본 연구는 2017년 수입식품 중 신규 기준설정 예정 농약인 tridemorph의 안전성 관리를 위한 공정시험법을 확 립하기 위하여 수행하였다. Tridemorph의 잔류물의 정의 는 모화합물로 규정하며, 확립된 시험법은 실험실내 및 실 험실간 검증을 통해 공정시험법으로의 유효성을 확인하였다. 대표 농산물 5종(감귤, 감자, 고추, 대두, 현미)에 대하여 잔류분석이 가능하도록 선택성과 감도가 우수한 LCMS/ MS를 사용하여 수용성 유기용매인 ACN로 추출 후 NH2 카트리지에 가장 회수율이 우수한 용매인 MeOH/DCM (1/99, v/v)를 정제조건으로 확립하여 시험법을 개발하였다. 개발된 tridemorph의 직선성은 결정계수(r2)가 0.99 이상으로 우수하였으며, 검출한계 및 정량한계는 각각 0.001 및 0.005 mg/kg으로 높은 감도를 나타내었다. 개발된 시험법의 평균 회수율은 75.9~103.7%였으며, 분석오차는 8.5% 이하로 정확성 및 재현성이 우수함을 확인할 수 있었다. 또한, 외부기관 검증 결과 평균 회수율은 87.0~109.2%이 었으며, 상대표준편차는 모두 7.8% 이하로 조사되어 국제적 잔류농약 분석 가이드라인 및 식품의약품안전평가원 가이드라인에 적합한 수준임을 확인하였다. 따라서 본 시험법은 농산물 중 tridemorph의 잔류검사를 위한 공정시 험법으로 활용할 수 있을 것으로 판단된다.
A reliable and selective liquid chromatography–ultraviolet detection method for determination of antiprotozoals (selamectin, doramectin and fenbendazol) has been described. HPLC separation of active constituents was achieved on various C18 columns using methanol, acetonitrile, 0.1% phosphoric acid, acetic acid and distilled water as mobile phase, with UV detection at 243, 245 and 224 nm. The analytical procedure has been successfully identified. The method was validated for specificity, linearity, accuracy, repeatability and intermediated precision. All calibration curves showed good linearity (R2 of 0.9999) within the concentrations ranges (0~200, 0~200 and 50~400 μg/mL). The accuracy and repeatability showed 99%, 100%, 100% and below 0.4%, 0.5%, 0.6%, respectively. The precision tests conducted for 3 days in three different concentrations with standard also revealed below 3.5%, 2.4% and 2.7%. The method has also been applied successfully to monitor post-market 5 veterinary products of which active ingredient are selamectin, doramectin and fenbendazol. There were no non-compliant products.
에너지 음료는 카페인을 주성분으로 타우린, 비타민 같은 다른 energy-enhancing 성분을 함유하고 있다. 미국과 유럽에서는 글루쿠로노락톤이 에너지 음료에 첨가될수 있으나, 국내에서 의약품으로는 허가되어 있다. 따라서 식품 첨가물로는 그 사용이 금지 되어 있어, 지속적으로 수입 및 유통 음료에서 시험검사를 하여 규제하고 있다. 현재 분석법으로 사용하는 LC-PDA 법은 복잡한 유도체화 과 정을 거치고, 음료 중에 당류들이 위양성 결과를 나타내 기도 한다. 이런 기존 방법의 단점을 개선하기 위해 HILICESI- MS/MS (hydrophilic interaction liquid chromatography coupled to electrospray ionization tandem mass spectrometry) 를 이용한 분석법을 개발하고, 선택성, 직선성, 검출한계, 정량한계, 정밀도, 정확성, 재현성에 대하여 분석법 유효성 검증을 수행했고, AOAC, EURACHEM 가이드라인에 부합되는 결과를 얻었다.
Formulation of the finite element method(FEM) for dynamic interaction between a pantograph and contact wire, including non-linear formulation associated with contact wire stagger in the railway overhead contact line is presented. Penalty method is chosen for modelling contact between a pantograph and contact wire. The formulation is validated by comparing the simulated contact forces with measurements taken from 300 km/h KTX operation.
The aim of this study was to investigate the method validation for the determination of loganin content in Cornus officinalis (CO). This medicinal plant reportedly mainly included loganin. The specificity, linearity, precision, accuracy, limit of detection (LOD), and quantification (LOQ) were measured by HPLC/DAD. Our results showed that high linearity in the coefficient of calibration correlation (R2) for loganin standard was 1. LOD and LOQ for loganin were 0.007 and 0.026 mg/mL, respectively. The recovery rate of loganin was revealed to be in the high range of 95.14-105.45%. The relative standard deviation of intra- and inter-day precision of loganin in CO was 1.8% and 2.3%, respectively. The loganin content of CO from Gurye, Uiseng, Ichoen, and China were 7.81, 3.41, 7.42, and 6.52 mg/g, respectively. In conclusion, these methods were validated for the detection of loganin in CO.
