PMID- 35903842 OWN - NLM STAT- MEDLINE DCOM- 20220801 LR - 20220915 IS - 1872-2059 (Electronic) IS - 1000-8713 (Print) IS - 1000-8713 (Linking) VI - 40 IP - 8 DP - 2022 Aug TI - [Determination of three oryzanols in rice by mixed-mode solid-phase extraction coupled with high performance liquid chromatography-tandem mass spectrometry]. PG - 746-752 LID - 10.3724/SP.J.1123.2021.12016 [doi] AB - Rice is a major dietary staple in many communities owing to its high nutritional value and characteristic aroma. Oryzanol, a mixture of ferulic acid esters of triterpene alcohols and phytosterols, is a major group of phytochemicals found in rice. 24-Methylenecycloartanyl ferulate (24MCA-FA), cycloartenyl ferulate (CA-FA), and campestanyl ferulate (Camp-FA) have been identified as the primary components of oryzanol. At present, for the quantification of oryzanol in rice and rice products, UV spectroscopy or high performance liquid chromatography (HPLC) is widely employed. However, these methods cannot differentiate individual oryzanols, resulting in higher measured values. To extract oryzanol, methods including liquid-liquid extraction, acidulation extraction, and direct solvent extraction have been typically employed, as they do not require specific extraction instrumentation. However, there has been no systematic study on the direct solvent extraction and purification conditions of oryzanol in rice. In this study, a rapid and accurate analytical method based on HPLC-MS/MS and mixed-mode anion exchange (MAX) solid-phase extraction was established to determine the content of three oryzanols (24MCA-FA, CA-FA, and Camp-FA) in rice. The MS parameters, such as the collision energy of three ion pairs of each oryzanol, were optimized. Further, the chromatographic separation conditions and response intensities of the oryzanols in different mobile phases were compared. The effects of different pretreatment conditions on the extraction efficiency of the three oryzanols in rice samples and different purification conditions on their recovery were investigated. Combined with the external standard method, the three oryzanols in rice were successfully quantified. The results showed that the baseline separation and highest response for the three oryzanols were achieved using the Agilent Eclipse XDB-C8 chromatographic column (150 mmx2.1 mm, 3.5 mum) when methanolratio acetonitrile in a 1ratio1 ratio (v/v) and an aqueous solution of 5 mmol/L ammonium acetate were used as the mobile phases for gradient elution. The extraction rate of the three oryzanols was highest when using 2.5 g of the sample, adding 20 mL of methanol, soaking for 12 h, ultrasonicating at a temperature of 40 ℃ for 20 min, and centrifuging the extracted solutions at 4500 r/min for 10 min. The samples were purified by MAX, and the sample matrix effect was found to be lesser than 1.6%-10.8%. Under the optimum conditions, the calibration curves of the three oryzanols showed good linearity (correlation coefficients r(2)>/=0.9983) within their respective linear ranges. The limits of detection were in the range of 0.5-1.0 mug/L, and limits of quantification were in the range of 2.0-3.5 mug/L. Accuracy and precision experiments were performed on rice samples spiked at three levels (2, 5, and 10 times the background concentration), with three replicates. The average recoveries of the three oryzanols ranged from 86.1% to 110.6%, and the relative standard deviations (RSDs) were between 0.9% and 3.2%. The method showed good performance when applied to the analysis of real samples. In conclusion, the developed method can determine the content of the three oryzanols in rice quickly and accurately, and can be used for the subsequent measurement of oryzanol compounds in rice. FAU - Li, Hongyan AU - Li H AD - China National Rice Research Institute, Rice Quality Inspection and Supervision Center, Ministry of Agriculture and Rural Affairs, Hangzhou 310006, China. FAU - Yang, Huan AU - Yang H AD - China National Rice Research Institute, Rice Quality Inspection and Supervision Center, Ministry of Agriculture and Rural Affairs, Hangzhou 310006, China. FAU - Ma, Chenyi AU - Ma C AD - China National Rice Research Institute, Rice Quality Inspection and Supervision Center, Ministry of Agriculture and Rural Affairs, Hangzhou 310006, China. FAU - Zhang, Wanyue AU - Zhang W AD - China National Rice Research Institute, Rice Quality Inspection and Supervision Center, Ministry of Agriculture and Rural Affairs, Hangzhou 310006, China. FAU - Xu, Qingyu AU - Xu Q AD - China National Rice Research Institute, Rice Quality Inspection and Supervision Center, Ministry of Agriculture and Rural Affairs, Hangzhou 310006, China. FAU - Chen, Mingxue AU - Chen M AD - China National Rice Research Institute, Rice Quality Inspection and Supervision Center, Ministry of Agriculture and Rural Affairs, Hangzhou 310006, China. FAU - Ma, Youning AU - Ma Y AD - China National Rice Research Institute, Rice Quality Inspection and Supervision Center, Ministry of Agriculture and Rural Affairs, Hangzhou 310006, China. LA - chi PT - Journal Article PL - China TA - Se Pu JT - Se pu = Chinese journal of chromatography JID - 9424804 RN - 0 (Phenylpropionates) RN - 0 (Solvents) RN - SST9XCL51M (gamma-oryzanol) SB - IM MH - Chromatography, High Pressure Liquid MH - *Oryza/chemistry MH - Phenylpropionates MH - Solid Phase Extraction MH - Solvents MH - *Tandem Mass Spectrometry PMC - PMC9404047 OTO - NOTNLM OT - high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) OT - mixed-mode solid-phase extraction OT - oryzanol OT - rice EDAT- 2022/07/30 06:00 MHDA- 2022/08/02 06:00 PMCR- 2022/08/08 CRDT- 2022/07/29 02:22 PHST- 2022/07/29 02:22 [entrez] PHST- 2022/07/30 06:00 [pubmed] PHST- 2022/08/02 06:00 [medline] PHST- 2022/08/08 00:00 [pmc-release] AID - 10.3724/SP.J.1123.2021.12016 [doi] PST - ppublish SO - Se Pu. 2022 Aug;40(8):746-752. doi: 10.3724/SP.J.1123.2021.12016.