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US 9,851,368 B2Granted
Kit and method for quantitative detection of steroids
AssigneeSwansea University
InventorsWilliam Griffiths, Yuqin Wang, Peter Crick, William Bentley
Life of this patent
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Abstract
The invention relates to a kit and methods for quantitative detection of steroids in a sample. The kit comprises quantitative charge tags and an oxidizing agent.
Claims16
A method for the quantitative detection of oxosteroids and hydroxy steroids in a sample, the method comprising:
- i. reacting a first portion of the sample with a first member of a pair of quantitative charge tags of formula (I) or formula (II);
- wherein R is hydrogen, C1-4 alkyl or aryl, OR1 or NR1R2,
- each R1 and R2 is C1-3 alkyl or R1 and R2 together with the nitrogen atom to which they are attached form a 5- or 6-membered heteroalkyl ring;
- each R3 is independently C1-3 alkyl;
- and
- X is a halide ion;
- ii. reacting a second portion of the sample with an agent capable of oxidising an OH group to a carbonyl group;
- iii. reacting the product of step (ii) with a second member of a pair of quantitative charge tags of formula (I) or formula (II);
- iv. combining the products of steps (i) and (iii);
- v. conducting mass spectrometry on the combined product of step (iv) and determining the quantities therein of compounds labelled with the first and second members of the pair of quantitative charge tags, thereby providing quantitative detection of both oxosteroids and hydroxy steroids in a single analysis.
7 more elements
A method for the quantitative detection of oxosteroids and hydroxy steroids in a sample, the method comprising:
- i. reacting the sample with a first member of a pair of quantitative charge tags of formula (I) or formula (II)
- wherein R is hydrogen, C1-4 alkyl or aryl, OR1 or NR1OR2;
- each R1 and R2 is C1-3 alkyl or R1 and R2 together with the nitrogen atom to which they are attached form a 5- or 6-membered heteroalkyl ring;
- each R3 is independently C1-3 alkyl;
- and X is a halide ion;
- ii. reacting a reference composition with a second member of a pair of quantitative charge tags of formula (I) or formula (II);
- iii. combining the products of steps (i) and (ii);
- iv. conducting mass spectrometry on the combined product of step (iii) and determining the quantities therein of compounds labelled with the first and second members of the pair of quantitative charge tags, thereby providing quantitative detection of both oxosteroids and hydroxy steroids in a single analysis.
5 more elements
A method for the quantitative determination of oxosteroids and hydroxy steroids in a sample comprising:
- i. reacting a first portion of the sample with a first member of a first pair of quantitative charge tags of formula (I) or formula (II);
- wherein R is hydrogen, C1-4 alkyl or aryl, OR1 or NR1R2;
- each R1 and R2 is C1-3 alkyl or R1 and R2 together with the nitrogen atom to which they are attached form a 5- or 6-membered heteroalkyl ring;
- each R3 is independently C1-3 alkyl;
- and
- X is a halide ion;
- ia. reacting a first portion of a reference composition with a first member of a second pair of quantitative charge tags of formula (I) or formula (II);
- ii. reacting a second portion of the sample with an agent capable of oxidising an OH group to a carbonyl group;
- iia. reacting a second portion of a reference composition with an agent capable of oxidising an OH group to a carbonyl group;
- iii. reacting the product of step (ii) with a second member of a first pair of quantitative charge tags of formula (I) or formula (II);
- iiia. reacting the product of step (iia) with a second member of a second pair of quantitative charge tags of formula (I) or formula (II);
- iv. combining the products of steps (i) and (iii);
- v. conducting mass spectrometry on the combined product of step (iv) and determining the quantities therein of compounds labelled with the first and second members of the pair of quantitative charge tags, thereby providing quantitative detection of both oxosteroids and hydroxy steroids.
10 more elements
Dependencies are inferred from the claim text.