Optimized glucuronide hydrolysis for the detection of psilocin in human urine samples

Kamata, Toshihide; Nishikawa, Mayumi; Katagi, Munehiro; Tsuchihashi, Hitoshi

doi:10.1016/j.jchromb.2003.08.030

Cited by 55 publications

(51 citation statements)

References 15 publications

(17 reference statements)

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“…The hydrolysis conditions optimized for the uri-nary metabolites of 5-MeO-DIPT are very different from those for urinary psilocin (4-hydroxy-N,Ndimethyltryptamine) glucuronide, the metabolite of psilocybin, established in our previous study, 6) although both drugs are psychotomimetic tryptamine derivatives. β-Glucuronidase was used for conjugated psilocin because only glucuronide was detected as its conjugate.…”

Section: Enzymatic Hydrolysis Of Conjugated Metabolitesmentioning

confidence: 72%

Urinary Excretion Profiles of 5-Methoxy-N,N-diisopropyltryptamine and Its Relevant Metabolites in Humans

Kamata

Katagi

Kamata

et al. 2007

JOURNAL OF HEALTH SCIENCE

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5-Methoxy-N,N-diisopropyltryptamine (5-MeO-DIPT), a psychotomimetic tryptamine derivative, and its relevant metabolites have been determined in eleven urine specimens from six 5-MeO-DIPT users, and their excretion profiles have been investigated by gas chromatography/mass spectrometry (GC/MS) and liquid chromatography/mass spectrometry (LC/MS). Three metabolites, 5-hydroxy-N,Ndiisopropyltryptamine (5-OH-DIPT), 6-hydroxy-5-methoxy-N,N-diisopropyltryptamine (6-OH-5-MeO-DIPT), and 5-methoxy-N-isopropyltryptamine (5-MeO-NIPT) were determined in the urine specimens. Urinary conjugated metabolites, both sulfates and glucuronides of 5-OH-DIPT and 6-OH-5-MeO-DIPT, were hydrolyzed completely by the use of Helix pomatia sulfatase/β-glucuronidase. Degradation of 6-OH-5-MeO-DIPT during incubation for hydrolysis was successfully prevented by the addition of ascorbic acid. The hydrolysis treatment increased the detection amounts of 5-OH-DIPT and 6-OH-5-MeO-DIPT in most of the specimens, and the increase in 6-OH-5-MeO-DIPT was more drastic than that in 5-OH-DIPT. The concentrations of 5-MeO-DIPT (< 1.7 µg/ml) and 5-MeO-NIPT (< 3.5 µg/ml) were lower than those of 5-OH-DIPT (0.01-47 µg/ml) and 6-OH-5-MeO-DIPT (< 69 µg/ml) detected after hydrolysis (the totals of their free and conjugated forms). These metabolites were detectable over longer periods post intake than the parent drug; 35 hr for 5-MeO-DIPT, 80 hr for 5-OH-DIPT, and

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Section: Enzymatic Hydrolysis Of Conjugated Metabolitesmentioning

confidence: 72%

Urinary Excretion Profiles of 5-Methoxy-N,N-diisopropyltryptamine and Its Relevant Metabolites in Humans

Kamata

Katagi

Kamata

et al. 2007

JOURNAL OF HEALTH SCIENCE

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“…As psilocin glucuronide is an important excretion product in urine, Kamata et al (2003) developed an optimized glucuronide hydrolysis method for the detection of psilocin by LC-MS-MS in human urine. Recently, Ramirez Fernandez et al (2007) reported a validated LC-MS-MS method for the simultaneous analysis of multiple hallucinogens, including psilocin, in urine of subjects that have ingested hallucinogenic mushrooms.…”

Section: Analysis Of Psilocybin and Psilocin In Human Fluids And Tissuesmentioning

confidence: 99%

“…As the average half-life of psilocin is reported to be about 2.5-3.0 hours, the psilocin concentration reaches the limit of quantification within around 6-7 hours (Hasler et al, 1997;Anastos et al, 2005). Psilocin is excreted in urine, mainly as glucuronide and unchanged psilocin Hasler et al, 2002;Kamata et al, 2003). Other metabolites found in lower quantities include 4-hydroxyindole-3-yl-acetaldehyde, 4-hydroxyindole-3-yl-acetic acid, and 4-hydroxytryptophol (Holzmann, 1995;Hasler et al, 1997).…”

