BACKGROUND: Nitroxoline is an antibiotic agent. It is used for the treatment of the second bacterial infection by the colibacillosis, salmonellosis and viral disease of the poultry. When the nitroxoline is indiscreetly used, the problem about the abuse of the antibiotics can occur. Therefore, this study presented the residue analytical method of nitroxoline in food for the safety management of animal farming products. METHODS AND RESULTS: A simple, sensitive and specific method for nitroxoline in chicken muscle by high performance liquid chromatograph with PDA was developed. Sample extraction with acetonitrile, purification with SPE cartridge (MCX) were applied, then quantitation by HPLC with C18 column under the gradient condition with 0.1 % tetrabutylammonium hydroxide-phosphoric acid and methanol was performed. Standard calibration curve presented linearity with the correlation coefficient ($r^2$) > 0.999, analysed from 0.02 to 0.5 mg/L concentration. Limit of quantitation in chicken muscle showed 0.02 mg/kg, and average recoveries ranged from 72.9 to 88.1 % in chicken muscle. The repeatability of measurements expressed as coefficient of variation (CV %) was less than 12 % in 0.02 and 0.04 mg/kg. CONCLUSION(S): Newly developed method for nitroxoline in chicken muscle was applicable to food inspection with the acceptable level of sensitivity, repeatability and reproducibility.
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.
Kim, Jeong-Il;Lee, Jae-Yong;Kim, Chun-Su;Park, Kwang-E.
Journal of Sericultural and Entomological Science
/
v.25
no.1
/
pp.1-20
/
1983
It has been known that the insect molting hormone and its analogues exist also in plant kingdom and their concentration has been found to be about 0.1~2.0% of dry matter, which is equivalent to $10^3{\sim}10^5$ times of those in insects. This study was carried out; 1) to isolate the phytoecdysones from Korean Achyranthes radix and characterize their physico-chemical properties. 2) to investigate the biological activity of this phytoecdysone on Bombyx mori larvae. The resuls were summarized as follows; 1. The extraction method of phytoecdysones was optimized by three consecutive reflux for 1hr using 200g of dried and milled radix per 1l methanol. 2. The purification from the crude extract was made by a series of steps such as precipitation of gum-type polymer with n-Butyl acetate, adsorption on technical grade silica and chromatography with neutral alumina. The conditions of each step were optimized and the resulting crude crystal was about 500mg per kg dry radix. 3. The crude crystal from the cultivated Achyranthes(Achyranthes japonia) contained ecdysterone (20-hydroxyecdysone) and inokosterone in the proportion of one to one. In order to separate these, a series of processes such as acetylation, separation by alumina column chromatography deacetylation by alcoholysis, deionization and crystallization were introduced and optimized 125mg of ecdysterone and 18mg of inokosterone per kg dry radix were thus obtained. 4. The wild Achyranthes (Achyranthes obtusifolia) radix was found to contain the ecdysterone only. A 285mg of ecdysterone was crystallized per kg dry radix. 5. Isolated ecdysterone, inodosterone and acetylated compounds were characterized by IR., UV., NMR spectroscopy, mp, TLC and densitometry. 6. Ligation experiment was undertaken to confirm the biological activity of the purified ecdysterone; the ecdysterone could induce larval-pupal metamorphosis in the ligated abdomen of 4th instar larvae injecting 0.5~1.0${\mu}g$. 7. By ecdysterone feeding experiment using artificial diet, it was elucidated that the critical time of feeding would be the first half of each instar resulting in increased weight of silk layer. 8. The ecdysterone was fed to 5th instar silkworm at the level of 1, 2, 3, 5ppm of dry feed of artificial diet containing 5% mulberry leaves for 72hrs. At 2ppm of the concentration. body weight and silk layer weight were arrived at maximum. But at higher concentrations body weight and silk layer weight decreased than the control group. At 2ppm of the concentration, body weight was increased by 12.5%. 9. Feeding 2ppm of ecdysterone at the later half of 5th instar, the duration of larvae was shortened.
The consumption of herbal medicines has been increasing with growing interest in health. However, due to recent climate change and the complex distribution process of herbal medicines with high import dependence the likelihood of contamination with mycotoxin has been increased. Mycotoxins are emerging as key indicators for ensuring safety of herbal medicines. A total of 498 herbal medicine samples were screened for mycotoxin contamination in this study. Aflatoxin in the herbal medicine samples was extracted by using immunoaffinity column, then the extracted aflatoxin was quantified via high performance liquid chromatography coupled with fluorescence detection (HPLC-FLD) method. The extraction method was verified by linearity, recovery, LOD and LOQ. Aflatoxins were detected in 39/498 samples in an average of $7.670{\mu}g/kg$ ($0.610-77.452{\mu}g/kg$ range). Although safety standards for Corydalis Tuber is not currently available in korea, five of the 39 samples had high concentration of aflatoxins (average of $14.9{\pm}4.1{\mu}g/kg$). In conclusion, it is urgent to establish safety criteria of aflatoxin in Corydalis Tuber. The results of the current study suggest that continuous monitoring is necessary for proactive management of herbal medicine safety.
