• Proceedings of the Ginseng society Conference
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• 2002.10a
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• pp.491-501
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• 2002

A cross-examination between KT&G Central Research Institute and Guangzhou Institute for Drug Control was carried out in order to select optimum conditions for extraction, separation and determination of ginsenosides in red ginseng and to propose a better method for the quantitative analysis of ginsenosides. The optimum extraction conditions of ginsenosides from red ginseng were as follows: the extraction solvent, $70\%$ methanol; the extraction temperature, $100^{\circ}C;$ the extraction time, 1 hour for once; and the repetition of extraction, twice. The optimum separation conditions of ginsenosides on the SepPak $C_{18}$ cartridge were as follows: the loaded amount, 0.4 g of methanol extract; the washing solvents, distilled water of 25 ml at first and then $30\%$ methanol of 25 ml; the elution solvent, $90\%$ methanol of 5 ml. The optimum HPLC conditions for the determination of ginsenosides were as follows: column, Lichrosorb $NH_2(25{\times}0.4cm,$ 5${\mu}m$, Merck Co.); mobile phase, a mixture of acetonitrile/water/isopropanol (80/5/15) and acetonitrile/water/isopropanol (80/20/15) with gradient system; and the detector, ELSD. On the basis of the optimum conditions a method for the quantitative analysis of ginsenosides were proposed and another cross-examination was carried out for the validation of the selected analytical method conditions. The coefficient of variances (CVs) on the contents of ginsenoside-$Rg_{1}$, -Re and $-Rb_1$ were lower than $3\%$ and the recovery rates of ginsenosides were $89.4\~95.7\%,$ which suggests that the above extraction and separation conditions may be reproducible and reasonable. For the selected HPLC/ELSD conditions, the CVs on the detector responses of ginsenoside-Rg, -Re and $-Rb_1$) were also lower than $3\%$, the regression coefficients for the calibration curves of ginsenosides were higher than 0.99 and two adjacent ginsenoside peaks were well separated, which suggests that the above HPLC/ELSD conditions may be good enough for the determination of ginsenosides.

• Korean Journal of Ecology and Environment
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• v.54 no.4
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• pp.265-271
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• 2021

Intertidal mud crab (Macrophthalmus japonicus) is an organism with a hard chitinous exoskeleton and has function for an osmotic control in response to the salinity gradient of seawater. Crustacean exoskeletons change in their natural state in response to environmental factors, such as changes in the pH and water temperature, and the presence of pollutant substances and pathogen infection. In this study, the ecotoxicological effects of irgarol exposure and heavy metal distribution were presented by analyzing the surface roughness of the crab exoskeleton. The exoskeleton surface roughness and variation reduced in M. japonicus exposed to irgarol. In addition, it was confirmed that the surface roughness and variation were changed in the field M. japonicus crab according to the distribution of toxic heavy metals(Cd, Pb, Hg) in marine sediments. This change in the surface roughness of the exoskeleton represents a new end-point of the biological response of the crab according to external environmental stressors. This suggests that it may affect the functional aspects of exoskeleton protection, support, and transport. This approach can be utilized as a useful method for monitoring the aquatic environment as an integrated technology of mechanical engineering and biology.

• Molecules and Cells
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• v.46 no.9
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• pp.545-557
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• 2023

Sphingomyelinase (SMase) catalyzes ceramide production from sphingomyelin. Ceramides are critical in cellular responses such as apoptosis. They enhance mitochondrial outer membrane permeabilization (MOMP) through self-assembly in the mitochondrial outer membrane to form channels that release cytochrome c from intermembrane space (IMS) into the cytosol, triggering caspase-9 activation. However, the SMase involved in MOMP is yet to be identified. Here, we identified a mitochondrial Mg²⁺-independent SMase (mt-iSMase) from rat brain, which was purified 6,130-fold using a Percoll gradient, pulled down with biotinylated sphingomyelin, and subjected to Mono Q anion exchange. A single peak of mt-iSMase activity was eluted at a molecular mass of approximately 65 kDa using Superose 6 gel filtration. The purified enzyme showed optimal activity at pH of 6.5 and was inhibited by dithiothreitol and Mg²⁺, Mn²⁺, Ni²⁺, Cu²⁺, Zn²⁺, Fe²⁺, and Fe³⁺ ions. It was also inhibited by GW4869, which is a non-competitive inhibitor of Mg²⁺-dependent neutral SMase 2 (encoded by SMPD3), that protects against cytochrome c release-mediated cell death. Subfractionation experiments showed that mt-iSMase localizes in the IMS of the mitochondria, implying that mt-iSMase may play a critical role in generating ceramides for MOMP, cytochrome c release, and apoptosis. These data suggest that the purified enzyme in this study is a novel SMase.

