Methdos for detecting estradiol by mass spectrometry
AssigneeQuest Diagnostics Investment Incorporated
InventorsMildred M. GOLDMAN, Nigel J. CLARKE, Richard E. REITZ
Life of this patent
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Abstract
Provided are methods for determining the amount of estradiol in a sample using mass spectrometry. The methods generally involve ionizing estradiol in a sample and detecting and quantifying the amount of the ion to determine the amount of estradiol in the sample.
Claims53
A method for determining the amount of estradiol in a test sample said method comprising:
- (a) purifying estradiol from said test sample by liquid chromatography;
- (b) ionizing the purified estradiol from said test sample to produce one or more ions detectable by tandem mass spectrometry;
- and
- (c) detecting the amount of the estradiol ion(s) by tandem mass spectrometry, wherein the amount of the estradiol ion(s) is related to the amount of estradiol in said test sample;
- wherein the limit of quantitation of the method is less than or equal to 80 pg/mL and estradiol is not derivatized prior to mass spectrometry.
A method for determining the amount of estradiol in a body fluid sample by tandem mass spectrometry said method comprising:
- (a) purifying estradiol from said body fluid sample by liquid chromatography;
- (b) generating a precursor ion of said estradiol having a mass/charge ratio of 255.07±0.5;
- (c) generating one or more fragment ions of said precursor ion, wherein at least one of said one or more fragment ions comprise an ion fragment having a mass/charge ratio of 133.20±0.5;
- and
- (d) detecting the amount of one or more of said ions generated in step (b) or (c) or both and relating the detected ions to the amount of said estradiol in said body fluid sample.
A method for determining the amount of estradiol in a body fluid sample by tandem mass spectrometry said method comprising:
- (a) purifying estradiol from said body fluid sample by liquid chromatography;
- (b) generating a precursor ion of said estradiol having a mass/charge ratio of 271.14±0.5;
- (c) generating one or more fragment ions of said precursor ion, wherein at least one of said one or more fragment ions comprise an ion fragment having a mass/charge ratio of 183.10±0.5;
- and
- (d) detecting the amount of one or more of said ions generated in step (b) or (c) or both and relating the detected ions to the amount of said estradiol in said body fluid sample.
A method for determining the amount of estradiol in a test sample said method comprising:
- (a) acidifying said test sample with an agent in an amount sufficient to free estradiol from a protein that may be present in said test sample;
- (b) purifying estradiol from said test sample by liquid chromatography;
- (c) ionizing the purified estradiol from said test sample to produce one or more ions detectable by tandem mass spectrometry;
- and
- (d) detecting the amount of the estradiol ion(s) by tandem mass spectrometry in positive ion mode, wherein the amount of the estradiol ion(s) is related to the amount of estradiol in said test sample.
A method for determining the amount of estradiol in a test sample said method comprising:
- (a) adding an agent to said test sample in an amount sufficient to free estradiol from a protein that may be present in said test sample;
- (b) purifying estradiol from said test sample by liquid chromatography;
- (c) ionizing the purified estradiol from said test sample to produce one or more ions detectable by tandem mass spectrometry;
- and
- (d) detecting the amount of the estradiol ion(s) by tandem mass spectrometry in negative ion mode, wherein the amount of the estradiol ion(s) is related to the amount of estradiol in said test sample.
Dependencies are inferred from the claim text.