Quantification of Carotenoid and Flavonoid Components in Brans of Some Thai Black Rice Cultivars Using Supercritical Fluid Extraction and High‐performance Liquid Chromatography‐mass Spectrometry

Abstract

A method employing supercritical fluid extraction (SFE) followed by high‐performance liquid chromatography‐electrospray ionization‐mass spectrometry (LC‐ESI‐MS) was developed and used for quantification of the antioxidants in groups of carotenoids and flavonoids in the bran of four Thai black rice cultivars: Kao Hom Nin BD, Kao Hom Nin BT, Kao Hom Nin BT no. 3 and 1000–11–2–26. Trans‐β‐carotene, quercetin and isorhamnetin were identified in the crude yellow extracts obtained from SFE using liquid chromatography‐tandem mass spectrometry (LC‐MS/MS). Their contents in the black rice bran samples were determined using LC‐ESI‐MS operated in the selected ion monitoring (SIM) mode. The response of the mass spectrometric detection in the SIM mode was linear, within the range of 1.00–40.00 µg/mL (r2 = 0.9997), 0.25–2.00 µg/mL (r2 = 0.9987) and 0.25–2.00 µg/mL (r2 = 0.9982); the limit of detection was at 0.10, 0.15 and 0.15 µg/mL, and the limit of quantitation was 0.80 g of rice bran for trans‐β‐carotene and 0.60 g of rice bran for both quercetin and isorhamnetin. Method precisions in terms of intraday and inter‐day coefficients of variation were under 3.67% (relative standard deviation). The accuracy of the method estimated by determination of known concentrations of trans‐β‐carotene in the certified reference material, Standard Reference Material 3276, was found to range from 92 to 101%. Trans‐β‐carotene, quercetin and isorhamnetin were present in the bran of all black rice cultivars within the range of 33.60–41, 1.08–2.85 and 0.05–0.83 µg/g, respectively. PRACTICAL APPLICATIONS Production and accumulation of trans‐β‐carotene, quercetin and isorhamnetin in the bran of black rice revealed in this study imply that more attention should be paid to the analysis for screening black rice that has high contents of these bioactive phytochemicals, especially those obtained in rice‐breeding programs. The developed supercritical fluid extraction and liquid chromatography‐mass spectrometry method for quantification of β‐carotene, quercetin and isorhamnetin in black rice bran samples proved to be selective, precise, linear and sensitive, and can efficiently be employed for such analytical purposes. Additionally, extraction of carotene components by supercritical CO2 prior to LC‐MS analysis not only makes the total analysis time much shorter than that of the conventional extraction methods but also eliminates a sample clean‐up step, thus offering a clean analytical procedure.

Kinetic Study of the Scavenging Reaction of the Aroxyl Radical by Eight Kinds of Vegetable Oils in Solution

“…values was estimated to be between 3.0% and 4.7% for each extract. Polyphenols are also included in rice bran (Ajitha, Mohanlal, Suresh, & Jayalekshmy, ; Nakornriab, Sriseadka, & Wongpornchai, ). These polyphenols may also contribute to ArO• scavenging.…”

Variations in Bran Carotenoid Levels within and between Rice Subgroups

“…Abdul-Hamid et al [10] found lycopene and β-carotene in the bran from bulk rice obtained from a commercial mill and Frei and Becker [7] measured β-carotene in colored and non-colored Asian landraces. Abdul-Hamid et al [10] identified β-carotene and lycopene in rice bran obtained from a local mill; Nakornriab et al [36] identified β-carotene in the brans of two Thai black rices.…”

Anthocyanin content and antioxidant capacity in bran extracts of some Thai black rice varieties

“…Extensive studies have been reviewed on biological functions, preclinical tests and the structural determination of naturally occurring anthocyanins Cooke et al, 2005;. Among these studies, some have focused on anthocyanins contained in bran of black-coloured rice (Nakornriab et al, 2008;Revilla et al, 2009;Finocchiaro et al, 2010), as well as on anthocyanins' biological activities, which were mainly antioxidation. The quantitative information concerning anthocyanin pigments in black-coloured rice based on the use of effective instrumental methods has, however, been limited.…”