From a 95% ethanolic extract of H. diffusa, four marker compounds (HD1~HD4) were isolated, which were relatively unique and exist in comparably high contents. The structures of marker compounds were identified as digitolutein (1), 2-hydroxy-3-methylanthraquinone (2), (E/Z)-6-O-p-coumaroyl scandoside methyl ester (4:1 mixture) (3), and (E/Z)-6-O-p-methoxycinnamoyl scandoside methyl ester (4:1 mixture) (4), respectively, on the basis of $^{13}C$ and $^1H$-NMR analyses. The calibration curves of marker compounds showed high linearity, as their correlation coefficient ($R^2$) were in the range of 0.9991~0.9999. In addition, the limit of detection (LOD) and the limit of quantification (LOQ) were $0.03{\sim}0.07{\mu}g/ml$ and $0.099{\sim}0.231{\mu}g/ml$, respectively. The intra-day/inter-day precision and accuracy were 0.23~2.00%/0.25~1.16% and 94.60~108.44%/94.73-110.23%, respectively. The optimal HPLC conditions for the simultaneous quantification of HD1~HD4 were as follows: stationary phase; Merck Chromolith RP-18e ($100{\times}4.6mm$, $5{\mu}m$), column temp.; room temperature, UV detection at 280 nm, flow rate; 2.0 ml/min, injection volume; $10{\mu}l$, mobile phase; start with the mixture of 80% solvent A ($H_2O$ containing 0.5% acetic acid) and 20% solvent B (methanol containing 0.5% acetic acid) and gradually decrease solvent A to 40% in 9 min., then retain this condition to 18 min. Under the HPLC condition, the four marker compounds 1~4 were successfully separated without any interference of other constituents. The results obtained in this study are expected to be helpful for the development of nutraceutics and natural medicines and for the quality control of this plant.
Laboratory scale experiments were performed to investigate the removal efficiency of the in-situ chemical oxidation method and the air-sparging method for diesel contaminated soil and groundwater. Two kinds of diesel contaminated soils (TPH concentration : 2,401 mg/kg and 9,551 mg/kg) and groundwater sampled at Busan railroad station were used for the experiments. For batch experiments of chemical oxidation by using 50% hydrogen peroxide solution, TPH concentration of soil decreased to 18% and 15% of initial TPH concentration. For continuous column experiments, more than 70% of initial TPH in soil was removed by using soil flushing with 20% hydrogen peroxide solution, suggesting that most of diesel in soil reacted with hydrogen peroxide and degraded into $CO_2$ or $H_2O$ gases. Batch experiment for the air-sparging method with artificially contaminated groundwater (TPH concentration : 810 mg/L) was performed to evaluate the removal efficiency of the air-sparging method and TPH concentration of groundwater decreased to lower than 5 mg/L (waste water discharge tolerance limit) within 72 hours of air-sparging. For box experiment with diesel contaminated real soil and groundwater, the removal efficiency of air-sparging was very low because of the residual diesel phase existed in soil medium, suggesting that the air-sparging method should be applied to remediate groundwater after the free phase of diesel in soil medium was removed. For the last time, the in-situ box experiment for a unit process mixed the chemical oxidation process with the air-sparging process was performed to remove diesel from soil and groundwater at a time. Soil flushing with 20% hydrogen peroxide solution was applied to diesel contaminated soils in box, and subsequently contaminated groundwater was purified by the air-sparging method. With 23 L of 20% hydrogen peroxide solution and 2,160 L of air-sparging, TPH concentration of soil decreased from 9,551 mg/kg to 390 mg/kg and TPH concentration of groundwater reduced to lower than 5 mg/L. Results suggested that the combination process of the in-situ hydrogen peroxide flushing and the air-sparging has a great possibility to simultaneously remediate fuel contaminated soil and groundwater.
