• Analytical Science and Technology
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• v.21 no.2
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• pp.84-92
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• 2008

A method for the determination of trace amount of caffeine in urine and various drink samples using hollow fiber-liquid phase microextraction (HF-LPME) and capillary gas chromatograph/nitrogen phosphorus detector (GC/NPD) has been established. HF-LPME method has been optimized with respect to several experimental parameters including the effects of the hollow fiber length, extraction solvent, stirring mode, pH and salt concentration for the determination of caffeine from aqueous samples. The correlation coefficient of calibration curve for caffeine was 0.9994. The average recovery was 102%(n=3). The established method is feasible for the determination of trace amounts of caffeine in several aqueous sample. The limit of detection (LOD) and the limit of quantitation (LOQ) have been found to be 2.5 and 10 ng/mL, respectively. The established HF-LPME method for the analysis of caffeine from aqueous sample can be used for the determination of biological, food and environmental samples.

• Analytical Science and Technology
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• v.37 no.2
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• pp.98-113
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• 2024

The current investigation entails the characterization of five degradation products (DPs) formed under different stress conditions of loteprednol using liquid chromatography-tandem mass spectrometry (LC-MS/MS). In addition, this study developed a stable high-performance liquid chromatography (HPLC) method for evaluating loteprednol along with impurities. The method conditions were meticulously fine-tuned which involved the exploration of the appropriate solvent, pH, flow of the mobile phase, columns, and wavelength. The method conditions were carefully chosen to successfully resolve the impurities of loteprednol and were employed in subsequent validation procedures. The stability profile of loteprednol was exposed to stress degradation experiments conducted under five conditions, and DPs were structurally characterized by employing LC-MS/MS. The chromatographic resolution of loteprednol and its impurities along with DPs was effectively achieved using a Phenomenex Luna 250 mm C18 column using 0.1 % phosphoric acid, methanol, and acetonitrile in 45:25:30 (v/v) pumped isocratically at 0.8 mL/min with 243 nm wavelength. The method produces an accurate fit calibration curve in 50-300 ㎍/mL for loteprednol and LOQ (0.05 ㎍/mL) - 0.30 ㎍/mL for its impurities with acceptable precision, accuracy, and recovery. The stress-induced degradation study revealed the degradation of loteprednol under basic, acidic, and photolytic conditions, resulting in the formation of seven distinct DPs. The efficacy of this method was validated through LC-MS/MS, which allowed for the verification of the chemical structures of the newly generated DPs of loteprednol. This method was appropriate for assessing the impurities of loteprednol and can also be appropriate for structural and quantitative assessment of its degradation products.

• Journal of Microbiology and Biotechnology
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• v.34 no.2
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• pp.436-456
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• 2024

Several thermostable proteases have been identified, yet only a handful have undergone the processes of cloning, comprehensive characterization, and full exploitation in various industrial applications. Our primary aim in this study was to clone a thermostable alkaline protease from a thermophilic bacterium and assess its potential for use in various industries. The research involved the amplification of the SpSKF4 protease gene, a thermostable alkaline serine protease obtained from the Geobacillus thermoglucosidasius SKF4 bacterium through polymerase chain reaction (PCR). The purified recombinant SpSKF4 protease was characterized, followed by evaluation of its possible industrial applications. The analysis of the gene sequence revealed an open reading frame (ORF) consisting of 1,206 bp, coding for a protein containing 401 amino acids. The cloned gene was expressed in Escherichia coli. The molecular weight of the enzyme was measured at 28 kDa using sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The partially purified enzyme has its highest activity at a pH of 10 and a temperature of 80℃. In addition, the enzyme showed a half-life of 15 h at 80℃, and there was a 60% increase in its activity at 10 mM Ca²⁺ concentration. The activity of the protease was completely inhibited (100%) by phenylmethylsulfonyl fluoride (PMSF); however, the addition of sodium dodecyl sulfate (SDS) resulted in a 20% increase in activity. The enzyme was also stable in various organic solvents and in certain commercial detergents. Furthermore, the enzyme exhibited strong potential for industrial use, particularly as a detergent additive and for facilitating the recovery of silver from X-ray film.

• Journal of Food Hygiene and Safety
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• v.31 no.2
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• pp.85-93
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• 2016

