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Kratom (Mitragyna speciosa Korth) is a tropical plant in Thailand, which an addictive effect. Kratom is classified as a Penal Drug Category-5 of the Thai Narcotics Act 2522 (1979), the least restrictive and punitive level. Mitragynine is a major alkaloid of Kratom and is responsible for its opioid effects, which has pharmacological action. Therefore may increase the use of kratom by pretending to be medical benefits. This research aims to study method validation to quantify Mitragynine from Kratom in serum and urine by using Online-SPE-LC-MS/MS. This method has six topics for validation. First, the linearity has r2 (0.995 - 1.000) in blood and urine equal to 0.9998 and 0.9992, respectively. Second, the specificity has not to be disturbed in the chromatogram of Mitragynine in the blood and urine. Third, the accuracy with %recovery (± 15%) in blood and urine equal to 95.25-109.12% and 94.98-111.20%, respectively. Forth, the precision shows that the Intraday-Precision and Interday-Precision values are %RSD (<15%) within acceptable levels in both blood and urine. Fifth, the detection limit (S/N ratio > 3) in the blood and urine is equal to 0.20, 0.15 ng/mL, respectively. And sixth, the limit of quantitation (S/N ratio > 10) in the blood and urine is equal to 1.0, 2.0 ng/mL, respectively. The obtained LOQ has to be in the right criteria. The determination of Mitragynine in the blood and urine using this method is appropriate. This method could analyze samples precisely and reliably.
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