Intestinal solubility is higher in fed state compared to fasted state. However, the dissolution rate does not increase to the same extent. Dog seems to be a good model for man with respect to dissolution in the small intestine after intake of a meal, whereas FeSSIF is a poorer means of determining intestinal saturation solubility in the fed state.

This article is available online at http://www.jlr.org implicated in several diverse diseases, such as type 2 diabetes, Alzheimer disease, ( 2 ) and cancer ( 3 ). Lipid analysis has been an important area of research for several decades, and due to technological advances, the fi eld has experienced a renaissance in the last decade. In a modern laboratory, a comprehensive lipid characterization can be performed that generates quantitative data of several hundreds of molecular lipids from several different lipid classes. This kind of analysis, often called lipidomics, is based on HPLC and mass spectrometry (MS) instrumentation. The analysis is performed unattended in the 96-well format and is fully automated.An important component for successful analysis is the quality of the lipid extract. It is important that the lipid extract that is injected on the HPLC or infused into the mass spectrometer is pure; therefore, it is important that interfering substances and particles are removed. Inability in removing these substances might result in a high chemical background, which will have an effect on both the sensitivity and selectivity of the analysis. In contrast to fully automated and high-throughput analysis, lipid extraction is still often performed manually, involving exhaustive and time-consuming pipetting steps and hazardous solvents such as chloroform. Thus, a fully automated, chloroformfree method that can be used with standard 96-well robots would signifi cantly improve sample throughput, as well as reduce the negative impact on health and environment. The aim of this study was to develop that method.Abstract Lipid extraction from biological samples is a critical and often tedious preanalytical step in lipid research. Primarily on the basis of automation criteria, we have developed the BUME method, a novel chloroform-free total lipid extraction method for blood plasma compatible with standard 96-well robots. In only 60 min, 96 samples can be automatically extracted with lipid profi les of commonly analyzed lipid classes almost identically and with absolute recoveries similar or better to what is obtained using the chloroformbased reference method. Lipid recoveries were linear from 10-100 µl plasma for all investigated lipids using the developed extraction protocol. The BUME protocol includes an initial one-phase extraction of plasma into 300 µl butanol:methanol (BUME) mixture (3:1) followed by twophase extraction into 300 µl heptane:ethyl acetate (3:1) using 300 µl 1% acetic acid as buffer. The lipids investigated included the most abundant plasma lipid classes (e.g., cholesterol ester, free cholesterol, triacylglycerol, phosphatidylcholine, and sphingomyelin) as well as less abundant but biologically important lipid classes, including ceramide, diacylglycerol, and lyso-phospholipids. This novel method has been successfully implemented in our laboratory and is now used daily. We conclude that the fully automated, highthroughput BUME method can replace chloroform-based methods, saving both human and environment...

Acetylenic bonds are present in more than 600 naturally occurring compounds. Plant enzymes that catalyze the formation of the Delta12 acetylenic bond in 9-octadecen-12-ynoic acid and the Delta12 epoxy group in 12,13-epoxy-9-octadecenoic acid were characterized, and two genes, similar in sequence, were cloned. When these complementary DNAs were expressed in Arabidopsis thaliana, the content of acetylenic or epoxidated fatty acids in the seeds increased from 0 to 25 or 15 percent, respectively. Both enzymes have characteristics similar to the membrane proteins containing non-heme iron that have histidine-rich motifs.