Neohesperidin Dihydrochalcone

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

C₂₈H₃₆0₁₅               612.58

1-Propanone, 1-[4-[[2-O-(6-deoxy-α-L-mannopyranosyl)-B-D-glucopyranosyl)oxy]-2, 6-dihydroxyphenyl]-3-(3-hydroxy-4-methoxyphenyl)-; 3,5-Dihydroxy-4-[3-(3-hydroxy-4-methoxyphenyl)propanoyl]phenyl 2-0-(-α-L-rhamnopyranosyl)-B-D-glucopyranoside    CAS RN^®: 20702-77-6.

1 DEFINITION

Neohesperidin Dihydrochalcone contains NLT 96.0% and NMT 102.0% of neohesperidin dihydrochalcone (C₂₈H₃₆0₁₅ ), calculated on the anhydrous basis. 

2 IDENTIFICATION

Change to read:

A. ^▲SPECTROSCOPIC IDENTIFICATION TESTS (197), Infrared Spectroscopy: _{▲197A (CN 1-MAY-2020)}

B. The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay.

3 ASSAY

3.1 PROCEDURE

Solution A: 0.5% (v/v) glacial acetic acid in water

Mobile phase: Acetonitrile and Solution A (20:80)

3.1.1 Diluent: Dimethyl sulfoxide

System suitability solution: Suspend 100 mg of USP Neohesperidin Dihydrochalcone RS in 10.0 mL of a 100-g/L solution of sulfuric acid in water. Heat the suspension for 5 min on a boiling water bath. Immediately dilute 1.0 mL of the resulting solution with Diluent to 50.0 mL. [NOTE-Neohesperidin dihydrochalcone related compound F and neohesperidin dihydrochalcone related compound G are prepared in situ in this solution.]

Standard solution: 1.0 mg/mL of USP Neohesperidin Dihydrochalcone RS in Diluent

Sample solution: 1.0 mg/mL of Neohesperidin Dihydrochalcone in Diluent

3.1.2 Chromatographic system

(See Chromatography (621), System Suitability.)

Mode: LC

Detector: UV 282 nm

Column: 3.9-mm x 15-cm; 4-µm packing 11, with a carbon loading of 7%

Column temperature: 30°

Flow rate: 1.0 mL/min

Injection volume: 10 µL

Run time: 5 times the retention time of neohesperidin dihydrochalcone

3.1.3 System suitability

Samples: System suitability solution and Standard solution

[NOTE-For the approximate relative retention times of related substances, see Table 1.1

3.1.4 Suitability requirements

Resolution: NLT 2.5 between the neohesperidin dihydrochalcone peak and the neohesperidin dihydrochalcone related compound F peak,

3.1.5 System suitability solution

Relative standard deviation: NMT 2.0%, determined from the neohesperidin dihydrochalcone peak, Standard solution

Chromatogram similarity: The chromatogram obtained from the Standard solution is similar to the reference chromatogram provided with the lot of USP Neohesperidin Dihydrochalcone RS being used.

3.1.6 Analysis

Samples: Standard solution and Sample solution

Calculate the percentage of neohesperidin dihydrochalcone (C₂₈H₃₆0₁₅) in the portion of Neohesperidin Dihydrochalcone taken:

                 Result = (r_U/r_S) x (C_S/C_U) × 100

r_U = peak area of neohesperidin dihydrochalcone from the Sample solution

r_S = peak area of neohesperidin dihydrochalcone from the Standard solution

C_S = concentration of USP Neohesperidin Dihydrochalcone RS in the Standard solution S

C_U = concentration of Neohesperidin Dihydrochalcone in the Sample solution

Acceptance criteria: 96.0%-102.0% on the anhydrous basis

4 IMPURITIES

4.1 RESIDUE ON IGNITION (281)

NMT 0.2%

4.2 ORGANIC IMPURITIES

Mobile phase, Diluent, System suitability solution, and Chromatographic system: Proceed as directed in the Assay.

Standard solution A: Use the Standard solution, as prepared in the Assay.

Standard solution B: 0.04 mg/mL of USP Neodiosmin RS in Diluent

Standard solution C: 0.02 mg/mL of USP Neohesperidin Dihydrochalcone RS from Standard solution A in Diluent

Sensitivity solution: 0.002 mg/mL of USP Neohesperidin Dihydrochalcone RS from Standard solution C in Diluent

Sample solution: 2.0 mg/mL of Neohesperidin Dihydrochalcone in Diluent

4.2.1 System suitability

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

[NOTE-For the approximate relative retention times of related substances, see Table 1.]

