Deoxycholic Acid Assay by HPLC – AppNote
August 18, 2021
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A
Reliable
Method for a Bile Acid
This Deoxycholic Acid Assay produces Run to Run consistency and Precision with Retention Time RSD values of 0.1%. A sensitive HPLC-UV Method that may be used for quantification in raw material and liquid pharmaceutical formulation is presented.
Column: Cogent Bidentate C18™, 4um, 100Å
Catalog No.: 40018-15P
Dimensions: 4.6mm x 150mm
Mobile Phase:
-- A: DI Water
-- B: 20:80 Acetonitrile / Buffer (50mM KH2PO4 pH adjusted to 7.5)
-- C: Acetonitrile
Injection vol.: 100μL
Flow rate: 1.0mL / minute
Detection: UV @ 205nm
Column Temperature: 35°C
Sample Preparation: Deoxycholic Acid 50μg / mL in DI Water
Note: Deoxycholic Acid also known as Cholanoic Acid and sometimes sold as Kybella™ or Belkyra™, is one of the secondary bile acids, which are metabolic byproducts of intestinal bacteria and can be used to determine intestinal fat adsorption. Another research application is when a micelle is formed with it, it can used as a mild detergent for the isolation of membrane proteins.
This Deoxycholic Acid Assay produces Run to Run consistency and Precision with Retention Time RSD values of 0.1%. A sensitive HPLC-UV Method that may be used for quantification in raw material and liquid pharmaceutical formulation is presented.
Figure 1.
Full Chromatogram
Figure 2.
Magnified Chromatogram to show peak of interest
Peak:
Deoxycholic Acid
Met hod Conditions:Column: Cogent Bidentate C18™, 4um, 100Å
Catalog No.: 40018-15P
Dimensions: 4.6mm x 150mm
Mobile Phase:
-- A: DI Water
-- B: 20:80 Acetonitrile / Buffer (50mM KH2PO4 pH adjusted to 7.5)
-- C: Acetonitrile
| Time (minutes ) | A | B | C |
|---|---|---|---|
| 0 | 55% | 20% | 25% |
| 2 | 55% | 20% | 25% |
| 2.05 | 48% | 20% | 32% |
| 8 | 48% | 20% | 32% |
| 8.05 | 20% | 20% | 60% |
| 11 | 20% | 20% | 60% |
| 11.05 | 55% | 20% | 25% |
| 15 | 55% | 20% | 25% |
Injection vol.: 100μL
Flow rate: 1.0mL / minute
Detection: UV @ 205nm
Column Temperature: 35°C
Sample Preparation: Deoxycholic Acid 50μg / mL in DI Water
Note: Deoxycholic Acid also known as Cholanoic Acid and sometimes sold as Kybella™ or Belkyra™, is one of the secondary bile acids, which are metabolic byproducts of intestinal bacteria and can be used to determine intestinal fat adsorption. Another research application is when a micelle is formed with it, it can used as a mild detergent for the isolation of membrane proteins.
his Method was developed by and is presented courtesy of ARL- Eutech Scientific Services.