Literature DB >> 26977432

Data on individual metabolites of synthetic cannabinoids JWH-018, JWH-073 and AM2201 by Cunninghamella elegans.

Shimpei Watanabe1, Unnikrishnan Kuzhiumparambil2, Zophia Winiarski3, Shanlin Fu1.   

Abstract

Synthetic cannabinoids JWH-018, JWH-073 and AM2201 were metabolised by the fungus Cunninghamella elegans. In this article, data on individual metabolites of their retention times, mass accuracies, major product ions and structures indicated by product ions are presented. The data in this article is related to "Biotransformation of synthetic cannabinoids JWH-018, JWH-073 and AM2201 by Cunninghamella elegans" [1].

Entities:  

Year:  2016        PMID： 26977432      PMCID： PMC4781962          DOI： 10.1016/j.dib.2016.02.039

Source DB:  PubMed          Journal:  Data Brief        ISSN： 2352-3409


Specifications Table Value of the data Chromatographic and mass spectrometric data on individual metabolites are provided for reference. Product ions indicative of the structures of metabolites are listed. The data can be compared with human or other in vitro metabolism.

Data

Table 1, Table 3, Table 5 list all the metabolites with biotransformation, retention time, observed accurate mass, formula and major product ions of JWH-018, JWH-073 and AM2201, respectively. Product ions representative of structures for JWH-018, JWH-073 and AM2201 metabolites are presented in Table 4, Table 6, respectively. Overlaid extracted ion chromatograms of all the metabolites of JWH-018, JWH-073 and AM2201 are shown in Fig. 2 of Ref. [1] with annotated metabolite identification (ID) names (Table 1, Table 2, Table 3, Table 4, Table 5, Table 6).
Table 1

Metabolites of JWH-018 after C. elegans incubation.

IDBiotransformationRT, minm/z [M+H]+Mass accuracy (ppm)FormulaMajor product ions
Ma1Dihydrodiol formation+N-dealkylation8.0306.1123−0.7C19H15NO3143, 144, 171, 189
Ma2Dihydrodiol formation+hydroxylation at pentyl side chain8.5392.1852−1.0C24H25NO4143, 144, 171, 189, 230, 374
Ma3Dihydrodiol formation+hydroxylation at pentyl side chain9.2392.1850−1.6C24H25NO4143, 144, 171, 189, 230, 374
Ma4Dihydrodiol formation+ketone formation at pentyl side chain9.3390.1694−1.4C24H23NO4143, 144, 171, 189, 228
Ma5Dihydrodiol formation+ketone formation at pentyl side chain10.3390.1695−1.1C24H23NO4143, 144, 171, 189, 228
Ma6Dihydroxylation at pentyl side chain and naphthalene moiety12.5374.1745−1.5C24H23NO3143, 144, 171, 230
Ma7Dihydroxylation at pentyl side chain12.8374.1745−1.6C24H23NO3127, 144, 155, 246
Ma8Dihydroxylation at pentyl side chain and naphthalene moiety13.1374.1746−1.3C24H23NO3143, 144, 171, 230
Ma9Dihydroxylation at pentyl side chain and naphthalene moiety14.1374.1746−1.3C24H23NO3143, 144, 171, 230
Ma10Ketone formation at pentyl side chain+hydroxylation at naphthalene moiety14.5372.1590−1.0C24H21NO3143, 144, 171, 228
Ma11Dihydroxylation at pentyl side chain and naphthalene moiety14.9374.1745−1.5C24H23NO3143, 144, 171, 230
Ma12Dihydrodiol formation16.1376.1901−1.6C24H25NO3143, 171, 189, 214
Ma13Carboxylation (N-pentanoic acid)a16.4372.1591−0.9C24H21NO3127, 144 155 244
Ma14Hydroxylation at pentyl side chain (5-hydroxypentyl)a17.2358.1796−1.6C24H23NO2127, 144, 155, 230
Ma15Hydroxylation at pentyl side chain (4-hydroxypentyl)a17.6358.1798−1.0C24H23NO2127, 144, 155, 230
Ma16Hydroxylation at pentyl side chain19.9358.1798−1.1C24H23NO2127, 144, 155, 230
Ma17Ketone formation at pentyl side chain20.2356.1640−1.6C24H21NO2127, 144, 155, 228
Ma18Ketone formation at pentyl side chain22.4356.1641−1.2C24H21NO2127, 144, 155, 228
Ma19Hydroxylation at naphthalene moiety24.3358.1797−1.3C24H23NO2143, 171, 214
Ma20Hydroxylation at naphthalene moiety24.8358.1797−1.2C24H23NO2143, 171, 214
Ma21Dehydrogenation28.7340.1692−1.1C24H21NO127, 144, 155, 212
JWH-01831.4342.1851−0.5C24H23NO127, 144, 155, 214

Position confirmed by the use of reference standards.