4.2.2 Suitability requirements

Resolution: NLT 2.5 between the neohesperidin dihydrochalcone peak and the neohesperidin dihydrochalcone related compound F peak, System suitability solution

Relative standard deviation: NMT 5.0%, determined from the neohesperidin dihydrochalcone peak, Standard solution C

Signal-to-noise ratio: NLT 10, determined from the neohesperidin dihydrochalcone peak, Sensitivity solution

Chromatogram similarity: The chromatogram obtained from the Standard solution A is similar to the reference chromatogram provided with the lot of USP Neohesperidin Dihydrochalcone RS being used.

4.2.3 Analysis

Samples: Standard solution B, Standard solution C, and Sample solution

Calculate the percentage of neodiosmin in the portion of Neohesperidin Dihydrochalcone taken:

                 Result = (r_U/r_S) x (C_S/C_U) × 100

r_U = peak area of neodiosmin from the Sample solution

r_S = peak area of neodiosmin from Standard solution B 

C_S = concentration of USP Neodiosmin RS in Standard solution B (mg/mL) 

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

Calculate the percentage of each impurity except neodiosmin in the portion of Neohesperidin Dihydrochalcone taken:

                 Result = (r_U/r_S) x (C_S/C_U) × 100

r_U = peak area of each individual impurity except neodiosmin from the Sample solution Tu

r_S = peak area of neohesperidin dihydrochalcone from Standard solution C s

C_S = concentration of USP Neohesperidin Dihydrochalcone RS in Standard solution C (mg/mL)

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

Acceptance criteria: See Table 1.

Table 1.

^a 1-14-[[2-0-(6-Deoxy-α-L-mannopyranosyl)-β-D-glucopyranosyl)oxy]-2,6-dihydroxyphenyljethanone (phloroacetophenone neohesperidoside).

^b 7-[[2-0-(6-Deoxy-α-L-mannopyranosyl)-β-D-glucopyranosyl)oxy]-5-hydroxy-2-(3-hydroxy-4-methoxyphenyl)-4H-1-benzopyran-4-one; 5-Hydroxy-2-(3-hydroxy-4-methoxyphenyl)-4-oxo-4H-chromen-7-yl 2-0-(-α-L-rhamnopyranosyl)-B-D-glucopyranoside.

^c (2RS)-7-[[2-0-(6-Deoxy-α-L-mannopyranosyl)-ẞ-D-glucopyranosyl)oxy]-5-hydroxy-2-(3-hydroxy-4-methoxyphenyl)-2,3-dihydro-4H-1-benzopyran-4-one (neohesperidin).

^d 1-[4-[[2-0-(6-Deoxy-α-L-mannopyranosyl)-B-o-glucopyranosyl)oxy]-2,6-dihydroxyphenyl)-3-(4-hydroxyphenyl)propan-1-one (naringin dihydrochalcone).

^e 1-[4-[[6-0-(6-Deoxy-α-L-mannopyranosyl)-B-D-glucopyranosyl]oxy]-2,6-dihydroxyphenyl]-3-(3-hydroxy-4-methoxyphenyl)propan-1-one (Hesperidin dihydrochalcone).

^f 1-[4-(B-D-Glucopyranosyloxy)-2,6-dihydroxyphenyl]-3-(3-hydroxy-4-methoxyphenyl)propan-1-one (hesperetin dihydrochalcone 7'-glucoside).

^g 3-(3-Hydroxy-4-methoxyphenyl)-1-(2,4,6-trihydroxyphenyl)propan-1-one (hesperetin dihydrochalcone).

5 SPECIFIC TESTS

WATER DETERMINATION (921), Method : NMT 12.0%

MICROBIAL ENUMERATION TESTS (61) and TESTS FOR SPECIFIED MICROORGANISMS (62): The total aerobic microbial count does not exceed 10³ cfu/g and the total combined molds and yeasts count does not exceed 10² cfu/g.

6 ADDITIONAL REQUIREMENTS

PACKAGING AND STORAGE: Preserve in tight, light-resistant containers.

USP REFERENCE STANDARDS (11)

USP Neodiosmin.BS

5-Hydroxy-2-(3-hydroxy-4-methoxyphenyl)-4-oxo-4H-chromen-7-yl 2-0-(-α-L-rhamnopyranosyl)-B-D-glucopyranoside.

C₂₈H₃₂0₁₅          608.55

USP Neohesperidin Dihydrochalcone RS

(NF 1-Dec-2019)