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Research Article

Development and validation of HPLC-DAD methodology for simultaneous qualitative and quantitative determination of thirteen substances with a steroid structure

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Article: 2209660 | Received 13 Feb 2023, Accepted 28 Apr 2023, Published online: 09 May 2023

Figures & data

Figure 1. The target AAS included in nutritional supplements for body building.

Figure 1. The target AAS included in nutritional supplements for body building.

Table 1. Preparation of standard stock solutions in methanol.

Table 2. Elution conditions.

Figure 2. Chromatogram of a mix of targeted compounds at λ = 254 nm, where 13.229 is trenbolone, 13.784 is nandrolone, 14.485 is androstenedione, 16.426 is methanedienone, 19.613 is testosterone, 25.020 is methyltestosterone, 26.073 is trenbolone acetate, 34.209 is nandrolone propionate, 36.748 is testosterone propionate, 46.508 is trenbolone enanthate, 49.498 is testosterone enanthate, 52.780 is nandrolone decanoate and 54.477 is boldenone undecylenate.

Figure 2. Chromatogram of a mix of targeted compounds at λ = 254 nm, where 13.229 is trenbolone, 13.784 is nandrolone, 14.485 is androstenedione, 16.426 is methanedienone, 19.613 is testosterone, 25.020 is methyltestosterone, 26.073 is trenbolone acetate, 34.209 is nandrolone propionate, 36.748 is testosterone propionate, 46.508 is trenbolone enanthate, 49.498 is testosterone enanthate, 52.780 is nandrolone decanoate and 54.477 is boldenone undecylenate.

Table 3. Coefficient of determination R2 and RSD.

Table 4. Linearity range for the target compounds.

Figure 3. Chromatogram of a standard solution—a mixture of all 13 target compounds obtained by optimizing the chromatographic conditions.

Figure 3. Chromatogram of a standard solution—a mixture of all 13 target compounds obtained by optimizing the chromatographic conditions.

Table 5. Method repeatability, precision and recovery of HPLC-DAD procedure for simultaneous quantification of 13 target AAS.

Supplemental material

Supplemental Material

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Data availability statement

All data that support the findings reported in this study are available from the authors Z.Z. and D.D. upon reasonable request.