Our Organization
Our Organization
MISSION STATEMENT
“The PNRI contributes to the improvement
of the quality of Filipino life through the
highest standards of nuclear research and
development, specialized nuclear services,
nuclear technology transfer, and efficient
and effective implementation of nuclear
safety practices and regulations”
VINEGAR
Vinegar being one of the oldest chemicals known to
man, has been used as as a condiment, and as a
natural preservative, salad dressing
dressing,, pickling solution,
etc., in the preparation of food.
Acetic acid, CH3COOH is the compound in vinegar
that gives vinegar its sour taste. Vinegar is 44-5%
acetic acid by volume.
UNCOVERING THE METHOD OF
PRODUCTION AND DETECTION OF
SYNTHETIC ACETIC ACID
ADULTERATION IN VINEGAR BY
TANDEM USE OF 14C LIQUID
SCINTILLATION COUNTING AND
13C/12C RATIO MASS
SPECTROMETRY.
Stefan Wechner, 2 AndreyVoropaev 2, Lorenz Eichinger,2 Flora L Santos 1, Soledad Castaneda1,Joseph Michael Racho1,
Preciosa Corazon Pabroa1, Ryan Morco1 and Raymond J. Sucgang,
Sucgang,1
Philippine Nuclear Research Institute, Diliman,
Diliman, Quezon City Philippines; email:[email protected]
HYDROISOTOP GmbHk Scweitenkirchen, Germany.
Fraud in one form or another has
accompanied commercial
transactions for centuries...
EXAMPLE 1: FRAUD IN THE DECLARATION OF BOTANICAL
ORIGIN:
Corn syrup sold as “Dalandan
“Dalandan Juice” or as Ponkan Juice
EXAMPLE 2: SUBSTITUTION WITH CHEAPER SYNTHETIC
RAW MATERIALS:
Adulteration of Vinegar With Synthetic Acetic Acid Solutions.
EXAMPLE 3: MISDECLARATION OF
GEOGRAPHICAL ORIGIN:
Carabao mangoes planted in Mexico and Sold in the U.S as
Guimaras Philippine mangoes
DETECTION OF NON BIOGENIC/ SYNTHETIC
ACETIC ACID ADULTERATION.
The Department of Health Administrative
Order 134 s 1970
1970,, which defines the quality
of vinegar for local consumption, prohibits
the sale of vinegar containing synthetic or
nonbiogenic acetic acid
acid..
The lack of
reliable analytical tools however, has
hampered the proper implementation of
these laws
laws..
C-12
C-14
C-12
SYNTHETIC ACETIC ACID
C-14
BIOGENIC ACETIC ACID
SUCCESS STORIES
14
C ANALYSIS OF VINEGAR
PNRI EXPERIENCE
Vinegar is an exact molecular match with
acetic acid from an industrial supply but
not isotopically.
Natural acetic acid as well as ethanol has a
measurable radioactivity which can be a useful
criterion for the differentiation of biogenic from
synthetic materials.
The method: LOW LEVEL LIQUID SCINTILLATION COUNTING
Figure 2. Variation of natural C-14 activity with time
25
C-14 activity (dpm/g C)
20
15
15.75
15.5
y = -0.2638x + 539.97
R2 = 0.9808
15.49
(PNRI) 15.7
15.21
Calculated C-14 activity
10
C-14 levels in Acetic Acid
(1970-1985)
C-14 levels in Acetic Acid
(1986 to date)
5
Linear (C-14 levels in
Acetic Acid (1970-1985))
0
1965
1970
1975
1980
1985
Year
1990
1995
2000
2005
METHODOLOGY for CC-14 analysis:
1
Vacuum distillation of vinegar, lower & higher
boiling fractions discarded
2
Treatment with CaCO3,
excess CaCO3 filtered off.
off.
3
4
5
Concentration of filtrate in rotary evaporator,
and drying of residue on water bath; final drying
of Ca acetate in vacuum oven.
Reaction of dried Ca acetate precipitate with
pyrophosphoric acid in rotary evaporator and
vacuum distillation of glacial acetic acid.
Preparation of 5 ml 90% acetic acid solution; addition
of 15 ml of scintillant; counting in lowlow-level LSC.
