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Development and Validation of the Analytical Method for Oxytetracycline in Agricultural Products using QuEChERS and LC-MS/MS (QuEChERS법 및 LC-MS/MS를 이용한 농산물 중 Oxytetracycline의 잔류시험법 개발 및 검증)

  • Cho, Sung Min;Do, Jung-Ah;Lee, Han Sol;Park, Ji-Su;Shin, Hye-Sun;Jang, Dong Eun;Cho, Myong-Shik;Jung, ong-hyun;Lee, Kangbong
    • Journal of Food Hygiene and Safety
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    • v.34 no.3
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    • pp.227-234
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    • 2019
  • An analytical method was developed for the determination of oxytetracycline in agricultural products using the QuEChERS (Quick, Easy, Cheap, Effective, Rugged and Safe) method by liquid chromatography-tandem mass spectrometry (LC-MS/MS). After the samples were extracted with methanol, the extracts were adjusted to pH 4 by formic acid and sodium chloride was added to remove water. Dispersive solid phase extraction (d-SPE) cleanup was carried out using $MgSO_4$ (anhydrous magnesium sulfate), PSA (primary secondary amine), $C_{18}$ (octadecyl) and GCB (graphitized carbon black). The analytes were quantified and confirmed with LC-MS/MS using ESI (electrospray ionization) in positive ion MRM (multiple reaction monitoring) mode. The matrix-matched calibration curves were constructed using six levels ($0.001{\sim}0.25{\mu}g/mL$) and coefficient of determination ($r^2$) was above 0.99. Recovery results at three concentrations (LOQ, $10{\times}LOQ$, and $50{\times}LOQ$, n=5) were from 80.0 to 108.2% with relative standard deviations (RSDs) less than of 11.4%. For inter-laboratory validation, the average recovery was in the range of 83.5~103.2% and the coefficient of variation (CV) was below 14.1%. All results satisfied the criteria ranges requested in the Codex guidelines (CAC/GL 40-1993, 2003) and the Food Safety Evaluation Department guidelines (2016). The proposed analytical method was accurate, effective and sensitive for oxytetracycline determination in agricultural commodities. This study could be useful for safety management of oxytetracycline residues in agricultural products.

Establishment of Analytical Method for Dichlorprop Residues, a Plant Growth Regulator in Agricultural Commodities Using GC/ECD (GC/ECD를 이용한 농산물 중 생장조정제 dichlorprop 잔류 분석법 확립)

  • Lee, Sang-Mok;Kim, Jae-Young;Kim, Tae-Hoon;Lee, Han-Jin;Chang, Moon-Ik;Kim, Hee-Jeong;Cho, Yoon-Jae;Choi, Si-Won;Kim, Myung-Ae;Kim, MeeKyung;Rhee, Gyu-Seek;Lee, Sang-Jae
    • Korean Journal of Environmental Agriculture
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    • v.32 no.3
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    • pp.214-223
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    • 2013
  • BACKGROUND: This study focused on the development of an analytical method about dichlorprop (DCPP; 2-(2,4-dichlorophenoxy)propionic acid) which is a plant growth regulator, a synthetic auxin for agricultural commodities. DCPP prevents falling of fruits during their growth periods. However, the overdose of DCPP caused the unwanted maturing time and reduce the safe storage period. If we take fruits with exceeding maximum residue limits, it could be harmful. Therefore, this study presented the analytical method of DCPP in agricultural commodities for the nation-wide pesticide residues monitoring program of the Ministry of Food and Drug Safety. METHODS AND RESULTS: We adopted the analytical method for DCPP in agricultural commodities by gas chromatograph in cooperated with Electron Capture Detector(ECD). Sample extraction and purification by ion-associated partition method were applied, then quantitation was done by GC/ECD with DB-17, a moderate polarity column under the temperature-rising condition with nitrogen as a carrier gas and split-less mode. Standard calibration curve presented linearity with the correlation coefficient ($r^2$) > 0.9998, analysed from 0.1 to 2.0 mg/L concentration. Limit of quantitation in agricultural commodities represents 0.05 mg/kg, and average recoveries ranged from 78.8 to 102.2%. The repeatability of measurements expressed as coefficient of variation (CV %) was less than 9.5% in 0.05, 0.10, and 0.50 mg/kg. CONCLUSION(S): Our newly improved analytical method for DCPP residues in agricultural commodities was applicable to the nation-wide pesticide residues monitoring program with the acceptable level of sensitivity, repeatability and reproducibility.

