Falsely Increased Serum Estradiol Results Reported in Direct Estradiol Assays

Leung, Yat-Sun; Dees, K.; Cyr, Richard C.; Schloegel, I. W.; Kao, Pai C.

doi:10.1093/clinchem/43.7.1250

Cited by 20 publications

(6 citation statements)

References 2 publications

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“…154 This is consistent with experience of quantitation of other sex steroids, where immunoassay can give an over estimation of steroid concentrations due to positive interference by other steroids. 160–164 The major molecular interference in the DHT assays is testosterone -which is removed by potassium permanganate treatment.…”

Section: Diagnostic Utilitymentioning

confidence: 99%

Clinical biochemistry of dihydrotestosterone

Marchetti

Barth

2013

Ann Clin Biochem

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Dihydrotestosterone (DHT) is the most potent natural androgen in humans. There has been an increasing interest in this androgen and its role in the development of primary and secondary sexual characteristics as well as its potential roles in diseases ranging from prostate and breast cancer to Alzheimer's disease. Despite the range of pathologies shown to involve DHT there is little evidence for measurement of serum DHT in the management of these diseases. In this review we describe the physiology of DHT production and action, summarize current concepts in the role of DHT in the pathogenesis of various disorders of sexual development, compare current methods for the measurement of DHT and conclude on the clinical utility of DHT measurement. The clinical indications for the measurement of DHT in serum are: investigation of 5a reductase deficiency in infants with ambiguous genitalia and palpable gonads; men with delayed puberty and/or undescended testes; and to confirm the presence of active testicular tissue. Investigation is aided by the use of human chorionic gonadotrophin stimulation. Due to paucity of published data on this procedure, it is important to follow guidelines prescribed by the laboratory performing the analysis to ensure accurate interpretation.Ann Clin Biochem 2013; 50: 95-107.

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Section: Diagnostic Utilitymentioning

confidence: 99%

Clinical biochemistry of dihydrotestosterone

Marchetti

Barth

2013

Ann Clin Biochem

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“…Analysis of estrogens and androgens in biological samples has been commonly performed using immunoassays for many years. However, these assays are suboptimal, as there is cross-reactivity with similar analytes, and they have moderate specificity and sen-sitivity [7,8]. Furthermore, there is a significant lack of agreement between the results of different immunoassays.…”

Section: Introductionmentioning

confidence: 99%

“…Moreover, utilizing the combination of highly specific mass transitions associated with these analytes and their respective internal deuterated standards provided a high degree of specificity to the identity of these hormones. sitivity [7,8]. Furthermore, there is a significant lack of agreement between the results of different immunoassays.…”

Section: Introductionmentioning

confidence: 99%

An LC-MS/MS Methodological Framework for Steroid Hormone Measurement from Human Serum

2022

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Estrogens and androgens are important regulators of sexual development and physiological processes in men and women, acting on numerous organs throughout the body. Moreover, they can contribute to a variety of pathologies, including osteoporosis, cancer, and cardiovascular and neurologic diseases. Analysis of estrogens and androgens in biological samples has been commonly performed using immunoassays for many years. However, these assays are suboptimal, as there is cross-reactivity with similar analytes, and they have moderate specificity and sensitivity. Thus, there is a clinical need to develop highly sensitive and specific methods for the accurate measurement of estrogen and androgen concentrations. Herein, we describe the development of three liquid chromatography coupled tandem mass spectrometry-based methods that incorporate the use of a Triple Quadrupole Mass Spectrometer for quantitative measurement of endogenous concentrations of various steroid hormones in human serum samples: (1) the simultaneous measurement of testosterone, androstenedione, and cortisol, (2) dehydroepiandrosterone (DHEA), and (3) 17β-estradiol (E2). The use of derivatizing reagents, Girard’s reagent P and dansyl chloride, allowed for significant gains in sensitivity in the analysis of DHEA and E2, respectively, relative to the underivatized analyte. These procedures proved efficient and adequately sensitive for steroid hormone analysis in extracted patient sera samples from older men and postmenopausal women, providing reliable data down to low nanogram/ml and sub-nanogram/ml levels. Moreover, utilizing the combination of highly specific mass transitions associated with these analytes and their respective internal deuterated standards provided a high degree of specificity to the identity of these hormones.

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“…Immunoassay is the mainstream method for steroid analysis in clinic in the past few decades, which has the advantages of high throughput and fast speed. However, this method is vulnerable to interference by body fluid matrix or structural analogues, resulting in the lack of specificity and accuracy [ 2 , 3 ]. Therefore, gas chromatography mass spectrometry (GC–MS) was later used as the most accurate method [ 4 , 5 ].…”

Section: Introductionmentioning

confidence: 99%

Quantitative-Profiling Method of Serum Steroid Hormones by Hydroxylamine-Derivatization HPLC–MS

Liu

Chi

Fan

et al. 2019

Nat. Prod. Bioprospect.

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A sensitive and rapid high performance liquid chromatography–mass spectrometry (HPLC–MS) method was developed and validated for simultaneous quantification of ten steroid hormones, including estrogens, androgens, progesterones, and corticosteroids four classes of steroids. The following ten steroid hormones were analyzed: progesterone, 21-deoxycortisol, estrone, 4-androstenedione, testosterone, dihydro-testosterone, androstenone, dehydroepiandrosterone, corticosterone and cortisone. Stable deuterated isotopes were used as internal standards for quantification. Sample preparation with and without derivatization were performed after liquid–liquid extraction, and the corresponding results were compared according to sensitivity and selectivity. Hydroxylamine derivatization was found to improve the ionization efficiency of the analytes for electrospray ionization MS analysis. The gradient of mobile phase and experimental parameters for HPLC separation were optimized. The lower limits of quantification were in the range of 0.05–5 ng mL −1 with wide linear range for the ten steroid hormones. The intra-day precision < 11.1% and recovery of 84.5–120% with negligible matrix effect were achieved, where within the acceptance limits of the FDA guideline. Total HPLC–MS analysis time was 6 min. This method enables simultaneous quantification of steroids in human serum. It will be helpful for the serum steroid profiling in order to understand various endocrinology diseases. Graphical Abstract

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Falsely Increased Serum Estradiol Results Reported in Direct Estradiol Assays

Cited by 20 publications

References 2 publications

Clinical biochemistry of dihydrotestosterone

Clinical biochemistry of dihydrotestosterone

An LC-MS/MS Methodological Framework for Steroid Hormone Measurement from Human Serum

Quantitative-Profiling Method of Serum Steroid Hormones by Hydroxylamine-Derivatization HPLC–MS

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