Testosterone Phenylpropionate Research Articles

A novel quadruple templates molecularly imprinted polymer electrochemical sensor assisted by second-order calibration methods for detection of Sustanon abuse

Sustanon (SUS) consists of testosterone propionate (TP), testosterone phenylpropionate (TPP), testosterone isocaproate (TIC) and testosterone decanoate (TD) which is generally used as a medication to treat the testosterone deficiency in men. Misuse of SUS to enhance the ability of the athletes can be a very serious risk for their health which is discouraged. Therefore, the main aim of this work was based on developing a novel biosensing system for determination of different types of testosterones in SUS to monitor it. A rotating glassy carbon electrode (GCE) was modified with fullerene C60-multiwalled carbon nanotubes-ionic liquid (C60-MWCNTs-IL) and used as the working electrode for recording second-order hydrodynamic square wave voltammetric (hydrodynamic SWV) data at different step potentials including 0.02, 0.025, 0.03, 0.035, 0.04 and 0.045 V as a sensitive method. Different types of testosterones were extracted and preconcentrated by quadruple templates molecularly imprinted polymers (QTMIPs) and they were determined by C60-MWCNTs-IL/GCE. The testosterones showed severely overlapped peaks at the surface of C60-MWCNTs-IL/GCE which were resolved by three-way calibration based on second-order hydrodynamic SWV data. Therefore, a novel sensor assisted by QTMIPs and second-order calibration methods was developed which had a very good performance for the analysis of synthetic and real matrices in simultaneous determination of TP, TPP, TIC and TD with aim of monitoring the SUS.

Multiplex lateral flow immunochromatographic assay for the qualitative and quantitative detection of six steroid hormone residues in chicken and pork

To ensure food safety from animal origins and to prevent potential hazards caused by steroid hormones, there is an urgent need to develop a rapid detection method for steroid hormones in foods from animal origins. Three highly sensitive monoclonal antibodies (mAb) against testosterone propionate (Tp), stazolol (STA) and estradiol benzoate (EB) were prepared, with IC50 values of 0.13, 0.14 and 0.19 ng/mL, respectively. Tp-mAb is a group-selective mAb that simultaneously detects Tp, testosterone (T), methyltestosterone (MT) and testosterone phenylpropionate (TPP), and their IC50 values were 0.13, 0.77, 0.90, and 1.25 ng/mL, respectively. A novel multiplex lateral flow immunochromatographic strip (ICS) was further prepared, which could simultaneously detect Tp, STA, and EB in chicken and pork samples with limits of quantification (LOQ) of 0.32, 0.51, 0.35 μg/kg and 0.36, 0.64, 0.43 μg/kg, respectively. Based on the broad spectrum of Tp-mAb, ICS has been further developed to detect Tp, T, MT and TPP simultaneously. This multiplex ICS rapid detection method was efficient, sensitive, and fast, for the simultaneous detection of multiple steroid hormones in actual samples. Its detection time is only 40 min, which it represents a favorable monitoring tool for the detection of steroid hormones in animal-derived foods from the marketplace.

Quantification of endogenous steroid sulfates and glucuronides in human urine after intramuscular administration of testosterone esters

For an effective detection of doping with pseudo-endogenous anabolic steroids, the urinary steroid profile is of high value. In this work, the aim was to investigate steroid metabolism disruption after exogenous intramuscular administration of different testosterone esters. The investigation focused on both sulfo – and glucoro conjugated androgens. A single intramuscular injection of either 1000 mg testosterone undecanoate (Nebido®) or a mixture of 30 mg testosterone propionate, 60 mg testosterone phenylpropionate, 60 mg testosterone isocaproate, and 100 mg testosterone decanoate (Sustanone®), was given to six healthy volunteers. Urine was collected throughout a testing period of 60 days. A LC-MS method was developed and validated for the analysis of eight conjugated steroids in their intact form. The results show that urinary changes in both sulfo – and glucuro conjugated steroid levels are prominent after the injection of testosterone esters. A promising potential marker for the intake of exogenous testosterone is the combined ratio of epitestosterone sulfate/epitestosterone glucuronide to testosterone sulfate/testosterone glucuronide ((ES/EG)/(TS/TG)) as a complementary biomarker for testosterone abuse. This represents a new piece of evidence to detect testosterone doping, representing a new approach and being independent from the metabolic connections of the markers in the steroid passport.

