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FDA/ORA/ORS
LIB #4578
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mitragynine.
Validation was performed by characterizing a dry leaf Kratom product and
extracting this material seven times a day over a three day period for a total number of extraction
n=21, refer to Table 4. Once the positive control was characterized, the same positive control
was extracted in duplicate (n=2) for every analytical batch. The UPLC/PDA data demonstrated a
mean value of 1.041% (n=21, 4.2%) and the LC-MS/MS 1.140% (n=14, 6.81%) for mitragynine
in the positive control.
Table 4 :
Validation Data of Mitragynine in Positive Control
Day
Mean % (RSD) Mean % (RSD)
UPLC
LC-MS/MS
1 (n=7)
1.061 (3.6%)
1.156 (5.95%)
2 (n=7)
1.056 (3.1%)
1.125 (7.82%)
3 (n=7)
1.004 (4.4%)
n/a
Overall (n=21, 14)
1.041 (4.2%)
1.1403 (6.81%)
The same positive control was used for all the analysis of products received by the Denver
Laboratory. Table 5 is a summary of three different chemists extracting the positive control
over a 1 year time frame and analyzed on the UPLC/PDA. The overall results demonstrate the
extraction procedure generates reproducible results and the positive control is homogenous.
Table 5: Positive Control over 2 years and different analysis
Batches
Analyst Control
% Mitragynine
1
A
Positive Control
1.177
Positive Control Duplicate
1.110
2
A
Positive Control
1.182
Positive Control Duplicate
1.110
3
B
Positive Control
1.092
Positive Control Duplicate
1.321
4
B
Positive Control
1.241
Positive Control Duplicate
1.207
5
C
Positive Control
1.101
Positive Control Duplicate
1.131
Mean ( n=10)
1.1672
sd
0.074
%RSD
6.31
The same set of samples were extracted and analyzed via both the UPLC/PDA and the LC-
MS/MS. The results for the amount of mitragynine are similar; hence the LC-MS/MS method
can be used for the confirmation and quantitation of mitragynine in Kratom.