Browsing by Subject "HPLC"
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Item Clinical pharmacokinetics and safety of zonisamide in apparently normal dogs following single and multiple dosing(Texas A&M University, 2004-11-15) Perkins, Jeremy DaneThe purpose of this study was to design a dosing regimen and evaluate the safety of zonisamide (ZNS) following multiple dosing and to determine appropriate monitoring methods. Clinical pharmacokinetics were studied in 8 adult dogs (4 male and 4 female) ranging from 3 to 4 years of age using a randomized crossover design following single intravenous (IV) and oral administration, 6.85 and 10.25 mg/kg, respectively. Samples were collected intermittently for 48 hours. Dogs were then dosed orally (10.17 mg/kg) twice daily for 8 weeks. Blood samples were collected weekly and at discontinuation of the drug. Additionally, urine was collected to determine 24 hour urine ZNS clearance following IV administration. Safety was based on clinical pathology, thyroid and urine testing during both studies. ZNS was measured using high performance liquid chromatography in serum, plasma, erythrocytes (RBC) and whole blood. Data were subjected to standard non-compartmental pharmacokinetic analysis using computer assisted linear regression (WinNonLin?). Comparisons were made in different compartments using one-way ANOVA to identify any differences. Safety parameters at study beginning and end were compared using a Student t-test. ZNS concentrations differed among blood compartments after single dosing, with oral maximum concentration (Cmax) being greatest in RBC (28.73?g/ml) and least (14.36?g/ml) in plasma. Volume of distribution also differed, being greater (1096.05ml/kg) in plasma and least in (379.23ml/kg) RBC. Clearance of ZNS was 57.55ml/hr/kg from plasma and 5.06ml/hr/kg from RBC. Elimination half- life in plasma was 16.4 hr in serum and 57.4 hr in RBC. Bioavailability was 126.8% for RBC and 189.6% for plasma. Following multiple dosing, at steady-state, Cmax averaged 65.8?g/ml with fluctuations of 17.2% between dosings. Accumulation of ZNS was 3.5 (plasma) and 4.3 (RBC). Concentrations did not differ among blood compartments at the end of multiple dosing. Although differences did occur across time in clinical pathology tests, all were within normal limits at study end except for T4. In conclusion, ZNS dosed at 10 mg/kg twice daily for dogs would maintain therapeutic levels (10 to 70?g/ml) recommended in human epileptic patients. Therapeutic monitoring would be best measured in serum or plasma accompanied with thyroid and urine testing.Item HPLC separation of amines with a zirconia-based column coupled to a gas- phase chemiluminescence nitrogen specific detector (CLND)(Texas A&M University, 2004-09-30) Salinas, Silvia AdrianaGas phase chemiluminescence nitrogen specific detector (CLND)is used for the direct analysis of underivatized nitrogen-containing components such as alkylamines that can not be detected by the so called universal HPLC detector, the UV detector. However, alkali metal hydroxides can not be used as mobile phase constituents with the CLND because they form non volatile particulate combustion products that foul the detector. Therefore, trimethylsulfonium hydroxide (TMSOH) has been selected as a strong base for use with the CLND, because its combustion products, CO2, H2O and SxOy are volatile. An alkali-stable zirconia-based column was used and coupled to the CLND. Zirconia-based columns are mechanically and hydrolytically more stable than silica-based columns, which have a working pH range from 3 to 8 only. Zirconia-based columns can be used at a pH from 1 to 14 and can be used at temperatures up to 200?C. The separation of amines was carried out at high pH values where the amino groups were deprotonated. Primary, secondary, tertiary and quaternary amines were separated using a pH=13.7 mobile phase that contained only TMSOH, methanol and water. Good peak shapes were observed for all, except n-alkylamines and samples that contained both amino groups and alcohol groups.Item Pharmacokinetics and cytoprotective evaluation of Caffeic Acid Phenethyl Amide and fluorinated derivatives against oxidative stress(2012-12) Yang, John; Stavchansky, Salomon; Bowman, Phillip D; Kerwin, Sean M; Williams, Robert O; McGinity, James WIschemic injury occurs when the flow of blood is reduced or blocked to an area of the body and can cause significant tissue damage by generation of reactive oxygen species (ROS), activation of apoptotic pathways and through induction of the inflammatory response. Restoration of blood flow and reperfusion of the blocked site, while essential, can generate a second injury that itself needs