1H NMR determination of adulteration of anabolic steroids in seized drugs

Counterfeiting and adulteration of pharmaceuticals is a prevalent problem worldwide and represents a major health risk to the population, with anabolic steroids being one of the main classes of drugs consumed and obtained from dubious sources. In this work, we propose the use of the 1H NMR technique to evaluate formulations containing anabolic steroids, with analysis of 40 samples of anabolic drugs that are used in injectable and capsule forms. The samples analyzed presented the following active ingredients: testosterone propionate, testosterone phenylpropionate, testosterone isocaproate, testosterone decanoate, testosterone cypionate, testosterone undecanoate, stanozolol, drostanolone propionate, trenbolone acetate, oxymetholone, and methandrostenolone. The 1H NMR spectroscopic measurements were performed using a 600 MHz Bruker Avance III spectrometer, with deuterated chloroform (CDCl3) containing 0.1% TMS as solvent. Of the 40 samples analyzed, eight did not show the presence of the active principle stated on the label. Three types of adulteration were found in the analyzed samples: absence of the active ingredient, adulteration with other substances, and concentration values below those indicated on the label. Sildenafil citrate was found in four samples. The GC–MS technique was used to confirm the adulteration results found using 1H NMR. Quantitative determination by NMR was performed using internal standard and ERETIC 2 methods, and the results obtained were statistically the same.

Psychosis in Kallmann Syndrome: a case illustration based on possible genetic liability

Kallmann Syndrome (KS) is an uncommon disorder caused by a deficient production or secretion of gonadotropin-releasing hormone (GnRH) and manifests with an absence of spontaneous puberty and impaired sense of smell. Multiple inheritance patterns have been recognized in the etiology. KS and psychosis have occasionally been con-sidered to originate in similar predisposing genetic profile. Defective KAL-X gene which had previously been attributed in the etiopathogenesis of schizophrenia was asserted to lead to an impairment in the migration of GnRH neurons and olfactory deficits in various severity and disruption in sexual developmental, consequently. Our case is a 19-year-old male previously diagnosed with KS at his 13, who has been under the monthly intramuscular treat-ment of testosterone propionate and testosterone phenylpropionate at the dose of 250 mg. He was admitted to our clinic with auditory and visual hallucinations that started two years ago. Persecutory delusions, social isolation, and suicidal thoughts also existed. We diagnosed him schizophrenia with comorbid KS following the hospitalization and a detailed psychiatric examination. Olanzapine was initiated as 5 mg/day and potentiated to 15 mg/day. His delu-sions, hallucinations and suicidal thoughts had improved with the treatment. In this illustration, we purposed to discuss probable common etiopathogenesis underlying this comorbidity.

Determination of testosterone esters in the hair of male greyhound dogs using liquid chromatography–high resolution mass spectrometry

The doping of greyhound dogs with testosterone is done in an attempt to improve their athletic performance, but such doping cannot easily be confirmed, especially in male dogs owing to the natural presence of endogenous testosterone. As testosterone is usually administered as its esters, their direct detection in hair would provide confirmatory evidence of the administration of a pharmaceutical product. This article demonstrates that the use of a liquid chromatography-high resolution mass spectrometry method with heated electrospray ionisation (HESI) combined with the use of amino solid-phase extraction (SPE) cartridges for sample clean-up, is suitable for the sensitive determination of propionate, phenyl propionate, isocaproate, decanoate, and enanthate esters of testosterone in greyhound hair. The method is linear over the range, 0.1μg/kg-10μg/kg, for all the testosterone esters analysed. The limits of detection (LOD) are 0.05μg/kg for testosterone phenyl propionate, isocaproate, and decanoate, 0.025μg/kg for testosterone propionate, and 0.25μg/kg for testosterone enanthate. This method was applied to hair samples collected from male greyhounds before and after a single administration of a product containing several testosterone esters, each of which could be detected up to 100days post-administration. The study also demonstrates that tail hair is the specimen of choice for the analysis of testosterone in dog hair and that washing of dogs does not impact the analysis of testosterone esters in hair. This method may be useful in racing regulation for the detection of illegitimate use of testosterone in all species.

