Scopolamine hydrobromide trihydrate and the acetylcholinesterase enzyme (from electric eel) were purchased from Sigma-Aldrich (now Merck). Donepezil hydrochloride monohydrate was received ex gratis from Wockhardt Research Center, Aurangabad, Maharashtra (India). Vanillin was purchased from LobaChemie Pvt. Ltd., Mumbai, Maharashtra (India).
Scopolamine hydrobromide trihydrate
Manufactured by Merck Group
Sourced in United States
Scopolamine hydrobromide trihydrate is a chemical compound commonly used in laboratory settings. It serves as a muscarinic antagonist, blocking the action of the neurotransmitter acetylcholine in the body.
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Lab products found in correlation
Nafion 117, by Merck Group (3 mentions)
Atropine sulfate, by Merck Group (2 mentions)
Lithium perchlorate, by Merck Group (2 mentions)
Acetonitrile, by Scharlab (2 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)
Trypsin edta, by Merck Group (1 mentions)
Sh sy5y, by American Type Culture Collection (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Tryptophan, by Merck Group (1 mentions)
Tyrosine, by Merck Group (1 mentions)
R sch 23390 hydrochloride, by Merck Group (1 mentions)
Sr lab software, by San Diego Instruments (1 mentions)
Butylated hydroxytoluene, by HiMedia (1 mentions)
β carotene, by HiMedia (1 mentions)
Linoleic acid, by HiMedia (1 mentions)
Rosmarinic acid, by Merck Group (1 mentions)
Huperzine a hu, by Enzo Life Sciences (1 mentions)
Pooled human serum, by Merck Group (1 mentions)
Lactic acid, by Merck Group (1 mentions)
Sodium chloride (nacl), by Avantor (1 mentions)
12 protocols using scopolamine hydrobromide trihydrate
1
Investigating Acetylcholinesterase Inhibition
2
Scopolamine-induced Memory Modification
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Scopolamine hydrobromide trihydrate (Sigma Aldrich), the mAChR antagonist, was dissolved in 0.9% physiological saline at a dose of 0.8 mg/kg, and was administered with intraperitoneal injections 20-min prior to the reactivation phase in the object memory modification task.
3
Neuroblastoma Cell Culture with Suffruticosol A
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The human neuroblastoma cell line, SH-SY5Y cells were purchased from ATCC. These cells were cultured under the following media condition: High glucose Dulbecco's modified Eagle's medium (DMEM; HyClone, S, Logan, UT, USA) supplemented with 10% fetal bovine serum (Gibco, Grand Island, NY, USA) and penicillin–streptomycin (10,000 U/mL; Grand Island, NY, Gibco). The cells were incubated in a condition of 5% CO2 humidified atmosphere at 37 °C. Cell subcultures were performed every 3–4 days using trypsin–EDTA (Sigma, Saint Louis, MO, USA) for cell detachment. For the experiments, 2 × 104 cells were seeded in a 96-well plate in 100 μL of culture medium and pre-incubated overnight to allow cell attachment. The next day, suffruticosol A (1, 5, or 20 μM) and scopolamine hydrobromide trihydrate (0 or 2 mM; Sigma, Saint Louis, MO, USA) was added to a final volume of 200 μL medium, followed by further incubation of 24 h.
4
Dipeptide and Neurotransmitter Synthesis
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The dipeptides WY, tyrosine-Tryptophan (YW), Tryptophan-methionine (WM), methionine-Tryptophan (MW), Tryptophan-valine (WV), Tryptophan-leucine (WL), and Tryptophan-phenylalanine (WF) (purity > 98%) were purchased from NARD Chemicals, Ltd. (Amagasaki, Japan). Tryptophan, tyrosine, (−)-scopolamine hydrobromide trihydrate, and R(+)-SCH-23390 hydrochloride were purchased from Sigma Aldrich Co. (St. Louis, MO, USA).
5
Startle Response and Prepulse Inhibition Assay
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PPI was measured using automated startle chambers (San Diego Instruments, San Diego, CA, USA), as described previously [37 (link)]. In brief, rats were placed in a transparent Plexiglas cylinder that sat within a sound-attenuating cabinet. Audiogenic stimuli were delivered with a speaker, and whole-body startle responses were measured using the automated SR-Lab software (San Diego Instruments). The 30-minute session consisted of 80 trials presented at variable intervals (8–27 s), 8 no tone (i.e., only background, 70 dB), 32 pulse-alone (4 blocks of 8 × 115 dB) trials, and 40 prepulse trials (8 of each: 2, 4, 8, 12, or 16 dB above background followed 100 ms later by the 115 dB pulse). Startle data were measured using all four blocks of pulse-alone trials. The percentage of PPI was calculated as the difference in amplitude between the startle response to the pulse-alone trials and the prepulse-pulse trials, divided by the response to the pulse-alone trial × 100%.