Physico-Chemical Properties of Aggregate By-Products as Artificial Soil Materials (골재 부산물의 용토재 활용을 위한 특성 분석)

  • Yang, Su-Chan;Jung, Yeong-Sang;Kim, Dong-Wook;Shim, Gyu-Seop
    • Korean Journal of Soil Science and Fertilizer
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    • v.40 no.5
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    • pp.418-428
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    • 2007
  • Physical and chemical properties of the aggregate by-products including sludge and crushed dust samples collected from the 21 private companies throughout the country were analyzed to evaluate possible usage of the by-products as artificial soil materials for plantation. The pH of the materials ranged from 8.0 to 11.0. The organic matter content was $2.85g\;kg^{-1}$, and the total nitrogen content and available phosphate content were low as 0.7 percents and $12.98mg\;kg^{-1}$, respectively. Exchangeable $Ca^{2+}$, $Mg^{2+}$, $K^+$, and $Na^+$ were 2.29, 0.47, 0.02 and $0.05cmol\;kg^{-1}$, respectively. Heavy metal contents were lower than the limits regulated by environmental law of Korea. Textural analysis showed that most of the materials were silt loam with low water holding capacity ranged from 0.67 to 7.41 percents, and with low hydraulic conductivity ranged from 0.4 to $2.8m\;s^{-1}$. Mineralogical analysis showed that the aggregate by product materials were mostly composed of silicate, alumina and ferric oxides except calcium oxide dominant materials derived from limestones. The primary minerals were quartz, feldspars and dolomites derived from granite and granitic gneiss materials. Some samples derived from limestone material showed calcite and graphite together with the above minerals. According to the result, it can be concluded that the materials could be used as the artificial soil material for plantation after proper improvement of the physico-chemical properties and fertility.

Studies on the Citric Acid Fermentation with Fungi (Part III) Citric Acid Fermentation with Selected Strains (사상균에 의한 구연산발효에 관한 연구 (제III보) 선정균에 의한 구연산발효)

  • 성낙계;김명찬;심기환;정덕화
    • Microbiology and Biotechnology Letters
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    • v.8 no.3
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    • pp.181-191
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    • 1980
  • For the purpose of studies on the citric acid production, some experiments were carried out with isolated strains. The results obtained were as follows. 1) The optimal culture media of the strain M-80 in surface culture contained 140g of sucrose, 3.0g of (N $H_4$)$_2$S $O_4$, 1.5g of K $H_2$P $O_4$, 0.2g of MgS $O_4$.7$H_2O$, 3.0mg of F $e^{++}$, 1.0mg of Z $n^{++}$, 0.5N HCI to a pH of 5.0 and distilled water to 1.0 liter; and that of the strain M-315 in surface culture contained 140g of sucrose, 2.0g of N $H_4$N $O_3$, 1.0g of K $H_2$P $O_4$, 0.25g of MgS $O_4$. 7$H_2O$, 2.0mg of F $e^{++}$, 2.0mg of Z $n^{++}$, 0.05mg of C $u^{++}$, 0.5N HCI to a pH of 4.5 and distilled water to 1.0 liter. While that of the strain M-315 in submerged culture contained 140g of sucrose, 2.5g of N $H_4$N $O_3$, 1.5g of K $H_2$P $O_4$, 0.3g of MgS $O_4$. 7$H_2O$, 3.0mg of F $e^{++}$, 0.1mg of C $u^{++}$, 0.5N HCI to a pH of 4.5 and distilled water to 1.0 liter. The optimal temperature and size of inoculum were mostly 28-3$0^{\circ}C$, 10$^{7}$ -10$^{8}$ spores/50ml, respectively. 2) Through the course of citric acid production, the growth of strains had nearly been completed, pH value was rapidly decreased below 2.0 and the content of sugar was also reduced, while the accumulation of citric acid in media was remarkably begun in about 3-4 days. The yields of citric acid generally reached the maximum level in 8-10 days in surface or submerged fermentation process. 3) Methanol was effective citric acid production when they were added to fermentation media. In the case of surface culture, by addition of 2% (strain M-80), 3% (strain M-315), the yields of citric acid was increased 6.5%, 20.6%, respectively and 5.0% yield was increased by addition of 3% methanol in submerged culture media of the strain M-315. 4) Chromatography analysis of culture broth after fermentation under optimal culture conditions detected that the majority of acid in media was citric acid. 72.1mg/ml, 98.1mg/ml, of citric acid were determined in surface culture media by strains of M-80, M-315, and 59.8 mg/ml of citric acid was contained in the submerged culture media by the strain M-315. strain M-315.