Detection and quantification of 12 anabolic steroids and analogs in human whole blood and 20 in hair using LC-HRMS/MS: application to real cases.

We developed and validated a method to detect and quantify 12 anabolic steroids in blood (androstenedione, dihydrotestosterone, boldenone, epitestosterone, mesterolone, methandienone, nandrolone, stanozolol, norandrostenedione, tamoxifene, testosterone, trenbolone) and eight more in hair samples (nandrolone phenylpropionate, nandrolone decanoate, testosterone propionate, testosterone benzoate, testosterone cypionate, testosterone decanoate, testosterone phenylpropionate, testosterone undecanoate) using liquid chromatography coupled to high-resolution mass spectrometry. This method used a benchtop Orbitrap mass spectrometer operating with an APCI probe under positive ionization mode. Analysis was realized in full scan experiment with a nominal resolving power of 140,000. After addition of the internal standard (testosterone-D3) and incubation in phosphate buffer pH=5 for hair, 200μL of blood and 30mg of hair samples were extracted with heptane. LOQ and LOD were determined at 5 and 1ngmL-1 in whole blood and 10 to 100pgmg-1 and 2 to 20pgmg-1 in hair according to the compounds, respectively. The method was linear in the 5-1000ngmL-1 range in whole blood and between 10 or 100pgmg-1 and 1000pgmg-1 in hair with correlation coefficients >0.99, and intra- and inter-day accuracy and precision were <14.8% for all compounds except for some esters in hairs (<19.9%) probably due to an important matrix effect for these compounds. This sensitive and specific method to detect anabolic steroids has been successfully applied to two real cases, for which various anabolic steroids in whole blood, urine, and hair were identified and quantified.

Detection of testosterone esters in blood.

Injections of synthetic esters of testosterone are among the most common forms of testosterone application. In doping control, the detection of an intact ester of testosterone in blood gives unequivocal proof of the administration of exogenous testosterone. The aim of the current project was to investigate the detection window for injected testosterone esters as a mixed substance preparation and as a single substance preparation in serum and plasma. Furthermore, the suitability of different types of blood collection devices was evaluated. Collection tubes with stabilizing additives, as well as non-stabilized serum separation tubes, were tested. A clinical study with six participants was carried out, comprising a single intramuscular injection of either 1000 mg testosterone undecanoate (Nebido(®)) or a mixture of 30 mg testosterone propionate, 60 mg testosterone phenylpropionate, 60 mg testosterone isocaproate, and 100 mg testosterone decanoate (Sustanon(®)). Blood was collected throughout a testing period of 60 days. The applied analytical method for blood analysis included liquid-liquid extraction and preparation of oxime derivatives, prior to TLX-sample clean-up and liquid chromatography-tandem mass spectrometry (LC-MS/MS) detection. All investigated testosterone esters could be detected in post-administration blood samples. The detection time depended on the type of ester administered. Furthermore, results from the study show that measured blood concentrations of especially short-chained testosterone esters are influenced by the type of blood collection device applied. The testosterone ester detection window, however, was comparable.

Use of dried blood spots in doping control analysis of anabolic steroid esters

Dried blood spot (DBS) sampling, a technique for whole blood sampling on a piece of filter paper, has more than 50-years tradition, particularly in the diagnostic analysis of metabolic disorders in neonatal screening. Due to the minimal invasiveness, straightforwardness, robustness against manipulation and fastness DBS sampling recommends itself as an advantageous technique in doping control analysis. The present approach highlights the development of a screening assay for the analysis of eight anabolic steroid esters (nandrolone phenylpropionate, trenbolone enanthate, testosterone acetate, testosterone cypionate, testosterone isocaproate, testosterone phenylpropionate, testosterone decanoate and testosterone undecanoate) and nandrolone in DBS. The detection of the intact esters allows an unequivocal proof of the administration of conjugates of exogenous testosterone and its derivatives. Precise, specific and linear conditions were obtained by means of liquid chromatography high resolution/high accuracy mass spectrometry. Sensitivity in the low ppb range was accomplished by the preparation of the methyloxime derivatives of the target compounds. Labeled internal standards (d3-nandrolone, d3-nandrolone caproate and d3-nandrolone undecanoate) were applied to compensate for the broad range in chain length of the esters.The assay presented here outlines the application of DBS for the analysis of anabolic steroid esters in doping controls for the first time providing great potential to simplify the proof of exogenous administration of testosterone.

