HPLC-PDA를 이용한 반하, 호장남성, 수반하의 분류 및 함량분석

조지은 ( Ji Eun Jo ),이아영 ( A Yeong Lee ),김효선 ( Hyo Seon Kim ),문병철 ( Byeong Cheol Moon ),최고야 ( Go Ya Choi ),지윤의 ( Yun Ui Ji ),김호경 ( Ho Kyoung Kim ) 대한본초학회 2013 大韓本草學會誌 Vol.28 No.5

Objectives: A quantitative method using high performance liquid chromatography with a photodiode array detector(HPLC-PDA) was established for the quantitative analysis of the four main compound and pattern analysis to classification Piiellia ternate, P. pedatisecta and Typhonium flagelliforme. Methods: The analytical procedure for the determination of P. ternata, together with the known main compounds uracil, uridine, guanosine and adenosine was established. Optimum HPLC-PDA separation of these P. ternata was possible on Luna C18(2) column material, using water and acetonitrile as mobile phase. The method was validated according to regulatory guidelines. In addition, this assay method were analyzed for the content of four main compound in P. ternata, P. pedatisecta and T. flagelliforme and by data obtained from the HPLC-PDA analysis was performed principal component analysis(PCA). Results: Validation results indicated that the HPLC method is well suited for the determination of the roots of P. ternata with a good linearity (r2> 0.999), precision and recovery rates. Analysis of HPLC-PDA, the average content of uracil, uridine, guanosine and adenosine was significantly higher in P. ternate>P. pedatisecta> T. flagelliforme order. The application of PCA to main compound data by HPLC-PDA permitted the effective discrimination among the three species. Conclusions: Analysis of both HPLC-PDA and PCA confirmed the fact that four main compound and pattern profiles of P. ternata, P. pedatisecta and T. flagelliforme were different from each other.

HPLC-PDA와 LC-MS를 이용한 식음료 중 Dimethyl Dicarbonate 분석법 개발 및 유효성 검증

신재욱,김정복 한국식품영양과학회 2019 한국식품영양과학회지 Vol.48 No.9

A sensitive and simple analytical method to identify derivatives of dimethyl dicarbonate (DMDC) residues in fruit beverage and wine was developed and then validated using HPLC-PDA. The calibration curve was obtained from 1 µg/mL to 200 µg/mL with a satisfactory correlation coefficient of 0.999. The limit of detection and limit of quantitation for a derivative of DMDC were 0.30 µg/mL and 0.92 µg/mL, respectively. The recoveries of derivatives of DMDC from spiked samples at levels of 10∼200.0 µg/mL ranged from 92.8 to 101.0% with the relative standard deviation between 0.9 and 1.9%. The measurement of uncertainty of the developed HPLC method for identifying derivatives of DMDC was based on the analytical process and quantification. These results show that the method developed in this study is appropriate for DMDC identification and it can be used to maintain the safety of food products that contain DMDC residues. 본 연구에서는 국내 미설정 보존료인 dimethyl dicarbonate(DMDC)가 보존료로 사용된 식품 유형들의 기준규격 설정과 관리를 위해 국내외 다양한 분석법을 조사하고 비교 검토하였다. 다양한 분석법을 적용하여 DMDC의 분석법을 조사한 결과 GC 장비로는 DMDC의 낮은 휘발성 때문에 분석이 불가능하였고 유도체화 방법을 사용하지 않고는 HPLC 장비로는 분석이 불가능하였다. 본 연구에서 matrix 특히 white wine과 red wine에서 matrix effect로 인해 정량분석이 불가능하였다. 이러한 문제를 해결하기 위해 benzyl amine을 이용한 유도체 방법을 선택하여 matrix effect가 없는 정량분석법을 확립하였으며 안정성 또한 12시간을 확보하였다. DMDC의 직선성은 1~200 μg/mL 범위에서 R2=0.999 이상의 좋은 직선성을 나타내었으며, 정밀도는 9.4% 이하, 정확도 97.4~116.7%로 확인하였다. 일내, 일간 정밀도는 1.2~8.6%, 정확도는 96.3~118.4%로 확인되었다. 액체 시료의 회수율은 92.8~101.0%(상대표준편차 1.9% 이하), 고체의 경우 99.4~111.4%로 확인하였다. 검출한계는 0.30 μg/mL, 정량한계는 0.92 μg/mL로 측정되었다. 측정 불확도는 25.25±0.73 mg/kg(신뢰수준 95%, K=2)으로 확인되었다. 시료는 총 107건을 수집하였으며, 미국 현지에서 구입한 시료는 33건, 서울・경기 대형마트 및 중소형 식품 도매점에서 74건인데 이중 액체 54건, 고체시료 20건을 수집, 분석한 결과 모든 시료에서 불검출을 확인하였다. 본 연구는 향후 국내 미설정 보존료인 DMDC를 사용한 식품에 대해 정량한계(0.92 μg/mL)까지 분석할 수 있는 분석법 확립과 국내에 비의도적으로 혼용될 수 있는 수입식품의 사전사후 관리에 기여할 수 있을 것으로 판단된다.

