
Allergies
Fatigue
High blood pressure
Gallstones
Loss of libido
Asthma
Numbness, tingling, itching of feet and hands
Heaviness of the chest
It is still controversial which of these side effects are due to the estrogen component as compared
to the progesterone component. Therefore, if side effects persist for a few months, the doctor will
often alter either the progesterone or the estrogen part of the hormone therapy (HT).
Contrary to common belief, recent research has confirmed that women who take commonly
prescribed doses of hormone therapy (HT) are no more likely to gain weight than women not
taking hormone therapy (HT). This is probably because menopause or aging itself is associated
with weight gain, regardless of hormone therapy.
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The more serious health concerns for women undergoing hormone therapy (HT) include:
Hormone therapy (HT) increases the risk of vein clots in the legs (deep vein thrombosis)
and blood clots in the lungs (pulmonary embolus) by about 2 or 3 fold. However, it is
important to remember that these conditions are extremely rare in healthy women. Thus,
the true increase in risk for healthy women is minimal. Women with a personal or family
history of these clots should review this issue when considering hormone therapy (HT).
Uterine Cancer (endometrial cancer): Research shows that women who have their
uterus and use estrogen alone are at risk for endometrial cancer. Today, however, most
doctors prescribe the combination of estrogen and progestin. Women without a uterus
(including women who have had a hysterectomy) have no risk of endometrial cancer.
Breast cancer: Recent research indicates that hormone therapy (HT), and especially
EPT, increases the risk of breast cancer, although the increase in risk is very small. For
example, the Women's Health Initiative, a reliable large study of hormone therapy (HT)
in menopausal women, predicted that there were approximately 8 extra cases per 10,000
women who took hormone therapy (HT) for 1 year, compared to women taking a placebo
pill. The increase in risk of breast cancer associated with hormone therapy (HT) likely
increases with duration of use and is especially increased with 5 or more years of use.
Heart disease: Even though hormone therapy (HT) lowers the bad LDL cholesterol and
raises the good HDL cholesterol, hormone therapy (HT) increases the risk of heart attacks
in women who already have heart disease, as well as in women who do not have known
heart disease. Hormone therapy (HT) does not prevent heart attack based on recent
research from the Women's Health Initiative.
Abnormal vaginal bleeding: Women on hormone therapy (HT) are more likely than
other postmenopausal women to experience abnormal vaginal bleeding. What is called
"abnormal bleeding" depends on the type of hormone therapy (HT). With cyclic therapy,
in which 5monthly bleeding is expected, bleeding is abnormal if it occurs when it is not
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expected or is excessively heavy or long in duration. With daily continuous therapy,
irregular bleeding can last for 6 months to a year, therefore, irregular bleeding that lasts
for more than a year is considered abnormal.
Stroke: Hormone therapy (HT) slightly increased the risk of stroke in women Because of
the possibility of increased breast cancer, stroke, and heart disease risks, women who
have no major menopause symptoms may choose to avoid hormone therapy (HT).
These adverse effects diverted the attention from conventional estrogens and increased
the interest in phytoestrogens. The result of Women’s health initiative demonstrated that
many of the ill-effects of conventional estrogen therapy are due to the late age (63 years)
of women taking this treatment. The great promise of phytoestrogen use is that they are
natural and basically derived from food products; they can be successfully used in all
stages of menopause, without any serious harmful effects reported so far. However, the
efficacy of conventional estrogens in treating acute vasomotor and psychometric
symptoms is much superior to that of the weaker phyoestrogens. The rule of the thumb is
to use conventional estrogen for the alleviation of greater efficacy. Conversely,
phytoestrogens should be used for long-term prevention as they are slow-acting, but safer
on extended use.
PHYTOESTROGENS:
Phytoestrogens are plant-derived xenoestrogens functioning as the primary female sexhormone
estrogen not generated within the endocrine system but consumed by eating phytoestrogonic
plants. Also called "dietary estrogens", are a diverse group of naturally occurring nonsteroidal
plant compounds that, because of their structural similarity with estradiol (17-β-estradiol), have
the ability to cause estrogenic or/and antiestrogenic effects.
