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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

Supelco

Dehydrated Alcohol, Pharmaceutical Secondary Standard; Certified Reference Material

Supelco

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

Supelco

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

Sigma-Aldrich

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

Sigma-Aldrich

(R)-(+)-Limonene, ≥93%

Sigma-Aldrich

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

Sigma-Aldrich

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

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

USP

Dehydrated Alcohol, United States Pharmacopeia (USP) Reference Standard

Supelco

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

Sigma-Aldrich

Acetonitrile, electronic grade, 99.999% trace metals basis

Sigma-Aldrich

Ethanol, for residue analysis

Sigma-Aldrich

Ethanol, tested according to Ph. Eur.

Supelco

Acetonitrile, analytical standard

Sigma-Aldrich

Acetonitrile, anhydrous, 99.8%

Sigma-Aldrich

Ethyl alcohol, Pure, 190 proof, ACS spectrophotometric grade, 95.0%

Sigma-Aldrich

Ultrapure Acetonitrile

Sigma-Aldrich

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

Supelco

Acetonitrile, Pharmaceutical Secondary Standard; Certified Reference Material

Sigma-Aldrich

Acetonitrile, ReagentPlus^®, 99%

Sigma-Aldrich

Acetonitrile, for preparative HPLC, ≥99.8% (GC)

Sigma-Aldrich

Ethyl alcohol, Pure, 200 proof, for molecular biology

Sigma-Aldrich

Ethyl alcohol, Pure, 190 proof, for molecular biology

Sigma-Aldrich

Acetonitrile, biotech. grade, ≥99.93%

Sigma-Aldrich

Acetonitrile, ≥99.5% (GC)

Sigma-Aldrich

Acetonitrile, suitable for DNA synthesis, ≥99.9% (GC)

Sigma-Aldrich

Ethyl alcohol, Pure, 200 proof, ACS reagent, ≥99.5%

Sigma-Aldrich

Ethyl alcohol, Pure, 200 proof, meets USP testing specifications

Sigma-Aldrich

Acetonitrile, ACS reagent, ≥99.5%

Sigma-Aldrich

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