Boldenone

Elucidation of the biosynthetic pathways of boldenone in the equine testis.[Pubmed:30951760]

Steroids. 2019 Apr 2;146:79-91.

Boldenone is an anabolic-androgenic steroid that is prohibited in equine sports. Urine from the uncastrated male horse contains Boldenone that is thought to be of endogenous origin and thus a threshold ('cut-off') concentration has been adopted internationally for free and conjugated Boldenone to help distinguish cases of doping from its natural production. The testis is likely to be a source of Boldenone. Qualitative analysis was performed on extracts of equine testicular homogenates (n=3 horses) incubated non-spiked and in the presence of its potential precursors using liquid chromatography tandem mass spectrometry (LC-MS/MS) and LC high resolution mass spectrometry (LC-HRMS). Samples were analysed both underivatised and derivatised to increase the certainty of identification. In addition to previously reported endogenous steroids, analysis of non-spiked testicular tissue samples demonstrated the presence of Boldenone and boldienone at trace levels in the equine testis. Incubation of homogenates with deuterium or carbon isotope labelled testosterone and androstenedione resulted in the matching stable isotope analogues of Boldenone and boldienone being formed. Additionally, deuterium and carbon labelled 2-hydroxyandrostenedione was detected, raising the possibility that this steroid is a biosynthetic intermediate. In conclusion, Boldenone and boldienone are naturally present in the equine testis, with the biosynthesis of these steroids arising from the conversion of testosterone and androstenedione. However, additional work employing larger numbers of animals, further enzyme kinetic experiments and pure reference standards for 2-OH androstenedione isomers would be required to better characterize the pathways involved in these transformations.

Determination of boldenone in postmortem specimens including blood and urine samples using LC-MS/MS.[Pubmed:30851513]

J Pharm Biomed Anal. 2019 May 30;169:111-115.

Boldenone (BOLD), one of androgenic anabolic steroids (AAS), although banned in humans, is still available illegally. AAS abuse has previously been associated with various cardiovascular adverse events including acute myocardial infarction, arrhythmia, and sudden death. In this study, the concentration of BOLD was determined in postmortem specimens from the corpse of a human male who intentionally injected BOLD undecylenate into his shoulder muscle. In addition, the endogenous levels of BOLD in the blood and urine samples of young human males have been reported. A liquid chromatography-tandem mass spectrometry (LC-MS/MS) method with solid-phase extraction (SPE) was developed and validated for the analysis of BOLD in blood, muscular tissue and urine samples. The validation parameters including linearity, accuracy, precision, matrix effect, and recovery were satisfactory. The concentrations of BOLD in the blood of 20 young human males who didn't take BOLD were under the limit of quantitation (LOQ, 0.5 ng/mL). Additionally, the mean level of BOLD in the urine samples was 3.19 +/- 1.65 ng/mL (range: 0.37 6.02 ng/mL). The concentrations of BOLD in the victim's blood from the femoral vein and heart were 140.44 and 25.74 ng/mL, respectively. On the other hand, those in the muscular tissue from the injection site and the urine sample were 142.3 ng/g and 3474 ng/mL, respectively.

Highly efficient synthesis of boldenone from androst-4-ene-3,17-dione by Arthrobacter simplex and Pichia pastoris ordered biotransformation.[Pubmed:30848360]

Bioprocess Biosyst Eng. 2019 Mar 8. pii: 10.1007/s00449-019-02092-y.

Boldenone (BD) is an important steroid hormone drug which is the derivative of testosterone. In this study, an ordered biotransformation method was proposed employing Arthrobacter simplex and recombinant Pichia pastoris with 17beta-hydroxysteroid dehydrogenase from Saccharomyces cerevisiae to produce BD from androst-4-ene-3,17-dione (AD) efficiently. To lower the oxidation towards BD in A. simplex, the transformation was conducted sequentially by C1,2 dehydrogenation in A. simplex and 17beta-carbonyl reduction in recombinant P. pastoris GS115. Moreover, the reaction system was inactivated before recombinant P. pastoris GS115 was added to further inhibit the oxidation of BD by A. simplex, and the productivity of BD was improved 10.6% compared with the control. Furthermore, by optimizing the conditions of transformation from AD to BD, 4.2 g/L BD was generated with 83% productivity from 5.0 g/L AD, which was the highest productivity reported by biological method. This study offers a promising method to produce BD by ordered biotransformation system, which can also be used to manufacture other steroidal compounds that are difficult to acquire directly.

