Simultaneous Analysis and Efficient Separation of Anabolic Androgenic Steroids in Dietary Supplement by a Validated HPTLC Method.

Background: Using sports supplements is common among athletes. The presence of anabolic steroids in sports supplements as a hormonal contaminant can increase production efficiency. Since anabolic steroids cause health problems and result in positive doping tests in athletes, it is important to investigate their presence in the supplement preparations consumed by athletes. Objectives: This paper aims to simultaneously determine ten anabolic steroids by high-performance thin-layer chromatography (HPTLC) method in sports supplements. Methods: Chromatographic analysis was conducted on glass silica gel 60F254 plates. The extracts loaded on silica gel plates are subjected to programed multiple development (PMD) to separate anabolic androgenic steroids (AASs). Densitometric scanning is carried out at the wavelength of 245 and 366nm. The method was validated according to the ICH guidelines. Results: Spots at retardation factor (Rf) 0.72 (elution system 1), 0.4 (elution system 1), 0.29 (elution system 2), 0.25 (elution system 2), 0.1 (elution system 1), 0.65 (elution system 2), 0.59 (elution system 1), 0.44 (elution system 1), 0.8 (elution system 3), and 0.82 (elution system 3) values were recognized as 19-nor androstenedione, 19-nortestosterone, methyl testosterone, clostebol, stanozolol, trenbolone enanthate, oxymetholone, oxandrolone, testosterone enanthate, and nandrolone decanoate, respectively. The linear ranges were 25 - 250 µg/mL for oxymetholone, 7 - 50 µg/mL for 19-nor androstenedione, 19-nortestosterone, and oxandrolone, and 3 - 20 µg/mL for methyl testosterone, clostebol, stanozolol, trenbolone enanthate, testosterone enanthate, and nandrolone decanoate. The developed method is validated by acceptable precision (CV < 20%) and good accuracy (94% < R < 114%). The value of limit of detection (LOD) for all derivatives was in the range of 0.02 - 0.16 µg/spot (20-160 µg/g of supplement), while limit of quantitation (LOQ) was found to be in the range of 0.06 - 0.5 µg/spot (60 - 500 µg/g of supplement). Fifty sports supplement samples as real sample were collected and analyzed. None of the samples screened positive using the HPTLC method. Conclusions: In the present study, the fast, cheap, and simple HPTLC method could be used for the multi-residue analysis of ten anabolic androgenic steroids in sports supplements.

Survey of protein-based sport supplements for illegally added anabolic steroids methyltestosterone and 4-androstenedione by UPLC-MS/MS.

There is some evidence that marketable supplements contain hormones not declared on the product label. The presence of these androgenic anabolic steroids (AAS) in sports supplements can be considered an adulteration and affect the health of consumers, who are predominantly athletes. This study aimed to measure anabolic hormones (methyltestosterone and 4-androstenedione) in sport supplements. Ultra Performance Liquid chromatography coupled mass spectrometry (UPLC-MS/MS) with electrospray ionization (ESI) in positive mode was employed under the Multiple Reaction Monitoring (MRM) ion program. To overcome matrix effects and quantify the selected analyte, the calibration curve was made using Matrix Match method. The LOQ and LOD were 1 ng/g and 0.3 ng/g for both analytes. The recovery of 4-androstenedione and methyltestosterone was in the range of 86.87-107.35 and 77.31-113.98, respectively. In terms of reproducibility, CV % for 4-androstenedione and methyltestosterone ranged from 6.56 to 16.87% and 1.45-15.12%, respectively. 4-androstenedione was found in 11 samples including 9 whey as 1.578 ±â€¯0.154 ng/g and 2 whey albumin samples with an amount of 1.134 ng/g and 1.474 ng/g. Consequently, continuous controlling of sport supplements comprising intentionally or unintentionally added androgens could be important for health and discuss in the context of compliance with anti-doping.

Probabilistic non-carcinogenic and carcinogenic risk assessments (Monte Carlo simulation method) of the measured acrylamide content in Tah-dig using QuEChERS extraction and UHPLC-MS/MS.

A modified "Quick, Easy, Cheap, Effective, Rugged, and Safe" QuEChERS in combination with Ultra-High Performance Liquid Chromatography/tandem mass spectrometry (UHPLC-MS/MS) method was optimized for the determination of acrylamide content in different types of tah-dig (rice, bread, and potato). Also, the non-carcinogenic and carcinogenic risks (target hazard quotient (THQ) and incremental lifetime cancer risk (ILCR)) due to ingestion of acrylamide via tah-dig in the adults and children were assessed by Monte Carlo simulation (MCS) method. The recoveries of acrylamide at five concentration levels (n = 3) ranged from 83.82% to 106.41%. The repeatability of the proposed method was demonstrated with RSD% in the range of 11.3-20%. In addition, the limits of detection and quantification were reported as 5 ngg-1 and 15 ngg-1, respectively. The mean levels of the acrylamide contents in rice tah-dig, bread tah-dig, and potato tah-dig were measured as 24.65 ngg-1, 39.48 ngg-1, and 714.11 ngg-1, respectively. The highest acrylamide content was determined in potato tah-dig (2100 ngg-1) and the lowest acrylamide in rice tah-dig (≤LOQ). Based on the conducted risk assessment, the P (95%) of cumulative probability in the MCS method, the lowest and highest THQ was observed in the adults (ingestion bread tah-dig: 1.29E-2), and children (ingestion potato tah-dig: 1.90E+00), respectively. Additionally, the lowest and highest ILCR were reported in adults (ingestion bread tah-dig: 1.29E-5) and children (ingestion potato tah-dig: 7.49E-3), respectively. The rank order of type tah-dig based on THQ and ILCR for all groups of consumers was potato tah-dig > rice tah-dig > bread tah-dig. There is a considerable non-carcinogenic risk for the children due to ingestion potato tah-dig (THQ > 1). Additionally, the significant carcinogenic risk for the Iranian adults and children due to consumption of rice, bread, and potato tah-dig (ILCR > 1.00E-5) was observed.

An applicable strategy for improvement recovery in simultaneous analysis of 20 pesticides residue in tea.

It is important to have a reliable method to analyze pesticides in tea, a beverage commonly consumed in Iran. A validated method was developed for the determination of 20 pesticides in tea based on QuEChERS sample preparation and capillary gas chromatography-quadrupole mass spectrometry in selective ion monitoring mode (GC-MS/SIM) using triphenyl methane (TPM) solution as an internal standard. We used fortified, extracted, and cleaned-up tea samples instead of calibration standards for quantitation, which substantially reduced adverse matrix-related effects and negative recovery affected by graphite carbon black (GCB) on pesticide analysis. The recovery of pesticides at 3 concentration (40, 60, and 240 ng/g) ranged from 79.5% to 111.4% (n = 3). The method had acceptable repeatability with RSDr < 20%. The limits of quantification (LOQ) for all pesticides were ≤20 ng/g. The analytical results of the proposed method were in good agreement with proficiency test results (FAPAS, 19116). The recoveries and repeatabilities were in accordance with the criteria set by SANCO Guideline. The validated method was suitable for the analysis of pesticides in tea.