buy am 2201 online/AM-2201

$270.00$3,600.00

Product Name:AM – 2201
IUPAC Name:1-[(5-Fluoropentyl)-1H-indol-3-yl]-(naphthalen-1-yl)methanone
Other Names:1-(5-fluoropentyl)-3-(1-naphthoyl)indole
Cas Number:335161-24-5
Molecular Formula:C24H22FNO
Molar Mass:359.44 g/mol
Effect:stimulant, psychedelic
Purity of the substance:99.9%
Physical properties:Crystals, Powder
Clear
N/A

Description

buy am 2201 online/AM-2201

Abstract For buy am 2201 online/AM-2201

buy am 2201 online/AM-2201 . Novel synthetic cannabinoids are appearing in recreational drug markets worldwide. Pharmacological characterization of these new drugs is needed to inform clinicians, toxicologists, and policy makers who monitor public health. [1-(5-Fluoropentyl)-1H-indol-3-yl](1-naphthyl)methanone (AM-2201) is an abused synthetic cannabinoid that was initially created as a research tool for investigating the endocannabinoid system. Here we measured the pharmacodynamic effects of AM-2201 in rats, and simultaneously determined plasma pharmacokinetics for the parent drug and its metabolites. Male Sprague-Dawley rats were fitted with surgically implanted temperature transponders and indwelling jugular catheters under pentobarbital anesthesia. One week later, rats received subcutaneous injection of AM-2201 (0.1, 0.3, and 1.0 mg/kg) or its vehicle, and serial blood specimens were withdrawn via catheters. Core temperatures and catalepsy were measured just prior to each blood withdrawal, and plasma was assayed for drug and metabolites using liquid chromatography-tandem mass spectrometry. We found that AM-2201 produced dose-related hypothermia and catalepsy that peaked at 2 hours and lasted up to 8 hours. AM-2201 plasma concentrations rose linearly with increasing dose and ranged from 0.14 to 67.9 µg/l. Concentrations of three metabolites, buy am 2201 online/AM-2201 N-(4-hydroxypentyl) (≤0.17 µg/l), naphthalen-1-yl-(1-pentylindol-3-yl)methanone (JWH-018) N-(5-hydroxypentyl) (≤1.14 µg/l), and JWH-018 N-pentanoic acid (≤0.88 µg/l) were detectable but much lower. Peak AM-2201, JWH-018 N-(5-hydroxypentyl), and JWH-018 N-pentanoic acid concentrations occurred at 1.3, 2.4, and 6.5 hours, respectively. Concentrations of AM-2201, JWH-018 N-(5-hydroxypentyl), and JWH-018 N-pentanoic acid were negatively correlated with body temperature, but, given the low concentrations of metabolites detected, AM-2201 is likely the major contributor to pharmacodynamic effects under our experimental conditions.

Introduction For buy am 2201 online/AM-2201

[1-(5-Fluoropentyl)-1H-indol-3-yl](1-naphthyl)methanone (AM-2201) is a synthetic cannabinoid that was first developed in 2000 as a pharmacological tool to study the endocannabinoid system (Fig. 1) (). buy am 2201 online/AM-2201 is a full agonist at cannabinoid-1 receptors (CB1Rs) (), producing psychoactive effects similar to the phytocannabinoid Δ9-tetrahydrocannabinol (), but with a binding affinity 40 times higher (). Similarly, the binding affinity of AM-2201 at cannabinoid-2 receptors, which are responsible for cannabinoid-mediated peripheral effects, is 14 times higher than that of Δ9-tetrahydrocannabinol (). Smoking is the predominant route of AM-2201 consumption, and typical smoked doses of 250 µg to 2 mg are estimated from online drug forums (e.g., https://drugs-forum.com/wiki/AM-2201). By contrast, a 5-mg AM-2201 oral dose was reported as a “comfortable entry dose,” whereas 10 mg would be “powerful” (https://drugs-forum.com/wiki/AM-2201). These dose differences indicate potential buy am 2201 online/AM-2201 degradation in the stomach, limited gastrointestinal absorption, or extensive gastrointestinal or hepatic metabolism. AM-2201 consumption induces typical cannabimimetic effects such as dry mouth, nausea, drowsiness, confusion, mydriasis, and tachycardia (). At higher doses, psychiatric and autonomic complications, such as extreme anxiety, acute psychosis, hyperemesis, and convulsions are reported ().

