(±)-Nicotine-d3 (N-methyl-d3)

Catalog NumberACM69980241-2

CAS Number69980-24-1

CategoryClassification by Isotopic Labels

Synonyms	(±)-3-(1-methyl-2-pyrolidinyl)pyridine
Molecular Weight	165.25
Functional Group	Amines & Amine Salts,Heterocycles
Isotopic Enrichment	99 atom % D
Packaging	0.1 g In Stock 0.25 g In Stock
Shipping Hazards	TOXIC
Stability	Stable if stored under recommended conditions (see section 7 of SDS). After three years, the compound should be re-analyzed for chemical purity before use.
Storage Conditions	Store refrigerated
CAS (Unlabeled)	22083-74-5

Case Study

Nicotine-D3 for the Determination of Nicotine in Dried Mushrooms by Extraction Method

Kang H H, et al. Food Additives & Contaminants: Part A, 2020, 37(10), 1687-1694.

This study reported the development of a GC-MS method for the quantitative analysis of nicotine in mushrooms, based on the QuEChERS extraction method. Recovery was 89.5-92.5%, intra-day precision was 0.32-0.85 %, inter-day precision was 0.73-2.36%, and limits of detection and quantification were 0.38 and 1.15 μg kg-1, respectively. The relative expanded uncertainty was 2.8-4.0%. The analysis method was carried out with 11 mushrooms successfully, containing nicotine values of 0.033-1.713 mg kg-1.
Extraction procedure
· A sample of 1-2 g of homogenized dried mushrooms was placed in a 50 mL tube, to which 10-20 mL of distilled water was added before vortexing.
· It was known that a higher pH correlates with improved recovery. Accordingly, this procedure employed 5 M NaOH to maintain the extract's pH in the range of 10-11. The pH was assessed using pH indicator strips with a scale of 0-14.
· Following this, 0.5 mL of the internal standard solution (nicotine-d3, 2000 μg kg-1) and 10 mL of the extraction solvent (ethyl acetate) were incorporated, and the mixture was vortexed for 1 minute. Subsequently, 6 g of MgSO4 and 1.5 g of NaCl were added, and the mixture was shaken for 15 minutes.
· The mixture was then centrifuged at 1590 g for 5 minutes, and the resulting supernatant was filtered through a 0.45 μm membrane filter to obtain the test solution.

Nicotine-D3 for Mass Spectrometry Analysis of Residual Erythrocyte Fragments

Wiencek J R, et al. American journal of clinical pathology, 2019, 151(5), 516-521.

For determining nicotine and nicotine metabolites concentrations per unit of erythrocytes, erythrocyte particles were analysed for nicotine, cotinine or trans-3-hydroxycotinine using liquid chromatography-tandem mass spectrometry (LC-MS/MS).
Mass spectrometry analysis procedure
· For LC-MS/MS measurements, 100 L portions of blank dialyzed plasma, RBC segment supernatant, calibrators and quality control samples were added to 96-well polypropylene plate. Each well received 500 L of a solution containing 0.1 M acetic acid and 1 μg/mL deuterium-labelled internal standards (nicotine-d3, cotinine-d3, 3-OH-cotinine-d3). Mixing was done to a full, samples loaded onto a 96-well solid phase extraction plate.
· An automated liquid dispenser applied positive pressure to extract samples through the columns at a flow rate of one drop every 4 seconds. The columns were washed, one at a time, with 1 mL of 0.1 M acetic acid and 1 mL of a 10:10:80 acetonitrile/methanol/water solution. The bound analytes were gravity flow eluted into 96-well plate with 1 mL of methanol/NH4OH (98:2) after 15 minutes of drying under high pressure.
· Subsequent addition of 50 μL of 0.12 M HCl in methanol and a 30-minute drying step at 40°C prepared the samples for reconstitution. Each well received 100 μL of a 95:5 mobile phase A/B mixture, followed by vortex mixing for 1 minute. The final reconstituted samples were then loaded onto the LC-MS/MS system for analysis.

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(±)-Nicotine-d3 (N-methyl-d3) (69980-24-1)

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