차 중에 있는 살균제 tridemorph의 잔류량을 검사하기 위해 LC-ESI-MS/MS를 이용한 정확하고 감도가 좋은 분석방 법을 개발하였다. Tridemorph 잔류물은 샘플을 수화한 후 acetonitrile로 추출하고, NaCl을 이용한 액-액 분배, NH2 카트리지 정제를 거쳐 기기분석을 수행하였다. 직선성은 0.02~1.0 μgmL−1 범위에서 상관계수(r2) 0.9999로 높은 직선 성을 보였다. 0.02와 0.05 mgkg−1 처리수준으로 회수율을 실험한 결과는 75.0~84.7% 이었으며, 상대표준편차는 10% 미만이었다. 분석방법의 검출한계와 정량한계는 각각 0.01 와 0.02 mgL−1 이었다. 이러한 결과들을 통해 확립된 분석법은 차 중 tridemorph의 잔류량 분석에 적합함을 확인할 수 있었다.
농산물 중에 있는 제초제 saflufenacil의 잔류량을 검사 하기 위해 HPLC-UVD와 LC-MS를 이용한 정확하고 감도가 좋은 분석방법을 개발하였다. Saflufenacil 잔류물은 acetone 추출, dichloromethane을 이용한 액-액 분배, silica 와 carbon 카트리지 정제를 거쳐 기기분석을 수행하였다. 검량선 작성을 위해 0.1~5.0 μgmL−1 범위로 표준품을 만들어 실험한 결과 상관계수(r2)는 0.999로 높은 직선성을 보였다. 0.02~0.5 mg kg−1 처리수준으로 회수율을 실험한 결 과는 80.5~110.2% 이었으며, 상대표준편차는 10% 미만이었다. 분석방법의 검출한계와 정량한계는 각각 0.005와 0.02 mg L−1 이었다. 확립된 시험법으로 본청, 부산지방식품의 약품안전청과 경인지방식품의약품안전청에서 실험실간 검증을 실시한 결과 만족스런 결과를 얻었다. 이러한 결과 들을 통해 확립된 시험법은 농산물 중 saflufenacil의 잔류 량 분석에 적합함을 확인할 수 있었다.
본 연구에서는 식품공전에 제시되어 있는 microwave digestion 전처리방법과 유도결합 플라즈마 방출분광기을 이용하여 식품 중의 9종의 무기질(Na, Ca, K, P, Mg, Fe, Cu, Mn 및 Zn)을 측정하는 분석법에 대한 직선성, 정밀성 및 정확성 등의 분석법 유효성 검증을 실시하였다. 본 연구에 사용된 표준시료는 Certificated reference material1849a 조제분유로 미국 national institute of standards & technology에서 구입하였다. 직선성은 표준품을 사용한 표준검정곡선 측정농도범위에서 상관계수 0.9999 이상의 양호한 결과를 나타내었다. 검출한계는 0.1005 mg/kg, 정량 한계는 0.3351 mg/kg 으로 각각 나타났다. 또한 정밀도는 상대적표준편차(relative standard deviation)가 일내(withinday, n=3), 반복측정의 경우 0.09~4.80%, 일간(between-day, n=12)의 경우 1.19~18.19%로 양호한 결과를 나타내었으며 정확성은 회수율 90.35-110.63%로 매우 양호한 결과를 나타내었다. 따라서 microwave 전처리방법과 유도결합 플라즈마 방출분광기 측정법은 식품 중 9종의 무기질을 측정하는데 매우 유용할 것으로 사료되며 국민 건강 증진을 위한 식품성분표 데이터베이스 구축, 유통식품의 품질평가 등 공익적 분석사업에 이용될 수 있을 것으로 생각된다.
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 μg/kg and 18.8 μg/kg, respectively. The calibration curve showed a good linearity, with correlation coefficients r² of 0.9999 in the concentration range from 20 to 500 μg/kg. Recoveries and Repeatabilities expressed as coefficients of variation (CV) spiked with 200 and 500 μg/kg were 82 ± 2.7% and 87 ± 1.3% in dried noodle, and 97 ± 1.6% and 91 ± 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 μg/kg and 500 μg/kg were estimated to be 163.8 ± 52.1 and 435.2 ± 91.6 μg/kg for dried noodle, and 194.3 ± 33.0 and 453.2 ± 91.1 μg/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.