Materials and Methods Of buy am 2201 online/AM-2201
Chemical and Reagents Of buy am 2201 online/AM-2201.
Analytical standards were purchased from Cayman Chemical (Ann Arbor, MI) and stored at −20°C until use. AM-2201 for administration to rats was provided by the National Institute on Drug Abuse (NIDA) Drug Supply Program (Rockville, MD). LC-MS–grade water, methanol, and formic acid (Optima LC/MS) were obtained from Thermo Fisher Scientific (Waltham, MA). LC-MS–grade acetonitrile, high-performance liquid chromatography–grade tert-butyl methyl ether, dimethyl sulfoxide, sodium metabisulfite, and Tween 80 were acquired from Sigma-Aldrich (St. Louis, MO), whereas sterile 0.9% NaCl (saline) was obtained from Hospira, Inc (Lake Forest, IL). Heparin saline (1000 IU/ml) was purchased from Thomas Scientific (Swedesboro, NJ), and sodium pentobarbital was furnished by the NIDA Intramural Research Program Pharmacy (Baltimore, MD). Distilled water was produced by an ELGA PURELAB Ultra Analytic Purifier (Siemens Water Technologies, Lowell, MA). BG-100 Red abalone enzyme solution from KURA Biotec (Puerto Varas, Chile) was diluted in distilled water to contain 15,625 U/ml glucuronidase and 1250 U/ml sulfatase. Ammonium acetate buffer was prepared with ammonium acetate salt (Sigma-Aldrich) dissolved in distilled water; pH was subsequently adjusted with glacial acetic acid (Thermo Fisher Scientific).

Subjects of buy am 2201 online/AM-2201.
Male Sprague-Dawley rats (weight, 300–400 g) were purchased from Harlan Laboratories (Frederick, MD). Subjects were double housed under conditions of controlled temperature (22 ± 2°C) and humidity (45% ± 5%) with ad libitum access to food and water. Lights were on between 7:00 AM and 7:00 PM. The NIDA Intramural Research Program Animal Care and Use Committee approved the animal experiments, and all procedures were carried out in accordance with the National Institutes of Health Guide for the Care and Use of Laboratory Animals. Vivarium facilities were fully accredited by the Association for Assessment and Accreditation of Laboratory Animal Care. Experiments were designed to minimize the number of animals included in the study.

Surgical Procedures.
Rats were anesthetized with sodium pentobarbital (60 mg/kg, i.p.), and catheters constructed of Silastic (Dow Corning, Midland, MI) and vinyl tubing were surgically implanted into the right jugular vein as previously described (Concheiro et al., 2014). Briefly, the proximal Silastic end of the catheter was advanced to the atrium while the distal vinyl end was exteriorized on the nape of the neck and plugged with a metal stylet. Immediately after catheter implantation, while still under anesthesia, rats received surgically implanted temperature transponders (model IPTT-300; Bio Medic Data Systems, Seaford, DE) to allow for the noninvasive measurement of body temperature (Elmore and Baumann, 2018). The temperature transponder emits radio frequency signals that are received by a compatible handheld reader system (DAS-7006/7r; Bio Medic Data Systems). Transponders are cylindrical in shape, 14 × 2 mm, and were implanted subcutaneously along the midline of the back posterior to the shoulder blades via a prepackaged sterile guide needle delivery system. Rats were single housed postoperatively and given at least 1 week to recover from surgery.

