Rapid simultaneous determination of amines and organic acids in citrus using high-performance liquid chromatography
2011
Uckoo, Ram M. | Jayaprakasha, Guddadarangavvanahally K. | Nelson, Shad D. | Patil, Bhimanagouda S.
Rapid analytical method for the simultaneous separation and determination of amines and organic acids is a vital interest for quality control of citrus and their products. In the present study, a simultaneous high performance liquid chromatography (HPLC) method for the rapid separation of three amines and two organic acids was developed. Chromatographic separation of compounds was achieved using Xbridge C₁₈ column at ambient temperature, with an isocratic mobile phase of 3mM phosphoric acid at a flow rate of 1.0mLmin⁻¹. A photodiode array (PDA) detector was used to monitor the eluent at 223nm and 254nm with a total analysis time of 10min. Extraction of amines and organic acids from citrus juice was optimized. The method was validated by tests of linearity, recovery, precision and ruggedness. The limit of detection (LOD) and limit of quantification (LOQ) for amines and ascorbic acid were determined to be 5ng and 9.8ng, respectively. All calibration curves showed good linearity (R²≥0.9999) within the test ranges. The recoveries of the amines and organic acids ranged between 84% and 117%. The identity of each peak was confirmed by mass spectral (MS) analysis. The developed method was successfully applied to analyze the content of amines and organic acids in six different species and two varieties of citrus. Results indicate that mandarin and Marrs sweet orange contain high level of amines, while pummelo and Rio Red grapefruit had high content of ascorbic acid (137–251μgmL⁻¹) and citric acid (5–22mgmL⁻¹). Synephrine was the major amine present in Clementine (114μgmL⁻¹) and Marrs sweet orange (85μgmL⁻¹). To the best of our knowledge, this is the first report on simultaneous separation and quantification of amines and organic acids in Marrs sweet orange, Meyer lemon, Nova tangerine, Clementine, Ugli tangelo and Wekiwa tangelo.
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