(T1130-05-31) Development and Validation of Quantitative Analysis Method for Testosterone Derivatives in Natural musk(Moschus) Using GC-MS

Purpose: It is necessary to develop an adequate quality control method to maintain herbal medicine’s quality above a certain level. Typically ‘Natural musk(Moschus, M. moschiferus)’ is one of the most expensive herbal medicine, it’s been required to develop an adequate quality control method for ‘Natural musk’ to prevent forgery and maintain its quality in regulatory authorities view. In this study, we’ve developed an analytical method that can simultaneously analyze the testosterone derivatives in the ‘Natural musk’, and the method has been validated in several parameters.

Methods: We’ve selected 6 reference standards among the testosterone derivatives reported to be contained in the ‘Natural musk’, referring the references. Analytical methods was developed using Agilent 7890B GC System and 7000C GC/MS Triple Quad mass spectrometer and the analysis mode was performed by EI technique. The developed method has been verified and validated in several parameters, in terms of linearity, limits of detection, limits of quantitation, accuracy, precision, and system suitability in accordance with the “test method validation guidelines for drugs” proposed by MFDS, Korea.

Results: As a result, the R² value was more than 0.99 and the limits of detection was 0.13 to 3.70 mg/mL, the limits of quantification was 0.75 ~ 7.5 mg/mL. Also, the range of recovery rate was 72.2% to 107.3 %, the range of relative standard deviation was 3.95 to 9.35 %, and the range of system suitability was confirmed as 1.23% to 3.19%. Therefore, the developed method satisfied all the requirements suggested in the guidelines.

Conclusion: It seems quite necessary to control the quality of expensive herbal medicines such as ‘Natural musk’ in regulatory authorities view. Thus, the analysis method verified in this study can be useful in proposing scientific data for establishing quantitative standards for musk in the future.

References: (1) The Korean Herbal Pharmacopoeia 4th edition, 2019
(2) Journal of Ethnopharmacology, Shuquan Lv et al., 2022, 284, 114799
(3) Journal of Steroid Biochemistry & Molecular Biology, Andreu Fabregat et al., 2013, SBMB-3939; No. of Pages 9
(4) J. of the Korean Society for Environmental Analysis, Chang-Kyu Kim, et al., 2009, 307~313

Acknowledgements: This research was supported by a grant (22201MFDS144) from Ministry of Food and Drug Safety in 2023.

Table. GC-MS Analysis condition.

Figure. Calibration curve of six reference standard.

Table. Results of method validation.