Blood Collection Procedures and buy am 2201 online/AM-2201 Injections.
On the day of an experiment, rats were brought into the laboratory in their home cages and allowed 1 hour to acclimate to the surroundings. Polyethylene extension tubes were attached to 1-ml tuberculin syringes, filled with sterile saline, and connected to the vinyl end of the catheters. The extension tubes were threaded outside the cage to facilitate blood sampling by an investigator remote from the animal. Catheters were flushed with 0.3 ml of 48 IU/ml heparin saline to facilitate blood withdrawal. To prepare drug solutions, each milligram of AM-2201 was diluted into 50 µl of dimethylsulfoxide and 50 µl of Tween 80, then sonicated for 1 minute to dissolve. To this solution, 900 µl of sterile saline was added to yield a 1 mg/ml stock solution of AM-2201. Aliquots of stock solution were diluted with vehicle consisting of dimethylsulfoxide/Tween 80/saline at 1:1:18 (v/v/v) to yield drug concentrations of 0.3 and 0.1 mg/ml. Groups of rats received subcutaneous injection of vehicle (control), or 0.1, 0.3, or 1.0 mg/kg AM-2201 on the lower back between the hips. The subcutaneous route of administration was chosen because our previous studies used this route when examining the effects of AM-2201 and other cannabinoids in rats (Schindler et al., 2017; Elmore and Baumann, 2018). Additionally, similar to the smoked route of administration, the subcutaneous route largely bypasses the first-pass metabolism in the liver. Rats were randomly assigned to each dose group. Blood specimens (300 µl) were withdrawn via catheters immediately before (t0) and at 0.25, 0.5, 1, 2, 4, 8, and 24 hours after subcutaneous injection. Specimens were collected into 1-ml tuberculin syringes, then transferred to 1.5-ml plastic tubes containing 5 µl of 250 mM sodium metabisulfite as a preservative and 5 µl of 1000 IU/ml heparin as an anticoagulant. Blood was centrifuged at 1000g for 10 minutes at 4°C. Plasma was decanted into cryovials and stored at −80°C until analysis. After each blood withdrawal, an equal volume of saline solution was infused via the intravenous catheter to maintain volume and osmotic homeostasis. Rats were free to move around the cage during the sampling procedure.

Measurement of Catalepsy and Body Temperature.
Catalepsy scores and body temperature were determined at each blood withdrawal. Rat behaviors were observed by an experienced rater for 1 minute just prior to the measurement of body temperature via a handheld reader sensitive to signals emitted by the surgically implanted transponder. The behavioral rater was blind to treatment conditions. On each test day, one investigator prepared drug solutions and administered the drug to rats, whereas another investigator performed the behavioral scoring without knowing the dose administered to each subject. During the 1-minute observation period, catalepsy behaviors were scored, as previously described, based on the following three overt symptoms: immobility, flattened body posture, and splayed limbs (Elmore and Baumann, 2018). Each symptom was scored as 1 = absent, 2 = present, or 3 = continuous or intense, at each time point. For each rat, catalepsy scores at each time point were summed, yielding a minimum score of 3 and a maximum score of 9. Blood samples were withdrawn immediately after temperature recording.

AM-2201 and Metabolites Quantification.
Plasma specimens were assayed using a fully validated analytical method capable of detecting and quantifying AM-2201 and 13 of its metabolites: buy am 2201 online/AM-2201 N-(4-hydroxypentyl), AM-2201 6′-hydroxyindole, AM-2201 7′-hydroxyindole, JWH-018 N-(2-hydroxypentyl), JWH-018 N-(3-hydroxypentyl), JWH-018 N-(4-hydroxypentyl), JWH-018 N-(5-hydroxypentyl), JWH-018 N-pentanoic acid, JWH-018 N-propanoic acid, JWH-073 N-(2-hydroxybutyl), JWH-073 N-(3-hydroxybutyl), JWH-073 N-(4-hydroxybutyl), and JWH-073 N-butanoic acid (Carlier et al., 2016). Briefly, 75-µl aliquots were hydrolyzed in 400 mM ammonium acetate buffer, pH 4.0, with β-glucuronidase and sulfatase enzymes. Samples were diluted in acetonitrile and ammonium acetate buffer then poured onto supported liquid extraction cartridges (1-ml ISOLUTE SLE+ cartridges; Biotage, Charlotte, NC). Analytes were eluted with tert-butyl methyl ether, and solvent was evaporated under a nitrogen stream. Residues were reconstituted in mobile phase before injection onto the chromatographic system.

