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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

Sigma-Aldrich

Ultrapure Acetonitrile

Sigma-Aldrich

Ethanol

Sigma-Aldrich

Acetonitrile, ≥99.9% (GC)

Sigma-Aldrich

Ethyl alcohol, Pure, 190 proof, meets USP testing specifications

USP

Residual Solvent Class 2 - Acetonitrile, United States Pharmacopeia (USP) Reference Standard

Sigma-Aldrich

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

Sigma-Aldrich

Acetonitrile, ≥99.5% (GC)

Sigma-Aldrich

Ethyl alcohol, Pure, 200 proof

Sigma-Aldrich

Ethyl alcohol, Pure 200 proof

Sigma-Aldrich

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

Sigma-Aldrich

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

Sigma-Aldrich

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

Sigma-Aldrich

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

Sigma-Aldrich

Ethanol, ACS reagent, prima fine spirit, without additive, F15 o¹

Sigma-Aldrich

Ethyl alcohol, Pure, 200 proof, anhydrous, ≥99.5%

Sigma-Aldrich

Ethanol, tested according to Ph. Eur.

Supelco

Ethanol, standard for GC

Sigma-Aldrich

Ethanol, BioUltra, ≥99.8%, (absolute alcohol, without additive, A15 o¹)

Sigma-Aldrich

Acetonitrile, biotech. grade, ≥99.93%

Sigma-Aldrich

Acetonitrile, anhydrous, 99.8%

Supelco

Acetonitrile, analytical standard

Sigma-Aldrich

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

Sigma-Aldrich

Acetonitrile, ReagentPlus^®, 99%

Sigma-Aldrich

Acetonitrile, HPLC Plus, ≥99.9%

Sigma-Aldrich

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

Sigma-Aldrich

Acetonitrile, ACS reagent, ≥99.5%

Sigma-Aldrich

Acetonitrile, electronic grade, 99.999% trace metals basis

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

Supelco

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

Sigma-Aldrich

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