Obreshkova Tribestan 2014

PHARMACIA, vol. 61, No. 3/2014 17Investigation of some food additives containing ...
INVESTIGATION OF SOME FOOD ADDITIVES CONTAINING
COMPOUNDS WIHT ANDROGENIC ACTIVITY AND THEIR ANALYTI-
CAL STUDY
St. Ivanova1
, D. Obreshkova2
, P. Atanasov3
, K. Ivanov1
, B. Zlatkov1
1
Medical University – Plovdiv, Faculty of Pharmacy
2
Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Medical University – Sofia
3
Clinic of THKI, UMHATEM „N.I.Pirogov”– Sofia
4
Faculty of Pharmacy, Medical University – Plovdiv
Abstract. The market of food additives is growing. There is a big demand of food supplements with andro-
genic compounds from extracts of Tribulus terrestris Linn., Rhaponticum carthamoides L. or tree Yohimbe.
Food additives with androgenic effects very often contain steroidal saponins, ecdysteroids, Yohimbine, De-
hydroepiandrosterone (DHEA) or combination of these substances. The steroids are forbidden substances
in food supplements. Recent studies show that non – hormonal additives such as vitamins, minerals and
aminoacids can contain a not declared on the labels of the products anabolic androgenic steroids: pro-
hormones of Testosterone or 19-Nortestosterone. These undeclared substances can cause health risks to
consumers and might lead to positive results in sports doping control, especially for Nandrolone metabolite
Norandrosterone. This paper reviews investigation of some food additives, containing compounds with
androgenic activity like extracts from Tribulus terrestris Linn., ecdysteroids, Yohimbine and DHEA and the
most used methods for their analysis.
Key words: additives, androgenic effects, Tribulus, analysis.
Introduction.
The consumption of food supplements and their
importance in people’s life are constantly increasing
[1]. This review is focused on some food additives
containing compounds with androgenic activity and
the applied methods for their analysis. In accordance
with the European Community Directive of 2002
(Directive 2002/46/EC) and Bulgarian legislation, is-
sued by the Ministry of Health 01.08.2005, food ad-
ditives are defined as substances, containing concen-
trated nutrients or other elements with a nutritional or
physiological effect, alone or in combination, distrib-
uted in certain dosage forms designed to enrich the
diet. The fact that food supplements are from plant
or animal origin, does not make them safe. The food
additives are tested for efficiency, which is support-
ed by scientific evidence, but in some supplements
the qualitative and quantitative composition of their
components is not declared [2].
The steroids are forbidden substances in food ad-
ditives. In 2009 and 2010, dietary supplements con-
taining the prohibited growth hormone – releasing
peptide – 2 are detected [3]. Some authors offer ana-
lytical strategies for the detection of non – labelled
anabolic androgenic steroids in nutritional supple-
ments. Recent studies showed that non – hormonal
supplements such as vitamins, minerals and amino
acids can contain undeclared anabolic androgenic
steroids, which can cause health risks to consumers
[4]. Food additives containing extracts from Tribulus
terrestris Linn., ecdysteroids, Yohimbine and DHEA
possess androgenic effect.
I. Food additives with extract from Tribulus ter-
restris Linn.
Extracts from Tribulus terrestris Linn. possess
antibacterial [5, 6], antifungal [5, 7], antihelmintic
(Tribulosin and β-sitosterol-D-glucoside) [8], analge-
sic [9], antihypertensive [10], diuretic [11], hypogly-
cemic [12], hypolipidemic [13] and cytotoxic (Meth-
ylprotodioscin) [14] activity and are applied for skin
care [15] and human hormones regulation [16, 17].

18 PHARMACIA, vol. 61, No. 3/2014 St. Ivanova, D. Obreshkova, P. Atanasov, K. Ivanov, B. Zlatkov
Active compounds in extracts from Tribulus ter-
restris Linn. are furostanol [18, 19] saponins and oth-
er steroidal saponins: Terrestrinins A and B [20], Ter-
restrosins A, B, C, D, E [21], Tribulosaponin B [22],
lignanamides (Tribulusamides A and B), Tribulosap-
onin B (Fig. 1), alkaloids, flavonoids rutin, quercetin
and kaempferol [23]. In the Bulgarian plant extracts,
the main components are Protodioscin, Dioscin (Fig.
1.), Metylprotodioscin, Prototribestin [24]. The first
standardized preparation is the bulgarian ”Tribestan”
(Sopharma). More products were developed lately as
preparations or food supplements and are used for
impotence disorders. The product UNEX is proposed
as diuretic [11].
