Nicotine

This article is compiled based on the United States Pharmacopeia (USP) – 2025 Edition

Issued and maintained by the United States Pharmacopeial Convention (USP)

DOWNLOAD PDF HERE

C₁₀H₁₄N₂          ▲162.24 (USP 1-Aug-2022)

3-(1-Methyl-2-pyrrolidinyl)pyridine;

β-Pyridyl-α-N-methyl pyrrolidine CAS RN^®: 54-11-5; UNII: 6M3C89ZY6R.

Change to read:

1 DEFINITION

Nicotine contains NLT ^▲98.0% _{▲(USP 1-Aug-2022)} and NMT ^▲102.0% _{▲(USP 1-Aug-2022)} of nicotine (C₁₀H₁₄N₂), calculated on the anhydrous basis.

2 IDENTIFICATION

A. SPECTROSCOPIC IDENTIFICATION TESTS (197), Infrared Spectroscopy: 197F

Standard: Transfer an amount of USP Nicotine Bitartrate Dihydrate RS equivalent to 100 mg of nicotine to a 100-mL glass-stoppered tube. Add 20 mL of 1 M ammonium hydroxide, 5 mL of 10 M sodium hydroxide, and 20 mL of n-hexane. Shake for 5 min, and allow the phases to separate. Transfer the upper hexane phase to an evaporating dish, and evaporate on a steam bath. FFICAL

Sample: Nicotine

Acceptance criteria: Meets the requirements

Change to read:

B. The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the ^▲Assay_{▲(USP 1-Aug-2022)}

3 ASSAY

Change to read:

PROCEDURE

^▲Solution A: Add 25 mL of 1 M glacial acetic acid to 900 mL of water, and then add 6 ml. of ammonium hydroxide. Adjust with either 2 M glacial acetic acid or 2 M ammonium hydroxide to a pH of 10.0, and dilute with water to 1000 mL.

Solution B: Acetonitrile

Mobile phase: See Table 1.

Table 1

System suitability solution: 1.5 mg/mL of USP Nicotine Bitartrate Dihydrate RS and 6 µg/mL each of USP Nicotine Related Compound A RS. USP Nicotine Related Compound B RS, USP Nicotine Related Compound C. RS. USP Nicotine Related Compound D. RS. USP Nicotine Related Compound E RS, USP Nicotine Related Compound F. RS, and USP Nicotine Related Compound G.RS in water. [NOTE-The concentration of each related compound is in terms of the free base.]

Standard solution: 2.4 mg/mL of USP Nicotine Bitartrate Dihydrate RS in water

Sample solution: 0.8 mg/ml. of Nicotine in water

Chromatographic system

(See Chromatography (621), System Suitability.)

Mode: LC

Detector: UV 254 nm

Column: 4.6-mm x 15-cm; 5-µm packing L1

Flow rate: 1.0 mL/min.

Injection volume: 20 µL

System suitability

Samples: System suitability solution and Standard solution

[NOTE-See Table 2 for the relative retention times.]

Suitability requirements

Resolution: NLT 2.5 between nicotine and nicotine related compound G, System suitability solution

Tailing factor: NMT 2.0 for nicotine, Standard solution

Relative standard deviation: NMT 0.73%, Standard solution

Analysis

Samples: Standard solution and Sample solution

Calculate the percentage of nicotine (C₁₀H₁₄N₂) in the portion of Nicotine taken:

                         Result = (r_U/r_S) × (C_S/C_U) × (M_r1/M_r2) ×100

r_U = peak response of nicotine from the Sample solution 

r_S = peak response of nicotine from the Standard solution

C_S = concentration of USP Nicotine Bitartrate Dihydrate RS on the anhydrous basis in the Standard solution (mg/mL) 

C_U = concentration of Nicotine in the Sample solution (mg/mL)

M_r1 = molecular weight of nicotine, 162.24

M_r2 = molecular weight of anhydrous nicotine bitartrate, 462.41 

_{▲(USP 1-Aug-2022)}

Acceptance criteria: ^▲98.0%-102.0%_{▲(USP 1-Aug-2022)} on the anhydrous basis

4 IMPURITIES

Change to read:

ORGANIC IMPURITIES

^▲Solution A, Solution B, Mobile phase, System suitability solution, Sample solution, and Chromatographic system: Proceed as directed in the Assay._{▲(USP 1-Aug-2022)}

Standard solution: 2.5 µg/mL of USP Nicotine Bitartrate Dihydrate RS in water

Sensitivity solution: 1.2 µg/mL of USP Nicotine Bitartrate Dihydrate RS in water, from the Standard solution

^▲_{▲(USP 1-Aug-2022)}

System suitability

Samples: System suitability solution, Standard solution, and Sensitivity solution

[NOTE-See Table 2 for the relative retention times.]

Suitability requirements

Resolution: NLT 2.5 between nicotine and nicotine related compound G, System suitability solution

Tailing factor: NMT 2.0 for nicotine, System suitability solution

Relative standard deviation: NMT 5.0%, Standard solution

Signal-to-noise ratio: NLT 10, Sensitivity solution

Analysis

Samples: Standard solution and Sample solution

Calculate the percentage of each impurity in the portion of Nicotine taken:

                         Result = (r_U/r_S) × (C_S/C_U) × (M_r1/M_r2) × 100

r_U = peak response of each impurity from the Sample solution

r_S = peak response of nicotine from the Standard solution

C_S = concentration of USP Nicotine Bitartrate Dihydrate RS on the anhydrous basis in the Standard solution (mg/mL)

C_U = concentration of Nicotine in the Sample solution (mg/mL)

M_r1 = molecular weight of nicotine, ^▲162.24_{▲(USP 1-Aug-2022)}

M_r2 = molecular weight of anhydrous nicotine bitartrate, 462.41 

Acceptance criteria: See Table 2. ^▲The reporting threshold is 0.05%. _{▲(USP 1-Aug-2022)}

Table 2

5 SPECIFIC TESTS

OPTICAL ROTATION (781S), Procedures. Specific Rotation

Sample solution: 20 mg/mL in alcohol

Acceptance criteria: 130° to 143

WATER DETERMINATION (921), Method /: NMT 0.5%

6 ADDITIONAL REQUIREMENTS

PACKAGING AND STORAGE: Store under nitrogen in well-closed containers below 25°, protected from light and moisture.

Change to read:

USP REFERENCE STANDARDS (11)

USP Nicotine Bitartrate Dihydrate RS

USP Nicotine Related Compound ARS

Anatabine;

1,2,3,6-Tetrahydro-2,3'-bipyridine.

C₁₀H₁₂N₂        160.22

USP Nicotine Related Compound B.RS

Nicotyrine;

3-(1-Methyl-1H-pyrrol-2-yl)pyridine.

C₁₀H₁₀N₂         158.20

USP Nicotine Related Compound C RS

Cotinine;

(S)-1-Methyl-5-(pyridin-3-yl)pyrrolidin-2-one.

C₁₀H₁₂N₂O       176.22

USP Nicotine Related Compound D.RS

Myosmine;

3-(4,5-Dihydro-3H-pyrrol-2-yl)pyridine fumarate.

C₉H₁₀N₂O · C₄H₄O₄           262.26

USP Nicotine Related Compound ERS

Nicotine N-oxide;

(1RS,2S)-1-Methyl-2-(pyridin-3-yl)pyrrolidine 1-oxide oxalate.

C₁₀H₁₄N₂O · C₂H₂O₄                 268.27

USP Nicotine Related Compound FRS

Nornicotine;

3-(Pyrrolidin-2-yl)pyridine.

C₉H₁₂N₂       ^▲148.21_{▲(USP 1-Aug-2022)}

USP Nicotine Related Compound G RS

Anabasine;

(S)-3-(Piperidin-2-yl)pyridine.

C₁₀H₁₄N₂    ^▲162.24_{▲(USP 1-Aug-2022)}