Sustanon Administration Induces Satellite Cell Proliferation and Giant Fiber Formation in Growing Skeletal Muscle without Exercise

Sustanon is one of the most commonly used anabolic androgenic drugs to increase skeletal muscle mass and strength. This drug is a blend of four esterized testosterone derivatives: testosterone propionate, testosterone phenylpropionate, testosterone isocaproate, and testosterone decanoate. Little is known about the effects of this drug on skeletal muscle at the cellular level. This study aimed to investigate the influence of Sustanon on the morphology of skeletal muscle fibers and the distribution of myogenic satellite cells (SCs) during postnatal growth. We hypothesized that Sustanon‐induced skeletal muscle hypertrophy is associated with an increase in the number of SCs. Robust immunocytochemical techniques and morphometric analyses were used to calculate the numbers of SCs and myonuclei within pectoralis muscle of the chicken. Sustanon significantly increased pectoralis mass and fiber size. All SC indices and the number of myonuclei increased significantly by Sustanon administration. This study indicates that Sustanon can induce hypertrophy and increases the numbers of SCs and myonuclei in skeletal muscle fibers without imposed exercise. Funds for this study were provided by a grant from the Deanship of Research at Jordan University of Science &amp; Technology.Grant Funding Source: Jordan University of Science &amp; Technology

Effective separation and simultaneous analysis of anabolic androgenic steroids (AAS) in their pharmaceutical formulations by a validated TLC-densitometry method.

BackgroundAnabolic androgenic steroids (AAS) are widely misused for the enhancement of performance in sports. Several drugs are available that contain different combinations or individual steroids in different dosage form. This paper describes a TLC densitometric method for simultaneous determination of four AAS of testosterone derivatives including testosterone propionate (TP), testosterone phenyl propionate (TPP), testosterone isocaproate (TI) and testosterone deaconate (TD) in their pharmaceutical products.ResultsSeparation was carried out on Al based TLC plates, pre-coated with silica gel 60F-254 using hexane and ethyl acetate (8.5:1.5, v/v). Spots at Rf 0.31 ± 0.01, 0.34 ± 0.01, 0.40 ± 0.01 and 0.45 ± 0.02 were recognized as TPP, TP, TI and TD, respectively. Quantitative analysis was done by densitometric measurements at λmax 251 nm for all derivatives. The developed method was validated as per ICH guidelines. Method was found linear over the concentration range of 200–1200 ng/spot with the correlation coefficient of 0.995, 0.993, 0.995 and 0.996 for TP, TPP, TI, TD, respectively. Limit of detection for all derivatives were in the range of 16.7-22.3 ng/spot while limit of quantitation were found to be in the range of 55.7-70.9 ng/spot.ConclusionsThe developed TLC method can be applied for the simultaneous routine analysis of testosterone derivatives in their individual and combined pharmaceutical formulations.

Effects of Sustanon on the Distribution of Satellite Cells and The Morphology of Skeletal Muscle Fibers During Maturation

Sustanon is one of the most commonly used anabolic androgenic drugs to increase skeletal muscle mass and strength. This drug is a blend of four esterized testosterone derivatives: Testosterone propionate, testosterone phenylpropionate, testosterone isocaproate and testosterone decanoate. Little is known about the effects of this drug on skeletal muscle at the cellular level. This study aimed to investigate the influence of Sustanon on the morphology of skeletal muscle fibers and the distribution of myogenic stem cells known as Satellite Cells (SCs) during postnatal growth. We hypothesized that Sustanon-induced skeletal muscle hypertrophy is associated with an increase in the number of SCs. Robust immunocytochemical techniques and morphometric analyses were used to calculate the numbers of SCs and myonuclei within the pectoralis muscle of chickens. Also, DNA concentration and Pax7 protein levels were measured to confirm immunocytochemical findings. Sustanon significantly increased pectoralis mass and fiber size. All SC indices and the number of myonuclei increased significantly by Sustanon administration. In addition, greater DNA concentration and Pax7 protein expression were found in Sustanon-treated birds. This study indicates that Sustanon can induce avian skeletal muscle hypertrophy and that this is correlated with increased numbers of SCs and myonuclei.

