Skip to Content

Ultra-High Performance Liquid Chromatography (UHPLC) Method for the Determination of Limonene in Sweet Orange (Citrus sinensis) Oil: Implications for Limonene Stability.

Ultra-High Performance Liquid Chromatography (UHPLC) Method for the Determination of Limonene in Sweet Orange (Citrus sinensis) Oil: Implications for Limonene Stability.

Journal of AOAC International (2015-04-11)

Matthew W Bernart

ABSTRACT

The citrus-derived bioactive monoterpene limonene is an important industrial commodity and fragrance constituent. An RP isocratic elution C18 ultra-HPLC (UHPLC) method using a superficially porous stationary phase and photodiode array (PDA) detector has been developed for determining the limonene content of sweet orange (Citrus sinensis) oil. The method is fast with a cycle time of 1.2 min, linear, precise, accurate, specific, and stability indicating, and it satisfies U.S. Pharmacopeia suitability parameters. The method may be useful in its present form for limonene processing, or modified for research on more polar compounds of the terpenome. A forced-degradation experiment showed that limonene is degraded by heat in hydro-ethanolic solution. PDA detection facilitates classification of minor components of the essential oil, including β-myrcene.

MATERIALS

Product Number

Brand

Product Description

Sigma-Aldrich

(R)-(+)-Limonene, 97%

Sigma-Aldrich

(R)-(+)-Limonene, technical, ~90% (sum of enantiomers, GC)

Supelco

(R)-(+)-Limonene, analytical standard

Sigma-Aldrich

(R)-(+)-Limonene, ≥93%

USP

Dehydrated Alcohol, United States Pharmacopeia (USP) Reference Standard

Sigma-Aldrich

3-(Benzyldimethylammonio)propanesulfonate, BioXtra, ≥99.0% (HPCE)

Sigma-Aldrich

Acetonitrile solution, contains 0.05 % (w/v) ammonium formate, 5 % (v/v) water, 0.1 % (v/v) formic acid, suitable for HPLC

Supelco

Residual Solvent - Acetonitrile, Pharmaceutical Secondary Standard; Certified Reference Material

(R)-(+)-Limonene, primary reference standard

Sigma-Aldrich

Ethanol Fixative 80% v/v, suitable for fixing solution (blood films)

Sigma-Aldrich

Acetonitrile solution, contains 0.1 % (v/v) trifluoroacetic acid, suitable for HPLC

Sigma-Aldrich

Acetonitrile solution, contains 0.05 % (v/v) trifluoroacetic acid

Sigma-Aldrich

Ethanol, JIS first grade, 94.8-95.8%

Sigma-Aldrich

Ethanol, JIS special grade, 94.8-95.8%

Supelco

Ethanol solution, certified reference material, 2000 μg/mL in methanol

Sigma-Aldrich

Acetonitrile solution, contains 0.1 % (v/v) formic acid, suitable for HPLC

Sigma-Aldrich

Ethyl alcohol, Pure, 190 proof, for molecular biology

Supelco

Acetonitrile, analytical standard

Sigma-Aldrich

Acetonitrile, anhydrous, 99.8%

Sigma-Aldrich

Acetonitrile, ≥99.5%, ACS reagent

Supelco

Acetonitrile, suitable for HPLC, gradient grade, ≥99.9% (GC)

Sigma-Aldrich

Acetonitrile, for residue analysis, JIS 5000

Sigma-Aldrich

Acetonitrile, for chromatography

Sigma-Aldrich

Acetonitrile, JIS special grade, ≥99.5%

Sigma-Aldrich

Acetonitrile, ≥99.8%, suitable for HPLC

Sigma-Aldrich

Acetonitrile, SAJ first grade, ≥99.0%

Sigma-Aldrich

Acetonitrile, ≥99.8%, for residue analysis, JIS 1000

Sigma-Aldrich

Acetonitrile, ≥99.8%, for residue analysis, JIS 300

Sigma-Aldrich

Acetonitrile, suitable for HPLC, gradient grade, ≥99.9%

Sigma-Aldrich

Acetonitrile, suitable for HPLC-GC, ≥99.8% (GC)