Avian influenza virus (AIV) is recognized as key to the emergence of pandemic influenza for humans; there are growing concerns that AIV H9N2 may become more efficient to transmit to humans in the near future, since the infection of poultry with AIV H9N2 has been common in recent years. In this study, we aimed to produce antisera recognizing the HA and NA proteins of AIV H9N2. Initially, coding sequences corresponding to the N-terminal regions of the HA and NA proteins of the Korean AIV H9N2 (A/Ck/Kr/MS96/96) isolated from a domestic chicken were amplified from the genomic RNA. Following cloning of the amplified cDNA fragments into pGEX4T-1 vector, two GST-fusion proteins (GST-HAln and GST-NAn) were expressed in E. coli BL21 and purified with glutathione sepharose columns; the recombinant GST-HAln and GST-NAn proteins were both used as immunogens in rabbits. The antigenicity of the rabbit antisera was analyzed by immunoblotting of the cell lysates prepared from AIV H9N2-infected MDCK cells. Overall, the recombinant HAln and NAn proteins fused to the C-terminus of GST and the rabbit antisera raised against the corresponding recombinant proteins would provide a valuable reagent for AIV diagnosis and basic research.
This study was performed to report a direct dose dependent stimulatory effect of the Flavonoid(F) on basal testosterone secretion and a dose dependent effect on LH induced testosterone production by Leydig cell of matured rats in vitro culture. F was obtained kom the Rhus vernicifua through aceton extraction and silica gel adsorption column chromatography. Leydig cells (1$\times$10$^{6}$ cells/well) from 12 weeks old rats were incubated with or without F(0, 20, 40, 80, 160 ng) or insulin-like growth factor-I(IGF-I) in the presence or absence of LH(10, 100ng). 1. The maximal stimulatory concentrations of testosterone in culture media were showed at 24hr of culture. but these testosterone level were decreased at 36 hr of culture. 2. Flavonoid(80ng) were significantly(P < 0.05) increased testosterone production compared with control groups for 12 hr culture. 3. Testosterone secretion by Leydig cells stimulated with LH(10, 100ng) for 6 hr and 12hr culture compared with 3 hr culture. 4. LH 10 ng augmented testosterone were increased by addition of F 40 ng for 12 hr culture. 5. F(0 and 40 ng) also enhanced LH 10 ng stimulated testosterone for 3 hr Leydig cells culture. 6. Addition of IGF-I 100 ng to the culture medium for 6 hr were increased the concentration of testosterone by Leydig cells stimulated with 100 ng LH. These results indicate that Flavonoid has a direct stimulatory effect on basal testosterone secretion in rat Leydig cells, and also modulates LH mediated testosterone. Therefore, Flavonoid may act as a modulator on gonadal development or gonadal steroidogenesis in direct or indirect.
Kim, Seon Hee;Kim, Jae-Hyun;Lee, Hwa Jung;Oh, Jae Myoung;Lee, Sung Hye;Bahn, Kyeong Nyeo;Seo, Il Won;Lee, Young Joo;Lee, Jin Hee;Kang, Tae Seok
Journal of Food Hygiene and Safety
/
v.30
no.2
/
pp.143-149
/
2015
This study was conducted to simultaneous analysis methods for water soluble vitamins B group (vitamin $B_1$, vitamin $B_2$, nicotinic acid, nicotinamide, vitamin $B_6$) which is used as health functional foods etc. Analytical methods of water-soluble vitamins B group by HPLC were established through instrumental analytical conditions, and the examination of data such as domestic and foreign reliable methods, and papers of journal. HPLC method analyzing water soluble vitamins B group was established using Capcell Pak C18 UG 120 column in 270 nm through test of columns. The validation has been performed on the method to determine linearity, accuracy, limits of quantification (LOQ) and repeatability for water soluble vitamins B group. An excellent linearity ($r^2=0.999$) was observed for vitamin $B_1$, vitamin $B_2$, nicotinic acid, nicotinamide, vitamin $B_6$ in the concentration range ($0.1{\sim}2{\mu}g/mL$). Observed recovery of vitamin $B_1$ was found to be between 100 and 103%, vitamin $B^2$ was found to be between 104 and 112%, nicotinic acid was found to be between 82 and 85%, nicotinamide was found to be between 121 and 124% and vitamin $B_6$ was found to be between 95 and 104%. LOQ of vitamin $B_1$ was found to be $0.04{\mu}g/mL$, vitamin $B_2$ was found to be $0.05{\mu}g/mL$, nicotinic acid was found to be $0.15{\mu}g/mL$, nicotinamide was found to be $0.08{\mu}g/mL$ and vitamin $B_6$ was found to be $0.63{\mu}g/mL$. Repeatability precision for vitamin $B_1$ was found to be 0.4%, vitamin $B_2$ was found to be 0.4%, nicotinic acid was found to be 0.5%, nicotinamide was found to be 0.7% and vitamin $B_6$ was found to be 0.4% relative standard deviation (RSD). Also, verify the accuracy of the simultaneous analysis methods, we monitored the labeled contents of the health functional foods and children's preferred foods.
