Evaluation of the short-term stability of 6α-chloro-testosterone, 6β-bromo-androstenedione and 6-oxo-androstenedione in dimethylsulfoxide and methanol using liquid and gas chromatography − mass spectrometry

IF 2.1 4区医学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY

Steroids Pub Date : 2025-03-09 DOI:10.1016/j.steroids.2025.109597

Dayamin Martínez_Brito , Patrizia Leogrande , Xavier de la Torre , Francesco Botrè

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引用次数: 0

Abstract

This work studied the short-term stability of 6α-chloro-testosterone (6-CT), 6β-bromo-androstenedione (6-BrAED) and 6-oxo-androstenedione (6-oxo-AED) in methanol (MeOH) and dimethylsulfoxide (DMSO) solutions by gas and liquid chromatography coupled to mass spectrometry.

Solutions of 6-CT, 6-BrAED and 6-oxo-AED were prepared in MeOH and DMSO. They were stored at room temperature, +4°C and −20 °C. Measurements were made at 0, 7, 30, 60 and 90 days after solutions preparation, by liquid chromatography-tandem mass spectrometry and gas chromatography-high resolution mass spectrometry.

Although the degradation of 6-CT and 6-BrAED was extensive, the most notable result was that the higher degradation occurred in DMSO instead of MeOH. The interaction of DMSO with halogenated species and secondary hydroxyl groups favored the degradation of these compounds by forming chemically related species. No degradation of 6-oxo-AED in either MeOH or DMSO was observed.

Pronounced degradation of the 6-CT and 6-BrAED, during the derivatization reaction for the gas chromatography-mass spectrometry analysis was observed. Because of the acidic condition of the reaction and depending on the reactant, it was favored the loss of the halogen molecule or the dehydration reaction to form the unsaturated (Δ6) steroid derivative.

Our finding suggests to take duly into account the possibility of degradation processes when performing quantitative determination of 6-CT, 6-BrAED and 6-oxo-AED by chromatographic-spectrometric techniques based on the use of reference solutions stored for sufficiently long times.

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液相和气相色谱-质谱法评价6α-氯睾酮、6β-溴雄烯二酮和6-氧雄烯二酮在二甲亚砜和甲醇中的短期稳定性。

采用气液相色谱联用质谱法研究了6α-氯睾酮（6-CT）、6β-溴雄烯二酮（6-BrAED）和6-氧-雄烯二酮（6-氧- aed）在甲醇（MeOH）和二甲基亚砜（DMSO）溶液中的短期稳定性。在MeOH和DMSO中制备6-CT、6-BrAED和6-oxo-AED溶液。分别保存于室温、+4°C和-20 °C。分别于溶液制备后0、7、30、60和90 d采用液相色谱-串联质谱法和气相色谱-高分辨率质谱法进行测定。虽然6-CT和6-BrAED的降解是广泛的，但最显著的结果是DMSO的降解率高于MeOH。DMSO与卤代物质和次生羟基的相互作用有利于这些化合物的降解，形成化学相关物质。在MeOH或DMSO中均未观察到6-oxo-AED的降解。在衍生化反应中，6-CT和6-BrAED明显降解，用于气相色谱-质谱分析。由于反应的酸性条件和根据反应物的不同，卤素分子的损失或脱水反应更有利于形成不饱和（Δ6）类固醇衍生物。我们的发现表明，在使用储存足够长时间的参比溶液进行6-CT、6-BrAED和6-oxo-AED定量测定时，色谱-光谱技术应充分考虑降解过程的可能性。

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来源期刊

Steroids 医学-内分泌学与代谢

CiteScore

5.10

自引率

3.70%

发文量

120

审稿时长

73 days

期刊介绍： STEROIDS is an international research journal devoted to studies on all chemical and biological aspects of steroidal moieties. The journal focuses on both experimental and theoretical studies on the biology, chemistry, biosynthesis, metabolism, molecular biology, physiology and pharmacology of steroids and other molecules that target or regulate steroid receptors. Manuscripts presenting clinical research related to steroids, steroid drug development, comparative endocrinology of steroid hormones, investigations on the mechanism of steroid action and steroid chemistry are all appropriate for submission for peer review. STEROIDS publishes both original research and timely reviews. For details concerning the preparation of manuscripts see Instructions to Authors, which is published in each issue of the journal.