to be controlled. Together the two injuries are termed ischemia/reperfusion (I/R) injury. This type of injury is frequently encountered in medicine and is a major medical problem. Therapeutic strategies to combat I/R injury include the introduction of compounds that can scavenge ROS or can induce metabolic pathways with the effect of inhibiting apoptosis. Caffeic Acid Phenethyl Ester (CAPE), a polyphenolic compound found in propolis, has been shown to protect a variety of cells types against ROS in vitro and has also been shown to induce a variety of genes including hemeoxygenase 1 (HMOX-1) , an enzyme that has been implicated in a cytoprotective pathway. Despite showing significant cytoprotection of cells against oxidant stress in vitro, CAPE is readily hydrolyzed in plasma and is also quickly removed from circulation. This result may explain the limited cytoprotective effects of CAPE in vivo. We have synthesized a series of CAPE amide derivatives, including Caffeic Acid Phenethyl Amide (CAPA), with the aim of improving CAPE’s stability properties while maintaining the cytoprotective effects of the parent compound. We found that CAPA, in addition to 2 other amide derivatives, were able to protect human umbilical vein endothelial cells (HUVEC) against ROS to a similar degree as CAPE. In addition, we have observed significant improvement in plasma stability of CAPA over CAPE at multiple temperatures. The elimination half-life of CAPA from the systemic circulation was also seen to be significantly improved over CAPE following intravenous administration to male Sprague-Dawley rats. The longer residence time of CAPA over CAPE in circulation may potentially result in greater cytoprotection in vivo.Item Production Systems and Processing Effect on Phytochemicals in Citrus Fruits and Their Analytical and Isolation Methods(2012-12-13) Uckoo, Ram 1980-The emerging scientific evidences on the role of food components in prevention of several chronic diseases are the momentum for shifting from a traditional focus on production to enhancement of nutritional quality. To further understand the role of these phytochemicals this dissertation describes the development of rapid analytical and isolation methods, and the effect of production systems and processing techniques on the levels of phytochemicals in citrus fruits. In the first study, a simultaneous high performance liquid chromatography (HPLC) method for the rapid analysis of amines and organic acids was developed. The simultaneous extraction and analysis of samples provides an economical method for analyzing a large number of samples. In the second study, rapid separation method of potent health beneficial phytochemicals such as polymethoxyflavones from citrus peels using flash chromatography was developed. Using the developed method, five polymethoxyflavones were separated and isolated with high purity in gram level quantity. In the third study, the levels of phytochemicals in organically and conventionally grown lemons and their storage at market simulated conditions were determined. Results suggest that organically produced citrus fruits have higher content of organic acids and flavonoids than conventionally produced. The fourth and fifth study determined the influence of household processing (blending, juicing, hand squeezing techniques) and emerging processing (high pressure processing [HPP], thermal processing) on the phytochemicals content of ?Rio Red? grapefruits. Fruits processed by blending had significantly higher levels of flavonoids, furocoumarins and limonin compared to juicing and hand squeezing, while HPP enabled in extending the shelf life of the processed juice without any adverse effects. Therefore, consuming grapefruit juice processed by blending may provide higher levels of health beneficial phytochemicals. The sixth study describes a rapid flash chromatography method for isolation of PMFs and furocoumarins from citrus industrial by products such as peel oil. In the seventh study the developed method was applied to isolate 10 different phytochemicals from an unexplored citrus species, Miaray mandarin (Citrus miaray TAN.). Among them, the 5,7,8,3',4' pentamethoxyflavone was isolated for the first time from the genus Citrus.Item Sample Preparation Methods and Pre-harvest Factors Influencing the Contents of Bioactive Compounds and Antioxidant Activity in Peppers(2012-02-14) Bae, Hae JinPeppers are a rich source of diverse bioactive compounds with potential health-promoting properties. The levels of bioactive compounds and antioxidant activity can be affected by analytical methods, pre-harvest factors, and the quality of peppers. In order to understand the nutrient composition and