Saline or 0.3 mg/kg scopolamine (Scopolamine hydrobromide trihydrate; Sigma-Aldrich, St. Louis, MO, USA) was administered subcutaneously 30 min prior to PPI testing. Scopolamine was dissolved in saline to a dose based on previous literature [25 (link),38 (link)] and administered in a volume of 1 ml/kg. Using a randomized, crossover protocol, rats received both treatments with 3–4 days allowed between each experiment.
Saline or 0.3 mg/kg scopolamine (Scopolamine hydrobromide trihydrate; Sigma-Aldrich, St. Louis, MO, USA) was administered subcutaneously 30 min prior to PPI testing. Scopolamine was dissolved in saline to a dose based on previous literature [25 (link),38 (link)] and administered in a volume of 1 ml/kg. Using a randomized, crossover protocol, rats received both treatments with 3–4 days allowed between each experiment.
6
Antioxidant Activity Evaluation
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Diazepam and piracetam were obtained from Cipla Ltd., Mumbai, India. β-Carotene, linoleic acid, and butylated hydroxytoluene (BHT) were purchased from Hi-Media (Mumbai, India). DPPH (2,2-Diphenyl-1-(2,4,6-trinitrophenyl)hydrazyl) and (-)-scopolamine hydrobromide trihydrate were purchased from Sigma-Aldrich (St. Louis, MO, USA). TLC-solvents were of HPLC grade obtained from Merck, Mumbai, India. All remaining reagents were used of analytical grade.
7
HPLC Analysis of Scopolamine, Rosmarinic Acid, and Huperzine A
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All reagents for HPLC analysis, scopolamine hydrobromide trihydrate (SC), and rosmarinic acid (RA) and reagents for biochemical analyses were purchased from Sigma-Aldrich (Poland). Huperzine A (HU) was obtained from Enzo Life Sciences AG (Alexis Corporation, Biomibo Distribution, Poland). Chemicals for gene expression analysis were obtained from Roche Diagnostic and ALAB (Poland). All chemicals and drugs were ex tempore prepared on the day of the experiment.
8
Electrochemical Biosensor Protocol
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Tris (2,2′-bipyridyl)dichlororuthenium(II) hexahydrate ([Ru (bpy) 3 ] 2+ ), (-)-scopolamine hydrobromide trihydrate, lithium perchlorate (LiClO 4 ), urea, lactic acid, human pooled serum, Surine™ and 117 Nafion (∼5% mixture of lower aliphatic alcohols and water) were purchased from Sigma-Aldrich. Absolute EtOH, acetic acid, aluminium chloride (AlCl 4 ) and NaCl were purchased from VWR Chemicals. All chemicals were used as received. All solutions were prepared in Milli-Q water (18 mΩ cm -1 ). Artificial saliva (Bioténe® oral balance gel) and synthetic skin were commercially purchased and used as received.
9
Scopolamine and Donepezil Pharmacology
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(-)Scopolamine hydrobromide trihydrate (scopolamine) was obtained from Sigma Aldrich, Steinheim, Germany and was dissolved in physiological saline. Donepezil hydrochloride was a kind gift from Emcure Pharmaceuticals Pvt. Ltd., India and was freshly prepared in physiological saline before administration.
Calcitriol (vitamin D 3 ) was a kind gift from Sun Pharmaceuticals Industries Ltd., India.
Calcitriol (vitamin D 3 ) was a kind gift from Sun Pharmaceuticals Industries Ltd., India.
10
Reagent Preparation for Electrochemical Analysis
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Reagents and Materials (-)-Scopolamine hydrobromide trihydrate, atropine sulfate, methamphetamine (MA), 3,4methylenedioxymethamphetamine (MDMA), lithium perchlorate (LiClO4) and 117 Nafion (~5% mixture of lower aliphatic alcohols and water) were purchased from Sigma-Aldrich. Acetonitrile (purchased from Scharlau) was distilled over calcium hydride under a nitrogen atmosphere and collected as required. The [Ir(bt)2(pt-TEG)] + and [Os(diars)2(bthp)] 2+ were prepared as chloride and hexafluorophosphate salts, respectively, according to previously published procedures. [51] [52] [53] [54] All aqueous samples were prepared with Milli-Q (18 mΩ cm -1 ).
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