Rapid measurement of estrogens and their metabolites in human serum by liquid chromatography-tandem mass spectrometry without derivatization

Guo, Tiedong; Gu, Jinlou; Soldin, Offie P.; Singh, Ravinder J.; Soldin, Steven J.

doi:10.1016/j.clinbiochem.2008.02.009

Cited by 97 publications

(79 citation statements)

References 16 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Positive atmospheric pressure chemical ionization and atmospheric pressure photoionization produced a good response for E 2 , but with higher background noise. The LOQs of E 2 assays using positive atmospheric pressure chemical ionization and atmospheric pressure photoionization were comparable with LOQs of assays using negative ESI (30)(31)(32). In this assay, negative ESI was used and in an effort to optimize the ESI response, mobile phase modification was needed.…”

Section: Discussionmentioning

confidence: 94%

“…In this assay, negative ESI was used and in an effort to optimize the ESI response, mobile phase modification was needed. Previous literature showed that adding ammonium hydroxide in the mobile phase could increase the [M-H] − ion intensity (30,31). Ionization suppression, a major concern in the LC-MS/MS method, is a compound-dependent effect and is more severe when operating in ESI, particularly for steroid estrogens (31).…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

High-Sensitivity Micro LC-MS/MS Assay for Serum Estradiol without Derivatization

Leung

Bridgman

et al. 2016

The Journal of Applied Laboratory Medicine

View full text Add to dashboard Cite

Background: There are considerable demands to accurately measure estradiol (E 2 ) at low concentrations (<20 pg/mL) in postmenopausal women, men, pediatric patients, and patients receiving breast cancer treatment. Most current high-sensitivity LC-MS/MS E 2 methods require large sample volumes and involve complex sample preparations with dansyl chloride derivatization. Our study aims to develop a high-sensitivity, underivatized method using micro LC-MS/MS to reliably measure E 2 concentrations below 5 pg/mL by the use of low sample volume. Methods: A total of 290 μL of sample was mixed with internal standard (IS), E 2 -d4, and extracted with a mixture of hexane/ethyl acetate (90/10) (v/v). After extraction, sample was separated by Eksigent Ekspert™ micro LC 200 system with a flow rate of 35 μL/min in a total run time of 3.5 min and detected by SCIEX QTRAP 6500 mass spectrometer in a negative mode using transitions: 271/145 (quantifier) and 271/143 (qualifier). In this method, it was crucial to use HPLC columns with stability at a pH >10. Results:The validation study demonstrated broad linear ranges (3.0 -820.0 pg/mL) with r 2 > 0.999. Total precision was below 15% at all QC levels, and limit of quantification (LOQ) was 3.0 pg/mL. Our method showed good correlation with E 2 RIA (r 2 = 0.96, bias = −1.0 pg/mL) and modest correlation with E 2 Roche Cobas automated immunoassay (r 2 = 0.86, bias = 6.0 pg/mL). Conclusions:In conclusion, we developed and validated a routinely applicable micro LC-MS/MS method without derivatization for E 2 in blood samples with an LOQ of 3.0 pg/mL. IMPACT STATEMENTPatients under assessment of puberty delays, pubertal growth, gynecomastia, and efficiency of aromatase inhibitor (AI) treatment will benefit from the information presented here. Evidence presented on high-sensitivity LC-MS/MS assay for E 2 will allow better characterization of E 2 concentration in blood in the low measurement range. Knowledge in the field of LC-MS/MS method development for E 2 will be advanced by the information presented.

show abstract

Section: Discussionmentioning

confidence: 94%

Section: Discussionmentioning

confidence: 99%

High-Sensitivity Micro LC-MS/MS Assay for Serum Estradiol without Derivatization

Leung

Bridgman

et al. 2016

The Journal of Applied Laboratory Medicine

View full text Add to dashboard Cite

show abstract

“…The method used a derivatization procedure to enhance the ionization efficiency, thus allowing uE 3 determination to an LOD of 0.05 ng/mL in serum. Although our method has a higher LOD than other LC-MS/MS methods, mainly because others have used more sensitive LC-MS/MS instrumentation (19,20 ), our method can still meet clinical needs in detecting serum uE 3 .…”