A rapid method using HPLC-MS/MS has been developed for quantitative determination of the metabolites of nitrofurans, namely 3-amino-2-oxazolidone (AOZ), 5-morpholinomethyl-3-amino-2-oxazolidinone (AMOZ), 1-ammino-hydantoin (AHD) and semicarbazide (SEM) in loach. The extraction procedure was founded on simultaneous acidic hydrolysis and derivatization using 2-nitrobenzaldehyde (2-NBA) for 1 hour at $50^{\circ}C$, followed by purification with liquid-liquid extraction. Recovery was evaluated by spiking standards into blank samples at three levels (0.5, 1.0 and $2.0{\mu}g/kg$), and the mean recovery was 75.1-108.1%. Precision values expressed as the relative standard deviation (%RSD) were ${\leq}8.7%$ and ${\leq}8.5%$ for intra-day and inter-day precision, respectively. Linearity was studied in the range of $0.2-20{\mu}g/Kg$ for NBAOZ, $0.8-20{\mu}g/Kg$ for NBAMOZ, $0.2-20{\mu}g/Kg$ for NBAHD, and $0.1-20{\mu}g/Kg$ for NBSEM, and the obtained coefficient correlations (r) were ${\geq}0.99$ for all compounds. Limits of detection (LODs) for the derivatized nitrofuran metabolites were established at $0.06{\mu}g/Kg$ for NBAOZ, $0.24{\mu}g/Kg$ for NBAMOZ, $0.06{\mu}g/Kg$ for NBAHD, and $0.03{\mu}g/Kg$ for NBSEM. Limits of quantification (LOQs) were established at $0.2{\mu}g/Kg$ for NBAOZ, $0.8{\mu}g/Kg$ for NBAMOZ, $0.2{\mu}g/Kg$ for NBAHD, and $0.1{\mu}g/Kg$ for NBSEM. This simplified rapid method for reducing the derivatization and hydrolysis times can be applied to the determination of nitro-furan residues in loach.

• Applied Biological Chemistry
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• v.22 no.2
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• pp.109-122
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• 1979

Present study was carried out to reduce residual toxicity of BHC insecticides inherent in the organochlorine pesticides. For This end, r-isomer, the most potent insecticidal component among the BHC stereoisomers, was isolated and thus fortified by means of solvent precipitation. In parallel, 3-(2,4,5-trichlorophenyl)-1-methyl urea was prepared in good yield from technical BHC via 1,2,4-trichlorobenzene, 1,2,4,-trichloronitrobenzene, and 2,4,5-trichloroaniline. In addition, certain merit of the compound which make it possible to use as a herbicide is discussed. The results are summarized as follows; 1. Recrystallizing technical BHC from methanol-water binary solvent system, r-isomer was enriched to 49.7% at 95% recovery of r-isomer. 2. By partitioning technical BHC in 85% of methanolic solution into chloroform, r-isomer was fortified to 89.6% at 90.5% recovery of r-isomer. 3. Yield of 1,2,4-trichlorobenzene from technical BHC was greatly dependent upon concentration of alkalies and to less degree on the alkalies. 4. Surfactants, in particular cationic a quartenary ammonium salt, increased yield of 1,2,4-trichlorobenzene from technical BHC by alkaline hydrolysis. 5. Conversion of 1,2,4-trichlorobenzene to 2,4,5-trichloronitrobenzene was effected almost quantitatively utilizing $HNO_3-H_2SO_4$ nitrating agent at low temperature. 6. Yield of 91.4% was observed for the synthesis of 2,4,5-trichloroaniline by reducing 2,4,5-trichloronitrobenzene in the presence of iron turning and hydrochloric acid. 7. Overall yield based on BHC of 3-(2,4,5-trichlorophenyl)-1- methyl urea was 60.8%. 8. Inhibition effects, both germination and growth, 3-(2,4,5-trichlorophenyl)-1-methyl urea on several crops were found comparable to or more potent than those of $linuron{\circledR}\;and\;diuron{\circledR}$. In addition, it was also noted that susceptibility to the prepared compound depended upon the crops as well as specific part (shoots, roots) of the plant exposed to the chemicals.

• Journal of the Korean Society of Food Science and Nutrition
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• v.46 no.4
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• pp.459-464
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• 2017

Biphenyl is used as an intermediate in the production of crop protection products, a solvent in pharmaceutical production, and as a component in the preservation of citrus fruits in many countries. Biphenyl is not authorized for use and also does not have standards or specifications as a food additive in Korea. National and imported food products are likely to contain biphenyl. Therefore, control and management of these products is required. In this study, a simple analytical method was developed and validated using HPLC to determine biphenyl in food. These methods are validated by assessing certain performance parameters: linearity, accuracy, precision, recovery, limit of detection (LOD), and limit of quantitation (LOQ). The calibration curve was obtained from 1.0 to $100.0{\mu}g/mL$ with satisfactory relative standard deviations (RSD) of 0.999 in the representative sample (orange). In the measurement of quality control (QC) samples, accuracy was in the range of 95.8~104.0% within normal values. The inter-day and inter-day precision values were less than 2.4% RSD in the measurement of QC samples. Recoveries of biphenyl from spiked orange samples ranged from 92.7 to 99.4% with RSD between 0.7 and 1.7% at levels of 10, 50, and $100{\mu}g/mL$. The LOD and LOQ were determined to be 0.04 and $0.13{\mu}g/mL$, respectively. These results show that the developed method is appropriate for biphenyl identification and can be used to examine the safety of citrus fruits and surface treatments containing biphenyl residues.