Table 3

Metabolites of JWH-073 after C. elegans incubation.

IDBiotransformationRT, minm/z [M+H]+Mass accuracy (ppm)FormulaMajor product ions
Mb1Dihydrodiol formation+N-dealkylation8.0306.1123−0.5C19H15NO3143, 144, 171
Mb2Dihydrodiol formation+hydroxylation at butyl side chain8.2378.1700−0.1C23H23NO4143, 144, 171, 189, 216
Mb3Dihydrodiol formation+hydroxylation at butyl side chain8.9378.1690−2.6C23H23NO4143, 144, 171, 189, 216
Mb4Dihydrodiol formation+ketone formation at butyl side chain9.0376.1543−0.1C23H21NO4143, 144, 171, 189, 214
Mb5Dihydroxylation at butyl side chain11.7360.15940.0C23H21NO3127, 144, 155, 232
Mb6Dihydroxylation at butyl side chain and naphthalene moiety12.1360.15950.1C23H21NO3143, 144, 171, 216
Mb7Dihydroxylation at butyl side chain and naphthalene moiety12.8360.1594−0.1C23H21NO3143, 144, 171, 216
Mb8Dihydrodiol formation13.7362.17510.1C23H23NO3143, 144, 171, 189, 200
Mb9Ketone formation at butyl side chain+hydroxylation at naphthalene moiety13.9358.14380.0C23H19NO3143, 144, 171, 214
Mb10Ketone formation at butyl side chain+hydroxylation at naphthalene moiety14.5358.14380.0C23H19NO3143, 144, 171, 214
Mb11Carboxylation (N-butanoic acid)a15.4358.1437−0.1C23H19NO3127, 144, 155, 230
Mb12Hydroxylation at butyl side chain (4-hydroxybutyl)a15.7344.1645−0.2C23H21NO2127, 144, 155, 216
Mb13Hydroxylation at butyl side chain (3-hydroxybutyl)a17.1344.16460.3C23H21NO2127, 144, 155, 216
Mb14Ketone formation at butyl side chain19.5342.14880.0C23H19NO2127, 144, 155, 214
Mb15Hydroxylation at naphthalene moiety21.5344.1645−0.1C23H21NO2143, 144, 171, 200
Mb16Hydroxylation at naphthalene moiety22.0344.16450.0C23H21NO2143, 144, 171, 200
Mb17Dehydrogenation25.7326.1539−0.1C23H19NO127, 155, 198
JWH-07328.8328.16970.3C23H21NO127, 144, 155, 200

Position confirmed by the use of reference standards.

Table 5

Metabolites of AM2201 after C. elegans incubation.