Hwang, Kyung Mi;Ham, Hyeon Suk;Lee, Hwa Jung;Kang, Yoon Jung;Yoon, Hae Seong;Hong, Jin Hwan;Lee, Hyoun Young;Kim, Cheon Hoe;Oh, Keum Soon
Journal of Food Hygiene and Safety
/
v.32
no.5
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pp.411-417
/
2017
This study was conducted to establish the analysis method for the contents of choline in infant formulas and follow-up formulas by ion chromatograph (IC). To optimize the method, we compared several conditions for extraction, purification and instrumental measurement using spiked samples and certified reference material (CRM; NIST SRM 1849a) as test materials. IC method for choline was established using Ion Pac CG column and 18 mM $H_2SO_4$ mobile phase. The parameters of validation were specificity, linearity, LOD, LOQ, recovery, accuracy, precision and repeatability. The specificity was confirmed by the retention time and the linearity, $R_2$ was over 0.999 in range of 0.5~10 mg/L. The detection limit and quantification limit were 0.14, 0.43 mg/L. The accuracy and precision of this method using CRM were 95%, 2.1% respectively. Optimized methods were applied in sample analysis to verify the reliability. All the tested products were acceptable contents of choline compared with component specification for nutrition labeling. The standard operating procedures were prepared for choline to provide experimental information and to strengthen the management of nutrient in infant formula and follow-up formula.
To study lipid components of Panax ginseng produced in Korea, the lipids of fresh ginsengs were extracted with the mixture of chloroform-methanol (2:1, v/v) and those of dried ginsengs were extracted with diethyl ether respectively. The lipid components extracted were separated and quantitated by column, thin layer and gas-liquid chromatographies. The results were summarized as follows : 1. Fresh ginseng contained 0.62% total lipid of which 45.28% were neutral lipids, 18.12% glycolipids, and 36.60% phospholipids. But dried ginseng contained 0.89% total lipids of which 86.48% were neutral lipids, 9.20% glycolipids, and 4.32% phospholipids. 2. Triglycerides (37.6 to 42.5% of the total neutral lipids) and sterol esters (16.5 to 19.6%) in all the fresh and dried ginseng were the major components among the neutral lipids. Monoglycerides, diglycerides, free fatty acids and free sterols were minor components. 3. Digalactosyl diglycerides (23.5% of the total glycolipids) in the fresh ginseng and steryl liglycosides (28.9%) in the dried ginseng were predominant components among the glycopids, respectively, Esterified steryl glycosides and monogalactosyl diglycerides were also identified, and four unknown spots in the fresh ginseng and two unknown spots in the dried ginseng were present. 4. Phosphatidyl cholines (31.3 to 31.9% of the total phospholipids) and phosphatidyl glycerols (34.8 to 36.7%) in all the fresh and dried ginseng were the major components among the phospholipids. Phosphatidyl inositols and phosphatidyl ethanolamines were also identified. 5. The major fatty acids in the fresh and dried ginseng were linoleic $(62.29{\sim}64.32%)$, palmitic $(13.16{\sim}15.63%)$, oleic $(5.73{sim}7.23%)$ and linolenic $(5.73{sim}7.23%)$. The fatty acid compositions in neutral lipid fraction was similar to the pattern in those of the total lipids. But glycolipid and phospholipid fractions contained a lower percent of linoleic acid and a higher percent of palmitic acid than the neutral lipid fraction.