In Table 1. are presented food supplements with
extracts from Tribulus terrestris Linn.
II. Food supplements with ecdysteroids.
Food supplements with ecdysteroids are consid-
ered to possess androgenic effect. Recent data shows
that in plants over 250 ecdysteroid analogs have been
identified. Rich of ecdysteroids is the plant Rhapon-
ticum carthamoides (Maral root), growing wild in
Southern Siberia, Kazakhstan, Altay, Eastern Eu-
rope. Animal studies indicate that Maral root may
Protodioscin Dioscin
Fig 1. Chemical structures of Protodioscin and Dioscin.
Table 1. Food additives with extracts from Tribulus ter-
restris Linn.
1. Bali Mojo caps. Not labelled
2.
Fountain Of Youth HGH
Complete,700 g powder
Not labelled
3 Irexis caps. Not labelled
4 Libilov (USA), 90 caps. Not labelled
5 Natrolex, 60 caps. Not labelled
6.
Natural-T (Top Secret Nutrition),
90 caps.
Not labelled
7. Virectin, 90 caps. Not labelled
8.
Test Booster (Blade Nutrition),
60 caps.
100 mg
9 Anotesten (Muscletech) 250 mg
10 Tribex (Biotest), 60 caps. 250 mg
11 Tribestan (Sopharma), 60 tabl. 250 mg
12 Tribosten (Thermolife), 60 tabl. 250 mg
13. MuscleTech TEST HD, 90 caps. 250 mg
14. Everlastin, 30 caps. 300 mg
15. Testogenix, 30 caps. 300 mg
16. Testosyn, 60 caps. 300 mg
17. TribuPlex (MRM), 60 caps. 400 mg
18. Tribestan (Sopharma), 60 tabl. 500 mg
19 Tribulus (Now), 100 caps. 500 mg
20 Trioxalon 500 (AST), caps. 500 mg
21.
Tropinol XP (iForce Nutrition),
100 caps.
563 mg
22. Tribulus (Optimum), 50 caps. 625 mg
23.
Tribulus Pro (Universal Nutrition),
100 caps.
625 mg
24. TribX (All Max Nutrition), 90 caps. 750 mg
25. Tribulus (MET-Rx), 90 caps. 750 mg
26.
Tribulus 2400 (iForce Nutrition),
90 caps.
800 mg
27. Tribulus (Now), 180 tabl. 1000 mg
28. Tribuvar, (S.A.N), 90 tabl. 1000 mg
29. Tribulus Terrestris, 100 caps. 1250 mg
have a beneficial effect on memory and learning in
rats [25], increases working capacity of skeletal mus-
cles and possesses anabolic and adaptogenic effects
in rats [26]. Ecdysteroids include insect moulting
hormones, which regulate insect development, and
similar chemical structures widely spread through-
out plants and affecting various biological functions
[27]. 20-Hydroxyecdysone is derivative of Ecdysone
(Fig. 2.) and is the most common naturally occur-
ring ecdysteroid, which controls the ecdysis (moult-
ing) and metamorphosis of arthropods. It is also a

Fig 2. Chemical structures of Ecdysone and 20-Hydroxy-
ecdysone. Ecdysone 20-Hydroxyecdysone
Table 2. Food additives with 20-Hydroxyecdysterone.
Fig 3. Enzymic interconversions of 3β-ecdysteroids, 3-dehydroecdysteroids and 3α- ecdysteroids.
phytoecdysteroid produced by various plants, includ-
ing Cyanotis vaga, where its purpose is presumably
to disrupt the development and reproduction of in-
sects. In arthropods the compound acts through the
ecdysone receptor. Although in mammals this recep-
tor is nor included, 20-Hydroxyecdysone may affect
mammalian biological systems. 20-Hydroxyecdysone
is an ingredient of some bodybuilding nutritional sup-
plements, which aim to enhance physical performance
[28], strength or athletic improvement [29].
On Fig. 3 is illustrated the enzymic pathway
for transformation of 3-dehydroecdysteroids to
3β-ecdysteroids in presence of 3-dehydroecdysteroid-
3β-reductase and the formation of 3α-ecdysteroids by
influence of 3-dehydroecdysteroids 3α-reductase.
Food supplements with 20-Hydroxyecdysterone
are summarized on Table 2.
