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This document specifies a method for the determination of six major oryzanol components and their total contents in rice bran oils by High-performance liquid chromatography (HPLC). This document is applicable to refined rice bran oils.
Rice is one of the major cereal crops of the world, according to statistics from the United Nations Food and Agriculture Organization (FAO), global rice production in 2023 reached 799.99 million metric tons. Rice bran, a primary by-product produced during the brown rice milling, accounts for 6%–8% of the total yield. Rice bran oil, extracted from rice bran, is recognized as a "healthy oil" by the World Health Organization (WHO), American Heart Association (AHA), and China Cereals and Oils Association (CCOA). Oryzanol, a key nutrient in rice bran oil, is considered one of the most important bioactive components.
Oryzanol (gamma oryzanol) is a mixture of ferulic acid esters of triterpene alcohols and sterols, primarily including Cycloartenyl ferulate, 24-Methylenecycloartenyl ferulate, Campesteryl ferulate, - Sitosteryl ferulate, etc. It exhibits physiological activities such as antioxidant effects, regulation of autonomic nervous function, reduction of serum cholesterol, and management of hypertension. Consequently, the oryzanol content in rice bran oil serves as a crucial indicator for evaluating its nutritional quality.
Currently, the only international method for the determination of oryzanol is specified in Appendix V5 of CXS 210-1999 by the Codex Alimentarius Commission. This method employs spectrophotometry to measure total oryzanols. However, due to the lack of sample purification steps, the results for lowconcentration samples tend to be overestimated. Therefore, this method is only applicable for determining total oryzanol in crude rice bran oil, failing to meet requirements for testing lowconcentration oryzanol samples, for example refined rice bran oil or analyzing individual oryzanol components. Notably, the International Organization for Standardization (ISO) has not yet established any standardized methods for the detection of oryzanol components or total oryzanols in refined rice bran oil.
Therefore, it is imperative to develop a high-performance liquid chromatography (HPLC) method for analyzing both total oryzanols and their individual components in refined rice bran oil. The proposed standards will provide technical support for scientifically evaluating the nutritional value of rice bran oil, and also offer essential methodologies for investigating how processing techniques affect oryzanol composition and content. This advancement will ultimately promote the sustainable and healthy development of the rice bran oil industry.
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