A Reliable Assay for Detection of a Bile Acid 

For routine analyses to quantify Deoxycholic Acid that has been used in manufacturing processes and since sample size is typically limited, sensitivity is crucial.  This Method provides the sensitivity needed and maintains consistent and reliable results as the Peak values were measured for week-long intervals.  The %RSD of Retention Times for both standards and sample were 0.1%.


Figure 1.  Full Chromatogram

Figure 2.  Chromatogram magnified to show peak of interest. 


Peak:
Deoxycholic Acid

Method Conditions:
Column: Cogent BDC18™, 4µm, 100Å
Catalog No.: 40018-15P
Dimensions: 4.6 x 150mm
Mobile Phase:
A. DI Water
B. 20:80 Acetonitrile / Buffer (Buffer: DI Water with 50mM K2PO4, pH adjusted to 7.5)
C. Acetonitrile
Gradient:

Time (minutes)  % A % B % C
0.00 55 20 25
2.00 55 20 25
2.05 48 20 32
8.00 48 20 32
8.05 20 20 60
11.00 20 20 60
11.05 55 20 25
15.00 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 in an injectable form is used to improve the appearance and profile of moderate to severe submental fat.  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. Also used in research, when a micelle is formed with it, it can used as a mild detergent for the isolation of membrane proteins.

This Method was developed by and is presented courtesy of Eutech Scientific Services