III. Food supplements containing Yohimbine.
1. Effects of Yohimbine.
Yohimbine is a mild monoaminooxidase inhibitor
with stimulant and aphrodisiac effects and blocks the
presynaptic and postsynaptic α2 receptors. Blockade of
postsynaptic α2 receptors causes minor corpus caverno-
sum smooth muscle relaxation. The majority of adreno-
ceptors in the corpus cavernosum are of the α1 type.
Blockade of pre – synaptic α2 receptors facilitates the
release of several neurotransmitters in the corpus caver-
nosum – nitric oxide and norepinephrine.Whereas nitric
oxide released in the corpus cavernosum is the major
vasodilator contributing to the erectile process, norepi-
nephrine is the major vasoconstrictor through stimula-
tion of α1 receptors on the corpus cavernosum smooth
muscle. Under physiologic conditions, nitric oxide at-
tenuates norepinephrine vasoconstriction. The clinical
use of Yohimbine in the management of impotence
shows a delayed action which is inconsistent with its
short apparent half life and thus its activity may reside
1. MMUSA T, 30 caps. Not labelled
2. Vemoherb, 90 caps. Not labelled
3. Leuzeia plus, 90 caps. 15 mg
4. Myprotein T Matrix, 180 tabl. 30 mg
5. USN 19 Anabol Testo, 90 caps. 50 mg
6.
Supreme Sports Enhancements
Omnibolic, 120 caps.
250 mg
7. Rhaponticum carthamoides solution 20 ml

in possible metabolites that can display a biological half
life longer than that of the parent drug. Continuous ad-
ministration of Yohimbine, as opposed to on – demand
administration, might result in less norepinephrine
output due to increased turnover or α1 receptors down
regulation via a feedback mechanism. α1 blockers pre-
vent vasoconstriction caused by norepinephrine [30].
Depending on dosage Yohimbine can either increase
or decrease systemic blood pressure (through vasocon-
striction or vasodilation, respectively): small amounts
of Yohimbine can increase blood pressure, while large
amounts can dangerously lower blood pressure. The
therapeutic index of Yohimbine is quite low. The range
between an effective dose and a dangerous dose is very
narrow [31]. Atypical dose for sexual dysfunction is 15
– 30 mg, whereas 100 mg is considered dangerous. This
may also lead to increasing of panic disorder type reac-
tions or heart attack. More serious adverse effects may
include seizures and renal failure.Yohimbine should not
be consumed by anyone with liver, kidney, heart disease
or a psychological disorder.
2. Methods for synthesis of Yohimbine.
Yohimbine and its isomers are monoterpenoid –
derived indole alkaloids formed from the initial con-
densation of tryptamine with secologanin in the pres-
ence of strictosidine synthase to form strictosidine.
The tryptamine portion of the alkaloid is derived
from the decarboxylation of tryptophan with trypto-
phan decarboxylase enzyme. Strictosidine, after de-
glycosylation with glycosidase, through a series of
reactive intermediates forms 4,21-dehydrocorynan-
theine aldehyde which undergoes isomerisation and
reduction to afford Yohimbine. A laboratory synthe-
sis of Yohimbine is illustrated on Fig. 3.
Probable biosynthetic pathway for the formation
of Yohimbe alkaloids is presented on Fig. 4. [32].
In studies aimed at measuring the actual content
of Yohimbine versus the one declared on the label, it
was found that Yohimbine content ranged from none
up to 18.8 mg/serving. Quantitative data for the other
Yohimbe alkaloids are not available but would also be
required in view of their known or potential biological
activity. Yohimbe bark extracts for use in food supple-
ments should be evaluated based on existing toxico-
logical data and data in the chemical specifications.
No data describing the absorption, distribution, me-
tabolism and excretion of Yohimbe alkaloids after oral
administration of Yohimbe bark preparations such as
extracts are available. When administered to humans,
Yohimbine is rapidly absorbed and its bioavailability
is reported to range widely from 7 % to 87 %. In the
liver Yohimbine undergoes oxidation to its pharmaco-
logically active metabolite 11-Hydroxy-yohimbine.
This metabolic pathway is dependent on the two cy-
tochrome P450 polymorphic variants CYP2D6 and
CYP3A4. The great interindividual variability in the
clinical effects may be attributable to the differences in
Fig 4. A Laboratory synthesis of Yohimbine. Fig 5. Biosynthesis of Yohimbine.

the hepatic metabolism. No short – term or subchronic
toxicity and carcinogenicity studies on yohimbe bark
or its preparations were available [32].
