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

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

Sigma-Aldrich

Acetonitrile, JIS special grade, ≥99.5%

Sigma-Aldrich

Ethanol, ≥99.5%, SAJ super special grade

Sigma-Aldrich

Ethanol, JIS 1000, ≥99.5%, for residue analysis

Sigma-Aldrich

Acetonitrile, ≥99.8%, suitable for HPLC

Sigma-Aldrich

Acetonitrile, SAJ first grade, ≥99.0%

Sigma-Aldrich

Ethanol, ≥99.5%

Sigma-Aldrich

Ethanol, JIS 300, ≥99.5%, for residue analysis

Sigma-Aldrich

Ethanol, ≥99.5%, suitable for absorption spectrum analysis

Sigma-Aldrich

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

Sigma-Aldrich

Ethanol, ≥99.5%, suitable for HPLC

Sigma-Aldrich

Ethanol, ≥99.5%, suitable for fluorescence

Sigma-Aldrich

Acetonitrile, HPLC Plus, ≥99.9%, poly-coated bottles

Sigma-Aldrich

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

Supelco

Acetonitrile, HPLC grade, ≥99.93%

Sigma-Aldrich

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

Sigma-Aldrich

Acetonitrile, ≥99.5%, ACS reagent

Sigma-Aldrich

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

Supelco

Ethanol standards 10% (v/v), 10 % (v/v) in H₂O, analytical standard

Sigma-Aldrich

Acetonitrile, for chromatography

Sigma-Aldrich

Acetonitrile, electronic grade, 99.999% trace metals basis

Sigma-Aldrich

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

Sigma-Aldrich

Ethyl alcohol, Pure, 200 proof, HPLC/spectrophotometric grade

Supelco

Acetonitrile, analytical standard

USP

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

Sigma-Aldrich

Acetonitrile, anhydrous, 99.8%

Sigma-Aldrich

Acetonitrile, for residue analysis, JIS 5000

Sigma-Aldrich

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

Sigma-Aldrich

Ethanol, purum, fine spirit, denaturated with 4.8% methanol, F25 METHYL¹, ~96% (based on denaturant-free substance)

Sigma-Aldrich

Ethanol, purum, absolute ethanol, denaturated with 2% 2-butanone, A15 MEK¹, ≥99.8% (based on denaturant-free substance)