• Journal of Wetlands Research
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• v.2 no.1
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• pp.31-40
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• 2000

The flora, vegetation structure and physico-chemical characteristics of surface sediment were investigated in Paksil-nup wetland of Hapcheon-gun, Gyeongsangnam-do, Korea in 1990~1998. Ranges of the surface sediment characteristics such as pH, conductivity, organic matter, total nitrogen, available phosphorus, exchangeable K, exchangeable Ca, exchangeable Mg, and exchangeable Na were 4.36~4.34, $19.0{\sim}1260.0\;{\mu}mho\;cm^{-1}$, 0.01~6.35%, 0.001~0.14%, 0.01~0.31 mg/l00g, 1.01~13.98 ppm, 16.75~143.80 ppm, 0.93~14.85 ppm and 0.21~3.86 ppm, respectively. Percentages of the particle size such as sand, silt, and clay were 13.0~93.3%, 5.4~71.7%, and 0.5~37.5%, respectively. The flora of the study area was composed of 72 families, 182 genera, 223 species, 36 varieties and 2 form or total 261 kinds, and those were 45 kinds of vascular hydrophytes and 216 kinds of vascular hygrophytes. The life form of vascular hydrophytes was classified as 27 kinds (60%) of emergent plants, 8 kinds (18%) of submerged plants, 6 kinds (13%) of free-floating plants and 4 kinds (9%) of floating-leaved plants, respectively. The importance value of Salix nipponica was highest as 123.78 in the shrub and tree layers, and that of Trapa japonica was highest as 16.69 in the herb layer. The vegetation type was divided into two groups according to the association analysis. The vegetation of the littoral zone was classified into 7 associations according to the cluster analysis based on the coverage data. These results showed significant differences with those of stand ordination by correspondence analysis based on the species composition and by PCA based on the sediment properties. Factors affecting the distribution of the vascular hydrophytes and hygrophytes were the gradient of particle size, altitude, and water depth.

• Korean Journal of Food Science and Technology
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• v.33 no.1
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• pp.33-39
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• 2001

To develop a method for separation process using Sep-pak $C_18$, simultaneous and systematic analysis of 8 permitted and 11 non-permitted synthetic food colors in Korea, optimization of analysis conditions for reverse phase ion-pair high performance liquid chromatography was carried out. For the best result of Sep-pak $C_18$ separation the pH of color standard mixture solution was $5{\sim}6$ and 0.1% HCl-methanol solution were set as eluent. The colors eluated from Sep-pak $C_18$ cartridge were determined and confirmed by high performance liquid chromatography with a photodiode array detector at 420 nm for yellow colors type, at 520 nm for red colors type, at 600 nm for blue and green colors type and at 254 nm for mixed colors. Conditions for HPLC analysis were as follows: column, Symmetry $C_18$ (5 m, 3.9 mm $i.d.{\times}150\;mm$); mobile phase, 0.025 M ammonium acetate (containing 0.01 M tetrabutylammonium bromide) : acetonitrile : methanol (65 : 25 : 10) and 0.025 M ammonium acetate(containing 0.01 M tetrabutylammonium bromide) : acetonitrile : methanol (40 : 50 : 10); flow rate, 1 mL/min. It takes 35 minutes for simultaneaus analysis and 18 minutes for systematic analysis. The detection limits range of each colors were $0.01{\sim}0.05\;{\mu}g/g$.

• Korean Journal of Ecology and Environment
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• v.47 no.spc
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• pp.48-56
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• 2014

Weekly changes of water environments and phytoplankton community with the salinity gradients were investigated at Gyoungpo Lake from April to November in 1998 and 2012. Underwater crossam in Gyoungpo Lake was removed in 2004. Thereafter, average salinity of Gyoungpo lake increased from 7.5 ppt in 1998 to 20 ppt in 2012. A total of 99 and 80 species of phytoplankton was observed from the sampled in 1998 and 2012, respectively. The number of common species during the 2 separate years was 40. Transparency, SS, $NO_3-N$ concentration and N/P ratio in 2012 were lower than those in 1998. During the period of water shortage (April, May) of 2012 transparency decreased due to decreased salinity and increased SS and Chl. a. Correlation coefficients between species and community scores of DCA ordination based on data matrix of the phytoplankton revealed larger variation among sampling seasons in 1998 than in 2012. The increase of seawater influx and conversion rates following the removal of the underwater crossbeam might explain such a differential variation. Gymnodium sp., Peridinium sp., Prorocentrum sp., Nitzschia longissima, Schroederia setigera, Lyngbya sp., Asterococcus limneticus, Asterococcus superbus and Cyclotella meneghiniana were found to well adapt at the high salinities in 2012. Comparatively, Asterrionella formosa, Nitzschia frustulum, Chlorella ellipsoidea, Scenedesmus bijuga and Scenedesmus ellipsoideus were observed at lower salinities in 1998. Two quite contrasting phytoplankton communities were found in the two seasons of a year, spring with limited precipitation and summer, the flood season.

