Year of Publication


Degree Name

Master of Science (MS)

Document Type

Master's Thesis


Agriculture, Food and Environment


Plant and Soil Sciences

First Advisor

Dr. David Hildebrand


This thesis includes two main parts:

I. Evaluation of techniques for oil (total lipid) quantification of chia and soybean seeds.

This study evaluated ten different methods of seed oil quantification, including some methods that have not been applied to oilseeds before. The main aim of this study was to find one or more techniques that are easy, inexpensive, safe and fast with a small amount of ground seeds. The Soxhlet method was used as a standard to compared between techniques of oil quantification. The oil extraction by the Soxhlet method was evaluated with two solvents petroleum ether and acetone. There is not a statistically significant difference between petroleum ether and acetone solvents. No significant differences for the amount of oil recovered via the Soxhlet method were found between Medium Moisture Content (MMC > 10%) and Low Moisture Content (LMC < 4.0 %). The Folch technique provided higher percentages of oil extraction than Bligh and Dyer and hexane-isopropanol techniques. There is not a statistically significant difference (P =0.0844) between Soxhlet method and Folch method but less than the Soxhlet method. A supercritical fluid extraction (HCH) method provided a lower yield of oil extraction compared with the Soxhlet method for three varieties of bias samples. A Direct Transesterification (DT) method with LMC and MMC provided a statistically significant difference than the Soxhlet method. The DT with LMC produced higher yield than DT with MMC between samples but lower than the Soxhlet method. A Double Direct Transesterification (DDT) following Griffiths protocol provided more accurate results with the stir bar technique than sonication technique. 11 oilseeds bias samples (10 bias samples of soybean and one of chia) covering an oil content range of 15.4 to 32.6% showed, there is a significant difference between the Soxhlet and DDTG method and high oil quantification found with DDTG. A Double Direct Transesterification following Qiao et al. (2015) provided similar oil extraction to the Griffiths et al. (2010) method and also there is a significant difference between the Soxhlet and DDTQ method and higher oil recovered with DDTQ. The Bead Beating Extraction (BBE) protocol showed, there is a statistically significant difference ( P< 0.001) than the Soxhlet method. The BBE provided high oil quantification comparing with the Soxhlet method. The BBE provided the best results since it is the easiest, cheapest and fastest oil quantification method. A Nile red fluorescence technique yield no clear results.

II. Characterization of oil and protein in mutant chia (Salvia hispanica L.) seeds.

The objectives of this study were to (1) measure heritability in chia plants of levels of oil and protein content and seed yield and (2) Compare differences between two locations over two years for oil and protein percentages in chia and seeds yield. A population of 180 M3 mutant individual chia plants was harvested and for which forty M4 chia seeds were planted based on seed composition characteristics, with six plants representing each characteristic (high and lower oil, protein, and density and high yield). The forty M4 chia seeds were planted, with two replications for two locations, have been chosen in Kentucky one on Spindletop farm and the other in Quicksand farm. The M5 progeny seeds from plants grown in Spindletop and Quicksand contained significantly (P < 0.05) more protein than did seeds from the M4 parents. There were significant differences (P < 0.05) between locations where Quicksand obtain higher oil content than Spindletop. There was no significant difference between M5 parents seeds and M4 progeny seeds and higher yield kg/ha at Quicksand than Spindletop.

Digital Object Identifier (DOI)