3. Yohimbine in food supplements.
Food additives containing Yohimbine are also con-
sidered to possess androgenic effects. Yohimbine, the
major alkaloid of Yohimbe bark and Raubasine, an-
other alkaloid occurring in lower concentrations in
the bark, are used as active ingredients in a number of
medicinal products for which adverse effects are de-
scribed as headache, nausea, increased urinary urge,
insomnia, anxiety, restlessness, irritability, increase of
blood pressure and pulse rate, palpitation, dizziness,
vomiting, anorexia, gastric complaints, diarrhoea,
flush, sweating, shivering, allergic reactions, nervous-
ness, hypotension, tremor, bronchospasm, dysuria, de-
creased urge, genital pains, exanthema.
Few states have registered medicinal products con-
taining Yohimbine for oral use (tablets). There is a sin-
gle registered medicinal product containing Raubasine
in Italy (peroral drops or coated pills). In Ireland, the
use of Yohimbe is not permitted in traditional herbal
medicinal products, as it would be controlled as a pre-
scription medicine under the terms of the Medicinal
Products Regulations. The United Kingdom does not
consider any products containing P. yohimbe or Yo-
himbine to be classified as foods. In the UK the sale
and supply of botanicals containing P. yohimbe are
controlled under the Medicines (Retail Sale or Sup-
ply of Herbal Plants Remedies) Order 1977 SI 2130.
The use of the bark of P. yohimbe is prohibited in the
production of foods in the Czech Republic. The use of
P. yohimbe and its alkaloids is not authorised as food
or food supplements in Belgium (Royal Decree of 29
August 1997). Food supplements with P. yohimbe
herbal preparations are not allowed on the Dutch mar-
ket based on the Warenwetbesluit Kruidenpreparaten
(Letter from EMA, personal communication, April
2013). The Austrian food codex (Codex Alimentarius
Austriacus, Österreichisches Lebensmittelbuch) in-
cludes the species P. johimbe in a list of plants not to be
used for the production of herbal additives. The Dan-
ish Drogelisten lists Yohimbe (Corynanthe johimbe K.
Schum) as a plant that is unacceptable as food regard-
less of amount [32]. On Bulgarian market are aviable
many food additives containing Yohimbine. Food sup-
plements with Yohimbine are presented on Table 3.
IV. Food supplements with (Dehydroepiandros-
terone (DHEA).
16-Dehydropregnenolone acetate with hydroxy-
lamine form oxime, which after reaction with hydro-
1.
VPX Redline Xtreme Shot, 90
ml solution
Not labelled
2.
BPI Sports 1.M.R Vortex, 150 g
powder
Not labelled
3.
Physique Enhancing Science
Alphamine, 150 g powder
Not labelled
4.
USP labs Jack 3 Advanced, 230
g powder
Not labelled
5. Cellucor Super HD, 10 caps. Not labelled
6.
Cellucor D4 Thermal Shock,
120 caps.
Not labelled
7. Evogen Lipocide, 60 caps. Not labelled
8.
HIT Supplements Torch, 135
caps.
Not labelled
9.
Muscle Meds Methyl Burn
Extreme, 60 caps.
Not labelled
10.
Yohimbine HCl (4 Dimension
Nutrition), 90 caps.
2.5 mg
11.
Fighter Diet FDXtreme Burn,
60 caps.
3 mg
12. Lipo-6 (Nutrex), 240 caps. 3 mg
13.
Hydroxycut Hardcore Elite
(MuscleTech) powder
28 mg
14.
Hydroxycut Hardcore Elite
(MuscleTech), 180 caps.
28 mg
15. Male Fuel (Twinlab), 120 caps. 400 mg
16. Yohimbe Bark (Natrol), 90 caps. 500 mg
17.
Kamagra (Borolla), 30 caps.
Yohimbine 50 mg
Tribulus terrestris 150 mg
Ginseng 150 mg
L-Arginine 250 mg
18
Kamasutra (Borolla) 30 caps.
Yohimbine 5 mg
Pyridoxine 6 mg
Zinc aspartate 7 mg
Extract of Ginkgo biloba 20 mg
α- Tocopherol 20 mg
Table 3. Food additives with Yohimbine.
chloric acid lead to intermediates which are trans-
formed to Dehydroepiandrosterone.
On Table 4. are summarized additives containing
Dehydroepiandrosterone.
For analysis of different compounds in food sup-
plements the very often applied are the following
methods: 1) HPLC with UV – detection for: a) deter-
mination of L – Arginine in Tonotyl® solution [33];

1.
