Urine cortisol

OH-cortisol (urine, 379 121 43-515 51-515 43-295 Quest Diagnostics (pers. comm.)... 

LC-MS with on-line SPE using a RAM pre-column with an internal ODS phase was described by van der Hoeven et al. (95) for the analysis of cortisol and prednisolone in plasma, and arachidonic acid in urine. The samples were injected directly and the only off-line pretreatment required was centrifugation. By using the on-line SPE-LC-MS system, cortisol and related compounds could be totally recovered and quantified in 100 p.1 plasma within 5 min with a typical detection of 2 ng/ml (Figure 11.6(b)). The RAM-type of sorbents, in which the outer surface of the particles is covered with aj-acid glycoprotein, also appear to be useful for direct SPE of... 

Hour urinary Collect urine over 24 Urinary free cortisol greater Easy to perform but should not be used... 

The presence of hypercortisolism can be established with a midnight plasma cortisol, late-night (11 PM) salivary cortisol, 24-hour urine free cortisol, and/or low-dose dexamethasone suppression test. 

The application from van der Hoeven et al. (1997) used an ADS cartridge online SPE to measure cortisol and prednisolone in plasma and arachidonic acid in urine. A precolumn packed with a C18 alkyl-diol support (LiChrosphere RP-18 ADS, 25 /an, Merck) was used. To reduce run time, column switching was programmed as heart-cut , diverting only the analyte fraction into the analytical column. Another LiChrosphere column (125 x 4 mm inner diameter, Merck) handled separation. After the injection of 100 fiL plasma, the lower limit of detection for prednisolone was 1 ng/mL while cortisol was readily quantitated at its endogenous level of 100 ng/mL. The run time was 5 min. For arachidonic acid, a Hypersil ODS column (200 x 3.0 mm inner diameter, 5 /.an) was used. The injection volume was 200 //I. and run time was 9.5 min. The detection limit was 1 ng/mL and recovery was 77%. 

Barrett Y.C. et al., 2005. Automated online SPE LC-MS/MS method to quantitate 6-beta-hydroxycortisol and cortisol in human urine. J Chromatogr B 821 159. 

Abu Ruz S, Millership J, Heaney L, McElnay J. 2003. Simple liquid chromatography method for the rapid simultaneous determination of prednisolone and cortisol in plasma and urine using hydrophilic lipophilic balanced solid phase extraction cartridges. J Chromatogr B Biomed Sci Appl 798(2) 193-201. 

Suppression tests For Cushing syndrome, give 1 mg at 11 pm. Draw blood for plasma cortisol determination the following day at 8 am. For greater accuracy, give 0.5 mg every 6 hours for 48 hours. Collect 24-hour urine to determine 17-hydroxycorticosteroid excretion. 

The initial report of sustained, lower urinary cortisol levels in PTSD highlighted the disassociation between cortisol and catecholamine levels in PTSD. Norepinephrine and epinephrine levels assayed from the same urine specimens revealed elevations in both of these catecholamines, while cortisol levels in PTSD fell within the normal range of 20-90 pg/day, indicating that the alteration was not in the hypoadrenal or endocrinopathologic range (Mason et al. 1986). This finding established the expectation that alterations in basal levels of cortisol might be subtle, and not easily differentiated from normal values (Mason et al. 1986). 

Denotes findings in which cortisol levels were significantly lower than comparison subjects, or, in the case of Kosten et al., from depression only. Denotes findings in which cortisol levels were significantly higher than comparison subjects. Results are from a 12-h rather than 24-h urine collection and are expressed as pg/12h. 

System (2) has been employed for the determination of cortisol and cortisone in urine [150]. Urine containing 6a-methyl prednisolone as an internal standard was applied to a Sep-Pak Cig cartridge, eluted with aqueous 90% acetonitrile, and the eluent analyzed by HPLC. The analytical column (25 cm x 4.6 mm) is packed with Altex Ultrasphere C s bonded silica (5 pm), and requires the use of a (5 cm x 5 mm) Hypercil ODS (5 pm) pre-column. The mobile phase used gradient elution with less than 100% acetonitrile, and detection is by UV absorbance at 260 nm. 

