감귤류의 껍질은 플라보노이드의 중요한 소스 중의 하나로서 동아시아에서 내장 및 염증성 질환을 치료하는 민간 의약품으로서 사용되어 왔다. 본 연구에서는 하귤 (C. natsudaidai) 껍질에 포함되어 있는 플라보노이드 성분을 고성능액체크로마토그래피-이중질량분석법 (HPLC-MS/MS)으로 10 개성분을 동정하였다. 플라바논, 플라본 및 쿠마린 유도체는 각각 hesperetin, noviletin 및 coumarin을 사용하여 유효화 하였으며 유효화된 방법으로 정량하였다. 상관계수 (r2)는 > 0.9970으로서 높은 값을 보여주었다. LOD는 >0.01 mg/L이었으며 LOQ는 >0.05 mg/L이었다. 플라보노이드의 총량은 9229.7 ± 0.5 mg/kg 이었다. Naringin의 량이 5010.0 ± 4.5 mg/kg으로 가장 많았으며 sinensetin의 량이 0.6 ± 0.1 mg/kg으로 가장 적었다. 항산화력을 25 μg/mL 에서 500 μg/mL의 농도범위에서 DPPH·, ABTS·+, NO· 소거능 및 FRAP의 항으로 분석하였다. 감귤 플라보노이드의 항산화 능력은 시료의 량이 증가하면 증가함을 알 수 있다.

This study was carried out to optimize the conditions for a chemotaxonomic classification of Ganoderma species. The mycelia of Ganoderma species were extracted with 100% MeOH, and the concentrated extracts were further purified and partitioned with column chromatography (HP20) and n-BuOH, respectively. From the result of high-performance liquid chromatography (HPLC), the constituents of the samples were efficiently separated with a Chemco Pak C18 column (250 mmX4.6 mm) by linear gradient elution using water and acetonitrile as mobile phase components at a flow rate of 0.8 ml/min and detector wavelength at 210 nm. However, the samples without purification or partition were not detected the characteristic peaks. This profile could be used to classify and identify the various Ganoderma species.

본 연구에서는 계면활성제 분석법의 개발을 위하여 High performance liquid chromatography-evaporative light scattering detection(HPLC-ELSD) 기기를 이용하여 음이온 및 비이온 계면활성제 중 다소비 계면활성제인 Alkyl polyglucoside(APG)와 Alpha olefin sulfonate(AOS)를 분석하였다. 실험 결과 APG와 AOS 모두 직선성의 상관관계 계수(R2)가 0.99 이상으로 직선성이 양호하였으며, 검출한계는 APG가 2.29 mg/L, AOS가 16.55 mg/L로 나타났고, 정량한계는 APG가 7.63 mg/L, AOS가 55.16 mg/L로 나타났다. 또한, 회수율 검증을 실시한 결과 APG는 99.29%, AOS는 96.11%로 나타나 회수율 100±10% 이내의 검증기준을 모두 만족하였다. 정밀성 및 정확성을 판단한 결과 APG의 정밀성은 상대표준편차 0.30%, 정확성은 0.26-0.52%로 분석되었으며, AOS의 정밀성은 상대표준편차 0.10%, 정확성은 0.01-0.52%로 분석되어 모두 상대표준편차 1.0% 이하의 검증기준에 적합하였다. 확립된 분석법을 주방세제에 적용하여 분석한 결과 샘플 3종 모두에서 APG와 AOS가 각각 검출되었으며, 표준사용량을 기준으로 APG, AOS의 평균 함유량은 각각의 LD50에 크게 미치지 못하는 것으로 나타났다. 따라서 본 연구를 통하여 확립된 음이온 및 비이온 계면활성제의 분석법은 모든 성분에 대해 감응함으로 한가지 검출기를 이용하여 여러성분을 동시에 측정할 수 있어 식기 등의 잔류물질 혹은 식품 중 혼입되어 있는 다양한 계면활성제 함유량 분석에 적용하여 계면활성제의 관리를 위한 기초자료로 활용될 것으로 본다.