Effects of Sexually Activated Rams or Ewes on Pulsatile LH Secretion in Anoestrous Sheep

home Acta Veterinaria Brno Journal of the University of Veterinary Sciences Brno, Czech Republic Search:

Determination of circular dichroism and ultraviolet spectral parameters of norgestimate- and other Delta(4)-3-ketosteroid oxime isomers via normal phase HPLC method.

The oxime formation reaction of therapeutical progestogen (levonorgestrel, levonorgestrel acetate, norethisterone), androgen (methyltestosterone, testosterone phenylpropionate) and anabolic (nortestosterone phenylpropionate) Delta(4)-3-ketosteroids has been investigated. The ketosteroid-hydroxylamine reaction was monitored by reversed phase HPLC system. It was established, that under the experimental conditions applied the oxime formation was complete within 2 h. The reaction leads to the formation of Z and E oxime isomers. The isomers of norgestimate (levonorgestrel 17-acetate oxime) and other Delta(4)-3-ketosteroid oximes have been separated by a new normal phase HPLC method. The identification (elution order assignation) and determination of the formation ratio of the isomers have been performed by 1H NMR spectroscopy on the basis of the chemical shift differences of 4-H signals. The on-line CD and UV spectra of the pure oxime isomers were recorded and then molar ellipticities and absorbances of the isomers were calculated in the wavelength range of 200-300 nm via parameter estimation method.

Binding of Dexamethasone Phosphate and Testosterone Phenyl Propionate to Bovine Serum Albumin: Drug?drug Interaction at the Binding Site

Binding of Dexamethasone Phosphate and Testosterone Phenyl Propionate to Bovine Serum Albumin: Drug?drug Interaction at the Binding Site

Determination of delta4-3-ketosteroids based on oxime formation by difference circular dichroism spectroscopy.

delta4-3-Ketosteroids exhibit an intensive negative Cotton effect on the circular dichroism (CD) spectra in the wavelength range for the n-pi* electronic transition (270-350 nm). With hydroxylamine hydrochloride, delta4-3-ketosteroid compounds can be transformed into oxime derivatives. Following oxime formation, positive ellipticity with low intensity can be registered in this wavelength range. The quantitative determination of delta4-3-ketosteroids is based on the considerable difference between the ellipticities before and after oxime formation. The difference ellipticity for the six ketosteroids examined (norethisterone, levonorgestrel, levonorgestrel acetate, methyltestosterone, testosterone phenylpropionate, nortestosterone phenylpropionate) varies linearly with the concentration in the interval 6 x 10(-6)-3 x 10(-3) mol/L. The method can be well applied to determination of delta4-3-ketosteroid contamination of norgestimate [(+)-13-ethyl-17-hydroxy-18,19-dinor-17alpha-pregn4-en-20-yn-3-one oxime acetate]; 0.02-10% impurity can be measured.

Gas chromatographic–tandem mass spectrometric determination of anabolic steroids and their esters in hair: Application in doping control and meat quality control

We have developed a powerful and simple sensitive method for testing hair for anabolic steroids and their esters. A 100-mg amount of powdered hair was treated with methanol in an ultrasonic bath for extraction of esters, then alkaline digested with 1 M NaOH for an optimum recovery of other drugs. The two liquid preparations were subsequently extracted with ethyl acetate, pooled, then finally highly purified using a twin solid-phase extraction on amino and silica cartridges. The residue was derivatized with N-methyl- N(trimethylsilyl)-trifluoracetamide (MSTFA) prior to injection. Analysis was conducted by gas chromatography coupled to a triple quadrupole mass spectrometer. The generally chosen parent ion was the molecular ion while two daughter ions were selected for each compound with collision energies ranging from −16 to −21 eV. Internal standards were nandrolone d 3 for non-esterified drugs and testosterone phenyl propionate for esters. The limits of detection calculated from an analysis of the blanks ( n=30) were 0.08 pg/mg for nandrolone, 6.20 pg/mg for boldenone, 0.07 pg/mg for methyl testosterone, 0.15 pg/mg for ethinyl estradiol, 2.10 pg/mg for metandienone, 0.86 pg/mg for testosterone propionate, 0.95 pg/mg for testosterone cypionate, 1.90 pg/mg for nandrolone decanoate, 3.10 pg/mg for testosterone decanoate and 4.80 pg/mg for testosterone undecanoate. Application to doping control has been demonstrated. In a series of 18 sportsmen, two tested positive for anabolic steroids in hair whereas urinalysis was negative for both of them. The first positive case was nandrolone and the second case concerned the identification of testosterone undecanoate. Measured in 10 white males aged between 22 and 31 years, the testosterone concentration was in the range 1.7–9.2 pg/mg (mean=5.0 pg/mg). The method was also applied in meat quality control. Of the 187 analyses realized based upon hair and urine sampling in slaughter houses, 23 were positive for anabolic steroids in hair: one case for boldenone, one case for metandienone, two cases for testosterone propionate, three cases for nandrolone, five cases for testosterone decanoate and 11 cases for methyl testosterone. In the meantime, urinalysis was always negative for these drugs or their metabolites.