antioxidant activity in peppers, determination of factors influencing the contents of bioactive compounds is important. The overall objectives were to determine the efficient conditions of sample preparation and the impact of pre-harvest factors affecting bioactive compounds and antioxidant activity. Optimal extraction procedures were developed, and HPLC methods were validated for bioactive compounds in peppers. The highest flavonoids were extracted in ethanol, while myricetin was extracted using N-N-dimethylformamide. Optimized conditions for flavonoids were obtained during 3 h of extraction time and hydrolysis in M HCl for 60 min at 95 degrees C. Capsaicinoids and ascorbic acid were simultaneously separated and extracted using a solvent mixture consisting of 3% metaphosphoric acid: ethanol (2:8) after 30 min of sonication. To determine the relationship between bioactive compounds and antioxidant activities in pepper extracts from different solvent properties, bioactive compounds were analyzed, and the antioxidant activities were assayed by 2,2,-Diphenyl-1-picryl hydrazyl (DPPH), reducing power, and degradation of deoxyribose. Hexane extracts had the highest levels of capsaicinoids and carotenoids, while methanol extracts had the highest levels of flavonoids. Strong DPPH scavenging activity and reducing power were found in lipophilic extracts, while hydrophilic extracts were appropriate for inhibition of deoxyribose degradation. Variation in content of ascorbic acid, capsaicinoids, and flavonoids was evaluated at immature and mature stages of pepper cultivars in different locations over two years. Mature peppers contained the highest levels of capsaicinoids and ascorbic acid. Flavonoids were variable at different maturity stages. Interactions between pre-harvest factors and bioactive compounds were highly significant. This study demonstrated the efficient sample preparation methods and simultaneous separation of bioactive compounds, which reduces analysis time and leads to reduced cost. The antioxidant properties were strongly associated with the concentration of bioactive compounds based on selective pepper extracts. The pepper quality can be improved by using appropriate pre-harvest conditions that increase the levels of bioactive compounds in peppers.Item Speciation and transport of anthropogenic 129Iodine and natural 127Iodine in surface and subsurface environments(Texas A&M University, 2005-02-17) Schwehr, Kathleen AnnIodine is a biophilic element with one natural long-lived isotope, 129I (t1/2= 15.6 million years), and one stable isotope, 127I. The inventory of 129I in surface environments has been overwhelmed by anthropogenic releases over the past 50 years. The objective of this study is to utilize the elevated concentration and biophilic nature of 129I and the isotopic ratio of iodine (129I/127I) as a tracer of water mass movement and organic matter. Additionally, the significantly elevated values of 129I/127I could provide a geochronometer, similar to the way 14C is used, particularly for terrestrial organic matter that is less than 50 years old. A series of laboratory experiments and field investigations were carried out to characterize the dominant chemical forms of dissolved iodine, i.e., iodide (I-), iodate (IO 3-), and organic iodine (DOI) in natural waters. Sensitive methods were developed for the analysis of nanomolar quantities of 127I species in a variety of environmental systems using high performance liquid chromatography (HPLC) and an organic iodine decomposition technique, dehydrohalogenation. The potential use of 129I/127I as a hydrological tracer was evaluated through measurements of 129I and 127I, which were carried out in wells in the artificially recharged ground water basin of Orange County, California. Literature values of aquifer ages based on 3H/3He and δ18O tracer data, as well as time-series data of chloride and Santa Ana River flow rates over the past decade were compared to values for 129I and 127I. The iodine isotopes demonstrated a conservative behavior in these aquifers, suggesting that the observed variations of these isotopes reflect past river flow conditions during the time of recharge. The feasibility of using 129I/127I ratios to trace terrestrial organic matter across an estuary was tested. A novel analytical technique to determine 129I/127I ratios in DOI was developed for this investigation. The results of a Galveston Bay transect clearly show that 129I/127I ratios in DOI can remain elevated up to salinity of about 15, but that 129I/127I values of inorganic iodine species do not show any trend with change in salinity gradient due to fast isotopic and chemical equilibration in the estuarine waters.