Nanospray liquid chromatography/tandem mass spectrometry analysis of steroids from gray whale blubber

Rationale Analysis of steroids from precious blubber biopsies obtained from marine mammals, especially endangered species, can provide valuable information on their endocrine status. Challenges with currently used ELISA methodology include lack of absolute quantitation and incompatibility with multi...

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Bibliographic Details
Published in:Rapid communications in mass spectrometry 2017-07, Vol.31 (13), p.1088-1094
Main Authors: Hayden, Mary, Bhawal, Ruchika, Escobedo, John, Harmon, Clinton, O'Hara, Todd M., Klein, David, San‐Francisco, Susan, Zabet‐Moghaddam, Masoud, Godard‐Codding, Céline A. J.
Format: Article
Language:English
Subjects:
Adipose Tissue - chemistry
Animals
Biopsy
Calibration
Chromatography, Liquid - methods
Compatibility
Data acquisition
Endangered & extinct species
Endangered species
Female
Hormones
Hydrocortisone
Ions
Limit of Detection
Linear Models
Liquid chromatography
Male
Marine mammals
Mass spectrometry
Mathematical analysis
Nanotechnology - methods
Quantitative analysis
Reproducibility of Results
Scientific imaging
Steroids
Steroids - analysis
Tandem Mass Spectrometry - methods
Whales - physiology
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Summary:Rationale Analysis of steroids from precious blubber biopsies obtained from marine mammals, especially endangered species, can provide valuable information on their endocrine status. Challenges with currently used ELISA methodology include lack of absolute quantitation and incompatibility with multiple steroids analysis due to limited biopsy mass. Development of a sensitive, accurate analytical method for this purpose is critical. Methods A nanospray liquid chromatography/tandem mass spectrometry (nanoLC/MS/MS) method was validated for sensitive, specific and quantitative analysis of three steroid hormones, without derivatization, extracted from 50 mg blubber samples. Data was acquired with an LTQ XL ion trap mass spectrometer in positive ion mode, using single reaction monitoring. All three steroids were analyzed in a single run. Cholic acid was used as a surrogate internal standard for quantitation due to its steroidal structure and lack of measurable endogenous levels in blubber. Results The lowest limits of quantitation for progesterone, testosterone, and hydrocortisone were significantly improved compared to previous studies using conventional LC/MS/MS. The lowest limit of detection was 7 fg/μL using a 1 μL injection volume. Calibration curves for steroid quantification showed good linearity (r2 >0.99) between 14 and 3620 fg/μL, and accuracy was
ISSN:0951-4198
1097-0231
DOI:10.1002/rcm.7884