• Resources Recycling
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• v.17 no.1
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• pp.80-87
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• 2008

Cementation and solvent extraction processes were studied to separate nickel and iron ions from the $H_2SO_4$ leaching solution with 47 g/L $Fe(Fe^{2+}/Fe^{3+}=1.03),$, 23.5 g/L Ni and 0.90M $H_2SO_4$ which leached from Fe-Ni alloy. Iron powder was used as a reducing agent for the cementation of Ni ion from the leaching solution. The reduction percentage of Ni ion was $17{\sim}20%$ by adding 4 times stoichiometric amount of iron powder at $60{\sim}80$. This may result from the fact that the cementation of Ni ion occurred after the reduction of $Fe^{3+}$ to $Fe^{2+}$ and the neutralization of $H_2SO_4$ with iron powder. The cementation process was proved to be unfeasible for the separation/recovery of Ni ion from the leaching solution including $Fe^{3+}$ as a major component. $Fe^{2+}$ present in the leaching solution was converted to $Fe^{3+}$ for solvent extraction of Fe ion using D2EHPA in kerosene as a extractant. The oxidation of $Fe^{2+}$ to $Fe^{3+}$ was completed by the addition of 1.2 times stoichiometric amount of 35% $H_2SO_4$. 99.6% $Fe^{3+}$ was extracted from the leaching solution (23.5 g/L $Fe^{3+}$) by 4 stages cross-current extraction using 20 vol.% D2EHPA in kerosene. $NiSO_4$ solution with 98.5% purity was recovered from the $H_2SO_4$ leaching solution of scrapped Fe-Ni alloy.

• Resources Recycling
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• v.17 no.6
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• pp.50-56
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• 2008

A polypropylene fraction collected from the stream of post-consumer plastics was pyrolyzed. The aim of this study is to observe the dependence of yield of BTEX-aromatics normally used as solvent on the reaction temperature. To reach the goal, three experiments were carried out at different temperature between 650 and $700^{\circ}C$, using a fluidized bed reactor that shows an excellent heat transfer. In the experiments, product gases were used as a fluidizing medium to maximize the amount of BTEX-aromatics at fixed flow rate and feed rate during the pyrolysis. Oil, gas and char were obtained as product fractions. Product gases were analyzed with GCs(TCD, FID) and with a GC-MS system for qualitative analysis. For an accurate analysis of product oil, the product oil was distilled under vacuum, and separated the distillation residues from oil fractions that were actually analyzed with a GC-MS system. As the reaction temperature went higher, the content of BTEX-aromatics increased. The maximal yield of BTEX-aromatics was obtained at $695^{\circ}C$ with a value of about 30%. The main compounds of product gas were $CH_4$, $C_2H_4$, $C_2H_6$, $C_3H_6$, $C_4H_{10}$ and the product gas had an higher heating value about 45MJ/kg. It could be used as a heat source for a pyrolysis plant or for other fuel applications.

• Journal of the Korean Society of Food Science and Nutrition
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• v.43 no.11
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• pp.1791-1795
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• 2014

The objective of this study was to compare two extraction methods for determination of vitamin K1 (phylloquinone) in vegetables. In addition, analytical method validation parameters such as accuracy, precision, limit of detection (LOD), limit of quantification (LOQ), and linearity were calculated to ensure the method's validity. Vitamin K1 was quantified by reversed-phase HPLC using post-column derivatization and fluorescence detection ($Ex{\lambda}=243nm$, $Ex{\lambda}=430nm$). Higher analytical values were observed using solvent extraction compared to those from the enzyme extraction method. The results from the method validation showed high linearity in the calibration curve with a coefficient of correlation ($R^2$) of 0.9994. The LOD and LOQ were 0.1335 and 0.2784 ng/injection volume ($50{\mu}L$), respectively. The inter-day precision and inter-day precision were 2.0% and 2.1%, respectively. Overall recovery was close to 100% (n=5). The phylloquinone contents ranged from 9.42 to $1,212.57{\mu}g/100g$. Our study provides reliable data on the phylloquinone contents in commonly consumed vegetables in Korea.

• Journal of the Korean Society of Food Science and Nutrition
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• v.43 no.11
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• pp.1737-1741
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• 2014

An HPLC method for determination of naringin was developed to standardize it as a marker compound in Goheung yuzu extract as a functional health food. Optimum results were obtained by C-18 column chromatography using solvent mixtures (A: 0.5% acetic acid, B: acetonitrile) as the stationary phase and mobile phase. The method was fully validated and sensitive with a limit of detection (LOD) of 0.0218 mg/L and limit of quantification (LOQ) of 0.0661 mg/L. The method showed high linearity (coefficient of correlation=0.9986) and high accuracy, as recovery rates of naringin at concentrations of 1, 0.5, 0.1, 0.05 mg/mL were in the ranges of 95.74~98.25%, 97.67~101.01%, 97.33~104.64%, and 95.53~106.82%, respectively. Intra-day and inter-day variation, which are measures of method precision, were 1.39~1.95% and 0.17~1.49%, respectively. Therefore, the method could be used without modification for determination of naringin as a marker compound in Goheung yuzu extracts.