Bile acids are physiologically important metabolites, which are synthesized in liver as the end products of cholesterol metabolism and then secreted into intestine. They are amphiphilic molecules which play a critical role in the digestion and absorption of fats and fat-soluble vitamins through emulsification. The concentration of bile acids is an indicator for liver function. Individual suffering from liver diseases has a sharp increase in bile acid concentrations. Hence, the concentration level of bile acids has long been used as a biomarker for the early diagnosis of intestinal and liver diseases. Conventional methods of bile acid detection such as chromatography-mass spectrometry and enzymatic reactions are complex and expensive. It is highly desired to have a simple, fast, and low-cost detection of bile acids that is available for self-testing or point-of-care testing. To achieve this goal, we develop a liquid crystal-based biosensor for the detection of bile acids. The sensor platform is based on the anchoring transition of liquid crystals (LCs) at the sodium dodecyl sulfate (SDS)-laden LC/aqueous interface for the detection of bile acids in aqueous solution. The first part of this dissertation focuses on the detection mechanism of bile acids. Our studies show that the displacement of SDS from the LC/aqueous interface by the competitive adsorption of bile acids induces a homeotropic-to-planar anchoring transition of the LC at the interface, providing an optical signature for the simple and rapid detection of bile acids. The adsorption of bile acids on the interface was found to follow Langmuir-Freundlich isotherm. The adsorption kinetics of different bile acids is compared. We find that both the number and position of hydroxyl groups of bile acids affect their adsorption kinetics. The different optical patterns of LC films formed by the adsorption of bile acids are also discussed. The second part of this dissertation studies the effect of solution conditions, surfactants, and liquid crystals on the detection limit of the LC-based biosensor for bile acids. Low pH and high ionic strength in the aqueous solution can reduce the electrostatic interaction between SDS and bile acids, which leads to a decreased detection limit. Surfactants with smaller headgroup and lower packing density also help to reduce the detection limit. To further reduce the detection limit, we investigate the effect of LC structures and find that LCs with a shorter chain length give lower detection limits. Also, by substituting a phenyl ring with a cyclohexane ring, we find that the detection limit is further reduced due to the decrease of the interaction between the phenyl rings of LCs. By mixing different LCs together, the detection limit can be linearly tuned from 160 μM to 1.5 μM, which is comparable to the traditional methods. But the LC-based biosensors have much simpler design and manufacture process. The third part of this dissertation is to apply this LC-based biosensor to the detection of urinary bile acids. We test the influence of several potential interfering species such as urea, creatinine, uric acid and ascorbic acid by conducting experiments in synthetic urine. By adjusting the concentration of SDS, we are able to eliminate the impact of those interfering species, and demonstrate that the LC-based biosensors can selectively detect urinary bile acids in human urine, suggesting its potential for screening liver dysfunctions. The final part of this dissertation is to investigate the application of LC-based biosensors in detecting the lipolysis process by porcine pancreatic lipase (PPL). It has been a long-standing argument over the role of bile salts on the activity of PPL. Thus, we study the time course of the hydrolysis of phospholipid L-dipalmitoylphosphatidylcholine (L-DPPC) by PPL at LC/aqueous interface. The hydrolysis of L-DPPC leads to a homeotropic-to-tilted anchoring transition of the LC at the interface, which allows the hydrolysis process to be monitored by a polarizing optical microscope. The microscopy image analysis reveals a lag-burst kinetics where a lag phase is followed by a burst phase. The effect of bile acids on these two phases is studied. We find that the activity of PPL both in the presence and absence of colipase can be improved by increasing the concentration of bile acids. The improvement becomes more distinct in the presence of colipase.
| Identifer | oai:union.ndltd.org:ucf.edu/oai:stars.library.ucf.edu:etd-1674 |
| Date | 01 January 2015 |
| Creators | He, Sihui |
| Publisher | STARS |
| Source Sets | University of Central Florida |
| Language | English |
| Detected Language | English |
| Type | text |
| Format | application/pdf |
| Source | Electronic Theses and Dissertations |
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