Analysis was performed by LC-MS/MS with a Shimadzu system consisting of an LC-30AD high-performance liquid chromatograph coupled to an LC-MS-8050 mass spectrometer (Shimadzu Corp., Columbia, MD). MS transitions were monitored as follows (quantification transition is in bold): 360 > 127 and 360 > 155 for AM-2201; 376 > 127.1 and 376 > 155.1 for AM-2201 N-(4-hydroxypentyl); 376 > 127 and 376 > 155 for AM-2201 6′-hydroxyindole; 376 > 127 and 376 > 155 for AM-2201 7′-hydroxyindole; 358 > 155 and 358 > 127 for JWH-018 N-(2-hydroxypentyl); 358 > 155 and 358 > 127 for JWH-018 N-(3-hydroxypentyl); 358 > 155 and 358 > 127 for JWH-018 N-(4-hydroxypentyl); 358 > 155 and 358 > 127 for JWH-018 N-(5-hydroxypentyl); 372 > 155 and 372 > 127 for JWH-018 N-pentanoic acid; 344 > 155 and 344 > 127 for JWH-018 N-propanoic acid; 344 > 155 and 344 > 127 for JWH-073 N-(2-hydroxybutyl); 344 > 127 and 344 > 155 for JWH-073 N-(3-hydroxybutyl); 344 > 127 and 344 > 155 for JWH-073 N-(4-hydroxybutyl); and 358 > 155 and 358 > 127 for JWH-073 N-butanoic acid. Data were processed with ASCENT software from Indigo BioAutomation (Indianapolis, IN). Gradient elution was performed at 700 µl/min on a Raptor LC biphenyl column (Restek, Bellefonte, PA) with mobile phase A (0.1% formic acid in water) and mobile phase B [0.1% formic acid in methanol/acetonitrile 50:50 (v/v)]. Two multiple reaction monitoring mass transitions were monitored for each analyte and internal standard.

Each sample batch was accompanied by seven calibrators, a blank; a blank fortified with internal standards; and low-, medium-, and high-quality controls (Carlier et al., 2016). The lowest limits of quantification were 0.1 µg/l for buy am 2201 online/AM-2201 N-(4-hydroxypentyl), JWH-018 N-propanoic acid, and JWH-073 N-(2-hydroxybutyl), and 0.05 µg/l for other analytes. Analyte recoveries and matrix effects were 58.4%–84.4% and −62.1% to −15.6%, respectively (n = 10). Interassay bias and imprecision were 88.8%–110.1% and 0.3%–11.9% CVs, respectively (n = 10).

Data Analysis and Statistics Of buy am 2201 online/AM-2201.
Pharmacodynamic and pharmacokinetic findings were statistically evaluated with GraphPad Prism version 7.04 (GraphPad Software, La Jolla, CA). Body temperature and catalepsy data were evaluated with two-way analysis of variance (dose × time) followed by Bonferroni post hoc tests. Pharmacokinetic data were analyzed using WinNonlin version 5.2 (Pharsight, Mountain View, CA) to calculate pharmacokinetic constants, including Cmax, time of maximum concentration (Tmax), area under the curve (AUC), and elimination half-life (t1/2) for each analyte. We examined the AUC from 0 to 24 hours postinjection for AM-2201 and its metabolites. AUC is the integral of the time-concentration profile for a given analyte and represents the total analyte exposure over time. The pharmacokinetic constants were subjected to one-way analysis of variance (dose) followed by Bonferroni post hoc tests to determine differences between dose groups. At least three data points on the terminal elimination phase were required for t1/2 determination. Relationships between analyte plasma concentrations and body temperature were assessed using a Pearson’s correlation analysis. P < 0.05 was used as the minimum statistical significance threshold for all comparisons.

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