역상 HPLC에 의한 polysorbate 20, polysorbate 40, polysorbate 60, polysorbate 80의 ethylene oxide(EO)분리 및 간편하고 빠른 정량분석 방법을 개발하였다. 분석조건으로 분리관은 YMC Pack Ph (250mm×4.6mm i.d., 5μm) 과 Phenomenex C4 (250mm×4.6mm i.d., 5μm)을 사용하였고, 검출기는 ELSD를 사용하였으며, 이동상은 water/acetonitrile의 기울기 용리에 의해 분석되었다. 이때 검량선의 상관계수(r2)는 180.2~980.5μg/mL 농도 범위에서 0.997이상 이었고, 검출한계, 정밀성이 우수하였다. 이 방법은 olysorbates의 산화에틸렌 분리분석 및 간편하고 빠르게 정량분석 가능함을 보여 주었다.

액체크로마토그래피법(HPLC)을 이용하여 화장품에 들어있는 자외선차단제류 등을 동시 분석하였다. 화장품 시료를 Tetrahydrofurane(THF)에 직접 용해시키고 0.45μm 필터로 여과하여 물/메탄올/THF를 이동상으로 하여 Extend C18의 비극성 컬럼을 사용하여 기울기용리조건에서 20분 안에 분리하여 UV/Vis detector방법으로 정량하였다. HPLC분석결과 검량선은 50~800μg/mL 농도범위에서 상관계수가 r2=0.9992 이상의 좋은 직선성을 나타내었으며 검출한계는 0.01μg/mL였다.

We analyzed the contents of γ-oryzanol, which is contained in brown rice of the nation rice varieties Chucheong, Heukjinju and Sindongjin, by HPLC. Furthermore we also performed experiments on its biological activity, to prove the effectiveness of rice bran. The contents of γ-oryzanol contained in hulled rice showed 1, 587 ppm for Heukjinju, followed by Chucheong(1, 038 ppm), and by Sindongjin(472 ppm). In anti-oxidative activity, we performed an experiment, by measuring the radical scavenging activity of DPPH. Heukjinju showed the best effect, and Chucheong showed the worst effect. In cholesterol lowering activity, Heukjinju showed the best activity and Sindongjin showed the worst effect. In anti-bacterial activity as well, Heukjinju showed the best activity, and Sindongjin showed the worst effect. Through these experiments, we compared the contents of γ-oryzanol, which is contained in hulled rice(Chucheong, Heukjinju, Sindongjin). Also, we found the anti-oxidation effect, cholesterol lowering effect, and anti-bacterial activity of the γ-oryzanol extracts. Based on our research, we expect that γ-oryzanol can work as a new drug, or nutritional supplement.

시중에 유통 중인 영양보충용 제품의 수용성 비타민 B1(thiamin), B2(riboflavin), B3(nicotinic acid and nicotine amide), B6(pyridoxine), C(ascorbic acid)의 신속한 동시분석 방법을 확립하기 위하여 본 연구를 실시하였다. 영양보충용 제품은 정제, 연질캅셀, 분말, 액상의 4가지 제형에 대해 27종의 제품을 구입한 후, Ion-pair 분리기법을 사용하여 HPLC-UVD를 이용한 동시분석 방법을 검토하였다. 비타민 B1, B2, B3, B6, C의 HPLC에 의한 동시분석 조건을 검토한 결과, 이동상은 0.02% triethylamine, 17.5% 메탄올, 5 μM sodium hexanesulfonic acid가 함유된 pH 3.5(acetic acid로 조절)의 수용액을 사용하였고, 용출시간은 다른 피크의 영향을 받지 않도록 30분으로 하였다. 수용성 비타민의 회수율은 96% 이상이었다. 본 연구에 의해 확립된 수용성 비타민의 동시분석 조건은 검량선의 직선성, 정밀성, 정확성, 기기적합성 등이 USP 및 ICH 기준에 적합하여 HPLC의 동시분석 방법으로 합당하였다. 수용성 비타민의 추출 용매는 제형에 따라 약간의 차이를 보이긴 했으나, 물이나 산성조건을 갖춘 HPLC의 이동상이 에탄올이나 메탄올보다 높은 추출 효율을 보였다. 초음파 추출기에 의한 추출 시간은 20분이 가장 적당하였다. 본 연구의 결과는 수용성 비타민의 신속한 추출 및 분석에 매우 효율적으로 이용될 것으로 기대된다.

