# US 11,079,329 B2 · Human waste water and human-derived pathogen scouting tool

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- Number: US 11,079,329 B2
- Document ID: US11079329B2
- Status: Granted
- Kind: B2
- Application number: 16/143,640
- Filed: 2018-09-27
- Published: 2019-03-28
- Granted: 2021-08-03
- Earliest claimed: 2017-09-27
- Assignees: University of North Carolina Wilmington
- Applicants: University of North Carolina Wilmington
- Inventors: Mary M. Turbe; Lawrence B. Cahoon
- Web page: https://patentgrep.com/patent/US11079329B2

## CPC

- G01N 21/643: {non-biological material}
- G01N 1/34: Purifying; Cleaning {(processes or apparatus for extracting or separating nucleic acids from biological samples C12N15/1003)}
- G01N 1/405: {by adsorption or absorption}
- G01N 1/4055: {by solubility techniques}
- G01N 21/64: Fluorescence; Phosphorescence
- G01N 33/18: Water
- G01N 2001/4061: {Solvent extraction}
- G01N 2021/6439: {with indicators, stains, dyes, tags, labels, marks}
- Y02W 10/37: using solar energy
- Y10T 436/255: including use of a solid sorbent, semipermeable membrane, or liquid extraction
- 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)
- Y02W: CLIMATE CHANGE MITIGATION TECHNOLOGIES RELATED TO WASTEWATER TREATMENT OR WASTE MANAGEMENT
- Y10T: TECHNICAL SUBJECTS COVERED BY FORMER US CLASSIFICATION

## Abstract

The present subject matter provides processes and kits for identifying the presence of waste water or human-derived pathogens, particularly those associated with the presence of human fecal matter, in surface or subsurface water sources.

## Claims

### Claim 1

A process for identifying a presence of human fecal matter in water samples, the process comprising:
collecting a water sample, the water sample containing optical brighteners;
running the water sample through a solid phase extraction cartridge;
reducing interference of naturally occurring fluorescing compounds by at least a first elution of the naturally occurring fluorescing compounds from the extraction cartridge while retaining at least a portion of the optical brighteners;
concentrating and enhancing a fluorescence signal corresponding to the optical brighteners, by at least a second elution of the optical brighteners from the extraction cartridge; and
determining a presence of the optical brighteners based on a detection of the fluorescence signal, wherein the presence of the optical brighteners indicates the presence of human fecal matter in the water sample.

### Claim 2 (depends on 1)

The process of claim 1, wherein a presence of pathogens associated with human fecal matter is also identified based on the fluorescence signal.

### Claim 3 (depends on 2)

The process of claim 2, further comprising filtering the water sample prior to running the water sample through the extraction cartridge.

### Claim 4 (depends on 3)

The process of claim 3, wherein the at least first elution is carried out using a dilute base.

### Claim 5 (depends on 4)

The process of claim 4, wherein the dilute base is sodium hydroxide.

### Claim 6 (depends on 3)

The process of claim 3, wherein the concentrating step is carried out using at least one of a polar organic solvent and a non-polar organic solvent.

### Claim 7 (depends on 6)

The process of claim 6, wherein the polar organic solvent is selected from the group consisting of methanol and ethanol.

### Claim 8 (depends on 6)

The process of claim 6, wherein the non-polar organic solvent is polyethylene glycol (PEG) 400.

### Claim 9 (depends on 3)

The process of claim 3, wherein the determining step is carried out using a fluorometric detector.

### Claim 10 (depends on 3)

The process of claim 3, wherein the reducing step is carried out using a dilute base, the concentrating step is carried out using a solvent selected from the group consisting of a polar organic solvent and a non-polar organic solvent, and wherein the determining step is carried out using a fluorometric detector.

### Claim 11 (depends on 10)

The process of claim 10, wherein the dilute base is sodium hydroxide, the polar organic solvent is selected from the group consisting of methanol and ethanol and the non-polar organic solvent is polyethylene glycol (PEG) 400.

### Claim 12

A process for identifying a presence of waste water in water samples, the process comprising:
collecting a water sample the water sample containing optical brighteners;
running the water sample through a solid phase extraction cartridge;
reducing interference of naturally occurring fluorescing compounds by at least a first elution of the naturally occurring fluorescing compounds from the extraction cartridge while retaining at least a portion of the optical brighteners;
concentrating and enhancing a fluorescence signal associated with the optical brighteners, by at least a second elution of the optical brighteners from the extraction cartridge; and
determining a presence of the optical brighteners based on a detection of the fluorescence signal, wherein the presence of the optical brighteners indicates the presence of waste water.

### Claim 13 (depends on 12)

The process of claim 12, wherein a presence of human pathogens associated with waste water is also identified based on the fluorescence signal.