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Studies on the Inheritance of Agronomic Characteristics in Upland Cotton Varieties (Gossypium hirsutum L.) in Korea (육지면품종의 유용형질의 유전에 관한 연구)

  • Bang-Myung Kae
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.21 no.2
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    • pp.281-313
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    • 1976
  • To obtain fundamental informations on cotton breeding efficiences for Korea, individual genetic relationships and interrelationships between the agronomic characteristics of Upland cotton were investigated. These experiments were couducted at the Mokpo Branch Station $(34^{\circ}48'N, $ $126^{\circ}23'E$ and altitude of 10m above sea level) from 1969 through 1972. Heterosis, combining ability, dominance and recessive gene action, genetic variance, and phenotypic and genotypic correlation were investigated by $F_1'S$ from an 11-parent partial diallel cross and the segregating $F_2$ and $F_3$ populations of the cross Paymaster times Heujueusseo Trice. The following points resulted from this study, 1. Heteroses for number of bolls per plant and lint yield were significant at 27, 84% and 37.26%, respectively. No other character had significant heteroses. 2. The GCA estimates for all studied characteristics were higher than the SCA estimates. Varieties with high GCA effects were Suwon 1 for earliness, Paymaster and Arijona for high lint percent, and Arijona for long fiber, etc, 3. SCA estimates for lint yield varied widely in crosses with Mokpo 4, Mokpo 6 and Heujueusseo Trice. Those crosses with the highest SCA effects were combinations with large characteristics differences, Example of these crosses are Mokpo 4 times Acala 1517W, Mokpo 4 times D. P. L. and Heujueusseo Trice aud Paymaster. 4. Early-maturing varieties were completely dominant to late-maturing varieties in some combinations while other crosses gave intermediate phenotypes. These results suggest additive genetic action by multi-genes. Heujueusseo Trice, Mokpo 6, and Suwon 1 showed highest degree of dominance for earliness. 5. There were no significant trends for inheritance of weight of boll and 100 seeds weight. 6. Long staple was partially to completely dominant to short staple. Though there were single gene ratios the rate of dominance decreased in the $F_2$ and $F_3$ populations in the cross between the long staple variety Paymaster and the short staple variety Heujueusseo Trice. Diallel cross $F_1$ hybrids showed complicated allelic gene action for staple length. Various dominance degree were shown by varieties. 7. Number of bolls per plant indicated strong over-dominance and small non-allelic additive gene action. 8. Lint Yield was characterized by over-dominance and by multiple non-allelic-gene action. High-yielding varieties were dominant to low-yielding ones. However, the low-yielding variety Heujueusseo Trice showed over-dominance, indicating different reactions according to the varieties and combinations. 9. Broad sense heritability for days to flowering was 34-39% while narrow sense heritability was 11%. Large variations of individual plants caused by Korean climatic conditions cause this situation. Heritability estimates for weight of boll was 30% for broad sense and 22% for narrow sense. 10. Heritability estimates for staple length and lint percent were very high suggesting strong selection effects. 11. Narrow sense heritability estimates for number of bolls per plant was 30% in the diallel cross $F_1$ hybrids and 36% in the $F_2$ population of the special cross. Broad sense heritability was estimated at 67% suggesting that. 12. Heritability estimates for lint yield was low due to high over-dominance in the diallel cross $F_1$ hybrids. Heritability estimates for yield was low in the $F_1$ hybrids but high in the $F_2$ and $F_3$ populations. 13. Phenotypic and genotypic correlations between lint percent and days to flowering and between staple length and days to flowering were high in the $F_1, $ $F_2$ and $F_3$ populations. Late-maturing varieties and individuals had long staple and high lint percent in general. As the correlation between days to flowering and lint yield was extremely low, the two traits were considered independent of each other. Days to flowering and number of bolls per plant were negatively correlated in the $F_3$ population, indicating early-maturing individual plants with many bolls may be readily selected. 14. Phenotypic and genotypic correlations between lint percent and staple length were high in $F_1, $ $F_2$ and $F_3$ populations. Accordingly, long staple varieties were high in lint percent. It was recognized that lint yield and lint percent were positively correlated in the diallel cross $F_1$ hybrids, and lint percent and staple length were positively correlated in the $F_2$ population, indicating that lint percent and staple length affect lint yield. 15. Lint yield was significantly and positively phenotypically correlated with number of bolls per plant in $F_1, $ $F_2$ and $F_3$ populations. A high genotypic correlation was also noted indicating a close genetic relationship. The selection efficiencies for a high-yielding variety can be increased when individual plants with many bolls are selected in later generations. The selection efficiencies for good fiber quality can be enhanced when individuals with long staple and high lint percent are selected in early generations.