IDBiotransformationRT, minm/z [M+H]+Mass accuracy (ppm)FormulaMajor product ions
Mc1Dihydrodiol formation+N-dealkylation8.0306.1122−1.0C19H15NO3143, 144, 171, 189
Mc2Dihydroxylation at pentyl side chain and naphthalene moiety+glucosidation8.2554.21850.0C30H32FNO8143, 144, 171, 248, 392
Mc3Dihydroxylation at pentyl side chain and naphthalene moiety+glucosidation8.4554.21860.2C30H32FNO8143, 144, 171, 248, 392
Mc4Oxidative defluorination+dihydrodiol formation (JWH-018 dihydrodiol-hydroxy)8.5392.18580.4C24H25NO4143, 171, 189, 230
Mc5Dihydrodiol formation+hydroxylation at pentyl side chain8.5410.17640.6C24H24FNO4143, 144, 171, 189, 248
Mc6Dihydrodiol formation+hydroxylation at pentyl side chain8.7410.1761−0.2C24H24FNO4143, 144, 171, 189, 248
Mc7Dihydroxylation at pentyl side chain and/or indole moiety+glucosidation9.0554.21860.3C30H32FNO8127, 155, 264, 392
Mc8Trihydroxylation at pentyl side chain, indole moiety and naphthalene moiety9.3408.16060.1C24H22FNO4143, 160, 171, 264
Mc9Dihydrodiol formation+dihydroxylation at indole moiety9.5426.17120.1C24H24FNO5143, 171, 176, 189, 264
Mc10Dihydrodiol formation+ketone formation at pentyl side chain9.7408.16060.0C24H22FNO4143, 171, 189, 246
Mc11Trihydroxylation at pentyl side chain, indole moiety and naphthalene moiety10.2408.16060.0C24H22FNO4143, 160, 171, 264
Mc12Hydroxylation at indole moiety+glucosidation10.5538.2235−0.1C30H32FNO7127, 155, 160, 248, 376
Mc13Dihydroxylation at pentyl side chain and/or indole moiety+glucosidation10.7554.2183−0.3C30H32FNO8127, 155, 264, 392
Mc14Hydroxylation at naphthalene moiety+glucosidation10.8538.2234−0.4C30H32FNO7143, 144, 171, 232, 376
Mc15Oxidative defluorination+hydroxylation at indole moiety (JWH-018 dihydroxy)11.1374.17510.2C24H23NO3127, 155, 160, 246
Mc16Dihydroxylation at naphthalene+glucosidation11.5554.2182−0.4C30H32FNO8187, 232, 392
Mc17Dihydroxylation at indole moiety and pentyl side chain11.5392.16570.0C24H22FNO3127, 155, 160, 264
Mc18Oxidative defluorination+hydroxylation at indole moiety (JWH-018 dihydroxy)11.8374.1748−0.8C24H23NO3127, 155, 160, 246
Mc19Dihydroxylation at indole moiety and pentyl side chain11.9392.1656−0.2C24H22FNO3127, 155, 160, 264
Mc20Oxidative defluorination to carboxylic acid+hydroxylation at naphthalene moiety12.0388.15471.0C24H21NO4143, 144, 171, 244
Mc21Oxidative defluorination+hydroxylation at pentyl side chain (JWH-018 dihydroxy)12.1374.17510.2C24H23NO3127, 144, 155, 246
Mc22Dihydroxylation at indole moiety and pentyl side chain12.2392.16580.5C24H22FNO3127, 155, 160, 264
Mc23Oxidative defluorination+hydroxylation at naphthalene moiety (JWH-018 dihydroxy)12.3374.1749−0.4C24H23NO3143, 144, 171, 230
Mc24Dihydroxylation at pentyl side chain and naphthalene moiety12.5392.16511.4C24H22FNO3143, 144, 171, 248
Mc25Dihydrodiol formation12.5394.1812−0.3C24H24FNO3143, 144, 171, 189, 232
Mc26Oxidative defluorination to carboxylic acid+hydroxylation at naphthalene moiety12.7388.15440.3C24H21NO4143, 144, 171, 244
Mc27Dihydroxylation at indole moiety and pentyl side chain12.7392.16570.0C24H22FNO3127, 155, 160, 264
Mc28Oxidative defluorination+hydroxylation at naphthalene moiety (JWH-018 dihydroxy)12.9374.1750−0.3C24H23NO3143, 144, 171, 230
Mc29Dihydroxylation at pentyl side chain and naphthalene moiety13.0392.16550.3C24H22FNO3143, 144, 171, 248
Mc30Dihydrodiol formation13.5394.18181.2C24H24FNO3143, 144, 171, 189, 232
Mc31Dihydroxylation at pentyl side chain and naphthalene moiety13.6392.1655−0.3C24H22FNO3143, 144, 171, 248
Mc32Dihydroxylation at pentyl side chain and naphthalene moiety14.4392.16590.6C24H22FNO3143, 144, 171, 248
Mc33Oxidative defluorination to carboxylic acid (JWH-018 N-pentanoic acid)a16.4372.1592−0.7C24H21NO3127, 144, 155, 244
Mc34Oxidative defluorination (JWH-018 N-(5-hydroxypentyl))a17.1358.18020.2C24H23NO2127, 144, 155, 230
Mc35Hydroxylation at pentyl side chain17.3376.17090.4C24H22FNO2127, 144, 155, 248
Mc36Hydroxylation at indole moiety17.7376.1707−0.2C24H22FNO2127, 155, 160, 248
Mc37Hydroxylation at pentyl side chain18.0376.17080.1C24H22FNO2127, 144, 155, 248
Mc38Dihydroxylation at naphthalene moiety+sulfation18.7472.1222−0.6C24H22FNO6S144, 158, 159, 186, 187, 232, 391, 392
Mc39Hydroxylation at pentyl side chain18.8376.17080.3C24H22FNO2127, 144, 155, 248
Mc40Hydroxylation at naphthalene moiety19.2376.17090.6C24H22FNO2143, 144, 171, 232
Mc41Hydroxylation at naphthalene moiety19.8376.1703−1.2C24H22FNO2143, 144, 171, 232
Mc42Dihydroxylation at naphthalene moiety+sulfation20.2472.1224−0.1C24H22FNO6S144, 158, 159, 186, 187, 232, 391, 392
Mc43Ketone formation at pentyl side chain20.4374.1549−0.5C24H20FNO2127, 144, 155, 246
Mc44Hydroxylation at naphthalene moiety20.6376.1706−0.3C24H22FNO2143, 144, 171, 232
Mc45Hydroxylation at naphthalene moiety21.3376.1707−0.1C24H22FNO2143, 144, 171, 232
Mc46Dihydroxylation at indole moiety+sulfation22.0472.1223−0.3C24H22FNO6S127, 155, 175, 176, 264, 391, 392
AM220125.6360.17590.2C24H22FNO127, 144, 155, 232
Mc47Defluorination+Demethylation (JWH-073)a28.8328.1695−0.3C23H21NO127, 144, 155, 200
Mc48Defluorination (JWH-018)a31.5342.18530.0C24H23NO127, 144, 155, 214

