Rapid analysis for cyanobacterial toxins

• US 10,338,067 B1
• Filed: 07/14/2013
• Issued: 07/02/2019
• Est. Priority Date: 09/06/2007
• Status: Active Grant

• Current Assignee: Abraxis LLC (Bristol-Myers Squibb Company)
• Original Assignee: Abraxis LLC (Bristol-Myers Squibb Company)
• Inventors: Rubio, Fernando Manuel, Kamp, Lisa Marie
• Primary Examiner(s): Zeman, Robert A
• Application Number:
  US13/941,524
• Time in Patent Office: 2,179 Days
• Field of Search: None
• CPC Class Codes: G01N 2021/495 the fluid being adsorbed, e.g. in porous medium
• CPC Class Codes: G01N 2021/775 Indicator and selective membrane
• CPC Class Codes: G01N 2021/7759 Dipstick; Test strip
• CPC Class Codes: G01N 2021/7769 Measurement method of reaction-produced change in sensor
• CPC Class Codes: G01N 2021/7793 Sensor comprising plural indicators
• CPC Class Codes: G01N 2035/00168 Manufacturing or preparing test elements
• CPC Class Codes: G01N 2035/00465 Separating and mixing arrangements
• CPC Class Codes: G01N 33/1813 Specific cations in water, e.g. heavy metals
• CPC Class Codes: G01N 33/182 Specific anions in water
• CPC Class Codes: G01N 33/1826 Organic contamination in water
• CPC Class Codes: G01N 33/56911 Bacteria
• CPC Class Codes: G01N 35/1002 Reagent dispensers

Abstract

Method and compositions using transition metal salts and/or ammonium chloride to liberate toxins and other molecules from cyanobacteria, useful for assaying for total cyanobacterial toxins in lakes, reservoirs and other waters.

Claims

1. A method of releasing a cyanobacterial toxin in a suspension comprising cyanobacteria suspended in an aqueous medium, wherein said method comprises adding a releasing agent to an aliquot of the suspension so as to release toxins from the bacteria into the medium, provided the cyanobacteria are not subjected to a freeze-thaw step before, during, or after the step where the releasing agent is added to the suspension, wherein the releasing agent is a metal salt that comprises a copper atom and which can dissociate in water to release the copper atom as a copper ion, and wherein the releasing agent is selected from the group consisting of copper acetate, copper benzoate, copper bromide, copper chloride, copper chlorate, copper fluoride, copper formate, copper nitrate, copper perchlorate, and copper sulfate.

2. A method of claim 1 wherein the metal salt is copper sulfate.

3. A method of claim 1 further comprising adding a detergent to the suspension subsequent to or simultaneously with adding the releasing agent.

4. A method of claim 3 wherein the detergent is Triton X-100 or Tween-20.

5. A method of claim 1 wherein the toxin, is selected from the group consisting of microcystins, nodularins, anatoxin-a, anatoxin-a(S), aplysiatoxins, cylindrospermopsins, lyngbyatoxin-a, and saxitoxins.

6. A method of claim 5 wherein the toxin is selected from the group consisting of a microcystin and a nodularin.

7. A method of claim 6 wherein the toxin is a microcystin.

8. A method of releasing a cyanobacterial toxin in a suspension comprising cyanobacteria suspended in an aqueous medium, wherein said method comprises:

(1) adding a releasing agent other than ammonium chloride to the suspension, and (2) adding ammonium chloride to the suspension so as to release toxins from the bacteria into the medium, provided the cyanobacteria are not subjected to a freeze-thaw step before, during, or after the step where the releasing agent is added to the suspension, wherein the releasing agent is a metal salt that comprises a copper atom and which can dissociate in water to release the copper atom as a copper ion, and wherein the releasing agent is selected from the group consisting of copper acetate, copper benzoate, copper bromide, copper chloride, copper chlorate, copper fluoride, copper formate, copper nitrate, copper perchlorate, and copper sulfate.

9. A method of claim 8 further comprising the step of adding a detergent.

10. A method of claim 9 wherein the detergent is Triton X-100 or Tween-20.

11. A method of releasing cyanobacterial toxin in a suspension comprising cyanobacteria suspended in an aqueous medium, wherein said method comprises adding a releasing agent to an aliquot of the suspension so as to release toxins from the bacteria into the medium, provided the cyanobacteria are not subjected to a freeze-thaw step before, during, or after the step where the releasing agent is added to the suspension, wherein the releasing agent is a metal salt comprises a copper atom further comprises an anion selected from the group consisting of acetate, benzoate, bromide, chloride, chlorate, fluoride, formate, nitrate, perchlorate, and sulfate, and wherein said method further comprises adding Tween and ammonium chloride to the aliquot of the suspension.