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ADVANTAGES OVER ESTROGENS:
These substances are estrogenic compounds which occur naturally in plants such as soy, black
cohosh, dong quai, licorice root, etc. In countries where large amounts of phyto-estrogens are
consumed, women suffer very few menopausal symptoms. Phyto-estrogens differ remarkably
from synthetic estrogens in that they are easily broken down, are not stored in the tissues and
spend very little time in the body, thus they are unlikely to cause cancer. And because humans
have spent millions of years eating plants and herbs, our bodies easily accept plant estrogens and
find them both effective and beneficial.
Phyto-estrogens appear to exert a natural balancing action in the body. If estrogen is low, phyto-
estrogens will increase estrogen activity in the body. When estrogen levels are high, phyto-
estrogens will compete for estrogen receptor sites, causing a decrease in the estrogen effects.
The result is a chemo protecting action. Numerous studies have indicated that high intake of
phyto-estrogenic substances may reduce the risk of breast cancer.
The US National Institutes of Health announced. In fact, years-long use of synthetic estrogens
and progestin increased otherwise healthy women's risk of a stroke by 41 percent, a heart attack
by 29 percent and breast cancer by 24 percent
STRUCTURE:
Phytoestrogens mainly belong to a large group of substituted natural phenolic compounds: the
coumestans, prenylated flavonoids and isoflavones are three of the most active in estrogenic
effects in this class. The best-researched are isoflavones, which are commonly found in soy and
red clover. Lignans have also been identified as phytoestrogens, although they are not
flavonoids.
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FOOD SOURCES:
Phytoestrogen can be also found in wide variety of food products (including herbs), even though
the level varies depending on the source. Below is a short list of phytoestrogen food sources.
Table 1. Foods high in phytoestrogen content.
Phytoestrogen food
sources
Phytoestrogen content
(µg/100g)
Flax seed 379380
Soy beans 103920
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Tofu 27150.1
Soy yogurt 10275
Sesame seed 8008.1
Flax bread 7540
Multigrain bread 4798.7
Soy milk 2957.2
Hummus 993
Garlic 603.6
Mung bean sprouts 495.1
Dried apricots 444.5
Alfalfa sprouts 441.4
Dried dates 329.5
Sunflower seed 216
Chestnuts 210.2
Olive oil 180.7
Almonds 131.1
Green bean 105.8
Peanuts 34.5
Onion 32
Blueberry 17.5
Corn 9
Coffee, regular 6.3
Watermelon 2.9
Milk, cow 1.2
Total phytoestrogen and lignin content in vegetables, fruits, nuts and drinks.
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Food items Lignan content
(µg/100g)
Total phytoestrogen
(µg/100g)
Vegetables
Soy bean sprouts 2.2 789.6
Garlic 583.2 603.6
Winter squash 113.3 113.7
Green beans 66.8 105.8
Collards 97.8 101.3
Broccoli 93.9 94.1
Cabbage 79.1 80
Fruits
Dried prunes 177.5 183.5
Peaches 61.8 64.5
Strawberry 48.9 51.6
Raspberry 37.7 47.6
Watermelon 2.9 2.9
Nuts and other legume seeds
Pistachios 198.9 382.5
Chestnuts 186.6 210.2
Walnuts 85.7 139.5
Cashews 99.4 121.9
Hazel nuts 77.1 107.5
Lentils 26.6 36.5
Beverages
Wine, red 37.3 53.9
Tea, green 12 13
Wine, white 8 12.7
Tea, black 8.1 8.9
Coffee, decaf 4.8 5.5
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Beer 1.1 2.7
Other
Black bean
source10.5 5330.3
Black licorice 415.1 862.7
Bread, rye 142.9 146.3
Epidomological studies give evidence that Asian women experience milder menopausal
symptoms and this because of high intake of soy products in the Asian diet.
Soy foods and flax seed have the highest amount of phytoestrogens however all pant foods
contain some.
EXTRACTION OF PHYTOESTROGENS FROM FOODS:
Several methods for the extraction of phytoestrogens, especially isoflavones, have been
proposed. Some methods are used to determine the various conjugated forms of phytoestrogens,
whereas others are used to determine total phytoestrogens within foods. To determine the
individual phytoestrogen composition in foods, extractions are done without a hydrolysis step.