• Journal of Food Hygiene and Safety
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• v.31 no.5
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• pp.327-334
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• 2016

This study was conducted to establish the standard method for the contents of biotin in milk formulas. To optimize the method, we compared several conditions for liquid extraction, purification and instrumental measurement using spiked samples and certified reference material (NIST SRM 1849a) as test materials. LC-MS/MS method for biotin was established using $C_{18}$ column and binary gradient 0.1% formic acid/acetonitrile, 0.1% formic acid/water mobile phase is applied for biotin. Product-ion traces at m/z 245.1 ${\rightarrow}$ 227.1, 166.1 are used for quantitative analysis of biotin. The linearity was over $R^2=0.999$ in range of $5{\sim}60{\mu}g/L$. For purification, chloroform was used as a solvent for eliminating lipids in milk formula. The linearity was over 0.999 in range of 5~60 ng/mL. The detection limit and quantification limit were 0.10, 0.31 ng/mL. The accuracy and precision of LC-MS/MS method using CRM were 103%, 2.5% respectively. Optimized methods were applied in sample analysis to verify the reliability. All the tested milk formulas were acceptable contents of biotin compared with component specification and standards for nutrition labeling. The standard operating procedures were prepared for biotin to provide experimental information and to strengthen the management of nutrient in milk formula.

• Korean Journal of Food Science and Technology
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• v.45 no.6
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• pp.693-699
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• 2013

A simple, sensitive, and specific method for quantifying the nicotine content of synthetic favoring substances (SFS) was developed using high performance liquid chromatography (HPLC) with a photo-diode array detector (PDA). Nicotine was extracted from SFS samples by using an acid-base liquid-liquid extraction method with dichloromethane and distilled water. The nicotine content was quantified by HPLC/PDA (261.9 nm) with a $C_{18}$ column under a gradient of 10% acetonitrile with 20 mM ammonium formate (ammonia solution adjusted to pH 8.7) to 100% acetonitrile. The calibration curve, analyzed from concentration standards between 0.1 to 2 mg/L, presented linearity with a correlation coefficient ($r^2$)>0.9999. The limit of quantitation (LOQ) of nicotine in SFS was 0.4 mg/kg, and the average recoveries ranged from 76.4% to 96.3%. The repeatability of measurements, expressed as the coefficient of variation (CV%), ranged from 1.74 to 5.12%. This newly developed method for nicotine quantification in SFS can be considered an analytical method with an acceptable level of sensitivity and repeatability.

• Korean Journal of Environmental Agriculture
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• v.36 no.2
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• pp.119-128
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• 2017

BACKGROUND: Miscellaneous cereal have been largely consumed in Korea as due to their physiological functions beneficial to human health. The cereals are currently a social concern because they have been found to contain heavy metals. Thus, monitoring heavy metals in the cereals is an important requirement for food safety analysis. In this study, we determined arsenic concentration in the cereals randomly harvested from different markets. METHODS AND RESULTS: Inorganic arsenic was determined by ICP-MS coupled with HPLC system. The HPLC-ICP-MS analysis was optimized based on the limit of detection and recover test to reach $0.13-1.24{\mu}g/kg$ and 94.3-102.1%, respectively. The concentrations of inorganic arsenic equivalent to daily exposure were levels of $19.91{\mu}g/day$ in mixed grain, $1.07{\mu}g/day$ in glutinous rice, $0.77{\mu}g/day$ in black brown rice, $0.13{\mu}g/day$ in barley and $0.11{\mu}g/day$ in soybeans. CONCLUSION: The levels of arsenic in miscellaneous cereals were found lower than the recommended The Joint FAO/WHO Expert Committee on Food Additives (JECFA) levels, suggesting that the cereals marketed in Korea are not potential concern in risk assessment.

• Journal of Food Hygiene and Safety
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• v.34 no.5
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• pp.413-420
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• 2019

In this study, a sample preparation method and a simultaneous determination method by ultra-high performance liquid chromatography coupled with tandem mass spectrometry for 9 isomers of chlorogenic acid and arbutin in fruits were developed. The samples were extracted using 90% methanol (pH 3.0), with the solutions being shaken and then sonicated for 10 min each. After centrifugation at 4,000 rpm for 10 min, the extraction was concentrated under a vacuum at $40^{\circ}C$ using a vacuum evaporator. The residue was dissolved in 5 mL of 5% methanol and filtered through a $0.45{\mu}m$ membrane before UHPLC-MS/MS analysis. The separations were performed on a C18 column with gradient elution of water (containing 0.1% formic acid) and methanol (containing 0.1% formic acid). The specificity, linearity, limit of detection, limit of quantification, accuracy, and precision of the proposed methods were also evaluated.