Laboratory of Veterinary Internal Medicine, Department of Clinical Sciences, Faculty of Veterinary Medicine, Hokkaido University, Kita 18-jo Nishi 9-chome, Kita-ku, Sapporo, Hokkaido 060-0818, Japan.
Laboratory of Toxicology, Department of Environmental Veterinary Sciences, Faculty of Veterinary Medicine, Hokkaido University, Kita 18-jo Nishi 9-chome, Kita-ku, Sapporo, Hokkaido 060-0818, Japan.
J Vet Med Sci. 2021 Oct 31;83(11):1634-1642. doi: 10.1292/jvms.21-0406. Epub 2021 Sep 17.
We developed an analytical method using an on-line column-switching liquid chromatography with triple quadrupole mass spectrometry (LC/MS/MS) for quantifying multiple steroids in serum. Using the developed method, we evaluated the serum concentration of nine steroids (cortisol, corticosterone, cortisone, 11-deoxycortisol, 21-deoxycortisol, deoxycorticosterone, progesterone, 17α-OH-progesterone and aldosterone) in dogs with hyperadrenocorticism (HAC). Serum was mixed with stable isotope internal standards and thereafter purified by the automated column-switching system. The limit of detection ranged 2-16 pg/ml for nine steroids. In the baseline samples, five steroids (cortisol, corticosterone, cortisone, 11-deoxycortisol, and 17α-OH-progesterone) were detected in all dogs. The concentrations of cortisone, 11-deoxycortisol, and 17α-OH-progesterone in dogs with HAC (n=19) were significantly higher those in dogs without HAC (n=15, P<0.02). After the adrenocorticotropic hormone stimulation test, six steroids (cortisol, corticosterone, cortisone, 11-deoxycortisol, 17α-OH-progesterone, and deoxycorticosterone) were above the limit of quantification in all dogs. Cortisol, corticosterone, cortisone, and deoxycorticosterone concentrations of dogs with HAC were significantly higher than those of dogs without HAC (P<0.02). In addition, 11-deoxycortisol and 17α-OH-progesterone concentration was higher in dogs with HAC than in dogs without HAC (P=0.044 and P=0.048, respectively). The on-line column-switching LC/MS/MS would be feasible for measuring multiple steroids in dog serum. The results suggest that cortisone, 11-deoxycortisol, and 17α-OH-progesterone would be related to HAC. Further studies are warranted to assess the clinical feasibility of steroid profile in dogs with HAC.
我们开发了一种使用在线柱切换液相色谱-串联三重四极杆质谱（LC/MS/MS）定量血清中多种类固醇的分析方法。使用所开发的方法，我们评估了患有库欣氏病（HAC）的犬的血清中 9 种类固醇（皮质醇、皮质酮、可的松、11-脱氧皮质醇、21-脱氧皮质醇、脱氧皮质酮、孕酮、17α-羟孕酮和醛固酮）的浓度。血清与稳定同位素内标混合，然后通过自动化柱切换系统进行纯化。9 种类固醇的检测限范围为 2-16 pg/ml。在基线样本中，所有犬均检测到 5 种类固醇（皮质醇、皮质酮、可的松、11-脱氧皮质醇和 17α-羟孕酮）。患有 HAC 的犬（n=19）的可的松、11-脱氧皮质醇和 17α-羟孕酮浓度明显高于未患有 HAC 的犬（n=15，P<0.02）。在促肾上腺皮质激素刺激试验后，所有犬的 6 种类固醇（皮质醇、皮质酮、可的松、11-脱氧皮质醇、17α-羟孕酮和脱氧皮质酮）的浓度均超过定量限。患有 HAC 的犬的皮质醇、皮质酮、可的松和脱氧皮质酮浓度明显高于未患有 HAC 的犬（P<0.02）。此外，患有 HAC 的犬的 11-脱氧皮质醇和 17α-羟孕酮浓度高于未患有 HAC 的犬（分别为 P=0.044 和 P=0.048）。在线柱切换 LC/MS/MS 可用于测量犬血清中的多种类固醇。结果表明，可的松、11-脱氧皮质醇和 17α-羟孕酮与 HAC 有关。需要进一步的研究来评估 HAC 犬类固醇谱的临床可行性。