To determine total phytoestrogen content, enzymatic hydrolytic with Aspergillums nigercellulase
is carried out.
Acidified Solvent Extraction of Phytoestrogens:
A general procedure that is suitable for the extraction of phytoestrogens. Raw soybean seeds (2
g) with their seed coats are ground, mixed with 2 ml 0.1N HCl and 10 ml of acetonitrile, stirred
for 2 h at room temperature, and filtered through Whatman No. 42 filter paper. The filtrate is
taken to dryness under vacuum at a temperature below 300c. The dried material is redissolved in
10 ml of 80% aqueous methanol and then filtered through a 0.45m filter unit. An aliquot of the
filtrate (20μ l) is analyzed by reversed phase HPLC. Although this extraction method preserves
the isoflavone conjugates, degradation of the O-malonylglycosides in aqueous methanol occur
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seven at room temperature. Coward et al. (1998) have shown that extraction of isoflavones at 40c,
where degradation is minimized, is possible if an internal standard such as fluorescein is used. In
this latter method, the extract is not concentrated prior to HLPC analysis, but instead analyzed
directly. Flavones and p-hydroxybenzoic acid have also been used as internal standards.
Solvent Extraction of Phytoestrogens:
Isoflavones in solid foods are extracted into 80% aqueous methanol (10 ml/g) by stirring for 1 h
at 600c. Other soy products (miso, soy milk, soy paste, and tofu) are freeze-dried and extracted
whole. The mixture is centrifuged at 2500g for 10 min and the supernatant is transferred to a
round bottom flask. The pellet is extracted twice more in 5 ml each and centrifuged.
Supernatants are combined in the round-bottom flask and evaporated to dryness in a rotary
evaporator. The dried extracts are resolubilized in 5 ml of 50% aqueous methanol and then
defatted by partitioning the neutral lipids into hexane (4 20 ml). The aqueous methanol is taken
to dryness in a rotary evaporator, and the resulting dried residue is resolubilized in 10 ml 80%
aqueous methanol. An aliquot is centrifuged at 14,000gfor 2 min and analyzed by HPLC as
described by Coward et al. (1993).While Coward et al. (1993) used aqueous methanol, Griffith
and Collison (2001) used acetonitrile for isoflavone extraction: soy foods (1 g) are dispersed into
a 10ml volume of acetonitrile followed by the addition of 6 ml double de ionized water and 0.5
ml apigenin [2000 g/ml in dimethylsulfoxide (DMSO)]. Samples are shaken to mix and extracted
on a rotary mixer for 2 h, after which the sample is recovered and the acetonitrile concentration
is adjusted to 50% (v/v) with the addition of deionized water. Samples are centrifuged at 2000g
for 10 min to pellet insoluble matter and eliminate foam. An aliquot is filtered through a 0.45 μ
m PVDF filter and analyzed by HPLC.
Extraction Followed by Enzymatic Hydrolysis :
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Phytoestrogen-containing materials (2.5 g) are mixed with a minimum of 5 ml of 80% aqueous
methanol. If the material under study is particularly enriched in phytoestrogens, then the amount
taken for analysis should be reduced, as phytoestrogens have a limited solubility in aqueous
methanol, which would thereby lead to an erroneously low result. Sonication for10 min
solubilizes the phytoestrogen glycosides in the methanol by breaking up cellular material. It is
followed by a further hour of soaking in the solvent. Insoluble material is removed by filtration
through a double layer of filter paper (Whatman No. 4 and then No. 1), and any adsorbed
phytoestrogens are washed through with fresh 80% aqueous methanol (>5ml). The methanol in
the filtrate is evaporated, and 100 Fishman units of cellulase from A. Niger are added to the
sample in 5 ml of 0.1 M sodium acetate buffer, pH 5. Samples are sonicated and subsequently
incubated overnight in a shaking water bath at 37. The hydrolyzed phytoestrogen aglycones
usually precipitate because of their reduced aqueous solubility. They are extracted from the
aqueous hydrolyzes by the addition of 100%ethyl acetate. Three 2 μ ml extracts with ethyl
acetate are combined; a 1 μ ml aliquot of the combined extract is pipetted into a separate vial and
taken to dryness at 600c under nitrogen prior to chromatographic analysis of the phytoestrogen
aglycones.