Analysis of isolated acetic acid
using a
Wallac 14141414-3 Guardian
Low – level LSC
LIQUID SCINTILLATION SPECTROMETRY (LSC)
Analytical Measurements Research
Philippine Nuclear Research Institute
Figure 1. Correlation between mean C-14 activity
and synthetic acetic acid concentration
Mean C-14 activity (dpm/g C)
18
16
15.74
14
12
11.69
10
y = -0.1574x + 15.676
8
7.76
2
6
R = 0.9999
4
3.84
2
0
0
-2
0
10
20
30
40
50
60
Percent synthetic acetic acid
70
80
90
100
Figure 11. Range of C-14 Activity in Commercial Vinegar Samples
18.0
16.0
C-14 Activity (dpm/g C)
14.0
12.0
10.0
8.0
6.0
4.0
2.0
0.0
Brand Code
Figure 11. Summary of CC-14 data grouped by brands (A to G)
(Metro Manila, Daet, Baguio City, Lucena, Isabela, and Iloilo)
Iloilo)
18
16
14
12
10
8
6
4
2
0
A A
A A
A A0 A1 B B
B B
B B B0 B1 C C C1 D D D D D D D D D1 DO E
E
E E
E E E1 EO F
F
F
F F1 FO G G G G
Figure 12. Summary of CC-14 data grouped by brands (G1 to Z)
(Metro Manila, Daet, Baguio City, Lucena, Isabela, and Iloilo)
Iloilo)
20
18
16
14
12
10
8
6
4
2
0
G1 H H H H H1 I
I
I1 J1 K K L L
L
L M N N
O P Q Q Q Q R S S T T
U V W Z
Ca
ne
_a
Ca
ne
_a
Ca
ne
_a
Ca
ne
_a
Ca
ne
_a
Ca
ne
_b
Ca
ne
_b
Ca
ne
_b
M
an
go
M
an
go
_a
Pi
ne
ap
pl
e
Pi
ne
ap
pl
e
Ri
ce
_a
Ri
ce
_b
Tu
ba
_a
Tu
ba
_a
Tu
ba
_a
Tu
ba
_a
Tu
ba
_a
Tu
ba
_b
C-14 content of Vinegar of known origin
18.00
16.00
14.00
12.00
10.00
8.00
6.00
4.00
2.00
0.00
SUCCESS STORIES
14
C ANALYSIS OF VINEGAR
PNRI EXPERIENCE
14C RADIOASSAY OF VINEGAR SAMPLES FROM
MAJOR CITIES THROUGHOUT THE PHILIPPINES
REVEALED THAT 7 OUT OF 10 BRANDS, WHICH
HAS PASSED THE LOCAL STANDARDS FOR
“NATURAL VINEGAR” ARE ACTUALLY
ADULTERATED WITH SYNTHETIC ACETIC ACID.
The method: LOW LEVEL LIQUID SCINTILLATION COUNTING
Radiocarbon assay of acetic acid
isolated from vinegar is an alternative
analytical technique which is based
on lower CC-14 levels in petroleumpetroleumderived synthetic acetic acid.
13C/12C
Ratio Determination by
Isotope Ratio Mass Spectrometry.
DETECTION OF GEOGRAPHICAL ORIGIN,
BOTANICAL ORIGIN AND PRODUCTION
PRACTICES.
Isotope ratio analysis
Mass
number
Atomic
number
12
1
1
16
6
1
8
13
2
17
6
1
8
14
3
18
6
1
8
C HO
C HO
C HO
Molecular Species of Water
of Practical Interest
Species
Mass
1H1H 16O
:
18
1H2H 16O
:
19
1H1H 18O
:
20
1H3H 16O
:
20
Examples of this phenomenon are:
1) 1H218O and 1H2H16O have lower vapour pressures than
1H 16O; they also evaporate less easily and
2
2) In most chemical reactions the light isotopic species
reacts faster than the heavy, e.g. Ca12CO3 dissolves faster
in an acid solution than does Ca13CO3.
• In an isotope equilibrium between two chemical
compounds the heavy isotope is generally more
concentrated in the compound having larger molecular
weight.
• At high temperatures the differences between binding
energies of isotopic molecules becomes smaller, resulting
in smaller and ultimately disappearing- isotope effects.
Isotopes in rain get more negative further inland and at higher
altitude
d18O = –21‰
d18O = –12‰
d2H = –158‰
d2H = –86‰
d18O = –11‰
d2H = –78‰
d18O = –3‰
d2H = –14‰
Continent
Ocean
d18O=0‰
d2H=0‰
SOURCE: PRESENTATION OF DR. A. HERCZERG
Ranges for δ 13C values in selected natural compounds
compounds..