Section: Discussionmentioning

confidence: 99%

Measurement of Unconjugated Estriol in Serum by Liquid Chromatography–Tandem Mass Spectrometry and Assessment of the Accuracy of Chemiluminescent Immunoassays

et al. 2014

View full text Add to dashboard Cite

BACKGROUND Unconjugated estriol (uE3) is routinely analyzed in clinical laboratories as risk assessment for Down syndrome. Immunoassays of various types are the most commonly used methods. The accuracies of RIAs and ELISAs for uE3 have been questioned, and to date there have been no independent studies investigating the accuracy of the relatively new chemiluminescent immunoassays. We developed and validated a liquid chromatography–tandem mass spectrometry (LC-MS/MS) method for uE3 measurements in serum. METHODS Serum samples from patients in the second trimester of pregnancy were used, and uE3 concentrations were measured by LC-MS/MS and the Beckman Coulter Access® 2 and Siemens IMMULITE 2000 automatic chemiluminescent immunoassay analyzers. RESULTS The LC-MS/MS method was validated and showed limit of detection 0.05 ng/mL; limit of quantification 0.2 ng/mL; linearity of response to 32 ng/mL; total imprecision of 16.2%, 10.4%, and 8.2% for uE3 at 1.10, 4.18, and 8.32 ng/mL, respectively; and analytical recoveries of 95.9%–104.2%. ANOVA of the correlation for LC-MS/MS results vs chemiluminescent immunoassays results showed R2 = 0.9678 (Access 2 = 0.9305 LC-MS/MS + 0.2177, Sy|x = 0.1786, P < 0.0001), and R2 = 0.9663 (IMMULITE 2000 = 0.8849 LC-MS/MS − 0.0403, Sy|x = 0.1738, P < 0.0001). Bland–Altman plots of uE3 results revealed concentration-dependent immunoassay biases. Mock risk analysis for Down syndrome showed no apparent difference in the risk assessment outcomes if the adjusted method-specific multiples of the median were used, and the assay imprecision was <10% CV. CONCLUSIONS Standardization of immunoassay methods for uE3 analysis is needed to improve the accuracy of the measurements.

show abstract

“…Earlier published LC-MS/MS methods for estrogens (Anari et al, 2002;Nelson et al, 2004) also lack the sensitivity and the specificity needed to measure low concentrations of estrogens. One of the approaches for enhancing the sensitivity was through the use of negative ion mode ESI on a more sensitive instrument API5000 (Applied Biosystems/ Sciex) (Guo et al, 2008). Additional enhancement in the sensitivity and the specificity for measurement of estrogens was achieved through the use of the derivatization with the use of the derivatization with dansyl chloride-an amine-containing sulfonyl halide (2), in combination with a multidimensional chromatographic separation .…”

Section: Lc-ms/ms In Clinical Diagnosticsmentioning

confidence: 99%

Liquid chromatography-tandem mass spectrometry applications in endocrinology

Kushnir

Rockwood

Bergquist

2009

Mass Spectrom. Rev.

139

View full text Add to dashboard Cite

Liquid chromatography-tandem mass spectrometry (LC-MS/ MS) has been recognized as a primary methodology for the accurate analysis of endogenous steroid hormones in biological samples. This review focuses on the use of LC-MS/MS in clinical laboratories to assist with the diagnosis of diverse groups of endocrine and metabolic diseases. Described analytical methods use on-line and off-line sample preparation and analytical derivatization to enhance analytical sensitivity, specificity, and clinical utility. Advantages of LC-MS/MS as an analytical technique include high specificity, possibility to simultaneously measure multiple analytes, and the ability to assess the specificity of the analysis in every sample. All described analytical methods were extensively validated, utilized in routine diagnostic practice, and were applied in a number of clinical and epidemiological studies, including a study of the steroidogenesis in ovarian follicles. #

show abstract

Rapid measurement of estrogens and their metabolites in human serum by liquid chromatography-tandem mass spectrometry without derivatization

Abstract: This method allows for the simultaneous measurement of four estrogens in human serum within 8 min. It can be routinely employed in a clinical environment and is attractive because of its simplicity in sample processing, micro sample requirement, and high throughput.

Cited by 97 publications

References 16 publications

High-Sensitivity Micro LC-MS/MS Assay for Serum Estradiol without Derivatization

High-Sensitivity Micro LC-MS/MS Assay for Serum Estradiol without Derivatization

Measurement of Unconjugated Estriol in Serum by Liquid Chromatography–Tandem Mass Spectrometry and Assessment of the Accuracy of Chemiluminescent Immunoassays

Liquid chromatography-tandem mass spectrometry applications in endocrinology

Contact Info

Product

Resources

About