KRA-01

FDA/ORA/ORS

LIB #4578 19 of 25

and needed to be within 10%; the ions found were in comparison m/z 174 hence the ion ratios are as follows: m/z 159 (68%) and 238 (61%). Mitragynine was confirmed in the positive control and the samples analyzed by the Denver Laboratory. The use of the TSQ provided the selectivity and sensitivity and mitragynine was not detected in the solvent blank and negative control. Figure 15 : a) Mitragynine Solvent Standard - Total Ion Chromatogram (TIC) b) Mitragynine Reconstructed Ion Chromatogram (RIC) Kraton_27Dec12_A2 - TIC - SM: 5 RT: 5.00 - 14.98 NL: 1.80E6

RT: 11.53

10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100 Relative Intensity

a

0 5

RT: 12.22

13.02 13.56

5.26 5.89

6.36

6.87

7.35

7.94

8.42

9.31

9.99

10.90

RT: 14.54

9

10

11

8

12

13

14

6

7

Time (min)

RT: 10.50 - 15.00

RT: 10.50 - 15.00

RT: 11.53

NL: 8.70E5 TIC F: + c ESI SRM ms2 399.200 [159.099-159.101, 174.099-174.101, 238.199-238.201] MS ICIS Kraton_28Dec12_A6

NL: 3.88E5 m/z= 173.90-174.10 F: + c ESI SRM ms2 399.200 [159.099-159.101, 174.099-174.101, 238.199-238.201] MS ICIS Kraton_28Dec12_A6

Sample Mitragynine TIC

100

10 20 30 40 50 60 70 80 90 100

b

95

90

SRM 399.2-> 174 Sample Mitragynine Rt=11.53

85

80

75

RT: 12.27

70

RT: 14.29

RT: 12.79 RT: 13.35

10 20 30 40 50 60 70 80 90 100 Relative Abundance 0

RT: 11.53

NL: 2.46E5 m/z= 159.00-159.20 F: + c ESI SRM ms2 399.200 [159.099-159.101, 174.099-174.101, 238.199-238.201] MS ICIS Kraton_28Dec12_A6

65

60

55

SRM 399.2-> 159.1 Sample Mitragynine Rt=11.53

50

Total Ion Chromatogram (TIC) Sample Mitragynine Rt=11.53

45

Relative Abundance

40

RT: 12.22 RT: 12.87 RT: 13.81

35

0 10 20 30 40 50 60 70 80 90 100 0

RT: 11.53

NL: 2.37E5 m/z= 238.00-238.20 F: + c ESI SRM ms2 399.200 [159.099-159.101, 174.099-174.101, 238.199-238.201] MS ICIS Kraton_28Dec12_A6

30

25

20

SRM 399.2-> 238.1 Sample Mitragynine Rt=11.53

15

10

5

RT: 12.39

RT: 12.79

RT: 12.44

0

15

13

14

12

15

11

14

13

11

12

Time (min)

Time (min)

Made with