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3 protocols using tripelennamine hydrochloride

1

Histamine Receptor Binding Assay

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The specific HRH1 antagonist [3H]pyrilamine was used in saturation and competitive binding experiments, at 20.0 Ci/mmol (Perkin-Elmer, Waltham, MA, United States), to label histamine receptors. The synthesis of protein labeled with carbon fourteen (14C) was produced using [14C]leucine (20.0 Ci/nmol) (Perkin-Elmer, Waltham, MA, United States). The following drugs were used as cold competitors: doxepin hydrochloride (Wako, Osaka, Japan), epinastine hydrochloride (Sigma–Aldrich, St. Louis, MO, United States), pyrilamine maleate (Sigma–Aldrich, St. Louis, MO, United States), ranitidine hydrochloride (Sigma–Aldrich, St. Louis, MO, United States), thioperamide (R&D Systems, Minneapolis, MN, United States), and alpha-methylhistamine (Sigma–Aldrich, St. Louis, MO, United States). Tripelennamine hydrochloride (pyribenzamine; Sigma–Aldrich, St. Louis, MO, United States) stabilized the HRH1 during the synthesis reaction.
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2

Measuring Neuronal Calcium Dynamics

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Chemicals used were imidazole, tetrodotoxin (Wako Pure Chemical, Osaka, Japan), atropine
sulfate salt monohydrate, tripelennamine hydrochloride, indomethacin, hemicholinium-3,
acetylcholine chloride, eserine (Sigma-Aldrich, St. Louis, MI, U.S.A.), fura2/AM (Dojindo
Laboratories, Kumamoto, Japan), and cremophor EL (Nacalai Tesque, Kyoto, Japan).
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3

Analysis of Seized Synthetic Cannabinoids

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Samples were test-purchased online from "legalhighs" websites prior to the 26 May 2016 UK blanket NPS ban. A set of e-liquid samples (6 brands, 9 samples) was also test-purchased from online retailers, prior to the ban. All had been sold as cannabinoid-containing mixtures, although the exact cannabinoid compound quoted varied between mixtures or, in some cases, was not disclosed. Three powder samples were supplied by TICTAC Communications Ltd. from street seizures. Finally, three blank A4 sheets of cannabinoid sprayed/impregnated blotting paper ("blotting paper 01-03") were obtained from an online retailer, the material was not accompanied by any specification about the type of drugs or their amounts. Samples were kept in a locked cabinet at room temperature prior to analysis. Methanol, acetonitrile, water, formic acid (FA) and trifluoroacetic acid (TFA), HPLC grade, dimethyl sulfoxide-d6, isotopic purity 99.9 atom %D, Methanol-d4, isotopic purity 99.8 atom %D, t-butylmethyl-ether (TBME), tripelennamine hydrochloride, and quinolone were all from Sigma-Aldrich (Dorset, UK).
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