Deoxynivalenol (DON) and zearalenone (ZEN) are mainly contaminated mycotoxins in feeds. The study was carried out to analyze and survey the contamination of DON and ZEN in one hundred thirteen samples of feeds. After cleaning all samples with immunoaffinity column, the mycotoxins were analysed by using high performance liquid chromatography/fluorescence with diode array detector (HPLC /FLD with DAD). The average recoveries of DON were 88.76 and 95.40% at the levels of 200 and 1,000 μg/kg and 87.09 and 98.40% of ZEN were recovered at the levels of 100 and 500 μg/kg, respectively. The limit of detection (LOD) were 6.0 and 3.0 μg/kg for DON and ZEN, respectively. The average concentrations of DON were 372.1, 324.0 and 990.9 μg/kg in chicken, pig and cattle feed, respectively. Those of ZEN were 76.1, 43.7 and 196.2 μg/kg for them, individually.

An effective method, use of a 2,4-dinitrophenylhydrazine (DNPH) derivatization and high-performance liquid chromatography (HPLC) with ultraviolet (UV) detection, has been used for separation carbonyl compounds. In particular, acetonitrile/water composition as a mobile phase could not separated acrolein/acetone and 2-butanone/ butyraldehyde. In this study, however, that"s compounds separated using acetonitrile/water/tetrahydrofuran as a mobile phase. The resolution was 1.48 and 1.46, respectively. Also, aldehydes and ketones was shown different decrease rate and increase resolution at increase tetrahydrofuran ratio. But butyraldehyde become coeluted methacrolein. Therefore, it is need to consider of mobile phase for separation of butyraldehyde.

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.

A method was established for the simultaneous determinatioin of sugar alcohols, erythritol, xylitol,sorbitol, inositol, mannitol, maltitiol, lactitol and isomalt by High Performance Liquid Chromatography (HPLC). The sugar alcohols were converted into strong ultraviolet (UV)-absorbing derivatives with p-nitrobenzoyl chloride (PNBC). HPLC was performed on Imtakt Unison US-C18 column, using acetonitrile : water (77:23) as a mobile phase and UV detection (260 nm). The calibration curves for all sugar alcohols tested were linear in the 10~200 mg/L range. The average recoveries of the sugar alcohols from three confectioneries spiked at 100 ppm of eight sugar alcohol standards ranged from 81.2 to 123.1% with relative standard deviations ranging fromo 0.2 to 4.9%. The limits of detection (LODs) were 0.5~8 μg/L and the limits of quantification (LOQs) were 2~17 μg/L. Reproducibility of 8 sugar alcohols was 0.28~1.97 %RSD. The results of the analysis of confectioneries showed that 89 samples of 130 were detected and the sugar alcohols content of samples investigated varied between 0.4 and 693.7 g/kg. A method for the simultaneous determination of eight sugar alcohols will be used as basic data for control of sugar alcohols in confectioneries, and qualilty control in food manufacturing.

A method based on high-performance liquid chromatography (HPLC) with ultraviolet detection has been developed to quantify coenzyme Q10 (CoQ10) in raw materials and dietary supplements. Single-laboratory validation has been performed on the method to determine linearity, selectivity, accuracy, limits of quantification (LOQ) and repeatability for CoQ10. An excellent linearity (r = 0.999) was observed for CoQ10 in the concentration range 15.625~500 μg/mL in dietary supplement. Observed recovery of CoQ10 was found to be between 98.33 and 99.38%. LOQ was found to be 250 μg/mL Repeatability precision for CoQ10 was between 0.15 and 0.21% relative standard deviation (RSD). Further, limited studies showed that some adulterants and degraded material could be satisfactorily separated from CoQ10 and identified.