Position confirmed by the use of reference standards.

Table 4

Key diagnostic product ions and their tentative structures used in elucidating biotransformation pathways of JWH-073 after C. elegans incubation.

BiotransformationIDKey diagnostic product ions (m/z) and tentative structures
Carboxylation at butyl side chainMb11144: unchanged indole, 230: carboxylated butylindole
DehydrogenationMb17198: dehydrogenated N-butylindole
Dihydrodiol formation at naphthalene moietyMb8189: naphthalene with dihydrodiol
Dihydrodiol formation at naphthalene moiety+hydroxylation at butyl side chainMb2, Mb3144: unchanged indole, 189: naphthalene with dihydrodiol, 216: hydroxylated N-butylindole
Dihydrodiol formation at naphthalene moiety+ketone formation at butyl side chainMb4144: unchanged indole, 189: naphthalene with dihydrodiol, 214: N-butylindole with ketone
Dihydrodiol formation at naphthalene moiety+N-dealkylationMb1171: hydroxylated naphthalene (resulting from naphthalene with dihydrodiol [2])
Dihydroxylation at butyl side chainMb5144: unchanged indole, 232: dihydroxylated N-butylindole
Dihydroxylation at butyl chain and naphthalene moietyM6, Mb7144: unchanged indole, 171: hydroxylated naphthalene, 216: hydroxylated N-butylindole
Hydroxylation at butyl side chainMb12, Mb13144: unchanged indole, 216: hydroxylated N-butylindole
Hydroxylation at naphthalene moietyMb15, Mb16171: hydroxylated naphthalene
Ketone formation at butyl side chainMb14144: unchanged indole, 214: N-butylindole with ketone
Ketone formation at butyl side chain+hydroxylation at naphthalene moietyMb9, Mb10144: unchanged indole, 171: hydroxylated naphthalene, 214: N-butylindole with ketone
Table 6

Key diagnostic product ions and their tentative structures used in elucidating biotransformation pathways of AM2201 after C. elegans incubation. Square brackets indicate phase II metabolism.