Extraction of Lignans:
Milder et al. (2004) developed an assay to extract lignans from a variety of foods, including
flaxseed, broccoli, whole wheat bread, and tea. Previous extraction methods allowed for the
quantitation of the major lignans secoisolarici resinol and matairesinol but were lacking for
pinoresinol and lariciresinol. To allow for quantification, the lignans need to be extracted from
their food matrices. This can be difficult due to their ability to oligomerize with 3-hydroxy-3-
methylglutaric acid through ester bonding. Therefore, lignan extractions require the use of
alkaline hydrolysis to release the lignan glycosides from the 3-hydroxy-3-methylglutaric acid.
There are many unknown lignan glycosides; therefore, lignan extraction methods typically have
a hydrolysis step to allow for the release of the glycosides. This results in an estimate of the total
lignan content in foods. The following describes the current extraction and analysis methods for
lignans from foods. Alkaline extraction of 1.0 g of dry food is performed with 24.0 ml of
methanol/water (70/30, v/v) containing 0.3Msodium hydroxide in a shaking water bath for 1 h at
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150 rpm and 600c. After extraction, the pH is adjusted to 5–6 with 750 μ l of 100% glacial acetic
acid and the extract is centrifuged at 4,500g for 10 min at 100c.An aliquot (1 ml) is transferred to
a pre weighed test-tube. Methanol is evaporated from this aliquot at 600c, under nitrogen, until
the residual weight is< 0.30 g. The volume is adjusted to approximately 1.2ml with 0.05 M
sodium acetate buffer, pH 5, and the extract is weighed again to calculate the dilution compared
to the original aliquot of 1 ml. A 1-ml aliquot of the weighed extract or 1 ml of beverage is
hydrolyzed by the addition of Helix pomatia β glucuronidase/sulfatase (0.83 mg in 1 ml0.05 M
sodium acetate buffer, pH 5). The samples are incubated overnight at 370c. Samples are extracted
twice with 2 ml diethyl ether, and the two organic phases are combined. The diethyl ether is
evaporated, and the dried samples are redissolved in 0.3 ml of methanol, mixed, and 0.7 ml water
is added. A 240- μ l aliquot of sample is added to 10 μ l of internal standard solution containing
50 ng of secoisolariciresinol-d8 and 50 ng of matairesinol-d6 in 30% aqueous methanol. Samples
are mixed and transferred to HPLC vials for LC-MRM-MS analysis.
MECHANISM OF ACTION:
Phytoestrogens exert their effects primarily through binding to estrogen receptors (ER) .There
are two variants of the estrogen receptor, alpha (ER-α) and beta (ER-β) and many phytoestrogens
display somewhat higher affinity for ER-β compared to ER-α.
The key structural elements that enable phytoestrogens to bind with high affinity to estrogen
receptors and display estradiol-like effects are:
The phenolic ring that is indispensable for binding to estrogen receptor
The ring of isoflavones mimicking a ring of estrogens at the receptors binding site
Low molecular weight similar to estrogens (MW=272)
Distance between two hydroxyl groups at the isoflavones nucleus similar to that
occurring in estradiol
Optimal hydroxylation pattern
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In addition to interaction with ERs, phytoestrogens may also modulate the concentration of
endogenous estrogens by binding or inactivating some enzymes, and may affect the
bioavailability of sex hormones by binding or stimulating the synthesis of sex hormone binding
globulin (SHBG).
Emerging evidence shows that some phytoestrogens bind to and transactivate peroxisome
proliferator-activated receptors (PPARs). In vitro studies show an activation of PPARs at
concentrations above 1 μM, which is higher than the activation level of ERs. At the
concentration below 1 μM, activation of ERs may play a dominate role. At higher concentrations
(>1 μM), both ERs and PPARs are activated. Studies have shown that both ERs and PPARs
influence each other and therefore induce differential effects in a dose-dependent way. The final
biological effects of genistein are determined by the balance among these pleiotrophic actions.
