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. 2019 Mar 4;6(1):35. doi: 10.3390/medicines6010035

Table 1.

Analytical methods for the identification and/or quantification of Mitragyna speciosa in biological samples.

Compounds Biological Sample (amount) Analytical Technique Internal Standard Extraction Process Linear Range
(ng/mL)
LOD
(ng/mL)
LOQ
(ng/mL)
Recovery
(%)
Reference
Mitragynine,
7-hydroxymitraginine, speciogynine, speciocilliatine, and paynantheine
Urine
(1 mL)
LC–ESI–MS–QTOF mitragynine-d3
7-hydroxymitraginine-d3
Solid-phase extraction
(PolyChrom ClinII cartridges)
2–500 0.25–1 0.5–1 96–63 [120]
Mitragynine,
5-desmethylmitragynine and 17-desmethyldihydro-Mitragynine, and
7-hidroxymitragynine
Urine
(0.2 mL)
UHPLC–ESI/MS–MS
and LC–ESI/MS–MS
Mitraphylline Enzymatic hydrolysis and liquid-liquid extraction (methyl tert-butyl ether) 1–500 - 1.00 78–94 [113]
Mitragynine Urine
(1 mL)
HPLC–DAD - BAµE (N-vinylpyrrolidone polymer) and back-extraction with methanol/acetonitrile (1:1, v/v) under sonication 0.6–24 0.1 0.33 103 [119]
Mitragynine Urine
(1 mL)
LC–ESI–MS
(QTRAP)
methyltestosterone Enzymatic hydrolysis and liquid-liquid extraction
(ethyl tert-butyl ether)
0.25–1.5 0.2 0.25 83 [115]
Mitragynine Peripheral blood, central blood, liver, vitreous, gastric content, and urine
(1 mL)
GC–MS
(EI)
mitragynine-d3 Solid-phase extraction
(Trace-J cartridges)
50–1000 30 50 - [45]
Mitragynine, paynantheine, speciogynine, speciociliatine, 16-carboxy-mitragynine, 9-O-demethyl-mitragynine, and 9-O-demethyl-16-carboxy-mitragynine Urine
(3 mL)
GC–MS
(EI)
- Enzymatic hydrolysis and solid-phase extraction
(HCX cartridge)
- 100 - - [110]
Mitragynine Urine
(2 mL)
LC–ESI/MS–MS Ajmalicine Liquid extraction;
liquid-liquid extraction
(Methyl tert-butyl ether)
0.01–5 0.02 0.1 81 [117]
Mitragynine Blood and urine (1 mL), tissues (liver, kidney, heart, spleen, lung −1g), bile, and vitreous humor LC–ESI/MS–MS Proadifen Enzymatic hydrolysis and liquid-liquid extraction
(n-butyl chloride)
1–10 0.25 1 103 [116]
Mitragynine Rat serum
(0.1 mL)
HPLC–UV
LC–ESI–MS
Acenapthene Liquid–liquid extraction
(diethyl ether)
100–10,000 30 100 85–84 [121]
Mitragynine Rat plasma
(0.1 mL)
HPLC–UV Mefloquine Solid-phase extraction
(MCX Oasis cartridges)
50–1000 25 50 96–98 [33]
Mitragynine Rat and human urine
(1 mL)
HPLC–DAD - Solid-phase extraction
(Oasis® HLB cartridge)
100–10,000 - 100 93–101 [118]
Mitragynine and metabolites Rat and human urine
(1 mL)
LC–ESI.LIT and
LC–ESI.Orbitrap MS
- Enzymatic hydrolysis and solid-phase extraction
(Isolute Confirm HCX and Isolute Confirm C18 cartridges)
- - - - [114]

BAµE: Bar adsorptive microextraction; DAD: Diode-array detection; EI: Electron ionization mode; ESI: Electrospray ionization; GC: Gas chromatography; HPLC: High-performance liquid chromatography; HPLC–UV: High-pressure liquid chromatography with ultraviolet detector; LC: Liquid chromatography; LD: Liquid desorption; LIT: Linear ion trap; MS: Mass spectrometry; LOQ: Limit of detection; LOQ: Limit of quantitation; MS/MS: Tandem mass spectrometry; MTBE: Methyl t-butyl ether; RP–HPLC: Reverse-phase high performance liquid chromatography; SIM: Selective ion mode; TBME: t-Butyl methyl ether; TOF: Time of flight; UHPLC: Ultra-high-performance liquid chromatography.