BiotransformationIDKey diagnostic product ions (m/z) and tentative structures
Defluorination (JWH-018)Mc48155: unchanged naphthalene, 214: unchanged N-pentylindole
Defluorination+Demethylation (JWH-073)Mc47155: unchanged naphthalene, 200: unchanged N- butylindole
Dihydrodiol formation at naphthalene moietyMc25, Mc30189: naphthalene with dihydrodiol
Dihydrodiol formation at naphthalene moiety+dihydroxylation at indole moietyMc9176: dihydroxylated indole, 189: naphthalene with dihydrodiol
Dihydrodiol formation at naphthalene moiety+hydroxylation at pentyl side chainMc5, Mc6144: unchanged indole, 189: naphthalene with dihydrodiol, 248: hydroxylated N-fluoropentylindole
Dihydrodiol formation at naphthalene moiety+ketone formation at pentyl side chainMc10144: unchanged indole, 189: naphthalene with dihydrodiol, 246: N-fluoropentylindole with ketone
Dihydrodiol formation at naphthalene moiety+N-dealkylationMc1189: naphthalene with dihydrodiol
Dihydroxylation at indole moiety [+sulfation][Sulfate Mc46]176: dihydroxylation at indole, [392: dihydroxylated AM2201]
Dihydroxylation at indole moiety and pentyl side chainMc17, Mc19, Mc22, Mc27160: hydroxylated indole, 264: dihydroxylated N-fluoropentylindole
Dihydroxylation at naphthalene moiety [+glucosidation and sulfation][Glucoside Mc16, sulfates Mc38, Mc42]187: dihydroxylated naphthalene, [392: dihydroxylated AM2201]
Dihydroxylation at naphthalene moiety and pentyl side chain [+glucosidation]Mc24, Mc29, Mc31, Mc32, [glucosides Mc2, Mc3]144: unchanged indole, 171: hydroxylated naphthalene, 248: hydroxylated N-fluoropentylindole, [392: dihydroxylated AM2201]
Dihydroxylation at pentyl side chain and/or indole moiety [+glucosidation][glucosides Mc7, Mc13]264: dihydroxylated N-fluoropentylindole, [392: dihydroxylated AM2201]
Hydroxylation at indole moiety [+glucosidation]Mc36, [glucoside Mc12]160: hydroxylated indole, 248: hydroxylated N-fluoropentylindole, [376: hydroxylated AM2201]
Hydroxylation at naphthalene moiety [+glucosidation]Mc40, Mc41, Mc44, Mc45, [glucoside Mc14]171: hydroxylated naphthalene, [376: hydroxylated AM2201]
Hydroxylation at pentyl side chainMc35, Mc37 and Mc39144: unchanged indole, 248: hydroxylated N-fluoropentylindole
Ketone formation at pentyl side chainMc43144: unchanged indole, 246: N-fluoropentylindole with ketone
Oxidative defluorination (JWH-018 N-(5-hydroxypentyl))Mc34144: unchanged indole, 230: hydroxylated N-pentylindole
Oxidative defluorination+dihydrodiol formation (JWH-018 dihydrodiol-hydroxy)Mc4189: naphthalene with dihydrodiol, 230: hydroxylated N-pentylindole
Oxidative defluorination+hydroxylation at indole moiety (JWH-018 dihydroxy)Mc15, Mc18160: hydroxylated indole, 246: dihydroxylated N-pentylindole
Oxidative defluorination+hydroxylation at naphthalene moiety (JWH-018 dihydroxy)Mc23, Mc28144: unchanged indole, 171: hydroxylated naphthalene, 230: hydroxylated N-pentylindole
Oxidative defluorination+hydroxylation at pentyl side chain (JWH-018 dihydroxy)Mc21144: unchanged indole, 246: dihydroxylated N-pentylindole
Oxidative defluorination to carboxylic acid (JWH-018 N-pentanoic acid)Mc33144: unchanged indole, 244: carboxylated pentylindole
Oxidative defluorination to carboxylic acid+hydroxylation at naphthalene moiety.Mc20, Mc26144: unchanged indole, 171: hydroxylated naphthalene, 244: carboxylated pentylindole
Trihydroxylation at indole moiety, naphthalene moiety and pentyl side chainMc8, Mc11160: hydroxylated indole, 171: hydroxylated naphthalene, 264: dihydroxylated N-fluoropentylindole
Table 2

Key diagnostic product ions and their tentative structures used in elucidating biotransformation pathways of JWH-018 after C. elegans incubation.

