Caprioli Giovanni, Genangeli Michele, Mustafa Ahmed M, Petrelli Riccardo, Ricciutelli Massimo, Sagratini Gianni, Sartori Stefano, Laus Fulvio, Vittori Sauro, Cortese Manuela
School of Pharmacy, University of Camerino, 62032 Camerino, Italy.
Department of Pharmacognosy, Faculty of Pharmacy, Zagazig University, Zagazig 44519, Egypt.
Pharmaceuticals (Basel). 2021 Apr 21;14(5):393. doi: 10.3390/ph14050393.
A simple and fast analytical method able to simultaneously identify and quantify 17 endogenous and exogenous steroidal hormones was developed in bovine and equine blood using UHPLC-MS/MS. A total amount of 500 µL of sample was deproteinized with 500 µL of a mixture of methanol and zinc sulfate and evaporated. The mixture was reconstituted with 50 µL of a solution of 25% methanol and injected in the UHPLC-MS/MS triple quadrupole. The correlation coefficients of the calibration curves of the analyzed compounds were in the range of 0.9932-0.9999, and the limits of detection and quantification were in the range of 0.023-1.833 and 0.069-5.5 ppb, respectively. The developed method showed a high sensitivity and qualitative aspects allowing the detection and quantification of all steroids in equine and bovine blood. Moreover, the detection limit of testosterone (50 ppt) is half of the threshold admitted in plasma (100 ppt). Once validated, the method was used to quantify 17 steroid hormones in both bovine and equine blood samples. The primary endogenous compounds detected were corticosterone (range 0.28-0.60 ppb) and cortisol (range 0.44-10.00 ppb), followed by androstenedione, testosterone and 11-deoxycortisol.
采用超高效液相色谱-串联质谱法（UHPLC-MS/MS）建立了一种简单快速的分析方法，可同时鉴定和定量牛血和马血中的17种内源性和外源性甾体激素。取500μL样品，加入500μL甲醇和硫酸锌的混合物进行脱蛋白处理，然后蒸发。混合物用50μL含25%甲醇的溶液复溶，注入UHPLC-MS/MS三重四极杆质谱仪。分析化合物校准曲线的相关系数在0.9932至0.9999之间，检测限和定量限分别在0.023至1.833和0.069至5.5 ppb范围内。所建立的方法具有高灵敏度和定性能力，可检测和定量马血和牛血中的所有类固醇。此外，睾酮的检测限（50 ppt）是血浆中允许阈值（100 ppt）的一半。经验证后，该方法用于定量牛血和马血样品中的17种类固醇激素。检测到的主要内源性化合物是皮质酮（范围为0.28至0.60 ppb）和皮质醇（范围为0.44至10.00 ppb），其次是雄烯二酮、睾酮和11-脱氧皮质醇。
