If you want to know more, please visit https://www.creative-proteomics.com/services/short-chain-fatty-acids-analysis-service.htm. Short chain fatty acids (SCFAs) are defined as fatty acids with two to six carbon atoms. SCFAs have a wide range of metabolic effects. And SCFA profiling has been a major topic in gut bacteria studies.

1. Short Chain Fatty Acids Analysis
Presented by Creative Proteomics

2. 01 Introduction
Dietary
Fiber
It has been studied that there are
associations between increased dietary fiber
intake and lower mortality from
cardiovascular diseases and certain types of
cancer.
Gut
Microflora
The gut microflora in the large intestine,
consisting more than 400 different species,
plays important physiological roles in human
physiology and health. Microflora has the
ability to produce hydrolytic enzymes that can
digest some of the complex carbohydrates.

3. 01 Introduction
S C F A s
When dietary fibers are fermented by the bacterial hydrolytic enzymes, the short chain
fatty acids (SCFAs) are the main products.
However, SCFAs may be generated from protein and amino acid decompositions as well.
Short Chain
Fatty Acids

4. 01 Introduction
Short chain fatty acids are defined as fatty acids with two to six carbon atoms. But the
definition varies and the upper limit may range between five and seven carbons in length.
Among SCFAs, three main types, acetic acid (C2), propionic acid (C3) and butyric acid
(C4), makes up 95% of all SCFAs.
Acetic acid Propionic acid Butyric acid

5. 01 Introduction
Function
01
02
They can be oxidized to provide energy and
have also been shown to affect the immune
system, colonic function, cholesterol
metabolism, satiety and oxidative stress.
A direct link between SCFAs (qualitatively
and quantitatively) and some human
pathological conditions, such as
inflammatory bowel disease (IBD), irritable
bowel syndrome (IBS), diarrhea and cancer
have been proposed.

6. 02 Methods
SCFAs have been measured in various biological materials such as blood plasma, serum, and feces.
Moreover, SCFAs have been detected in different environmental samples, food, waste leachates and even in
asphaltene.
In the following parts, we will focus on the SCFAs analytical methods.

7. Gas Chromatography
(GC)
• The principle of GC relies on a carrier gas
that serves as a mobile phase where sample
compounds are separated by differential
interaction with the column stationary phase.
• The pretreatment is important for the
successful detection of SCFAs by GC. There
are various methods, such as ultrafiltration,
centrifugation, acidification, or simple sample
dilution, which has its own advantages or
disadvantages.

8. Gas Chromatography
(GC)
• The flame ionization detector (FID), is
the most used conventional detector for
SCFAs detection in GC.
• It is sensitive to molecules that are
ionized in a hydrogen–air flame,
including most carbon-containing
compounds, and produces a current that
varies proportionally to the amount of
organic species in a sample.

9. Gas Chromatography
(GC)
• In addition to a conventional detector, GC can be bound to MS, causing a better
sensitivity and selectivity of the analysis.
• By using a GC-MS-MS instrument, quantitation of short-chain fatty acids that are
present in low concentrations in complex biological samples are available.
• One critical step in the GC-MS analysis of FAs is their conversion into suitable
volatile derivatives by derivatization.

10. High Performance Liquid Chromatography
(HPLC)
• The greatest advantage of the HPLC over the GC technique is the use of lower
running temperatures. And the resolved fatty acids are not destroyed during their
detection, which enables further analyses to be performed.
• Like GC, a successful SCFAs analysis by HPLC needs suitable combinations of
pretreatments, columns, running conditions and detectors.
• Due to the higher pressure, the mobile phase carrying analytes travels and the small
stationary phase particles with a larger area allow for a better interaction.
• The most commonly used technique is a reverse phase HPLC (RP-HPLC), where the
stationary solid phase (column) is hydrophobic (non-polar) and the mobile liquid
phase is hydrophilic (polar, watery).

11. Other Methods
Nuclear magnetic
resonance (NMR)
Capillary
Electrophoresis (CE)
Enzymatic detection
of SCFAs
• Advantages: fast, stable
and reprducible
• Drawbacks:
instrumentation cost and
sensitivity
• Advantages: speed and
minimal sample pretreatment
procedure.
• Drawbacks: low repeatability
and reproducibility.
• It can differentiate between
optical isomers, so the
lactate can be found in both
D- and L-form.
In addition to GC and HPLC, there are some other methods.

12. At Creative Proteomics, we have
developed a professional platform for
the identification and quantification of
short chain fatty acids by GC-MS. The
short chain fatty acids we can quantified
include acetic acid, propionic acid,
butyric acid, isobutyric acid, valeric acid,
isovaleric acid, and caproic acid.
Short Chain Fatty Acids
Analysis Services
03 Our Services

13. Thank you
Creative Proteomics
Please contact us for more information
Web:
Email:
www.creative-proteomics.com
info@creative-proteomics.com