BiotransformationIDKey diagnostic product ions (m/z) and tentative structures
Carboxylation at pentyl side chainMa13144: unchanged indole, 244: carboxylated pentylindole
Dehydrogenation at pentyl side chainMa21144: unchanged indole, 212: dehydrogenated N-pentylindole
Dihydrodiol formation at naphthalene moietyMa12189: naphthalene with dihydrodiol
Dihydrodiol formation at naphthalene moiety+hydroxylation at pentyl side chainMa2, Ma3144: unchanged indole, 189: naphthalene with dihydrodiol, 230: hydroxylated N-pentylindole
Dihydrodiol formation at naphthalene moiety+ketone formation at pentyl side chainMa4, Ma5189: naphthalene with dihydrodiol, 228: N-pentylindole with ketone
Dihydrodiol formation at naphthalene moiety+N-dealkylationMa1189: naphthalene with dihydrodiol
Dihydroxylation at pentyl side chainMa7144: unchanged indole, 246: dihydroxylated N-pentylindole
Dihydroxylation at pentyl side chain and naphthalene moietyMa6, Ma8, Ma9, Ma11144: unchanged indole, 171: hydroxylated naphthalene, 230: hydroxylated N-pentylindole
Hydroxylation at naphthalene moietyMa19, Ma20171: hydroxylated naphthalene
Hydroxylation at pentyl side chainMa14 – Ma16144: unchanged indole, 230: hydroxylated N-pentylindole
Ketone formation at pentyl side chainMa17, Ma18144: unchanged indole, 228: N-pentylindole with ketone
Ketone formation at pentyl side chain+hydroxylation at naphthalene moietyMa10144: unchanged indole, 171: hydroxylated naphthalene, 228: N-pentylindole with ketone

Experimental design, materials and methods

Chemicals

JWH-018 and JWH-073 were synthesized in-house following previously reported methods and characterized by mass spectrometry (MS) and 1D, 2D nuclear magnetic resonance (NMR) spectroscopy [3], [4]. AM2201 (purity 99.4%) was obtained from the National Measurement Institute (North Ryde, NSW, Australia). Reference standards JWH-018 N-(4-hydroxypentyl), JWH-018 N-(5-hydroxypentyl), JWH-018 N-pentanoic acid, JWH-073 N-(3-hydroxybutyl), JWH-073 N-(4-hydroxybutyl) and JWH-073 N-butanoic acid were obtained from PM separations (Capalaba, QLD, Australia). Reagent grade dichloromethane, methanol, KH2PO4, NaCl and LC grade acetonitrile and methanol were obtained from Chemsupply (Gilman, SA, Australia). Potato dextrose agar, glucose, peptone, and yeast extract were purchased from Oxoid Australia (Adelaide, SA, Australia).

Microbial culture and biotransformation conditions

Cultures of C. elegans ATCC 10028b (Cryosite Ltd, South Granville, NSW, Australia) were propagated on potato dextrose agar plates at 27 °C for 5 days. The mycelia from five plates were then transferred to 20 mL of sterile physiological saline solution and homogenized for 5 min. Approximately 3 mL aliquots of the homogenate were used to inoculate 250 mL Erlenmeyer flasks containing 100 ml of growth media. The cultures were incubated for 48 h at 26 °C on an Infors HT Multitron rotary shaker (in vitro Technologies, Noble Park North, VIC, Australia) operating at 180 rpm. After 48 h, 10 mg of JWH-018, JWH-073 or AM2201 dissolved in 0.5 mL of methanol was added to the culture and incubated for further 72 h [5]. Control experiments consisted of cultures without cannabinoids and flasks containing only media and cannabinoid [6], [7].