DHEA (Blade Nutrition Test Booster),
180 caps.
25 mg
2. DHEA (MRM), 90 caps. 25 mg
3. DHEA (Natrol), 300 tabl. 25 mg
4. DHEA Fast Dissolve (Natrol), 30 tabl. 25 mg
5. DHEA MAX (Nutraceutics), 60 tabl. 25 mg
6. DHEA (Ultimate Nutrition), 100 caps. 25 mg
7. DHEA (Healthy 'N Fit), 100 caps. 50 mg
8. DHEA (MRM), 60 caps. 50 mg
9. DHEA (Natrol), 60 tabl. 50 mg
10. DHEA (S.A.N.), 90 caps. 50 mg
11. DHEA (Ultimate Nutrition), 100 caps. 50 mg
12. DHEA (PharmaFreak), 28 caps. 60 mg
13. DHEA (Maximum Nutrients), 60 caps. 100 mg
Table 4. Additives containing Dehydroepiandrosterone.
Fig 6. Synthesis of Dehydroepiandrosterone.
b) control of aminoacids in organic foods and food
supplements [34]; c) quality control of content of
Vitamin C and Vitamin B6 in food supplements and
drugs [3]; 2) gas chromatography with flame ioniza-
tion detector for: determination in food supplements
– tablets of total content of aminoacids [36] and of
free aminoacids [37] and determination of total con-
tent and for quantity of free aminoacids in liquid food
additives [38]; 3) TLC – densitometric method for
simultaneous analysis of L – Leucine and L – Valine
in additives [39].
V. Methods for analysis of extracts from Tribu-
lus terrestris Linn. in food supplements.
For the determination of steroidal saponins from
extracts from Tribulus terrestris Linn. the most ap-
plied methods are HPLC [40] and GC [41].
Tribulus extracts from roots and fruits after extrac-
tion with of acetonitrile are analyzed by reverse phase
HPLC at the following conditions: gradient elution
with mobile phase: phosphoric acid buffer with pH = 3
and acetonitrile, injection volume: 20 μl, flow rate: 1.0
ml/min., UV – detection at λ = 203 nm [11].
For determination of glycosidic furostanolic and
spirostanolic saponins from dietary supplements of
Tribulus terrestris Linn. is applied an HPLC-ESI-
MS method using Protodioscin as external standard
[42]. Steroidal saponins in Tribulus terrestris Linn.
extracts are quantified by RP HPLC with evaporative
light scattering detection [43].
Chromatograms of comparative analytical inves-
tigation of Tribulus terrestris Linn. preparations from
different manufacturers [44] are shown on Fig 5.
Analysis of methanol extracts of Tribulus ter-
restris Linn. are analyzed by GC – MS, with capil-
lary column with dimethyl polysiloxane (30 m x 0.25
mm, 1 μm), oven temperature: 280o
C, injection port
temperature: 250o
C, helium flow rate as 1 ml/min.,
ionization voltage: 70 eV. On Table 5 are presented
name, retention time and peak areas of the analyzed
by GC – MS components in the methanolic extract of
Tribulus terrestris Linn.
In Tribulus terrestris Linn. extracts spirostanol sap-
onin Diosgenin and furostanol saponins are determined
by TLC – densitometric analysis after derivatization
with an anisaldehyde sulfuric acid, stationary phase:
precoated silica gel 60 F254
plates, mobile phase: tolu-
ene : ethyl acetate : methanol = 7: 3 : 1 v/v, detection
at 430 nm [46]. In fruits of Tribulus terrestris Linn.
steroidal saponins Protodioscin and Prototribestin are

N: tR Name of compound
Peak area
[%]
1. 14.52
3,7,11,15-tetramethyl-
2-hexadecan-1-ol
1.04
2. 16.27 n-Hexadecadienoic acid 8.83
3. 16.58
n-Hexadecadienoic acid
ethyl ester
0.74
4. 18.55 Phytol 0.99
5. 18.86 9,12-Octadecadienoic acid 1.86
6. 18.95
9,12,15-Octadecadienoic
acid
8.58
7. 19.26 Octadecanoic acid 2.95
8. 24.82
1.2-benzenedicarboxylic
acid, diisoocthyl ester
9.27
9. 31.46 α-Amyrin 65.73
quantified through HPTLC on precoated silica gel 60
F254
aluminum plates and mobile phase of n – butanol :
glacial acetic acid : water = 80 : 6 : 20 v/v [47].
