Liquid Chromatography–Tandem Mass Spectrometry Analysis of Urinary Free Cortisol

Kushnir, Mark M.; Rockwood, Alan L.; Nelson, Gordon J.; Terry, Alan H.; Meikle, A.W.

doi:10.1373/49.6.965

Cited by 64 publications

(35 citation statements)

References 5 publications

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“…24-h UFC reflects overall daily cortisol production and, compared to serial blood sampling, is relatively easy to collect in a large study population. An important advantage of this method is its reliability [36], although it is crucially dependent on the quality and performance of the particular analytical procedure applied. Indeed, several conditions may result in falsely elevated values such as alcoholism, high fluid intake, pregnancy, psychiatric disorders, or in falsely low levels such as cyclic CS or severe renal function [37,38].…”

Section: Discussionmentioning

confidence: 99%

The degree of urinary hypercortisolism is not correlated with the severity of cushing’s syndrome

et al. 2016

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Cushing syndrome (CS) is characterized by increased morbidity and mortality compared to the general population. However, there are patients who have more clinical aggressive forms than others. Aim of the study is to evaluate whether the degree of hypercortisolism, defined by the number of times urinary free cortisol (UFC) levels exceed the upper limit of the normal range (ULN), is related to the worsening of phenotypic features, as well as metabolic and cardiovascular parameters, in a cohort of CS patients. A cross-sectional study was conducted on 192 patients with active CS, consecutively presenting at the outpatients' clinic of the University Hospitals of Ancona, Naples, and Palermo. Patients were grouped into mild (UFC not exceeding twice the ULN), moderate (2-5 times the ULN), and severe (more than 5 times the ULN) hypercortisolism. Thirty-seven patients (19.3 %) had mild, 115 (59.8 %) moderate, and 40 (20.9 %) severe hypercortisolism. A significant trend of increase among the three groups was demonstrated for 8-, 16-, and 24-h serum cortisol levels (p \ 0.001) and serum cortisol after low dose of dexamethasone suppression test (p = 0.001). No significant trend of increase was found regarding phenotype and comorbidities. The degree of hypercortisolism by itself does not appear to be a sufficient parameter to express the severity of CS. Therefore, estimating the severity of CS according to biochemical parameters remains a challenge, while the clinical phenotype and the associated comorbidities might be more useful to assessing the severity of the CS.

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Section: Discussionmentioning

confidence: 99%

The degree of urinary hypercortisolism is not correlated with the severity of cushing’s syndrome

et al. 2016

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“…A introdução na rotina, de maneira pioneira, por vários laboratórios de referência internacional da nova metodologia na dosagem de cortisol livre urinário tem várias razões (25)(26)(27). Dentre estas, além da concentração relativamente elevada, a presença de inúmeros interferentes potenciais torna a medida altamente propícia a ganhos em especificidade metodológica.…”

Section: Cortisol Livre Urin Rio G/24h Lc-ms/msunclassified

“…Com a evolução dos equipamentos e das técnicas de introdução de amostras para leitura, a concretização de todo o potencial dessa metodologia está se tornando realidade. No caso da determinação do cortisol urinário, devido à sua relativa alta concentração, os recursos técnicos já estão disponíveis, e laboratórios de referência nos EUA e Europa têm recentemente publicado metodologias baseadas em cromatografia líquida e espectrometria de massa com este objetivo (25)(26)(27). Neste trabalho, apresentamos nossa experiên-cia no desenvolvimento de um método de alta especificidade para a medida de cortisol e cortisona livres na urina, baseado em cromatografia líquida e espectrometria de massa em tandem (LC-MS/MS).…”

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Dosagem de cortisol e cortisona livres urinários empregando cromatografia líquida associada a espectrometria de massa em tandem (LC-MS/MS)

Vieira

Nakamura

Carvalho

2005

Arq Bras Endocrinol Metab

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A dosagem de cortisol livre na urina (CLU) é um método útil na triagem de pacientes suspeitos de síndrome de Cushing. Os imunoensaios atuais apresentam limitações que dificultam sua aplicação e a comparação de resultados por diferentes ensaios. No presente método para dosagem de cortisol e cortisona livres urinários, baseado em cromatografia líquida com detecção por espectrômetro de massa em tandem (LC-MS/MS), uma alíquota de urina 24h (200µL) é misturada com solução contendo quantidade conhecida de cortisol deuterado e extraída on-line em fase sólida (C 18 ). O eluato é transferido para uma segunda coluna C 18 (Phenomenex Luna, 3µ, de 50 x 2mm), e o eluato obtido em modo isocrático é aplicado diretamente no MS/MS modelo Quattro Micro, operando no modo positivo de ionização química a pressão atmosférica (APCI). Todo o processo é automatizado e a quantificação é feita por diluição isotópica com base nas razões das áreas dos picos dos analitos e do padrão interno deuterado. O estudo de especificidade mostrou que nenhum esteróide testado apresenta reatividade cruzada superior a 1% com cortisol ou cortisona. A sensibilidade funcional é <1µg/L para os dois esteróides, e o CV interensaio <8%. Os estudos de recuperação e de linearidade foram satisfatórios, e a comparação entre os resultados obtidos para cortisol em 98 amostras de rotina, pelo ensaio em estudo e por um RIE, mostraram correlação de r= 0,838, sendo os valores obtidos por LC-MS/MS significativamente mais baixos (medianas de 22,0 e 49,4µg/24h, respectivamente) (P<0,0001). Os valores de referência para cortisol foram definidos entre 11 e 43µg/24h, compatíveis com métodos semelhantes recentemente descritos. A dosagem concomitante de cortisona livre urinária permite o estudo da atividade da enzima 11β-HSD2 e a pesquisa da síndrome do excesso aparente de mineralocorticóides. O método descrito representa a primeira descrição, em nosso meio, de uma nova geração de métodos para a medida de esteróides hormonais, baseados em processos preparativos automatizados e leitura por espectrometria de massa em tandem. Free urinary cortisol (UFF) measurement is one of the most useful screening tests for Cushing's syndrome. Immunoassays employed today by most clinical laboratories present limitations, specially concerning specificity. These limitations restrain a widespread application of the method, as well as the comparison of results obtained by the use of different methods. We present the development and characterization of a UFF and cortisone method based on liquid chromatography and tandem mass spectrometry (LC-MS/MS). A 200µL aliquot from a 24h urine sample

