this slide presentation was done during industrial training at als company at shah alam

1. BENSOR TEST
PREPARED BY: SITI RUBIAH
MOHAMAD
• Benzoic Acid
• Sorbic Acid

2. WHAT ARE THEY??
Type of chemical preservatives
According to Food Act & Regulations 1983
(Regulation 20) preservative means any
substance that, when added to food , is
capable of inhibiting, retarding or arresting
the process of decomposition, fermentation
or acidification of such food but shall not
include herb, spices, vinegar or wood
smoke.

3. BENZOIC ACID
 Commonly found in most fruits especially berries
(naturally)
 Compounds-Benzoic acid, benzoates (Na/K)and
benzoic acid esters
 For commercial purposes, it is prepared chemically
from toluene (synthetic)
 High concentration give sour taste thus limits it
application
 Usually used in fruit juices, pickles, carbonated
beverages.

4. SORBIC ACID
 This family of compounds is available as
 sorbic acid
 potassium sorbate (much more soluble in water than the acid)
 sodium sorbate or calcium sorbate.
 Give no taste or flavor to products
 Highly preferred as it are physiological
harmlessness and organoleptic neutrality
 Used in beverages, dairy products, fish and
seafood, fat-based products, baked goods and
confectionery products.

5. METHOD OF BENSOR EXTRACTION
Weighed sample (spike:1mL)
Extraction buffer added until 40 mL
Vortex and checked pH to 4.6
Marked up until 50 mL with extraction buffer and
centrifuged
Filtered into vials and sent to HPLC for analyse

6. REFERENCES METHOD:
1) Chapter 29(non-alcoholic beverages and
concentrates) AOAC 16th Edition (1995),
Method 978.08
2) Pearson’s Composition and Analysis of
Foods, pg 89.

7.  MOBILE PHASE OF BENZOIC / SORBIC ACID
1. Weighed and dissolved ammonium acetate in 1000 mL of
deionized water.
2. Adjust pH of the solvent to 4.6.
3. The solvent was filtered
4. Sonicate the solvent.
 EXTRACTION BUFFER BENZOIC/SORBIC ACID
1. Step 1-3 as prepare for its mobile phase.
2. 300 mL from solution were removed and replaced by 300
mL of methanol to get ratio 70: 30 (ammonium acetate :
methanol).

8. SERIAL DILUTION
 Blank = buffer solution
 Standard 1 = 100 ppm
 Standard 2 = 50 ppm
 Standard 3 = 25 ppm
 Standard 4 = 2.5 ppm
 Standard 5 = 0.25 ppm (0.1 ml standard 4 + 0.9 mL buffer)
 Quality check (QC) = 10 ppm (0.1 stock marked up in 10
ml)

9. Figure 1: A basic HPLC system configuration

10. min0 2 4 6 8 10 12 14
mAU
0
5
10
15
20
25
VWD1A,Wavelength=230nm(C:CHEM321DATA140625-BENSOR1406252014-06-2512-19-59027-0801.D)
Area:606.357
Area:718.841
QC
6.101-SodiumBenzoate
9.528-PotassiumSorbate
Figure 2: Example of chromatogram for quality check (QC)
(QC recovery range = 80~120)

11. min0 2 4 6 8 10 12 14
mAU
0
5
10
15
20
25
30
VWD1A,Wavelength=230nm(C:CHEM321DATA140625-BENSOR1406252014-06-2512-19-59035-1801.D)
samplenotdetectedwithbenzoic&sorbicacid
Figure 3: Example of chromatogram for sample product

12. min0 2 4 6 8 10 12 14
mAU
0
10
20
30
40
50
VWD1A,Wavelength=230nm(C:CHEM321DATA140625-BENSOR1406252014-06-2512-19-59038-1901.D)
Area:1144.94
Area:1399.33
samplespikewithbenzoic&sorbicacid
6.026-SodiumBenzoate
9.359-PotassiumSorbate
Figure 3: Example of chromatogram for sample spike

13. CALCULATION INVOLVED
1) conc. of sodium benzoate in sample
=HPLC reading x dilution factor x final volume
sample weight
2) conc. of benzoic acid
= conc. of sodium benzoate x conversion factor
Molecular weight of acid/ molecular
weight of it salt

14. According to Food Act and Regulation 1983 :
SCHEDULE SIXTH (REGULATION 20)
Food
Preservative
[Maximum permitted proportion in milligram per
kilogram (mg/kg)]
Benzoic acid (or sodium
benzoate calculated as
benzoic acid)
Sorbic acid (or its sodium,
calcium or potassium salts
calculated as sorbic acid)
Soya sauce, oyster
sauce, fish sauce, etc
1000 Nil
Fruit- candied, dried,
dried candied etc
350 500
Table 1: Maximum rate of preservatives that allowed in certain food products that
contain in schedule sixth (regulation 20)
Nil means the substance is prohibited in that food

15. SCHEDULE SIXTH (REGULATION 20)
Table 2: The substances specified in table II schedule sixth (regulation
20)
Food Preservative
Bread Propionic acid and its sodium,
potassium , calcium salts.
Flour confection Benzoic acid
Sorbic acid and its sodium,
potassium and calcium salts
Propionic acid andt its sodium,
potassium and calcium salts

16. EXAMPLES OF SAMPLES THAT DONE BENSOR
TEST
ND= Non Detected
Benzoic acid
Sorbic acid and its salts
Propionic and its salts

17. IMPLICATION OF BENZOIC/SORBIC
 Good effect:
 Shelf life of product more longer
 Inhibit microorganism from growth easily
 Bad effect:
 Allergic reaction, nausea, diarrhea
 Change taste and flavor to product
 In some people benzoic acid may liberate histamine
and thus cause pseudo-allergic reactions.
 Affect liver and kidney

18. THANK YOU FOR
LISTENING…