Extraction, isolation, and identification of metabolites

After 72 h of incubation, the contents of each flask, including the controls, were filtered through Buchner funnel into a separating funnel and extracted with three aliquots of dichloromethane (3×50 mL). The combined organic extracts were evaporated to dryness under vacuum at 40 °C using a Buchi rotary evaporator (in vitro Technologies, Noble Park North, VIC, Australia) and placed under high vacuum to remove traces of moisture. The residue was dissolved in acetonitrile to prepare 1 mg/mL stock solution and was filtered through 0.22 µM syringe filter before analysis. Cannabinoid parent drugs and metabolites were chromatographically separated using an Agilent Zorbax Eclipse XDB-C18 analytical column (150×4.6 mm, 5 μm). Mobile phases consisted of 0.1% formic acid in water (mobile phase A) and acetonitrile (mobile phase B). The gradient used consisted of 30% B (0 to 2 min), linear gradient from 30% B to 50% B (2 to 5 min), 50% B to 90% B (5 to 30 min, hold for 5 min) and 90% B to 30% B (35 to 40 min) run at 0.4 mL/min. MS data were acquired on an Agilent 6490 Triple Quadrupole mass spectrometer with an electrospray ionization source (ESI) source (positive ion mode), interfaced with an Agilent 1290 LC system. Samples prepared were injected in 2 µL volume to obtain full scan and product ion scan spectra. Product ion scan experiments were conducted on precursor ions that were presumed to be metabolites based on the comparison of full scan spectra of the samples and controls. A fragmentor voltage of 380 V with discrete collision energy of 10, 20, 30 and 40 eV (for product ion scan) was applied. The scanning mass range was set at m/z 100–1000 (scan time=500 ms). The sheath gas temperature and flow were set to 250 °C and 11 L/min, respectively. The capillary and nozzle voltages were 3000 V and 1500 V, respectively. High resolution quadrupole Time-of-Flight mass spectrometry (HRQToFMS) experiments were carried out on an Agilent 6510 Accurate Mass QToF Mass Spectrometer, equipped with ESI source operated in positive ion mode, in order to determine accurate masses of the metabolites. The LC system and conditions used were the same as above. The following operation parameters were used: injection volume 2 µL (full scan) and 10 µL (product ion scan); capillary voltage 3500 V; nebulizer pressure 40 psi (275790 Pa); drying gas 10.0 L/min; gas temperature 350 °C; fragmentor voltage 160 V; collision energy 10, 20 and 40 eV; skimmer voltage 60 V. HRQToFMS accurate mass spectra were recorded across the range from m/z 100 to m/z 1000. The mass axis was calibrated using the mixture provided by the manufacturer over the m/z 50–3200 range. A second orthogonal sprayer with a reference solution was used as a continuous calibration using the following reference masses: m/z 121.0509 and m/z 922.0098. The chromatographic conditions and column used were same as described above. The controls were subjected to the same analysis. Analysis of the chromatographic and mass spectrometric data was conducted using MassHunter Workstation Software Qualitative Analysis (version B.06.00, Agilent). Peaks present in the fungus sample, but not in the controls, were manually identified and their fragmentation patterns and accurate masses were examined to identify the metabolites. The signal-to-noise ratio of all the identified metabolites was greater than 5.
Subject areaPharmacology
More specific subject areaDrug Metabolism
Type of dataTable
How data was acquiredLiquid chromatography -tandem mass spectrometry (Agilent 1290 LC system coupled to Agilent 6490 Triple Quadrupole mass spectrometer), high resolution quadrupole Time-of-Flight mass spectrometry (Agilent 6510 Accurate Mass QToF Mass Spectrometer)
Data formatAnalysed
Experimental factorsSamples were extracted by dichloromethane.
Experimental featuresLiquid chromatography-tandem mass spectrometry analysis of fungal metabolites of synthetic cannabinoids JWH-018, JWH-073 and AM2201
Data source locationSydney, Australia
Data accessibilityData are available with this article
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6.  Detection of urinary metabolites of AM-2201 and UR-144, two novel synthetic cannabinoids.

Authors:  Tim Sobolevsky; Ilya Prasolov; Grigory Rodchenkov
Journal:  Drug Test Anal       Date:  2012-10-05       Impact factor: 3.345
  6 in total
  2 in total

1.  Metabolic Profile of Synthetic Cannabinoids 5F-PB-22, PB-22, XLR-11 and UR-144 by Cunninghamella elegans.

Authors:  Shimpei Watanabe; Unnikrishnan Kuzhiumparambil; My Ann Nguyen; Jane Cameron; Shanlin Fu
Journal:  AAPS J       Date:  2017-04-28       Impact factor: 4.009

2.  In vitro metabolism of synthetic cannabinoid AM1220 by human liver microsomes and Cunninghamella elegans using liquid chromatography coupled with high resolution mass spectrometry.

Authors:  Shimpei Watanabe; Unnikrishnan Kuzhiumparambil; Shanlin Fu
Journal:  Forensic Toxicol       Date:  2018-05-24       Impact factor: 4.096
  2 in total

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