METABOLISM:
The two main isoflavonoids (genistien and diadzen) are present in all soy bean foods either as
aglycone or as beta-glycoside(conjugated). The malonyl and acetyl glycosides are also present,
but they are susceptible to heat and are readily converted into more stable beta-glycoside. These
glycosides are readily hydrolysed to estrogenically active aglycones either as result of processing
and preparation of soy food or as result of the metabolism by intestinal microflora. Genistien is
further metabolized in gut to p-ethyl phenol, diadzein to equol, dihydrodiadzien to o-
methylangolensin.
The diadzein metabolite equol may exert significant biological effects, having greater antioxidant
activity than other phytoestrogens, and is often found in highest concentration in the blood and
urine in humans. Further metabolism of isoflavonoids takes place by conjugating the aglycone
with glucouronic acid and to lesser extent sulfuric acid. Coumestol is found in medicago sativa,
clovers and in low levels in beans and peas. It is least studied for biological activity and
metabolism. Linseed(flax seed) is a rich source of lignans. The two precursors of lignans namely
secoisolariciresinol and matairesinol occur in the aleuronic layer of the grain close to the fibre
layer. After fermentation by colonic microflora, two diphenols namely enterodiol and
enterolactone are formed, which structurally similar to estradiol.
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After absorption these are excreted in urine. Urinary isoflavonoid excretions range from about
0.3-30 μM/day. Though phytoestrogens can be measured in urine , plasma, feces, semen, bile,
saliva, and breast milk, the concentration of their metabolites differ widely among individuals
even after administration of controlled quantity of food supplements. This is due to gut
microflora, antibiotic use, gender difference and concamitent dietary intake. Urinary secretions
of vegetarians may contain 1000 times higher phytoestrogens than total urinary steroid estrogens.
CLINICAL USES:
Phytoestrogen supplements have a number of uses which mainly focus on increasing estrogen
levels and balancing hormone levels within the body.
Menopause :
Menopause can cause a number of uncomfortable side effects and there are a range of treatments
available for women during this time. Hormone replacement therapy (HRT) and Estrogen
replacement therapy (ERT) can both relieve menopausal symptoms and are recommended for
women who are post menopausal. As well as reducing hot flashes and mood swings, both HRT
and ERT can also help prevent cardiovascular disease and osteoporosis.
As both treatments include prescription estrogens, many people view them as being unnatural
and therefore potentially dangerous. Phytoestrogen supplements are currently being used as a
natural alternative and are proving to be extremely popular among menopausal women.
Osteoporosis :
Osteoporosis is a common condition which affects people as they mature and the result is
painful and debilitating swelling of the joints. Low estrogen levels are thought to contribute to
osteoporosis as post menopausal women often see their bones weaken. Phytoestrogen for
osteoporosis can be an effective treatment.
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Phytoestrogens Reduce Risk for Stroke:
Stroke is the third most common cause of death for middle-aged and older women. A 50-year-
old woman has about a one in five chance of suffering a stroke in her remaining lifetime. One of
the most common causes of stroke is blockage due to atherosclerosis of the carotid arteries in the
neck the arteries responsible for a major portion of the blood flow to the brain. Phytoestrogens
have been shown to improve a women's lipid profile to prevent strokes, similar to Premarin
without the harmful side-effects associated with such drugs.
Phytoestrogens Reduce Risk for Cancer :
In a study published in Environmental Health Perspectives, researchers reported that, in addition
to their benefits for the glandular system, phytoestrogens have been shown to have antioxidant
activity and can influence intracellular enzymes, protein synthesis, growth factor action, and cell
proliferation in a way that makes them "strong candidates for a role as natural cancer-protective
compounds."
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PHYTOESTROGEN PRODUCTS:
Emerita PhytoEstrogen Cream:
Emerita phytoestrogen cream is a remedy for:
Hot Flashes
Night Sweats
Mood Swings
Vaginal Dryness
Urinary Incontinence
Skin Conditions
Low Libido
Hair Loss
Sleeping Difficulties
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Instructions :
Massage 1/4 to 1/2 teaspoon of Emerita phytoestrogen cream into skin.
Suggested application sites of Emerita phytoestrogen cream include the palms, inner
arms, abdomen, chest, insides of the thighs, and backs of the knees.