Section: Pharmacokineticmentioning

confidence: 99%

Occurrence and use of hallucinogenic mushrooms containing psilocybin alkaloids

Andersson¹,

Gry²,

Kristinsson³

2009

TemaNord

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“…Psilocin glucuronide is a major psilocin metabolite in humans (Sticht and Käferstein, 2000;Kamata et al, 2003, and we have now examined the glucuronidation of psilocin, as well as 4-hydroxyindole, a structural analog of psilocin that lacks the N,N-dimethylaminoethyl side chain of this recreational drug (Fig. 1).…”

Section: Discussionmentioning

confidence: 99%

“…However, later pharmacokinetic and forensic studies provided evidence that psilocin is mostly eliminated by conjugative metabolism as psilocin glucuronide (Sticht and Käferstein, 2000;Grieshaber et al, 2001). It was found that addition of ␤-glucuronidase to the detection assays for psilocin in body fluids yielded additional psilocin, presumably released from psilocin glucuronide (Hasler et al, 2002;Kamata et al, 2003). Moreover, direct analysis by liquid chromatographymass spectrometry (LC-MS) of serum samples 5 h after mushroom intoxication showed that up to 80% of the psilocin was present as the glucuronide conjugate .…”

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confidence: 99%

Glucuronidation of Psilocin and 4-Hydroxyindole by the Human UDP-Glucuronosyltransferases

Manevski

Kurkela²,

Höglund³

et al. 2009

Drug Metab Dispos

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ABSTRACT:We have examined the glucuronidation of psilocin, a hallucinogenic indole alkaloid, by the 19 recombinant human UDP-glucuronosyltransferases (UGTs) of subfamilies 1A, 2A, and 2B. The glucuronidation of 4-hydroxyindole, a related indole that lacks the N,N-dimethylaminoethyl side chain, was studied as well. UGT1A10 exhibited the highest psilocin glucuronidation activity, whereas the activities of UGTs 1A9, 1A8, 1A7, and 1A6 were significantly lower. On the other hand, UGT1A6 was by far the most active enzyme mediating 4-hydroxyindole glucuronidation, whereas the activities of UGTs 1A7-1A10 toward 4-hydroxyindole resembled their respective psilocin glucuronidation rates. Psilocin glucuronidation by UGT1A10 followed Michaelis-Menten kinetics in which psilocin is a low-affinity high-turnover substrate (K m ‫؍‬ 3.8 mM; V max ‫؍‬ 2.5 nmol/min/mg). The kinetics of psilocin glucuronidation by UGT1A9 was more complex and may be best described by biphasic kinetics with both intermediate (K m1 ‫؍‬ 1.0 mM) and very low affinity components. The glucuronidation of 4-hydroxyindole by UGT1A6 exhibited higher affinity (K m ‫؍‬ 178 M) and strong substrate inhibition. Experiments with human liver and intestinal microsomes (HLM and HIM, respectively) revealed similar psilocin glucuronidation activity in both samples, but a much higher 4-hydroxyindole glucuronidation rate was found in HLM versus HIM. The expression levels of UGTs 1A6-1A10 in different tissues were studied by quantitative real-time-PCR, and the results, together with the activity assays findings, suggest that whereas psilocin may be subjected to extensive glucuronidation by UGT1A10 in the small intestine, UGT1A9 is likely the main contributor to its glucuronidation once it has been absorbed into the circulation.

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Optimized glucuronide hydrolysis for the detection of psilocin in human urine samples

Cited by 55 publications

References 15 publications

Urinary Excretion Profiles of 5-Methoxy-N,N-diisopropyltryptamine and Its Relevant Metabolites in Humans

Urinary Excretion Profiles of 5-Methoxy-N,N-diisopropyltryptamine and Its Relevant Metabolites in Humans

Occurrence and use of hallucinogenic mushrooms containing psilocybin alkaloids

Glucuronidation of Psilocin and 4-Hydroxyindole by the Human UDP-Glucuronosyltransferases

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