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The retrospective analysis of endogenous steroid hormones in nails can be used to elucidate endocrine diseases and thus help with their diagnosis and treatment. A liquid chromatographytandem mass spectrometry (LC-MS/MS) based method was developed for the simultaneous identification and quantification of 12 steroid hormones (aldosterone, cortisone, cortisol, corticosterone, 11deoxycortisol, androstenedione, testosterone, dehydroepiandrosterone (DHEA), 17-hydroxyprogesterone (17-OHP), dihydrotestosterone (DHT) and progesterone) in human fingernails. Steroid hormones were extracted from 0.5 mg -10 mg pulverized nail clippings by methanolic extraction, followed by a liquid-liquid extraction. The analysis was conducted with LC-MS/MS in electrospray ionization positive mode. The method was validated in terms of linearity, limit of detection (LOD), limit of quantification (LOQ), precision, accuracy, matrix effect, recovery and robustness. It was successfully applied for steroid profiling in nails of mothers and their infants where cortisol, cortisone, testosterone, progesterone, androstenedione and 11-deoxycorticosterone could be detected. Furthermore, it could be shown that there is no significant difference in concentrations between left and right hand for cortisol, cortisone and progesterone. A positive linear correlation between cortisol and cortisone in nails was found. In conclusion, it could be shown that nails are a suitable matrix for the retrospective monitoring of cumulative steroid hormone levels.Abstract: The retrospective analysis of endogenous steroid hormones in nails can be used to elucidate endocrine diseases and thus help with their diagnosis and treatment. A liquid chromatography-tandem mass spectrometry (LC-MS/MS) based method was developed for the simultaneous identification and quantification of 12 steroid hormones (aldosterone, cortisone, cortisol, corticosterone, 11-deoxycortisol, androstenedione, 11-deoxycorticosterone, testosterone, dehydroepiandrosterone (DHEA), 17α-hydroxyprogesterone (17-OHP), dihydrotestosterone (DHT) and progesterone) in human fingernails. Steroid hormones were extracted from 0.5 mg -10 mg pulverized nail clippings by methanolic extraction, followed by a liquid-liquid extraction. The analysis was conducted with LC-MS/MS in electrospray ionization positive mode. The method was validated in terms of linearity, limit of detection (LOD), limit of quantification (LOQ), precision, accuracy, matrix effect, recovery and robustness. It was successfully applied for steroid profiling in nails of mothers and their infants where cortisol, cortisone, testosterone, progesterone, androstenedione and 11-deoxycorticosterone could be detected. Furthermore, it could be shown that there is no significant difference in concentrations between left and right hand for cortisol, cortisone and progesterone. A positive linear correlation between cortisol and cortisone in nails was found. In conclusion, it could be shown that nails are a suitable matrix for the retrospective monitoring of cumula...

This study aimed to investigate correlation between hair cortisol levels and perceived stress scale (PSS) in addition to a range of demographic and physiological factors in a sample of older adults. ExperimentalHair cortisol concentrations were established from 42 older adults aged between 60 and 80 years old. Age, sex, hair colour, smoking status, employment status, daytime sleeping, medication, waist to hip ratio (WHR) and PSS scores were assessed through a questionnaire. Hair cortisol concentration was assessed through liquid chromatography coupled to tandem mass-spectrometry (LC-MS/MS). ResultsAmongst the older adult group there was no statistically significant correlation between hair cortisol concentration and age, employment status, daytime sleep duration, WHR or PSS. When compared to previous data assessing hair cortisol in toddlers (7 months to 3 years old), adolescents (12-17 years old) and adults (18-60 years old) it is observed that there is a trend for higher hair cortisol in older adults (60-80 years old). Hair cortisol concentrations were significantly higher in males (n = 20) than in females (n = 22) and in participants with dark brown hair (n = 8). No relationship was investigated between hair cortisol concentration and smoking status or medication intake. ConclusionsThe results confirm that hair samples are a useful alternative to the current mediums that are used to analyse biological cortisol. The results also validate the use of LC-MS/MS as an effective analytical method for the quantitation of hair cortisol concentrations.

Hair samples are increasingly used for measuring the long-term stress mediator cortisol. However, hair is not always available and nails (finger or toe), as a keratinized matrix, may be an alternative to hair. In order to measure cortisol and cortisone in the nail matrix, an LC-MS/MS method has been developed and validated using C-labeled surrogate analytes. Both analytes were measured in ESI negative mode as formic acid adducts. Different sample preparation techniques were assessed, and single-step extraction in methanol was established for determination of cortisone and cortisol in the nail matrix. The method was successfully validated with limits of detection (LOD) and limits of quantification (LOQ) of 0.5 and 1.0 pg/mg for cortisol and cortisone, respectively. The calibration curve was linear up to a concentration of 500 pg/mg. Recovery was good for both analytes and showed values over 50%. Matrix effects with ion suppression occurred for both substances but could be corrected by the use of internal standard. Accuracy and precision were in the accepted range of ± 20% for both substances. The method was successfully applied to determine cortisol and cortisone concentrations in authentic nail samples. Cortisol and cortisone concentrations varied significantly among different fingernails, being highest in the little fingernails and lowest in the thumbnails. It could be shown that even in only 1 mg nail sample cortisol and cortisone can be reliably quantified. No correlation between hair and nail cortisol and cortisone concentrations could be found. Furthermore, cortisol and cortisone concentrations were significantly higher in hair. Graphical abstract.