Emerita Phytoestrogen Cream may be used daily. Individual needs may vary.
Cautions :
Emerita phytoestrogen cream is not recommended when pregnant or breastfeeding unless
your health care provider specifically recommends it.
PHYTOESTROGEN PLUS ( capsules):
Contains plant source ingredients with known phytoestrogen activity, including black cohosh,
chaste berry, dong quai, red clover, and soy isoflavones. All these plant source ingredients are in
the form of standardized extracts in order to guarantee potency. The ingredients are contained in
a preservative-free gelatin capsule.
Phytoestrogen Plus also contains gamma oryzanol (a substance derived from rice bran that has
the proven ability to reduce hot flashes) and Vitamin E, providing a broader range of nutrients
that can support body function.
Phytoestrogen Plus should be used primarily to support women's health during perimenopause
and menopause, and also may be helpful for women experiencing post menopausal symptoms.
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Chattem - Rejuvex (Disc), 30 caplets:
The unique Rejuvex formula combines dong quai (a Chinese herb), magnesium, boron and
glandular powders to support a woman's health during and after the menopause years. These
ingredients, combined with synergistic levels of other vitamins and minerals, provide a woman
with nutrients she needs for a comfortable menopause and continuation of a full, active life.
Healthy bones
Rejuvex supplies 500mg of magnesium - essential for the support of strong healthy bones.
PHYTOESTROGEN &IPRIFLAVONES:
PhytoEstrogen & Ipriflavones Dosage:
Use only as directed. Take two capsules daily with a meal or glass of water. Store in a cool, dry
place.
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PhytoEstrogen & Ipriflavones Caution:
Do not use if safety seal is broken or missing. Keep out of reach of children.
ESTROSOY REPLENISH LIFT
Gives forty-plus skin a new lease on life. Phytoestrogen works to combat the effects of hormonal
changes. Brings back a youthful firmness, resilience and smoothness.
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SIDE EFFECTS OF USING PHYTOESTROGEN:
The side effects of phytoestrogens differ depending on the plant from which the chemical is
sourced. Soy, for instance, may have different phytoestrogenic side effects to black cohosh
Phytoestrogen acne: is one possible side effect and although not life threatening, it can be
embarrassing and uncomfortable.
Soy intoxication: If a person starts to eat soy beans as a way of introducing phytoestrogens into
their diet it important to be aware that the plant chemicals in these beans can be toxic and act as
anti-nutrients. The beans must be cooked slowly or fermented to ensure that these harmful
chemicals are not ingested.
Lost of Zinc absorption: Phytate, found in phytoestrogen, affects the body by binding to Zinc to
estrogen receptors. This can cause Zinc absorption in the body to decrease, potentially damaging
cell functions in the bones, skin, muscles, eyes, and kidneys.
Phytochemicals found in soy: Some phytochemicals slow down the enzymes that digest
protein into simple building blocks of amino acids, which does not occur in phytoestrogen-free
products.
Other hormone medication: Such as the contraceptive pill or HRT, should not be taken
alongside phytoestrogens. Side effects may include these treatments becoming ineffective.
Genistein inhibits an enzyme called GLUT-1: This enzyme transports glucose used in brain
cells and red blood cell.
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CONCLUSION
Because of the adverse effects caused by harmone replacement therapy now a days there is
diversion in attention from conventional estrogens and increase in interest in phytoestrogens. The
great promise of phytoestrogen use is that they are natural and basically derived from food
products; they can be successfully used in all stages of menopause, without any serious harmful
effects. Today more and more people are turning to these naturally-occurring compounds as an
alternative to man-made and potentially dangerous hormone replacement treatments.
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REFRENCES
WWW.WIKIPEDIA.ORG
WWW.MENOPAUSE-METAMORPHOSIS.COM
WWW.DIETARYFIBERFOOD.COM
WWW.JKSCIENCE.ORG
WWW.IJP-ONLINE.COM
WWW.NCBI.NLM.NIH.GOV
WWW.TOTALITYOFBEING.COM
WWW.INTEGRATEDHEALTH.COM
WWW.HERBS-FOR-MENOPA USE .COM
WWW.MENOPA USE RX.COM
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