A series of tests were conducted on the reesterification of rice bran oil containing high free fatty acids (acid value=119.7) with theoretical equivalent of glycerol. Test results showed that reaction rate (in terms of decrease in acid value) was increased as the reaction temperature was increased regardless of the presence of the catalyst and reaction time (42.7, 21.5 and 10.0 at $170^{\circ}C,\;210^{\circ}C\;and\;250^{\circ}C$, respectively) and as the reaction time was increased regardless of the temperature and the presence of the catalyst (31.1 vs 18.3 for 3 hrs vs 6 hrs). The presence of the catalyst (0.2% tin chloride) also accelerated the rate regardless of the reaction temperature and time (36.9 vs 12.5). Analysis by column chromatography showed that content of triglyceride in the oil was increased to 72.9% and 61.1% from 10.4% and content of free fatty acids in the oil was decreased to 1.4% and 6.1% from 60.2%, when the degummed oil was esterified at $250^{\circ}C$ for 6 hrs in the presence of and in the absence of the catalyst, respectively. The results estimated from the iodine values indicate that polymer formation was not significant, when the oil was esterified for 6 hrs at temperatures up to $210^{\circ}C$. However, it was somewhat significant for the oil esterified at $250^{\circ}C$ for 6 hrs. The catalyst did not affect the polymer formation. Analysis by high performance liquid chromatography showed that oleic acid (42.5%), linoleic acid (29.0%) and palmitic acid (20.3%) were the major fatty acid components of the rice bran oil.
Both fish community and water quality in Lake Doam were investigated from September 2004 to August 2005. The turbidity of Lake Doam located in the upper region of the Songchun River in the South River system, Korea was high whole year due to the effects of distributed non point source pollutions in the watersheds. During the experimental periods, mean concentration of chlorophyll-a in epilimnetic layer (0 ${\sim}$ 5 m) was 18.5 ${\mu}g\;L^{-1}$ and transparency ranged from 0.3 m to 2.4 m. Average TP and TN concentrations were 111 ${\mu}g\;L^{-1}$ and 4.1 $mg\;L^{-1}$, respectively. Lake was classified as eutrophic state based on the nutrient concentrations suggested by U.S.EPA (1976). Total number of fish collected in Lake Doam was 9,600 individuals in 26 species of 6 family. Both dominant and subdominant species in the lake were P. herzi (34.9%) and Z. platypus (22.5%), respectively. Occurrence of water column species was high at upper region of the lake, whereas benthic type of species highly ,appeared in downstream area. The different fish assemblage between upper and lower area would be considered as the difference of bottom substrate and concentrations of suspended solids. In addition high appearance of Comat type of fish that is hybrid between gold fish (C, auratus) and C. auratus was found in the lake. It was unclear the reasons that high proportion of mutant species appeared in the lake. More researches are required in this area in future.
Kim, Tae-Hoon;Ahn, Tae-Woong;Jung, Jae-Hoon;Choi, I-Song;Oh, Jong-Min
Korean Journal of Ecology and Environment
/
v.43
no.2
/
pp.263-270
/
2010
This is a research on development of water purification equipment called artificial floating island (=AFI) for the stagnant water area which can secure exuberant landscape and water-friendility. The equipment devised in this study is designed to make up the weakness of conventional AFIs and improves the removal efficiency of pollutants using the mixture of media and plants. The air compressor positioned at the inlet releases air with inflow continuously, the water pump at the outlet sprays as a form of fountain with causing a disturbance on stable water column, then, both of them contribute improvement of water quality over a large area. We applied Bio-stone as a media in this system and performed an experiment of pre-efficiency test, and we concluded that the higher pollutants concentration of inflow, the higher removal efficiency we obtained. At the result of lab-scale experiment, in the case of high-concentration inflow, in the removal efficiency of SS is 62.2%, BOD is 50.2%, COD is 55.1%, T-N is 31.6%, T-P is 38.4%. In addition, to evaluate the field application, we set up the facilities in Sin-gal lake located in Yongin-Si Gyeonggi-Do, and researched on the removal efficiency of outflow relative to the inflow. As a result, SS is 53.5%, BOD is 32.8%, COD is 36.9%, T-N is 22.6%, T-N is 33.2%.
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