Department of Clinical Chemistry, VU University Medical Center, PO Box 7057, Amsterdam 1007MB, The Netherlands.
Ann Clin Biochem. 2010 May;47(Pt 3):248-52. doi: 10.1258/acb.2010.009171. Epub 2010 Apr 20.
Differentiation between subtle changes in low serum testosterone concentrations, common in women and children, is not possible with current commercially available assays. The objectives of the study were to develop a method based on stable isotope dilution-liquid chromatography-tandem mass spectrometry (ID-LC-MS/MS) with adequate sensitivity and specificity and to investigate the applicability of this assay in serum samples from pre- and postmenopausal women.
For 16 women, testosterone levels were measured in blood samples drawn two years before and after physiological menopause, and for eight women in samples drawn before and after bilateral oophorectomy. Testosterone was extracted from serum, derivatized and analysed on an LC-MS/MS.
The developed ID-LC-MS/MS method allowed for specific and reproducible measurement of testosterone. Comparison with stable isotope dilution-gas chromatography coupled to mass spectrometry detection by Deming regression analysis gave a slope of 1.025 and an intercept of 0.055 nmol/L (r = 0.9998). A significant decrease was found in testosterone concentrations before and after bilateral oophorectomy (P = 0.02), whereas no significant difference was found before and after natural menopause (P = 0.4).
The ID-LC-MS/MS assay measures serum testosterone with acceptable accuracy and is useful in female samples, supporting the conclusion that the postmenopausal ovary contributes to circulating testosterone. To our knowledge, our analytical method compares favourably to similar published methods in terms of sensitivity. The sensitivity and specificity of this method comply with the reference method for measurement of testosterone in serum samples of women, children and men suffering from hypogonadism and can also be used for men with testosterone in the reference range.
目前市售的检测方法无法区分血清睾酮浓度的细微变化，而这种变化在女性和儿童中很常见。本研究的目的是开发一种基于稳定同位素稀释-液相色谱-串联质谱法（ID-LC-MS/MS）的方法，该方法具有足够的灵敏度和特异性，并探讨该方法在绝经前和绝经后妇女血清样本中的适用性。
对 16 名女性在生理绝经前两年和绝经后两年的血样进行了睾酮水平检测，对 8 名女性在双侧卵巢切除术前和术后的血样进行了检测。从血清中提取睾酮，衍生化后在 LC-MS/MS 上进行分析。
所开发的 ID-LC-MS/MS 方法可特异性和可重复地测量睾酮。通过 Deming 回归分析与稳定同位素稀释-气相色谱-质谱联用检测方法进行比较，得到的斜率为 1.025，截距为 0.055 nmol/L（r = 0.9998）。双侧卵巢切除术前和术后的睾酮浓度显著下降（P = 0.02），而自然绝经前后的睾酮浓度无显著差异（P = 0.4）。
ID-LC-MS/MS 检测法可准确测量血清睾酮，对女性样本具有实用性，支持绝经后卵巢可产生循环睾酮的结论。据我们所知，就灵敏度而言，我们的分析方法与类似的已发表方法相比具有优势。该方法的灵敏度和特异性符合女性、儿童和患有性腺功能减退症的男性血清睾酮检测的参考方法，也可用于处于参考范围内的男性睾酮检测。