Background : Korean mountain ginseng adventitious roots culture extract fermentation product (KGEF) is increased the content of low molecular weight ginsenoside Rk1 and Rg5 by purifying, steaming, and fermentation of the wild ginseng adventitious roots culture. In Ministry of Food and Drug Safety, the analysis method of low molecular weight ginsenoside (Rk1, Rg5, Rh2, compound K, etc.) has not been proven, therefore we conducted validation to confirm the suitability of the qualitative and quantitative analysis method for Rk1 and Rg5.
Methods and Results : Quantitative analysis was performed at a maximum absorption wavelength of 203 ㎚ (specificity). It was confirmed that the retention time of each peak of Rk1 and Rg5 was separated by chromatogram. The separation degree of Rk1 and Rg5 was 2.15 more as 1.5 a result of calculation according to the formula to evaluate the separation limit. (accuracy). The recovery rate was 101.5% of Rk1 and 103.9% of Rg5 in KGEF. (repeatability). The area value of ginsenoside Rk1 and Rg5 showed high reproducibility with relative standard deviation Rk1 0.86% and Rg5 0.68%. Retention time was also reproducible with relative standard deviation Rk1 of 0.054% and Rg5 of 0.09%. (linearity). The correlation coefficients were 0.999 of Rk1 and 0.999 of Rg5. The reproducibility of retention time in linearity was also high, with relative standard deviation Rk1 0.0017% and Rg5 0.0017% (limit of quantification, limit of detection). The quantitative limit of Rk1 was 53.73 ㎍/㎖ and the detection limit was 17.73 ㎍/㎖ and the detection limit of Rg5 was 259.03 ㎍/㎖ and detection limit was 85.48 ㎍/㎖.
Conclusion : In this study, we validated ginsenoside Rk1 and Rg5 for identification and content testing. It will be enables to verify physicochemical differentiation and analytical methods, and to be a research-based data of low molecular weight ginsenosides.
Aralia elata Seemann (AE) has long been used as a folk medicine for the treatment of various diseases including diabetes mellitus, anti-arthritic, and anti-gastric ulcer agent in Korea, Japan, and China. This study was performed to establish a simple and reliable HPLC/UV analytical method for determination of most active anti-hypertensive compound, a 3-O-α-L-rhamnopyranosyl(1→2)-α-L-arabinopyranosyl hederagenin 28-O-β-D-xylopyranosyl(1→6)-β-D-glucopyranosylester (HE) for the standardization of the shoot extract of AE as a health functional food ingredient. The quantitative analytical method of HE was optimized by HPLC analysis using reverse-phase C18 column at 40°C with H2O and acetonitrile (70:30, v/v) as an isocratic mobile phase at a flow rate of 1.0 mL/min and detection wavelength of UV 205 nm. This HPLC/UV analytical method showed good specificity and high linearity in the tested range of 0.03125-2.0mg/ml with excellent coefficient of determination (R2) of 0.9999. The limit of detection and limit of quantification were 12.0 μg/mL and 36.5 μg/mL, respectively. Relative standard deviation (RSD) values of data from intra- and inter-day precision were less than 0.2% and 0.1%, respectively. These results indicate that the established HPLC/UV analytical method is very simple, specific, precise, accurate, and reproducible and thus can be useful for the quantitative analysis of HE as a functional anti-hypertensive compound in AE extract.
Background : Acanthopanax sessiliflorus (Rupr. et Maxim) Seem, belonging to the Araliaceae family, is widely distributed in Korea, China, and Japan. The plants belonging to Acanthopanax species are traditionally used in Korea as anti-rheumatoid arthritis, anti-inflammatory and anti-diabetic drugs and are recognized to have ginseng-like activities. A simple and sensitive high-performance liquid chromatographic (HPLC) method was developed and validated for independent analysis of major compounds and chlorogenic acid in A. sessiliflorus fruits. Chlorogenic acid was reported that prevent cancer and cardiovascular disease in vivo. Also, it has antioxidant effect in vitro test. In the previous experiment, chlorogenic acid were found in A. sessiliflorus fruits. This study was performed to identification of the major compounds and investigate the method validation for the determination of chlorogenic acid in A. sessiliflorus fruits. Methods and Results : Three major compounds were recorded on a Varian Unity Inova AS-400 FT-NMR spectrometer and analyzed by the new HPLC analysis method. HPLC analysis was carried out using an Waters e2695 and PDA detector. The new analyasis method was validated by the measurement of intra-day, inter-day precision, accuracy, limit of detection (LOD, S/N=3), and limit of quantification (LOQ, S/N=10) of chlorogenic acid. The results showed that the correlation coefficient (R2) for the calibration curves of chlorogenic acid was 0.997 in terms of linearity. The limit of detection (LOD) and limit of quantification (LOQ) were 0.565 ㎍/ml and 2.88 ㎍/ml, respectively. There was no interfering peak observed each other and HPLC system was suitable for analysis showing goodness of peak and high precision. Conclusion : This method is suitable to detect and quantify major compounds in A. sessiliflorus fruits. Furthermore, the result will be applied to establish chlorogenic acid as an standard compound for A. sessiliflorus fruits.
