FAME analysis involves transesterifying lipids to fatty acid methyl esters (FAMEs) which are then analyzed by gas chromatography (GC). This technique is used to identify bacteria and characterize microbial communities based on their unique FAME profiles. The procedure involves growing bacteria, extracting fatty acids, derivatizing to FAMEs, and analyzing by GC. Peak patterns are compared to a library using Sherlock software to identify bacteria. FAME analysis allows for strain-level identification and is useful in clinical, environmental, and food applications.

2. FAME Analysis Technique
Sarfaraz Hussain
Roll no. 2054
Department of Bioinformatics and Biotechnology, GCUF
Responsible teacher: Dr Shahid

3. Contents
 Fame
 Objectives of Fame analysis
 Techniques
 Procedure
 Sherlock Software

4. FAME ?
 FAME is stand for Fatty Acid Methyl Ester.
FAME Production:
FAME is produced by transesterification. In the transesterification
process a glyceride reacts with an alcohol in the presence of a
catalyst, forming a mixture of fatty acids esters and an alcohol.

5. Objective of FAME Analysis
 This is widely used in characterizing
new species of bacteria, and is useful
for identifying pathogenic strains.
 Organisms have different ,Quality
and Quantity of Fatty Acid

6. FAME Analysis By Different Technique
There are different technique which are used for Fame analysis
1: FAME analysis by capillary GC
2: FAME analysis by gas chromatography

7.  Gas chromatography (GC) USED due to the volatility and thermal
stability of the FAME.
 Gas chromatography has become an important technique in fats
and oils analysis because accurate results can be obtained for
complex, as well as simple, sample matrices.
Chromatography (GC)

8.  The FAMEs are injected on a gas chromatography (GC)
system for separation, and are subsequently detected
by a flame ionization detector (FID). Analyte identity is
verified by an additional injection on a GC where the
analytes are separated and identified by a mass
spectrometric detector (MSD).
 Sherlock software is use to in this technique.

9.  Chromatograph shows fatty acid contents(Methyl esters)in the form
of peaks.
 Y-axis shows amount of fatty acids.
 X-axis shows time.
Results

10. FAME Analysis Technique
 Fatty acid methyl ester analysis by gas chromatography
(GC-FAME) is use to identify microbes in environmental
and clinical settings.
 It is specific to growth conditions, including the media and
temperature used.
 In addition to identification, FAME analysis is often
specific enough to allow for strain or sub-species level
comparisons.

11.  This is the specified method for determining total
fat content for nutritional labeling purposes.
After quantifying the total FAMEs present in the
derivatized sample, the amount of fat (as
triglycerides) in the sample is calculated, based
on initial sample weight.
FAME Analysis Technique

12. FAME Analysis Procedure
 The Sherlock System requires that bacteria to be grown
in culture.
 The fatty acids are extracted ,followed by derivatization
with dilute hydrochloric acid/methanol solution to give
the respective methyl esters (FAMEs).
 The FAMEs are then extracted from the aqueous phase by
the use of an organic solvent.

13. Sherlock software
 The Sherlock software automates all
analytical operations and uses a sophisticated
pattern recognition algorithm to match the
unknown FAME profile to the stored library
entries for identification.

14.  Sherlock Instant FAME (I-FAME)
 Sherlock Quality FAME (Q-FAME)
Sherlock FAME Analysis

15. Sherlock System Library
 The Sherlock System is capable of identifying a wide
range of microorganisms library was carefully
developed by collecting well characterized strains of
reference cultures from microbiologists specializing in
many areas, including: clinical, environmental, industrial,
drinking/waste water, and food.
 The current Sherlock System libraries have over 1,500
bacterial species, along with 200 species of yeast.

16. Fatty acid methyl ester (FAME) analysis

17.  describes the determination of total fat content based on the fatty
acid content.
 This is the specified method for determining total fat content for
nutritional labeling purpose.
 quantifying the total FAMEs present in the derivative sample, the
amount of fat (as triglycerides) in the sample is calculated
Applications