This is a protocol preprint, a preliminary version of a manuscript that has not completed peer review at a journal. Research Square does not conduct peer review prior to posting preprints. The posting of a preprint on this server should not be interpreted as an endorsement of its validity or suitability for dissemination as established information or for guiding clinical practice.
Method Article
Julián Andrés Castillo Vargas
Universidade Federal Rural da Amazônia
Corresponding Author
ORCiD: https://orcid.org/0000-0001-5163-5127
License:
This work is licensed under a CC BY 4.0 License. Read Full License
In ruminant animals, volatile fatty acids (VFA) or short-chain fatty acids (SCFAs) are derived from the protein and carbohydrate fermentation by rumen microorganism. Hence, the VFA determination in rumen fluid allows the evaluation of the nutritional quality of a diet, as well as its potential impact on the chemical composition of ruminant milk and meat. Thus, we developed a protocol to extract, identify, and quantify acetic, propionic, butyric, valeric, and caproic acids in ruminal fluid samples using RP-HPLC-DAD. Despite literature findings had shown that the most suitable column for VFA chromatographic separation under HPLC-DAD is an ion-exchange column, our protocol showed that a C18 column also allows an efficient VFA separation of the aforementioned acid, except for butyric and iso-butyric acids. This condition may constitute a limitation of the Hypersil GOLD C18 column use for VFA determination. However, considering that the concentration of iso-butyric acid is significantly lower than that of butyric acid, a good estimation of butyric acid can be obtained.
Keywords
fatty acid extraction, HPLC, rumen fluid, short-chain fatty acid
Figure 1
Figure 2
This is a list of supplementary files associated with this preprint. Click to download.
- Table1.pdf
Table 1. Quantities of reagents to be used to prepare the calibration curve
- Table2.pdf
Table. 2. Gradient used in the chromatographic run.
Loading...
Version History
CURRENT STATUS: Posted
Version 1
Posted 05 Sep, 2020
No community comments so far
Metrics
Comments: 0
HTML Views: ...
Method Article
Julián Andrés Castillo Vargas
Universidade Federal Rural da Amazônia
Corresponding Author
ORCiD: https://orcid.org/0000-0001-5163-5127
Version History
CURRENT STATUS: Posted
Version 1
Posted 05 Sep, 2020
No community comments so far
Metrics
Comments: 0
HTML Views: ...
License:
This work is licensed under a CC BY 4.0 License. Read Full License
In ruminant animals, volatile fatty acids (VFA) or short-chain fatty acids (SCFAs) are derived from the protein and carbohydrate fermentation by rumen microorganism. Hence, the VFA determination in rumen fluid allows the evaluation of the nutritional quality of a diet, as well as its potential impact on the chemical composition of ruminant milk and meat. Thus, we developed a protocol to extract, identify, and quantify acetic, propionic, butyric, valeric, and caproic acids in ruminal fluid samples using RP-HPLC-DAD. Despite literature findings had shown that the most suitable column for VFA chromatographic separation under HPLC-DAD is an ion-exchange column, our protocol showed that a C18 column also allows an efficient VFA separation of the aforementioned acid, except for butyric and iso-butyric acids. This condition may constitute a limitation of the Hypersil GOLD C18 column use for VFA determination. However, considering that the concentration of iso-butyric acid is significantly lower than that of butyric acid, a good estimation of butyric acid can be obtained.
Figure 1
Figure 2
Loading...