Isolation and Identification of Molecular Species of Phosphatidylcholine and Lysophosphatidylcholine from Jojoba Seed Meal (Simmondsia chinensis) FABIAN LE Ä ON, ² MAURITS VAN BOVEN,* ,‡ PETER DE WITTE, ² ROGER BUSSON, § AND MARNIX COKELAERE # Laboratory of Pharmaceutical Biology and Pharmacology and Laboratory of Toxicology and Food Chemistry, Katholieke Universiteit Leuven, Van Evenstraat 4, B-3000 Leuven, Belgium; Laboratory of Medicinal Chemistry, Katholieke Universiteit Leuven, Rega Institute, B-3000 Leuven, Belgium; and Interdisciplinary Research Center, Katholieke Universiteit Leuven Campus Kortrijk, Universitaire Campus, B-8500 Kortrijk, Belgium A mixture of lysophosphatidylcholine (LPC) and phosphatidylcholine (PC) has been isolated by column chromatography from a jojoba meal (Simmondsia chinensis) extract. The molecular species of both classes could be separated and isolated by C18 reversed phase HPLC. The two major compounds were identified by 1D and 2D 1 H and 13 C NMR, by MS, and by GC-MS as 1-oleoyl-3-lysophosphati- dylcholine and 1,2-dioleoyl-3-phosphatidylcholine. Eight other molecular species of LPC and four other molecular species of PC could be assigned by comparison of the mass spectra of the isolated compounds with the spectra of the two major compounds. Complete characterization of the individual molecular species was achieved by GC and GC-MS analysis of the fatty acyl composition from the isolated compounds. The PC/LPC proportion in the phospholipid mixture from three different samples is 1.6 ( 0.1. LPC is considered to be an important bioactive compound; the results of this study suggest further research for the evaluation of potential health benefits of jojoba meal phospholipids. KEYWORDS: Simmondsia chinensis; jojoba seed meal; phospholipids INTRODUCTION The jojoba plant [Simmondsia chinensis (Link) Schneider] is a dioecious evergreen shrub native to the Sonora Desert of the southwestern United States and northern Mexico. The plant is now cultivated in many arid and semiarid places around the world because of its oil-containing seeds (1). The “wild” plant is, however, today replaced by high-yielding cultivars (2, 3). The oil commonly referred to as jojoba oil is a liquid wax. The wax consists of esters of mono-unsaturated long-chain alcohols and fatty acids of 18-24 carbons; the esters have a chain length of 38-46 carbons (4). Only minor quantities of triacylglycerols are present in jojoba oil (5). The oil is mainly used in cosmetics and pharmaceuticals; derivatives of the oil are used in the wax and polish industries (6). Jojoba seed meal, the fraction remaining after winning the oil, contains ∼10-20% of a group of unique compounds, known as simmondsins (7, 8). Among them, simmondsin and sim- mondsin 2′-ferulate exhibit biological activity as food intake inhibitors (9, 10). Three patents have been registered for the use of jojoba meal and for simmondsin as a supplement in pet food (11-13). Other interesting characteristics of yet unidenti- fied jojoba components are described, as, for example, the influence of jojoba meal on oviduct development in chickens (14). The present study deals with the isolation and identification of biologically active compounds such as phospholipids and lysophospholipids from jojoba seed meal. Phospholipids serve as structural components of cell membranes and subcellular organelles. Lysophospholipids (LPL) are recognized as impor- tant cellular signaling molecules and are involved in important processes such as cell proliferation, cell survival, cell migration, diabetes, angiogenesis, inflammation, and cancer, mediated by LPL-specific G-protein-coupled receptors (15, 16). MATERIALS AND METHODS Jojoba Seed Meal and Solvents. Jojoba seed meal samples from three different clones were supplied by R. Holser (New Crops and Process Research, USDA-ARS-NCAUR, Peoria, IL) and defatted by continuous extraction with n-hexane. All solvents used were of analytical and HPLC grade (Sigma-Aldrich, Bornem, Belgium). Refer- ence fatty acid methylesters were obtained from Merck (Darmstadt, Germany). Extraction of Jojoba Seed Meal. Defatted jojoba meal samples (50 g) were extracted by means of extraction tubes with 0.5 L of * Author to whom correspondence should be addressed (telephone +32 16 323439; fax +32 16 323439; e-mail maurits.vanboven@pharm. kuleuven.ac.be). ² Laboratory of Pharmaceutical Biology and Pharmacology. ‡ Laboratory of Toxicology and Food Chemistry. § Laboratory of Medicinal Chemistry. # Interdisciplinary Research Center.