산화방지제인 아스코르빈산(AA), 에리쏘르빈산(isoAA), 아스코르빌파르미테이트(AP), 아스코르빌스테아레이트(AS)에 대한 ‘식품 중 식품첨가물 분석법’을 검증하고 미비점을 개선하였다. 고속액체크로마토그래피 자외선검출기법을 이용하여 검출한계(LOD), 정량한계(LOQ), 상관계수(R2) 등을 측정하였고 돼지기름(lard), 사이다를 이용한 모델식품에서의 회수율과 재현성을 측정하였다. AA와 isoAA의 검출한계는 각각 0.46, 0.48 μg/mL이었으며, 회수율은 각각 86.35-94.78, 84.76-95.02%를 나타내었고 상관계수는 모두 0.999이상을 보였다. AP와 AS의 현 분석법은 메탄올을 용매로 사용하지만 메탄올 용매에서 AP와 AS는 불안정하였다. 냉장온도에서 에탄올을 용매로 사용 시 다른 용매에 비해 유의적으로 높은 안정성을 나타내어 기존 용매의 불안정성을 개선할 수 있었다.

Tomato fruits(Lycoperisicon esculentum) synthesize the glycoalkaloids dehydrotomatine and α-tomatine, possibly as defense against bacteria, fungi and insects. We developed a new effective method to prepare and purify dehydrotomatine and α-tomatine that exists in tomato fruits using alumina column chromatography and high performance liquid chromatography (HPLC). The tomato glycoalkaloids(TGA) in tomato was extracted with 2% acetic acid, and then precipitated with ammonium hydroxide(pH=10.5). The dry precipitate substance was applied on alumina column, and then fractionated with water saturated n-butylalcohol. The TGA(Fr. No. 26~36) were collected and dried under reduced pressure. The TGA was performed on a reverse phase HPLC(Inertsil ODS-2, 5 ㎛), eluted with acetonitrile/20mM KH2PO4(24:76, v/v) at 208 ㎚. Two peaks were detected on HPLC, and individual peak was collected by repeating HPLC. Furthermore, to confirm the identity dehydrotomatine and α-tomatine, each peak isolated was hydrolyzed with 1N HCl into sugar and aglycone tomatidine. The sugars were converted to trimethylsilyl ester derivatives. The nature and molar ratios of sugars were identified by gas-liquid chromatography(GLC) and the aglycone by high-performance liquid chromatography(HPLC). The first peak (Rt=17.5 min) eluted from HPLC was identified as dehydrotomatine, and second peak(Rt=21.0 min) was as α-tomatine. This technique has been used effectively to prepare and isolate dehydrotomatine and α-tomatine from tomato fruits.

A HPLC-MS/MS method was developed for simultaneous determination of six sweeteners (acesulfame-K, cyclamate, saccharin, sucralose, stevioside, aspartame) in children's favorite foods. The procedure involves an extraction of the six sweeteners with 50% methanol solution, sample clean-up using the Carrez clearing reagent and filtering with cartridge filter. The HPLC separation was performed on a Hypersil Gold (150 mm × 2.1 mm 5 um) column using the water/acetonitrile mobile phase (95:5). Mass spectrometric analysis was carried out using the TSQ Quantum Ultra operated in negative and positive ESI/SRM. With this method, good linear relationship, sensitivity and reproducibility were obtained. The spike recoveries of six sweeteners for 2 kinds of foods spiked into 0.4 mg/kg ranged from 87.4 to 114.7%. The detection limits were above 0.02 mg/kg. The method has been applied to determination of six sweeteners in children's favorite foods.

This study was conducted to establish a method to analyze azodicarbonamide (ADA) in wheat flour. A new method using high performance liquid chromatography (HPLC) was developed for the determination of ADA in wheat flour. The recovery rate was 91.93~97.54%. The limit of detection for ADA was 0.02 mg/kg and the limit of quantification was 0.05 mg/kg. The monitoring results for ADA contents using the established methods showed that it was detected as the low value of 0.95 mg/kg in one of 51 flour samples (detection rate : 2%), but not detected in 59 bakery samples. The detected ADA level was suitable to its usage standard, compared to the standard (45 mg/kg). Although the detection rate was very low, the established analytical method of ADA will contribute to the management of ADA in processed foods such as wheat flour and bakery.