Kim, Jae-Young;Lee, Sang-Mok;Chang, Moon-Ik;Cho, Yoon-Jae;Lee, Han-Jin;Chae, Young-Sik;Rhee, Gyu-Sik
Korean Journal of Food Science and Technology
/
v.45
no.6
/
pp.693-699
/
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.
An isocratic high performance liquid chromatography (HPLC) method for routine analysis of deoxynivalenol in noodles was validated and estimated the measurement uncertainty. Noodles (dried noodle and ramyeon) were analyzed by HPLC-ultraviolet detection using immunoaffinity column for clean-up. The limits of detection (LOD) and quantification (LOQ) were 7.5 ${\mu}g$/kg and 18.8 ${\mu}g$/kg, respectively. The calibration curve showed a good linearity, with correlation coefficients $r^2$ of 0.9999 in the concentration range from 20 to 500 ${\mu}g$/kg. Recoveries and Repeatabilities expressed as coefficients of variation (CV) spiked with 200 and 500 ${\mu}g$/kg were $82{\pm}2.7%$ and $87{\pm}1.3%$% in dried noodle, and $97{\pm}1.6%$ and $91{\pm}12.0%$ in ramyeon, respectively. The uncertainty sources in measurement process were identified as sample weight, final volume, and sample concentration in extraction volume as well as components such as standard stock solution, working standard solution, 5 standard solutions, calibration curve, matrix, and instrument. Deoxynivalenol concentration and expanded uncertainty in two matrixes spiked with 200 ${\mu}g$/kg and 500 ${\mu}g$/kg were estimated to be $163.8{\pm}52.1$ and $435.2{\pm}91.6\;{\mu}g$/kg for dried noodle, and $194.3{\pm}33.0$ and $453.2{\pm}91.1\;{\mu}g$/kg for ramyeon using a coverage factor of two which gives a level of statistical confidence with approximately 95%. The most influential component among uncertainty sources was the recovery of matrix, followed by calibration curve.
File fish Navodon modestus was dehydrated in cabinet type hot-sir drier at $48-50^{\circ}C$ for 11 hours and also yellowfin puffer Fugu xanthopterum was dried in open air at $26-28^{\circ}C$ for 30 hours. Nucleotides and their related compounds were collected by extraction with cold perchloric acid and their amounts were determined by anion exchange column chromatography. The contents of ADP, IMP, ATP and hypoxanthine in fresh file fish muscle were 22.9, 12.1, 4.9, and 3.2 ${\mu}mole/g,$ dry wt. respectively. AMP and inosine were 0.9 ${\mu}mole/g,$ dry wt. equally. In fresh yellowfin puffer muscle, the contents of ADP, ATP, AMP, inosine and hypoxanthine were 25.6, 2.4, 1.6, 0.3, 0.6, and 0.4 ${\mu}mole/g,$ dry wt. respectively. In the case of file fish, ADP and ATP tended to degrade rapidly during hot-air dehydration. The contents of IMP were decreased slightly while AMP and inosine were increased. And another case of yellowfin puffer, ADP also tended to degrade rapidly during sun drying while AMP, IMP, inosine and hypoxanthine were increased. Especially, in both case of file fish and yellowfin puffer, inosine was increased twenty five and thirty five times during drying respectively.
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.
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
/
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.
The fucoidan purified from Korean brown seaweed, Ecklonia stolonifera was characterized on molecular structure and blood anticoagulant activities. Extraction was conducted at $100^{\circ}C$ with water and repeated twice. The crude fucodian was 151.1g out of 20.0 kg of Ecklonia stolonifera. The Fucoidan-1, which was purified from crude fucoidan using calcium chloride and cetyl pyridium chloride (CPC), was 35.2% against crude fucoidan. Fucoidan-5 was obtained approximately 28.1% from Fucoidan-1 through DEAE-Toyopearl 650 M ion-exchange column chromatography and showed one band by cellulose acetate electrophoresis. The molecular weight of Fucoidan-5 was estimated to be about 21,000∼23,000 dalton by Sephacryl S-300 gel filtration chromatography. Fucoidan-5 consists of 35.7% of fucose and 4.3% of galactose and the molar ratio of fucose and sulfate was about one to one. IR spectrum of Fucoidan-5 showed absorption at $1240\;cm^{-1}\;and\;850\;cm^{-1}$ and specific rotation value, $[\alpha]$, was $[\alpha]$. These results suggests that the sulfate maybe bind at $C_{4}$ carbon on ${\alpha}-L-fucose$. Gas chromatograph of methyl alditol acetate revealed that Fucoidan-5 is a fucose containing sulfated polysaccharide with $({\alpha}l-2)\;or\;({\alpha}l-2)$ glycosidic linkage. Anti-thrombin activity of the Fucoidan-5 was estimated as 1.4 time stronger than heparin. From above results, the purification methods using CPC and ion exchange chromatography is effective tools for obtaining highly purified fucoidan from Gompi, Ecklonia stolonifera.
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