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“…Liquid chromatography-mass spectrometry (LC-MS) and liquid chromatography-tandem mass spectrometry are specific and offer simpler approaches to sample preparation without sample derivatization steps. Recently, several LC-MS-based methods that use different ion sources were reported for the determination of the following steroid hormones: testosterone [37][38][39], cortisol [40][41][42][43][44], 11-deoxycortisol [45], androstenedione [38,39], DHEA [46], DHEA-S [38,46], progesterone [47], 17-hydroxyprogesterone [48], estriol [49], and estradiol [38,49].…”

Section: Methods Of Measurementmentioning

confidence: 99%

“…The ESI source is considered to be more sensitive than the APCI source for polar compounds. For the nonpolar or low polar compounds, such as most steroid molecules, the sensitivity that is provided by the ESI source is less satisfactory [44,48] than the APCI source. The more recently introduced APPI source is significantly more sensitive than APCI for certain compounds [50].…”

mentioning

confidence: 99%

Steroid hormones: relevance and measurement in the clinical laboratory

Holst

Soldin

Guo

et al. 2004

Clinics in Laboratory Medicine

139

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The steroid hormones are synthesized in the adrenal cortex, the gonads, and the placenta; are all derived from cholesterol and many are of clinical importance. Steroid hormones are synthesized in the mitochondria and smooth endoplasmic reticulum. Because they are lipophilic, they cannot be stored in vesicles from which they would diffuse easily and are therefore synthesized when needed as precursors. Upon stimulation of the parent cell, steroid hormone precursors are converted to active hormones and diffuse out of the parent cell by simple diffusion as their intracellular concentration rises.Because all steroid hormones are derived from cholesterol, they are not soluble in plasma and other body fluids. As a result, steroids are bound to transport proteins that increase their half-life and insure ubiquitous distribution. The protein-bound steroids are in equilibrium with a small fraction of free steroids, which are 'active.' Steroids can act quickly, by binding to cell surface receptors, or slowly, by binding to cytoplasmic or nucleic receptors and ultimately activate gene transcription.The adrenal glands are composed of the adrenal medulla and the adrenal cortex. The adrenal cortex is divided into three major anatomic zones: the zona glomerulosa, which produces aldosterone; and the zonae fasciculata and reticularis, which together produce cortisol and adrenal androgens. The medulla synthesizes catecholamines. More than 30 steroids are produced in the adrenal cortex; they can be divided into three functional categories: mineralocorticoids, glucocorticoids, and androgens.The steroids that are made almost exclusively in the adrenal glands are cortisol, 11-deoxycortisol, aldosterone, corticosterone, and 11-deoxycorti-costerone. Most other steroid hormones, including the estrogens, are made by the adrenal glands and the gonads [ MineralocorticoidsThe mineralocorticoids are formed in the zona glomerulosa. The main function of the mineralocorticoids is to promote tubular reabsorption of sodium and secretion of potassium and hydrogen ions at the collecting tubule, distal tubule, and collecting ducts [2]. When sodium is reabsorbed, water is absorbed simultaneously. The absorption of sodium and water increases fluid volume and arterial pressure.Aldosterone is the most potent mineralocorticoid and accounts for about 90% of the total mineralocorticoid activity. Mineralocorticoid potency in descending order is: aldosterone, 11-deoxycorticosterone, 18-oxocortisol, corticosterone, and cortisol [1]. Although cortisol has mainly glucocorticoid activity, it also has some mineralocorticoid activity. Cortisol has 1/400 of the potency of aldosterone, but its concentration is about 80 times greater than that of aldosterone [4]. The adrenal production of cortisol is approximately 25 mg/day and that of aldosterone is 100μg/day. Corticosterone has mainly glucocorticoid activity and some mineralocorticoid activity.Aldosterone secretion is regulated primarily by the renin-angiotensin system; it also is stimulated by increased serum potas...

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Liquid Chromatography–Tandem Mass Spectrometry Analysis of Urinary Free Cortisol

Cited by 64 publications

References 5 publications

The degree of urinary hypercortisolism is not correlated with the severity of cushing’s syndrome

The degree of urinary hypercortisolism is not correlated with the severity of cushing’s syndrome

Dosagem de cortisol e cortisona livres urinários empregando cromatografia líquida associada a espectrometria de massa em tandem (LC-MS/MS)

Steroid hormones: relevance and measurement in the clinical laboratory

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