# US 5,700,654 A · Method and compositions to assess oxidative stress in vivo

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- Number: US 5,700,654 A
- Document ID: US5700654A
- Status: Granted
- Kind: A
- Application number: 08/304,147
- Filed: 1994-09-12
- Granted: 1997-12-23
- Earliest claimed: 1991-06-14
- Assignees: Vanderbilt Unversity
- Inventors: L. Jackson Roberts; Jason D. Morrow
- Web page: https://patentgrep.com/patent/US5700654A

## CPC

- G01N 33/88: involving prostaglandins {or their receptors}
- G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q)

## Abstract

This invention relates to a method to assess oxidative stress in vivo by quantification of prostaglandin F.sub.2 -like compounds and their metabolites produced by a noncyclooxygenase free radical catalyzed mechanism.

## Claims

### Claim 1

A method to assess oxidative stress in vivo comprising:
(a) measuring the amount of noncyclooxygenase derived prostanoids in a biological sample before the ex vivo development of prostanoids in the sample;
(b) comparing the measured amount of the noncyclooxgenase derived prostanoids with a control; and
(c) assessing oxidative stress in vivo based on the comparison in step c.

### Claim 2 (depends on 1)

A method as set forth in claim 1 farther includes the step of storing the biological sample prior to said measuring step.

### Claim 3 (depends on 2)

The method of claim 2 wherein the stored sample is maintained at -70.degree. C.

### Claim 4 (depends on 1)

A method as set forth in claim 1 wherein the sample is urine.

### Claim 5 (depends on 1)

A method as set forth in claim 1 wherein the sample is a fresh sample of tissue.

### Claim 6 (depends on 1)

A method of claim 1 wherein said measuring step is further defined as
(a) subjecting phospholipids in the biological sample to reverse phase solid chromatography extraction using a C-18 cartridge;
(b) eluting prostanoids retained on the C-18 cartridge with a 1:1 mixture of ethyl acetate:heptane;
(c) maintaining the eluate of the phospholipids from the fresh sample of lipid containing biological fluid at -20.degree. C.;
(d) measuring the amount of noncyclooxygenase derived prostanoids present in the eluate of the phospholipids from the fresh sample of lipid containing biological fluid maintained at -20.degree. C.;
(e) comparing said measured amount of prostanoids with a control; and
(f) assessing oxidative stress in vivo based on the comparison in step b.

### Claim 7 (depends on 1)

The method of claim 1 wherein measurement is by mass spectroscopy.

### Claim 8 (depends on 1)

The method of claim 1 wherein measurement is by immunoassay.