VI. Methods for analysis of 20-Hydroxyecdysone
in additives.
20-Hydroxyecdysone is determined by the fol-
lowing methods: 1) TLC with detection at λ = 254
nm or at λ = 366 nm [48]; 2) spraying the TLC plate
with vanillin - sulfuric acid reagent to obtaining tur-
quoise fluorescenting spots at λ = 366 nm [48]; 3)
densitometric evaluation at λ = 250 nm in reflectance/
absorbance mode [49]; 4) HPLC [50, 51]; 5) micellar
electrokinetic chromatography [48].
Table 5. GC – MS analysis of Tribulus terrestris Linn. [45].
Fig 7. Chromatogram of preparation with Tribulus
terretris Linn.
VII. Methods for analysis of Yohimbine in
supplements.
Different methods were adopted for analyzing of
Yohimbine hydrochloride, including spectroscopic,
voltammetric, polarographic chemiluminometric
methods, TLC, HPLC and gas chromatography [52].
For a qualitative and quantitative analysis Yo-
himbine in commercial supplements are reported: 1)
HPLC-UV-MS technique with atmospheric pressure
chemical ionisation and electrospray ionisation [53];
2) ultra performance liquid chromatography using
Waters Acquity BEH C18
column, gradient elution
with mobile phase: 0.1 % v/v aqueous ammonium
hydroxide and 0.1 % ammonium hydroxide in meth-
anol and mas detection [54, 55]; 3) non – aqueous
capillary electrophoresis [56] and gas chromatogra-
phy with mass spectrometry [56, 57]. Metabolites
10- and 11-Hydroxy-yohimbine are analyzed by the
normal phase HPLC [58].
According to European Food Safety Authority
(EFSA) there are no official inter-laboratory validat-
ed methods for the determination of Yohimbine and
accompanying alkaloids in food supplements [32].
VIII. Methods for analysis of food supplements
with Dehydroepiandrosterone.
For the determination of Dehydroepiandroster-
one are applied HPLC with stationary phase: Zorbax
Rx C18
column, mobile phase: acetonitrile : 0.025 M
phosphate buffer = 60 : 40 v/v, or 75 : 25 v/v, UV
detection at λ = 292 nm [59]. Androstenedione, De-
hydroepiandrosterone and Testosterone are analysed
by liquid chromatography – mass spectrometry on
stationary phase Gemini C18
column (100 mm x 2.0
mm, 3 μm), injection volume: 50 μl, oven tempera-
ture: 30 °C [60]. A HPLC with electrospray ioniza-
tion - mass spectrometric (LC–ESI-MS/MS) method
is reported for the simultaneous quantification of Tes-
tosterone and its precursor, Dehydroepiandrosterone,
after derivatized with 2-hydrazino-1-methylpyridine,
is developed [61].
A gradient RP-HPLC method is developed for the
separation of Dehydroepiandrosterone, its sulfate es-
ter (DHEA-S), its three C7-oxidized metabolites, and
its biosynthetic derivates (Androstenedione, Testos-
terone, Estradiol, Pregnenolone) [62].
GC-MS analysis of the trimethylsilyl derivatives
of the steroids is developed. The limit of detection
was improved by the addition of a mixture of 1-N,N-
diisopropylamino-n-alkanes to the final extract. The
limits obtained limits of detection are demonstrated
to be 0.1ng/μg for Dehydroepiandrosterone and

estr-4-ene-3β,17β-diol, 0.7 ng/g nfor 5α-androstane-
3β,17β-diol and androsta-1,4-diene-3,17-dione, 1
ng/g for estr-5-ene-3β,17β-diol, estr-4-ene-3,17-
dione, 19-nortestosterone, androst-4-ene-3, 17-di-
one and testosterone, and 2 ng/g for androst-4-ene-
3β,17β-diol and androst-5-ene-3β,17β-diol [4].
Conclusion.
Food supplements containing extracts from Tribu-
lus terrestris Linn., Yohimbine or Ecdysteroids pos-
sess androgenic and anabolic effects, improving
sexual performance, memory and learning and in-
creasing working capacity of skeletal muscles. Low
consumer`s awareness of food additives in many
cases makes this type of product risk.
For quality control of this food additives the most
applied methods are: HPLC – MS, GC – MC, TLC
– densitometric method after derivatization, HPTLC.
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Corresponding author
Danka Obreshkova
Faculty of Pharmacy, Medical University - Sofia
2 Dunav Str. 1000 Sofia
e-mail: phddanka@yahoo.com
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