It is sometimes necessary to suppress the production of ACTH to identify the source of a particular hormone or to establish whether its production is influenced by the secretion of ACTH. In these circumstances, it is advantageous to use a very potent substance such as dexamethasone because the use of small quantities reduces the possibility of confusion in the interpretation of hormone assays in blood or urine. For example, if complete suppression is achieved by the use of 50 mg of cortisol, the urinary 17-hydroxycorticosteroids will be 15-18 mg/24 h, since one third of the dose given will be recovered in urine as 17-hydroxycorticosteroid. If an equivalent dose of 1.5 mg of dexamethasone is used, the urinary excretion will be only 0.5 mg/24 h and blood levels will be low. 

In patients in whom the diagnosis of Cushing s syndrome has been established clinically and confirmed by a finding of elevated free cortisol in the urine, suppression with large doses of dexamethasone will help to distinguish patients with Cushing s disease from those with steroid-... 

The metabolism of aldosterone is similar to that of cortisol, about 50 mcg/24 h appearing in the urine as conjugated tetrahydroaldosterone. Approximately 5-15 mcg/24 h is excreted free or as the 3-oxo glucuronide. 

Metyrapone is commonly used in tests of adrenal function. The blood levels of 11-deoxycortisol and the urinary excretion of 17-hydroxycorticoids are measured before and after administration of the compound. Normally, there is a twofold or greater increase in the urinary 17-hydroxycorticoid excretion. A dose of 300-500 mg every 4 hours for six doses is often used, and urine collections are made on the day before and the day after treatment. In patients with Cushing s syndrome, a normal response to metyrapone indicates that the cortisol excess is not the result of a cortisol-secreting adrenal carcinoma or adenoma, since secretion by such tumors produces suppression of ACTH and atrophy of normal adrenal cortex. 

The measurement of these compounds in urine is essential for the study of Cushings disease, glucocorticoid remediable aldosteronism (GRA), apparent mineralocorti-coid excess syndrome (AME), and related conditions. While RIA has been generally satisfactory for serum cortisol assay, urine contains many crossreacting steroids, rendering RIA unreliable. 

Taylor and colleagues [98] at the Mayo Clinic published a method for the simultaneous analysis of urinary cortisol and cortisone. They used 2H4 cortisol as an internal standard and took a 0.5-ml urine sample. An API 2000 with Turboion-spray source was used in the positive-ion mode. Chromatography was conducted on a standard-bore C18 column with Q8 precolumn filter. MRM was conducted in the positive-ion mode monitoring m/z 363—>121 for cortisol, 367—>121 for 2TL, cortisol, and 361— -121 for cortisone. Cortisol and cortisone were separated and both were eluted within 2 min. Inter- and intra-assay variation for both compounds was < 9% for amounts above 2 pig/dl. The values obtained agree well with those of other studies, such as ours (Table 5.3.2) [62]. They found a range for cortisol for adult males of 4.2-60 pg/24 h and for adult females 3.0-43 pg/24 h. In summary, the 3-min run time of their method has allowed the Mayo group to completely transfer their cortisol and cortisone workload from RIA and HPLC to MS/MS. 

Quest Diagnostics use an LC-MS/MS panel for diagnosing cortisol-related disorders by urine analysis. This panel was designed to diagnose Cushings syndrome and the hypertensive conditions AME and GRA. The panel quantifies cortisone, cortisol, 6/j-hydroxycorlisol and 18-hydroxycortisol (18-OHF). The Quest analysis uses 2H4 cortisol as an internal standard and HTLC for on-line extraction. This panel has replaced the RIA and HPLC methods previously used by this commercial laboratory. Another recent publication describes MS/MS of cortisone and cortisol in serum using APPI and similar conditions and MRM transformations to those listed above [43]. 

OHF is almost exclusively excreted without undergoing -ring reduction and conjugation and can be measured in urine by HPLC-MS. A useful diagnostic ratio is that of 18-OHF UFF, which is normally about 5 1 and rises to 50 1 in GRA. Serum analysis is used to measure aldosterone (elevated in the disorder) and cortisol, but not typically for 18-OHF. 

Palermo M, Gomez-Sanchez C, Roitman E, Shackleton CH (1996) Quantitation of cortisol and related - -4-ene steroids in urine using gas chromatography/mass spectrometry with stable isotope-labeled internal standards. Steroids 61 583-589... 

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