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Method Article
Extraction, identification, and quantification of volatile fatty acids (VFA) in rumen fluid samples using Reverse Phase High-Performance Liquid Chromatography with Diode Array Detector (RP HPLC-DAD)
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
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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.
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Method Article
Extraction, identification, and quantification of volatile fatty acids (VFA) in rumen fluid samples using Reverse Phase High-Performance Liquid Chromatography with Diode Array Detector (RP HPLC-DAD)
Julián Andrés Castillo Vargas
Universidade Federal Rural da Amazônia
Corresponding Author
ORCiD: https://orcid.org/0000-0001-5163-5127
Badges
Prescreen
This manuscript passed our Prescreen assessment
Complete author information
Appropriate declaration statements
Potential risks to human health
Prescreen
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