1.0 PURPOSE
The purpose of the document is to establish a general and uniform method for the determination of fatty acid profiling.
2.0 SCOPE
This standard testing procedure is suitable for the determination of fatty acid profiling.
3.0 RESPONSIBILITY
The expert and trained chemist are responsible for following this procedure. The technical manager is responsible for implementing this procedure.
4.0 REQUIREMENTS
4.1 Apparatus
4.1.1 Make/Model: GC-FID
4.1.2 Analytical Balance
4.1.3 Shaker
4.1.4 Vortex mixer
4.1.5 Heating mantle
4.2 Glassware
4.2.1 Measuring Cylinder -25 ml
4.2.2 Separating Funnels-125 ml
4.2.3 RB Flask -50ml &100ml
4.2.4 Volumetric Flask-5 ml &10 ml
4.2.5 Pipette-5 ml
4.3 Reagent
4.3.1 Methanol
4.3.2 BF3
4.3.3 n-Hexane
4.3.4 sodium chloride
4.3.5 Methyl red
4.3.6 Sodium sulphate(Anhydrous)
4.3.7 Methanolic KOH(2N)
5.0 PROCEDURE
5.1 FOR SOAP SAMPLE
5.1.1 Weigh about 200 mg of crushed sample in 100ml RB flask.
5.1.2 Add 5ml BF3 13-15%methanol reagent.
5.1.3 Attach a condenser and boil for 10 min over the heating mantle.
5.1.4 Wait till the sample cool down.
5.1.5 Add 5 ml of n-hexane through the condenser and boil for another 1 min. and again wait till the sample cools down.
5.1.6 Remove flask from condenser and Add 5ml n-hexane & shake gently.
5.1.7 Now add 15ml of saturated sodium chloride solution in the flask and shake vigorously for 15 sec.
5.1.8 Transfer the contents in a separating funnel (capacity 125 ml).
5.1.9 Remove the aqueous layer present on the lower side of the separating funnel.
5.1.10 Wash the n-hexane layer 3 times with D.M. water gently.
5.1.11 Add 3 drops of methyl red solution in the final washing (i.e. in the aqueous layer).
5.1.12 Wash till the color of the washing should become orange.
5.1.13 Pass the n-hexane through anhydrous sodium sulfate.
5.1.14 Dilute 1ml of this solution to 10 ml with n-hexane (for Bruker GC instrument)
5.1.15 Further dilute 1 ml of the above-diluted solution to 5 ml with hexane (for Shimadzu GC instrument).
5.1.16 Pass this solution with sodium sulfate (anhydrous)
5.1.17 Inject sample in GC instrument.
5.2 FOR OIL SAMPLE
5.2.1 Take a 200 mg sample.
5.2.2 Add 2 ml n-hexane.
5.2.3 Add 100 µl 2N methanolic KOH
5.2.4 Shake for 5 minutes on a vortex mixer.
5.2.5 Add 2 ml n-Hexane and shake again.
5.2.6 Take 0 5 ml and makeup To 10 ml.
5.2.7 Filter with sodium sulfate anhydrous
5.2.8 Inject sample in GC instrument.
5.3 Method
5.2.1 Column : Supelco SPTM-2560 Fused Silica Capillary column 75m x 0.18mm x 0.14um film thickness (Part No. 23348-U)
5.2.2 Injector: 1177 Split/Splitless
5.2.3 Temp 2500C
5.2.4 Split Ratio 1:100
5.2.5 Carrier Gas: Helium at flow rate of 1 ml/min
5.2.6 Oven Programming.
S.No. | Rate (0C/Min) | Step (0C) | Hold Time (Min) | Total Time (Min) |
1. | Initial | 100 | 1.00 | 1.00 |
2. | 4.0 | 150 | 15.00 | 28.50 |
3. | 3.0 | 230 | 10.00 | 65.17 |
5.2.7 Detector: : FID
5.2.8 Temp. : 2600C
5.2.9 Make Up (N2) Gas. : 25ml/Min
5.2.10 Combustion (H2) Flow. : 30ml/Min
5.2.11 Combustion (Air) Flow. : 300ml/Min
5.2.12 For the operation of GC-FID refer to the instrument operating procedure of GC-FID
6.0 CALCULATION
|
Individual Fatty acid content (g/100g) |
= |
% Area of
individual fatty acid from GC chromatogram |
X |
The total fat content of the sample |
|
100 |
7.0 REVISION/CHANGE HISTORY: NA
8.0 ATTACHMENTS
· Appendix-1: FLOW CHART FOR FALLY ACID PROFILE
9.0 REFERENCES:
· AOAC Official Method Ce2-66 Reapproved 2009.
You may like to read these links:
1. List of All SOPs and Documents for the Chemistry Laboratory
2. List of All SOPs and Documents for Quality Control
3. List of All SOPs and Documents For Quality Assurance Dept.
4. List of SOPs and Documents Required to Achieve NABL Accreditation
Hi. I’m Writer of Researchsop.com. ’ ’ Please share these SOPs to all concern pharma people for their development. I like to fullfill the need of curious people. These things inspire me to make things looks better.
0 comments:
Post a Comment