FAME Analysis TechniqueSarfaraz Hussain

Roll no. 2054Department of Bioinformatics and Biotechnology, GCUF

Responsible teacher: Dr Shahid

Contents Fame Objectives of Fame analysis Techniques Procedure Sherlock Software

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.

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

FAME Analysis By Different TechniqueThere are different technique which are used for Fame analysis

1: FAME analysis by capillary GC 2: FAME analysis by gas chromatography

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)

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.

Chromatograph shows fatty acid contents(Methyl esters)in the form of peaks.

Y-axis shows amount of fatty acids. X-axis shows time.

Results

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.

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

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.

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.

Sherlock Instant FAME (I-FAME) Sherlock Quality FAME (Q-FAME)

Sherlock FAME Analysis

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.

Fatty acid methyl ester (FAME) analysis

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

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