John Korth

Care regimen and lens material influence on silicone hydrogel contact lens deposition

Purpose. To quantitatively detect proteins and cholesterol extracted from worn silicone hydrogel contact lenses and determine the effect of various lens care solutions on deposit accumulation. Methods. Contact lenses, made from different polymers and worn on a daily wear schedule with different lens care solutions, were collected. Lipid and protein deposits were extracted by methanol:chloroform (1:1, v/v) and protein extraction solution (containing urea and surfactant), respectively. Lipid extracts were separated and cholesterol quantified using thin layer chromatography. Protein extracts were quantified using standard techniques. Results. Among all lenses tested, Balafilcon A lenses exhibited greatest extracted cholesterol (4.1 to 8.2 μg/lens) and total protein (5.4 to 23.2 μg/lens). AQuify was the most effective solution in reducing extracted deposits, especially extracted protein, from Balafilcon A lenses. AQuify and Opti-Free RepleniSH solutions were most effective in reducing extracted cholesterol from Senofilcon A and Galyfilcon A lenses, respectively. Use of Opti-Free Express solution resulted in more extracted protein from Lotrafilcon B lenses than use of other solutions. Generally, Lotrafilcon B, Senofilcon A, and Galyfilcon A lenses accumulated relatively low amount of proteins. Lotrafilcon B lenses accumulated the least amount of cholesterol deposit among all lenses tested regardless of solution used. Conclusions. Lens polymer (possibly associated with surface characteristics) is a prominent factor affecting lipid and protein accumulation. Within a lens polymer type, lens care solutions exhibit varying effectiveness in reducing protein and lipid accumulation.

Identification of volatile compounds released by roots of an invasive plant, bitou bush (Chrysanthemoides monilifera spp. rotundata), and their inhibition of native seedling growth

Allelopathy has been suggested as a mechanism promoting the monoculture formation of some invasive exotic plants. Previous studies have shown that hydrophobic extracts of the roots and soil of exotic bitou bush (Chrysanthemoides monilifera spp. rotundata (DC.) T. Norl.) inhibited the seedling growth of five Australian native plants, including the dominant acacia (Acacia longifolia var. sophorae (Labill.) F. Muell.). Based on this finding, we compared the hydrophobic root and soil chemical profiles of bitou bush and acacia to determine whether bitou bush roots release allelopathic compounds that are novel to the invaded system. We detected three compounds that were exclusive to the bitou bush root and soil, and seven compounds that were common to the bitou bush and acacia roots but only present in the bitou bush soil. The compounds unique to the bitou bush invaded soil were all sesqui- and diterpenes. Several of these compounds were found to inhibit the seedling growth of a native sedge, Isolepis nodosa (Rott.) R. Br. Of particular interest are the sesquiterpenes: β-maaliene, α-isocomene, β-isocomene, δ-cadinene, 5-hydroxycalamenene and 5-methoxycalamenene which were found in high concentrations in the bitou bush root and soil extracts and exhibited phytotoxic activity. Therefore, we present evidence to suggest that bitou bush exudes low molecular weight volatile compounds into the soil which inhibit native plant seedling growth. The reduced establishment of native plants via allelopathy is likely to create space and contribute to the invasion of bitou bush on the eastern Australian coast.

Headspace gas analysis as a measure of rancidity in corn chips

Headspace analysis by gas chromatography (GC) and gas chromatography-mass spectrometry (GC-MS) has been used to identify and quantify volatiles formed in the oxidative deterioration of corn chips. The effect of light and temperature on the formation of propanal, pentanal and hexanal in stored chips was examined and the data correlated with the results of sensory evaluation. The procedure is suitable for determining concentrations of pentanal and hexanal below, one ppm, which is below the level where rancidity based on sensory evaluation is detected. Problems associated with solute partitioning and sample decomposition in the analysis also were investigated.

Rapid and Quantitative Blood Analysis for Free Fatty Acids by Chemical Ionization Mass Spectrometry

Abstract A quantitative analysis of fatty acids in micro-samples of dried blood spots by chemical ionization mass spectrometry has been developed. Isotope determination was used as the quantitating technique using the corresponding deuterium labeled internal standards. The procedure yields excellent precision and accuracy as demonstrated by the analysis of known fatty acid mixtures and of both C12:0 to C18:0 acids and phytanic acid in the blood from patients.

Rapid quantification of free cholesterol in tears using direct insertion/electron ionization-Mass spectrometry

PURPOSE To establish a simple and rapid analytical method, based on direct insertion/electron ionization-mass spectrometry (DI/EI-MS), for measuring free cholesterol in tears from humans and rabbits. METHODS A stable-isotope dilution protocol employing DI/EI-MS in selected ion monitoring mode was developed and validated. It was used to quantify the free cholesterol content in human and rabbit tear extracts. Tears were collected from adult humans (n = 15) and rabbits (n = 10) and lipids extracted. RESULTS Screening, full-scan (m/z 40-600) DI/EI-MS analysis of crude tear extracts showed that diagnostic ions located in the mass range m/z 350 to 400 were those derived from free cholesterol, with no contribution from cholesterol esters. DI/EI-MS data acquired using selected ion monitoring (SIM) were analyzed for the abundance ratios of diagnostic ions with their stable isotope-labeled analogues arising from the D6-cholesterol internal standard. Standard curves of good linearity were produced and an on-probe limit of detection of 3 ng (at 3:1 signal to noise) and limit of quantification of 8 ng (at 10:1 signal to noise). The concentration of free cholesterol in human tears was 15 ± 6 μg/g, which was higher than in rabbit tears (10 ± 5 μg/g). CONCLUSIONS A stable-isotope dilution DI/EI-SIM method for free cholesterol quantification without prior chromatographic separation was established. Using this method demonstrated that humans have higher free cholesterol levels in their tears than rabbits. This is in agreement with previous reports. This paper provides a rapid and reliable method to measure free cholesterol in small-volume clinical samples.

Rapid quantification of free cholesterol in tears using direct insertion/electron ionization-mass spectrometry.

PURPOSE To establish a simple and rapid analytical method, based on direct insertion/electron ionization-mass spectrometry (DI/EI-MS), for measuring free cholesterol in tears from humans and rabbits. METHODS A stable-isotope dilution protocol employing DI/EI-MS in selected ion monitoring mode was developed and validated. It was used to quantify the free cholesterol content in human and rabbit tear extracts. Tears were collected from adult humans (n = 15) and rabbits (n = 10) and lipids extracted. RESULTS Screening, full-scan (m/z 40-600) DI/EI-MS analysis of crude tear extracts showed that diagnostic ions located in the mass range m/z 350 to 400 were those derived from free cholesterol, with no contribution from cholesterol esters. DI/EI-MS data acquired using selected ion monitoring (SIM) were analyzed for the abundance ratios of diagnostic ions with their stable isotope-labeled analogues arising from the D6-cholesterol internal standard. Standard curves of good linearity were produced and an on-probe limit of detection of 3 ng (at 3:1 signal to noise) and limit of quantification of 8 ng (at 10:1 signal to noise). The concentration of free cholesterol in human tears was 15 ± 6 μg/g, which was higher than in rabbit tears (10 ± 5 μg/g). CONCLUSIONS A stable-isotope dilution DI/EI-SIM method for free cholesterol quantification without prior chromatographic separation was established. Using this method demonstrated that humans have higher free cholesterol levels in their tears than rabbits. This is in agreement with previous reports. This paper provides a rapid and reliable method to measure free cholesterol in small-volume clinical samples.