Background: In the present study, we established an HPLC (high performance liquid chromatography)-analysis method for the determination of marker compounds as a part of the material standardization for the development of health-functional foods from Salvia plebeia R. Br. extract.
Methods and Results: The quantitative determination method of hispidulin as a marker compound was optimized by HPLC analysis using a YMC hydrosphere C18 column with a gradient elution system. This method was validated using specificity, linearity, accuracy, and precision tests. It showed a high linearity in the calibration curve with a coefficient of correlation (r2) of 0.999995. The method was fully validated, and was sensitive, with the limit of detection (LOD) at 0.09 ㎍• ㎖−1 and limit of quantification (LOQ) at 0.27 ㎍• ㎖−1. The relative standard deviation (RSD) values of the data from intra- and inter-day precision were 0.05 - 0.22% and 0.32 - 0.42%, respectively, and the intra- and inter-day accuracy of hispidulin were 99.5 - 102.3% and 98.8 - 101.5%, respectively. The average content of hispidulin in Salvia plebeia R. Br. extract was 3.945 ㎎• g−1 (0.39%).
Conclusions: These results suggest that the developed HPLC method is very efficient, and that it could contribute to the quality control of Salvia plebeia R. Br. extracts as a functional ingredient in health functional foods.
본 연구에서는 곡류 중 트리코테센류 곰팡이독소인 T-2 독소 및 HT-2 독소의 LC-MS/MS 분석방법을 검증하고 국내 유통 곡류 중 T-2 독소 및 HT-2 독소의 오염실태를 파악하였다. 곡류 중의 T-2 독소 및 HT-2 독소를 분석하기 위해, 염화나트륨을 포함한 90% 메탄올 용액으로 추출, 원심분리, 여과, 4% 염화나트륨용액으로 희석하고, 원심분리한 후, 여과한 후 면역친화성칼럼에 의해 정제한 시료를 LC-MS/MS 동시정량 분석하였다. T-2 독소 및 HT-2 독소의 검출한계 및 정량한계는 각각 0.5 μg/kg 및 1.5 μg/kg 얻었다. matrix-matched 표준 검량식에서 상관계수 0.99 이상의 직선성을 얻었으며, T-2독소와 HT-2 독소 2배에서 10배의 정량한계로 표준용액을 첨가한 시료에서 회수율은 T-2독소와 HT-2 독소 각각 100.6±7.2 %, 96.8 ± 9.4 %로 EU 가이드라인에서 제시하는 유효성 기준을 만족하였다. LC-MS/MS 정량법을 이용하여 국내 곡류 9품목 115건에 대해 T-2 독소와 HT-2 독소의 오염도를 조사하여 본 결과, 전체 곡류 115건 중에서 T-2 독소는 83건, HT-2 독소는 93건 검출되었으며 오염도는 T-2 독소는 N.D~37.1 ug/kg, HT-2 독소는 N.D~5.4 ug/kg 으로 낮은 수준이었으며, 오염도는 유럽기준치(100 μg/kg)이내 이었다. 본 연구에서 개발된 곡류 중 T-2 독소 및 HT-2 독소에 대한 분석법은 향후 우리나라 곡류 중 곰팡이독소 안전관리를 위한 시험법으로 활용가능하며, 오염도 자료는 안전성 평가의 기초자료로 활용이 가능할 것으로 사료된다.
Genetically modified (GM) papaya (Carica papaya L.) line 55-1 (55-1), which is resistant to papaya ringspot virus infection, has been marketed internationally. Many countries such as the European Union, Japan, and Korea have a mandatory safety assessment, approval and labeling regulations for GM foods. Thus, there is a need for specific methods for detecting 55-1. In this study, we established a real-time PCR detection method applicable to 55-1 for a variety of papaya products. The limit of detection was possible for fresh papaya fruit up to dilutions of 0.005% and 0.01% (weight per weight [w/w]) for homozygous SunUp and heterozygous Rainbow cultivars, respectively, in non-GM papaya. The 55-1 event-specific detection method observed parallelism (r2>0.99) between the concentration of line 55-1 cultivars and Ct values obtained in amplification plots at concentrations of 0.005-10% for SunUp DNA and 0.01-10% for Rainbow DNA. The method was applicable to the qualitative detection in various types of processed products (cocktail fruit, dried fruit, juice, etc.) containing papaya as a main ingredient. Monitoring papaya products for the presence of GM papaya were demonstrated using a P35S and T-nos real-time PCR detection method but no amplification signals were detected.