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Relative Standard Deviation (RSD %)
Analyte
Level 1
Level 2
Liquid Matrix
Semi-solid Matrix
Solid Matrix
mg/kg
mg/kg
Level 1
Level 2
Level 1
Level 2
Level 1
Level 2
Thujone
0.1
1
6
5
8
5
17
17
Menthofuran
0.1
1
14
5
2
15
19
21
Estragole
0.1
1
6
4
9
8
17
13
Pulegone
0.1
1
3
6
7
5
8
16
Methyl eugenol
0.1
1
7
6
3
4
9
12
Coumarin
1
10
13
2
13
3
13
7
β
-Asarone
0.1
1
8
11
6
5
4
10
Table 4: RSD (%) of 6 spiked samples at 2 levels
Flavoring
Retention
Molecular
Precursor
Quantifier
Qualifier
Ion Ratio
Collision
Substance
Time (min)
Weight
Ion
Ion
Ion 1
Qual/Quant
Energy (V)
Thujone
5.86
152.23
110.03
95.02
67.05
0.20
10
Menthofuran
6.66
150.22
107.94
79.01
77.00
0.52
15
Estragole
7.16
148.20
147.98
91.06
115.10
0.82
25
Pulegone
7.74
152.23
152.01
81.03
137.04
0.44
10
Methyl eugenol
9.18
178.23
177.98
147.03
163.05
0.72
15
Coumarin
9.50
146.14
145.92
117.99
89.93
0.68
20
coumarin-d
4
9.49
150.17
149.92
122.02
93.98
0.14
15
Dicyclohexylmethanol
10.42
196.33
112.27
79.05
81.05
0.90
10
β
-Asarone
10.43
208.26
207.99
165.08
193.11
0.91
15
Table 3: GC-MS/MS parameters for selected reaction monitoring of flavorings
Concentrations of
Concentration of
Volume of Working
Volume of Working
Volume of Standard
6 Flavoring Standards
DCHM & coumarin-d
4
Standard Solution
Internal Standard
Solution of
Volume of Water
+ Coumarin (mg/kg)
Standard (mg/kg)
Added (µL)
DCHM Added (µL)
coumarin-d
4
Added (µL)
Added (µL)
0.01 and 0.1
1 and 10
1
10
10
9979
0.05 and 0.5
1 and 10
5
10
10
9975
0.1 and 1
1 and 10
10
10
10
9970
0.5 and 5
1 and 10
50
10
10
9930
1 and 10
1 and 10
100
10
10
9880
1.5 and 15
1 and 10
150
10
10
9830
2 and 20
1 and 10
200
10
10
9780
DCHM = dicyclohexylmethanol internal standard
Table 1: Preparation of matrix matched standards for semi-solid and solid matrices
Concentrations of
Concentration of
Volume of Working
Volume of Working
Volume of Standard
6 Flavoring Standards
DCHM & coumarin-d
4
Standard Solution
Internal Standard
Solution of
and Coumarin (mg/kg)
Standard (mg/kg)
Added (µL)
DCHM Added (µL)
coumarin-d
4
Added (µL)
0.01 and 0.1
1 and 10
1
10
10
0.05 and 0.5
1 and 10
5
10
10
0.1 and 1
1 and 10
10
10
10
0.5 and 5
1 and 10
50
10
10
1 and 10
1 and 10
100
10
10
1.5 and 15
1 and 10
150
10
10
2 and 20
1 and 10
200
10
10
DCHM = dicyclohexylmethanol internal standard
Table 2: Preparation of matrix matched standards for liquid samples