Phytochemical profile, anti-oxidant, anti-inflammatory, and anti-proliferative activities of Pogostemon deccanensis essential oils.[Pubmed:30622869]

3 Biotech. 2019 Jan;9(1):31.

Essential oils (EOs) obtained from aerial parts of Pogostemon deccanensis were analyzed for GC-MS profiling, and evaluated for antioxidant, anti-inflammatory, and anti-proliferative activities. GC-MS analysis revealed a total of 47 constituents, establishing the EOs rich in sesquiterpene with > 20 sesquiterpenes constituting around 77% of the total EO yield. Major constituents included Curzerene (Benzofuran, 6-ethenyl-4,5,6,7-tetrahydro-3,6-dimethyl-5-isopropenyl-, trans-) (26.39%) and epi-Cadinol (22.68%), Ethanone, 1-(2,4,6-trihydroxyphenyl) (6.83%, Acetophenones), and Boldenone (3.47%, anabolic steroid). EOs found to be rich in phytochemicals attributed for antioxidant potentials of aromatic/medicinal plants, viz., flavonoids (2.71 microg quercetin equivalents g(-1) EO), total phenols (3.94 microg gallic acid equivalents (GAE) g(-1) EO), carotenoids (14.3 microg beta-carotene equivalents g(-1) EO), and ascorbic acid (2.21 microg ascorbic acid equivalents g(-1) EO). P. deccanensis EOs exhibited striking antioxidant activities assessed by wide range of assays including ferric reducing antioxidant potential (FRAP, 255.3 GAE at 2 microg mL(-1) EO), total antioxidant activity (TAA, 264.3 GAE at 2 microg ml(-1)) of EO, DPPH (65% inhibition at 2 microg mL(-1)), and OH (58% inhibition at 2 microg mL(-1)) scavenging. Interestingly, EOs showed considerably higher anti-lipid peroxidation activity than the standard antioxidant molecule ascorbic acid, with 50% protection by 1.29 microg mL(-1) EO against 20.0 microg mL(-1) standard. EOs showed strong anti-inflammatory activity with 50% inhibition at 1.95 microg mL(-1) EO. The anti-proliferative activity of EOs was tested against mouse cancer cell line and the EOs proved a potent anti-proliferative agent with only 2.1% cell survival at 2 microg mL(-1) EO, whereas the EOs were largely non-toxic-to-normal (non-cancerous) cells with approximately 80% cell survival at the 2 microg mL(-1) EOs. This being the first attempt of phytochemical profiling and wide array of biological activities of P. deccanensis EOs holds significance as the striking activities were observed at very low concentrations, in some cases at lower than the commercial standards, and has, therefore, great potential for pharmaceutical or commercial exploration.

Grape Seed Proanthocyanidin Ameliorates Cardiac Toxicity Induced by Boldenone Undecylenate through Inhibition of NADPH Oxidase and Reduction in the Expression of NOX2 and NOX4.[Pubmed:30116498]

Oxid Med Cell Longev. 2018 Jul 5;2018:9434385.

The effect of anabolic androgenic steroids on the cardiovascular system is poorly understood. Increased production of free radicals is coupled to the pathophysiology of many alterations within the circulatory system. The only function of the enzyme family NADPH oxidases (NOXs) is the generation of reactive oxygen species (ROS). Therefore, this study investigated the beneficial role of grape seed proanthocyanidin extract (GSPE) in ameliorating cardiac toxicity induced by the anabolic steroid Boldenone in male rats through NOX inhibition and reduction in the expression of NOX2 and NOX4. This study was conducted on forty male rats which are divided into four groups (normal control, positive control or GSPE, Boldenone, and posttreatment Boldenone with GSPE). A significant increase in relative body weight, relative heart weight, and hemodynamic parameters, as well as serum concentrations of lactate dehydrogenase, creatine kinase, creatine kinase-muscle brain, myoglobin, cholesterol, low-density lipoprotein cholesterol, risk factor 1/2, K(+), and Cl(-), in treated rats with Boldenone when compared with control. We also noted a significant increase in the levels of cardiac malondialdehyde, H2O2 generation in heart tissues, mRNA expression of NOX2 and NOX4, and immunoreactivity to proliferating cell nuclear antigen (PCNA). Posttreated rats with Boldenone and GSPE ameliorated cardiac toxicity via inhibition of NOX and a reduction in alteration of the expression of NOX2, NOX4, and PCNA induced by Boldenone. These novel insights into the antioxidative activity of GSPE should serve as a basis for the development of improved chemopreventive or therapeutic strategies for cardiac toxicity.