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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

Ethanol, for residue analysis

Sigma-Aldrich

Ethanol, tested according to Ph. Eur.

Supelco

Ethanol, standard for GC

Supelco

Acetonitrile, analytical standard

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Acetonitrile, electronic grade, 99.999% trace metals basis

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

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

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Acetonitrile, anhydrous, 99.8%

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

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

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

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Acetonitrile, for HPLC, for UV, ≥99.9% (GC)

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

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

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

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Acetonitrile, HPLC Plus, ≥99.9%

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

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

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

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

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Acetonitrile, biotech. grade, ≥99.93%

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Ethyl alcohol, Pure 200 proof, for molecular biology

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Acetonitrile, ReagentPlus^®, 99%

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Acetonitrile, ACS reagent, ≥99.5%

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Acetonitrile, ≥99.5% (GC)

USP

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

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Ultrapure Acetonitrile

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

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

Sigma-Aldrich

(R)-(+)-Limonene, ≥93%