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The Radioprotective Effect and Mechanism of Captopril on Radiation Induced-Heart Damage in Rats (방사선 조사 후 발생한 흰쥐 심장손상에서 Captopril의 방어역할과 기전)

  • Chang Seung-Hee;Lee Kyung-Ja;Koo Heasoo
    • Radiation Oncology Journal
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    • v.22 no.1
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    • pp.40-54
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    • 2004
  • Purpose : Captopril (angiotension converting enzyme inhibitor) is known to have a radioproptective effect in the lungs, intestines and skin, but its effect in the heart is unclear. To investigate the radioprotectlve efiect and mechanism of captopril on the heart, the histopathological changes and immunohistochemical stains were compared with radiation alone, and radiation combined with captopril, in the rats. Materials and Methods : The histopathological changes and immunohistochemical stains ($TNF{\alpha}$, $TGF{\beta}1$, PDGF and FGF2) were examined in the radiation alone and the combined captopril and radiation groups, 2 and 8 weeks after irradiation. Each group consisted of 8 to 10 rats (Sprague-Dawley). Irradiation (12.5 Gy) was given to the left hemithorax in a single fraction. Captopril (50 mg/Kg/d) mixed with water, was given orally and continuously from the first week prior to, up to the 8th week of the experiment. Results : In the radiation alone group, the ventricle at 2 weeks after irradiation showed prominent edema (p=0.082) and fibrin deposit (p=0.018) compared to the control group. At 8 weeks, the edema was decreased and fibrosis increased compared to those at 2 weeks. The histopathological changes of the combined group were similar to those of the control group, due to the reduced radiation toxicity at 2 and 8 weeks. The endocardial fibrin deposit (p=0.047) in the atrium, and the interstitial fibrin deposit (p=0.019) and edema (p=0.042) of the ventricle were reduced significantly in the combined group compared to those in the radiation alone group at 2 weeks. The expressions of $TNF-{\alpha}$, $TGF-{\beta}1$, PDGF and FGF-2 in the radiation alone group were more increased than in the control group, especially in the pericardium and endocardium of the atrium at 2 weeks. At 8 weeks, the pericardial $TNF-{\alpha}$ and $TGF-{\beta}1$ in the radiation alone group continuously increased. The expressions of $TNF-{\alpha}$, $TGF-{\beta}1$ and PDGF were decreased in the combined group at 2 weeks. At 8 weeks, the expressions of $TNF-{\alpha}$ in the atrial and ventricular pericardia were markedly reduced (p=0.049, p=0.009). Conclusion : This study revealed that the early heart damage induced by radiation can be reduced by the addition of captopril in a rat model. The expressions of $TNF-{\alpha}$, $TGF-{\beta}1$ and PDGF were further decreased in the combined compared to the radiation alone group at both 2 and 8 weeks. From these results, it may be concluded that these cytokines probably play roles in the radioprotective mechanism of captopril from the radiation-induced heart toxicity, similarly to in other organs.