Especially noteworthy is the spread in 13C seen in
different plant groups and resulting soil CO2
Commonly used feeds in the acetous fermentation
manufacture of vinegar include sugar cane, rice wine,
pineapple or mango shavings, and coconut juice or sap, etc.
The quality and taste are characteristic of the alcoholic
liquor from which the condiment is made.
Fraud may also be in the form of misrepresentation of the fine
vinegars, with the cheaper
and lower quality products, i.e.
substitution of apple cider and balsamic vinegar with cane or other
cheaper vinegars.
RESULTS
SAMPLE
DESCRIPTION
CODE
C-14
Delta values
(dpm/g C)
VI-0044
commercial cane vinegar
0.00
-30
VI-0109
commercial cane vinegar
0.11
-37,8
VI-0116
commercial vinegar
0.89
-33
VI-0140
commercial vinegar
14.33
-7,9
VI-0142
commercial vinegar
0.00
-38,6
VI-0152
commercial vinegar
14.89
-9,5
VI-0158
commercial vinegar
2,83
-32,7
VI-0130
Sugar Cane Acetator
14.31
-10,2
VI-0131
Natural Sugar Cane
13.22
-15,2
VI-0135
Fresh Pineapple Juice
14.28
-12,9
VI-0136
Fresh Mango Natural
15.67
-20,1
ISOTOPIC RATIOS CAN GIVE
INDICATIONS ABOUT THE
BOTANIC AND GEOGRAPHIC
ORIGIN OF PRIME MATTERS AND,
IN CERTAIN CASES, ON THE
TECHNOLOGICAL PROCESSES
USED IN FOOD PRODUCTION.
Ranges in the δ 34S content of sulphur and sulphur
compounds in different materials and environments.
Ranges in the δ 15N in natural materials
What may be analyzed:
PRODUCT
COMMON ADULTERATION
Fruit Juice Addition of /Dilution with Water
DETECTION METHOD
18
O IRMS
Fruit Juice
Addition of Sugar
13
Honey
Addition of Corn or Cane Sugar
13
Vanilla
Addition of Artificial Vanillin
or p-Hydroxybenzaldehyde
Mislabelling of Botanical Origin
13
Spirit
Vegetable
Oil
Addition of Cheaper Seed Oil
C IRMS
C IRMS
C IRMS
13
C IRMS
Mass Spectrometry
Wine
Dilution with Water
(D/H) and
Wine
Chaptalization
Wine
Mislabeling of Age
14
Vinegar
14
Vinegar
Addition of or Substitution with
Synthetic Acetic Acid
Mislabeling of Botanical Origin
Milk &
Products
Misdeclaration of
Geographical Origin
18
O IRMS
13
C IRMS
C Scintillation Counting
C Scintillation Counting
13
C IRMS
13
C IRMS, (D/H) ,18O,
34
S, 85Sr IRMS
Wine
Dilution with Water
(D/H) and
Wine
Chaptalization
Wine
Mislabeling of Age
14
Vinegar
14
Vinegar
Addition of or Substitution with
Synthetic Acetic Acid
Mislabeling of Botanical Origin
Milk &
Products
Misdeclaration of
Geographical Origin
18
O IRMS
13
C IRMS
C Scintillation Counting
C Scintillation Counting
13
C IRMS
13
C IRMS, (D/H) ,18O,
34
S, 85Sr IRMS
What may be analyzed:
PRODUCT
COMMON ADULTERATION
Fruit Juice Addition of /Dilution with Water
DETECTION METHOD
18
O IRMS
Fruit Juice
Addition of Sugar
13
Honey
Addition of Corn or Cane Sugar
13
Vanilla
Addition of Artificial Vanillin
or p-Hydroxybenzaldehyde
Mislabelling of Botanical Origin
13
Spirit
Vegetable
Oil
Addition of Cheaper Seed Oil
C IRMS
C IRMS
C IRMS
13
C IRMS
Mass Spectrometry
Wine
Dilution with Water
(D/H) and
Wine
Chaptalization
Wine
Mislabeling of Age
14
Vinegar
14
Vinegar
Addition of or Substitution with
Synthetic Acetic Acid
Mislabeling of Botanical Origin
Milk &
Products
Misdeclaration of
Geographical Origin
18
O IRMS
13
C IRMS
C Scintillation Counting
C Scintillation Counting
13
C IRMS
13
C IRMS, (D/H) ,18O,
34
S, 85Sr IRMS