Effects of testosterone on the rat renal medullary vasopressin receptor concentration and the antidiuretic response.

The renal concentrating ability declines with age in humans and animals. Studies suggest that the concentrating defect is due to a decrease in renal vasopressin sensitivity. With ageing, expression of the renal vasopressin V2 receptor in rat is impaired; the normal receptor expression is restored by testosterone treatment. The effect of testosterone on the renal sensitivity to vasopressin was investigated in young rats. Male rats after orchidectomy and chronic antiandrogen cyproterone acetate treatment, and female rats after chronic testosterone phenylpropionate treatment, were used. The plasma arginine-vasopressin (AVP) and testosterone concentrations, and the antidiuretic responses to AVP and the V2 agonist deamino-[8-D-arginine]-vasopressin (dDAVP) after volume loading were measured, and the renal [3H]AVP binding density was determined. The plasma AVP level decreased slightly, but not significantly, in male rats after orchidectomy and cyproterone acetate treatment, but did not alter in female rats after testosterone treatment. The AVP and dDAVP sensitivities decreased in male rats after orchidectomy and cyproterone acetate administration, and increased in female rats treated with testosterone, as compared with the animals with a normal gonadal function. [3H]AVP binding to the renal inner medullary membranes was decreased following orchidectomy or antiandrogen treatment in male rats, and increased in testosterone-treated female rats. The results suggest that testosterone may play a physiological role in maintenance of the V2 vasopressin receptor expression and hence in the normal urinary concentrating ability in rat.

Treatment of Constitutional Delayed Puberty with a Combination of Testosterone Esters

Thirteen boys who had constitutional delayed puberty (CDP) were treated with a combination of short- and long-acting testosterone esters (testosterone propionate, testosterone phenylpropionate, testosterone isocaproate). Mean age at the onset of treatment was 14.9 +/- 0.6 years and bone age delay was -2.7 +/- 0.9 years. An intramuscular dose of 200 mg testosterone was administered 4 times at 3-week intervals and the treated CDP boys were followed for 2 years. All boys with CDP entered puberty after the last dose (testicular volume > or = 4 ml) and growth rate increased from 4.5 +/- 0.5 cm/year pretreatment to 8.4 +/- 1.6 cm/year posttreatment after the 2-year follow-up period. Height for bone age SD score did not alter significantly from a mean of -1.1 pretreatment to -1.3 posttreatment as well as predicted height pretreatment (173.5 +/- 6.6 cm) and posttreatment (173.3 +/- 4.9 cm). A combination of testosterone esters in a given dose and schedule is a safe and effective treatment for prepubertal boys with CDP.

The action of trenbolone acetate, a synthetic anabolic steroid, on ovarian function in the guineapig

The action of trenbolone acetate, a synthetic anabolic steroid, on ovarian function was investigated in the guinea pig. Certain comparisons were made with testosterone, the naturally occurring androgen, administered as the phenylpropionate ester. Two milligrams trenbolone acetate per kg given subcutaneously on alternate days for 20 days blocked oestrous cyclicity and ovulation in 9 of 10 animals. A similar effect was shown by 2.2 mg of testosterone phenylpropionate. Treatment of trenbolone acetate-treated animals with exogenous gonadotrophins suggested that the production of follicle-stimulating hormone had been suppressed. Signs of abnormality were seen in the livers of animals receiving 2 mg trenbolone acetate and 2.2 mg testosterone phenylpropionate.