Developmental Capacity of Mouse Oocytes within Preantral Follicles Cultured in Medium Supplemented with Gonadotroplhins (성선자극호르몬이 첨가된 배양액에서 체외배양된 생쥐 Preantral Follicles 내 난자의 발생능력)

  • Kim, D.H;Kang, H.G.;Kim, M.K.;Han, S.W.;Chi, H.J.;Lee, H.J.;Lee, H.T.;Chung, K.S.
    • Korean Journal of Animal Reproduction
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    • v.24 no.4
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    • pp.395-406
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    • 2000
  • The present study was conducted to examine the developmental capacity of mouse oocytes within prenatal follicles cultured various concentrations of FSH and LH and the expression of cytochrome P450 cholesterol side-chain cleavage enzyme (P450scc) and cytochrome P450 17 $\alpha$ -hydroxylase (P450)$_{17{\alpha}}$ mRNA, as luteinization and atretic marker, in these culture conditions. In addition, we investigated the concentrations of progesterone and testosterone in culture medium. The developmental potential up to blastocyst of the oocytes grown in vitro was higher in the FSH alone (30.2%) and 10 $m\ell$U/$m\ell$ LH and 100 $m\ell$U/$m\ell$ FSH treated (28.0%) groups than in the 100 $m\ell$U/$m\ell$ LH and 100 $m\ell$U/$m\ell$ FSH treated group (22.0%). And the mean numbers of cell per blastocyst was higher in the FSH alone (50.9$\pm$26.1) and 10 $m\ell$U/$m\ell$ LH and 100 $m\ell$U/$m\ell$ FSH treated (51.0$\pm$21.1) groups when compared to the 100 $m\ell$U/$m\ell$ LH and 100 $m\ell$U/$m\ell$ FSH treated group (45.2$\pm$15.1). The expressions of P450scc and P450$_{17{\alpha}}$ mRNA in the oocyte -cumulus complexes were increased with increasing of LH concentration, and also the secretions of progesterone and testosterone were increased. Especially, in the 100 $m\ell$U/$m\ell$ LH and 100 $m\ell$U/$m\ell$ FSH treated group, the expression of P450scc and P450$_{17{\alpha}}$ were significantly increased, and the secretion of progesterone and testosterone were significantly increased. Therefore, these data show that gonadotrophins are essential for the in vitro culture of preantral follicles, but that increasing of LH concentration is reduced the developmental capacity of oocytes. The cause of these findings may be due to increasing of progesterone and testosterone secretion by the enhance of P450scc and P450$_{17{\alpha}}$ mRNA expressions, as markers of luteinization and atresia. Conclusively, this study suggest that supplementation of 100 $m\ell$U/$m\ell$ FSH or 10 $m\ell$U/$m\ell$ LH and 100 $m\ell$U/$m\ell$ FSH may be optimal condition for the culture of mouse pre antral follicles.

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Metabolizing analysis according to the sawdust media of the known anticancer trees by Pleurotus ostreatuss (느타리버섯의 항암수목자원 배지속 함유성분의 분해능 평가)

  • Shin, Yu-Su;Yang, Bo-Hyun;Kang, Bo-Yeon;Kim, Hyun-Soo;Lee, Ji-Hyun;Hong, Yoon-Pyo;Lee, Sang-Won;Lee, Chan-Jung;Kim, Seung-Yoo
    • Journal of Mushroom
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    • v.9 no.4
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    • pp.186-189
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    • 2011
  • The transitivity of Chemical constituents by Pleurotus ostreatus cultivated in different raw sawdusts, which are Juglans mandchurica, Cudrania tricuspidata and Lindera glauca, was investigated. The HPLC chromatography patterns on the chemical constituents of P. ostreatus showed the similar chromatography patterns in all different raw sawdusts and control sawdust. The unknown chemical constituents of P. ostreatus cultivated in the 10%, 20% mixed medium added 10 %, 20% different raw sawdusts, respectively, were increased. But the significance results in the mixed medium added 50% different raw sawdusts were not showed. The chromatography patterns of mycelia grown in media added the 80% MeOH extracts of three tree species showed the similar patterns in comparison with control mycelia. In the results, the secondary metabolites of functional media were not degrade and changed to other derivatives compounds by P. ostreatus.

