FAME (Fatty Acid Methyle ester) analysis

FAME is a technique which is use to identify microbes on the basis of Fatty Acid present in there cell membraine and in cytoplasm.

1. FAME Analysis Technique
Athar Hussain Mutahari
BS(Hons)
Department of Bioinformatics and Biotechnology, Government
College University, Faisalabad
Pakistan

2. What is 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.

3. 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

4. Sources
FAMES, USUALLY OBTAINED FROM VEGETABLE
OILS, ANIMAL FATS OR WASTE COOKING
OILS BYTRANS ESTERIFICATION.
ALGA IS VERY IMPORTANT TO EXTRACT
BIODIESEL.

5. Thisisthespecifiedmethodfordeterminingtotal
fatcontentfornutritionallabelingpurposes.
AfterquantifyingthetotalFAMEspresentinthe
derivatizedsample,theamountoffat(as
triglycerides)inthesampleiscalculated,based
oninitialsampleweight.
FAME Analysis Technique

6. FAME Analysis Technique
Fattyacidmethylesteranalysisbygaschromatography
(GC-FAME)isusetoidentifymicrobesinenvironmental
andclinicalsettings.
Itisspecifictogrowthconditions,includingthemediaand
temperatureused.
Inadditiontoidentification,FAMEanalysisisoften
specificenoughtoallowforstrainorsub-specieslevel
comparisons.

7. FEATURES OF FAME ANALYSIS
Technique
FAME ANALYSIS Technique many advantages.The technology
is very:
I.Accurate
II.Inexpensive technique
III.Fatty acid analysisis an effective tool for strain
tracking, an advantage over genetic identification
systems.
IV.All microbes Catalyze with Same procedure.
V.FAMEs are more volatile than their respective fatty
acids and therefore more suitable to GC analysis.

8. FAME Analysis Procedure
TheSherlockSystemrequiresthatbacteriatobegrown
inculture.
Thefattyacidsareextracted,followedbyderivatization
withdilutehydrochloricacid/methanolsolutiontogive
therespectivemethylesters(FAMEs).
TheFAMEsarethenextractedfromtheaqueousphaseby
theuseofanorganicsolvent.

9. Continue---------
TheFAMEsareinjectedonagaschromatography(GC)
systemforseparation,andaresubsequentlydetected
byaflameionizationdetector(FID).Analyteidentityis
verifiedbyanadditionalinjectiononaGCwherethe
analytesareseparatedandidentifiedbyamass
spectrometricdetector(MSD).
Sherlocksoftwareisusetointhistechnique.

10. FAME Analysis Using Capillary GC
CapillaryGCisespeciallyusefulfordeterminingtotal
fatcontent
Fattyacidcontentinfoods.
Thechoiceofcapillarycolumndependsonthe
informationrequired.
ForpolyunsaturatedFAMEsanalysis,aFAMEWAX™
columnallowsfast,accuratequantification.

11. 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.

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

13. Sherlock Instant FAME(I-FAME)
In2007SherlockInstantFAME
TM
(I-FAME),.I-FAME
ismostusefulforGram-positiveenvironmental
aerobesandFAMEanalysisofnon-microbes
(e.g.animaltissue,edibleoils,biofuel
feedstocks,etc.).Theturnaroundtime(TAT)for
IFAMEislessthan15minutes.

14. Sherlock Quality FAME(Q-FAME),
In2011SherlockQ-FAME
TM
(Q-FAME),Q-FAMEis
mostusefulforAerobicbacteriaofimportance
topharmaceuticalQualityControlleaswellas
clinicalreferencelaboratories.Theturnaround
time(TAT)forQ-FAMEislessthan25minute.

15. Sherlock System Library
TheSherlockSystemiscapableofidentifyingawide
rangeofmicroorganisms librarywascarefully
developedbycollectingwellcharacterizedstrainsof
referenceculturesfrommicrobiologistsspecializingin
manyareas,including:clinical,environmental,industrial,
drinking/wastewater,andfood.
ThecurrentSherlockSystemlibrarieshaveover1,500
bacterialspecies,alongwith200speciesofyeast.

16. Fatty acid methyl ester (FAME) analysis