Traditional Korean diet can alter the urine organic acid profile, which may reflect the metabolic influence of the diet (한식의 체내 대사에 미치는 영향에 대한 연구: 소변 유기산 분석을 통한 한식의 효과)

  • Shin, Phil-Kyung;Chun, Sukyung;Kim, Myung Sunny;Park, Seon-Joo;Kim, Min Jung;Kwon, Dae Young;Kim, KyongChol;Lee, Hae-Jeung;Choi, Sang-Woon
    • Journal of Nutrition and Health
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    • v.53 no.3
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    • pp.231-243
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    • 2020
  • Purpose: To determine the metabolic influence of the traditional Korean diet (K-diet), which has been regarded as a healthy diet, we investigated the profile of urine organic acids that are intermediates of various types of metabolism including energy metabolism. Methods: Ten women aged 50-60 years were recruited and randomly divided into 2 diet groups, K-diet and control diet, the latter of which is a Westernized Korean diet that is commonly consumed by Koreans nowadays. Before and after the 2-week intervention, 46 urine organic acids were determined using LC/MS/MS, along with clinical parameters. Results: The average concentrations of succinate (4.14 ± 0.84 ㎍/mg creatinine vs. 1.49 ± 0.11, p = 0.0346) and hydroxymethylglutarate (3.67 ± 0.36 ㎍/mg creatinine vs. 2.97 ± 0.29, p = 0.0466), both of which are intermediates of energy metabolism, decreased in the K-diet group after the 2-week intervention, but these were not observed in the control diet group. In particular, the average concentration of succinate in the K-diet group was lower than that in the control group (3.33 ± 0.56 ㎍/mg creatinine vs. 1.49 ± 0.11, p = 0.0284) after 2 weeks. The concentrations of two tryptophan metabolites, 5-hydroxyindolacetate (3.72 ± 0.22 ㎍/mg creatinine vs. 3.14 ± 0.21, p = 0.0183) and indican (76.99 ± 8.35 ㎍/mg creatinine vs. 37.89 ± 10.06, p = 0.0205) also decreased only in the K-diet group. After the 2-week intervention, the concentration of kynurenate, another tryptophan metabolite, was lower in the K-diet group than that in the control diet group (3.96 ± 0.51 ㎍/mg creatinine vs. 2.90 ± 0.22, p = 0.0356). Interestingly, the urine level of kynurenate was positively correlated with BMI (r = 0.61424, p = 0.0003) and total cholesterol (r = 0.46979, p = 0.0088), which decreased only in the K-diet group (239.40 ± 15.14 mg/dL vs. 198.20 ± 13.25, p = 0.0163). Conclusion: The K-diet alters the urinary excretion of organic acids involved in energy metabolism and tryptophan metabolism, suggesting the influence of the K-diet on these types of metabolism. Urine organic acids changed by the K-diet may serve as biomarkers in future studies.

Development of analytical method for determination of spinetoram residues in livestock using LC-MS/MS (LC-MS/MS를 이용한 축산물 중 Spinetoram 공정시험법 개발 및 검증)

  • Ko, Ah-Young;Kim, Heejung;Do, Jung Ah;Jang, Jin;Lee, Eun Hyang;Ju, Yun Ji;Kim, Ji Young;Chang, Moon-Ik;Rhee, Gyu-Seek
    • Analytical Science and Technology
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    • v.29 no.2
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    • pp.94-103
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    • 2016
  • An analytical method was developed to determine the amount of spinetoram (spinetoram J and spinetoram L) in livestock samples. The spinetoram was extracted with acetonitrile and purified through a primary secondary amine (PSA) sorbent. The spinetoram residues were then quantified and confirmed using a liquid chromatography–tandem mass spectrometer (LC-MS/MS) in the positive ion mode using multiple reactions monitoring (MRM). Matrix-matched calibration curves were linear over the calibration ranges (0.005-0.5 mg/kg) into a blank extract with r2 > 0.994. The limits of detection and quantification were 0.002 and 0.01 mg/kg, respectively. The recovery results of spinetram ranged between 81.9-106.4% at different concentration levels (LOQ, 10LOQ, 50LOQ, n=5) with relative standard deviations (RSDs) less than 10%. All values were consistent with the criteria ranges requested in the Codex guidelines (CAC/GL40, 2003). An interlaboratory study was conducted to validate the method. The proposed analytical method proved to be accurate, effective, and sensitive for spinetoram determination. The method will be used as an official analytical method in Korea.