Simultaneous Determination of the Seven Phenylpropanoids in Xanthii Fructus Using a HPLC-PDA and LC-MS

서창섭,신현규 한국생약학회 2018 Natural Product Sciences Vol.24 No.3

Xanthii Fructus has been traditionally used for the treatment of rhinitis, rheumatoid arthritis, and eczema. In this study, a high-performance liquid chromatography-photodiode array (HPLC-PDA) method was developed and then used for the simultaneous analysis of eight phenylpropanoids in Xanthii Fructus. The analytical column used for this separation was a SunFireTM C18 column, maintained at 40?C. The mobile phase used was 1.0% acetic acid in distilled water and 1.0% acetic acid in acetonitrile with gradient elution. For identify of each component, the mass spectrometer (MS) was used a Waters triple quadrupole mass spectrometer requipped with electrospray ionization (ESI) source. The HPLC-PDA method showed good linearity: correlation coefficients were ? 0.9996. The limits of detection and quantification of the eight compounds were 0.02 - 0.04 and 0.06 - 0.14 mg/mL, respectively. The extraction recoveries ranged from 97.51 to 108.67%. The relative standard deviation values of intra- and inter-day precision were 0.06 - 1.55 and 0.09 - 1.68%, respectively. The validated HPLC-PDA method was applied to simultaneously analyse the amounts of eight phenlypropanoids in Xanthii Fructus.

HPLC-PDA을 이용한 이진탕 중 6종 성분의 동시분석

김성실,김정훈,신현규,서창섭,Kim, Seong-Sil,Kim, Jung-Hoon,Shin, Hyeun-Kyoo,Seo, Chang-Seob 대한한의학방제학회 2013 大韓韓醫學方劑學會誌 Vol.21 No.1

Objectives : Yijin-tang has been used in the treatment of gastrointestinal diseases such as irritable bowel syndrome, gastritis, and gastric ulcer. In this study, a high-performance liquid chromatography (HPLC) method was established for simultaneous analysis of six compounds, liquiritin, glycyrrhizin, hesperidin, 6-gingerol, homogentisic acid, and 3,4-dihydroxybenzaldehyde in Yijin-tang, a traditional Korean herbal medicinal preparation. Methods : A Gemini C18 column was used for the separation of six constituents at $40^{\circ}C$. The mobile phase using gradient elution consist of two solvent systems, 1.0% acetic acid in water (A) and 1.0% acetic acid in acetonitrile (B). The flow-rate was 1.0 mL/min and injection volume was $10{\mu}g$. The detector was a photodiode array (PDA) detector set at 254 nm and 280 nm. Results : The calibration curves of six compounds showed good linearity in various concentration ranges ($R^2{\geq}0.9997$). The limits of detection (LOD) and limits of quantification (LOQ) were 0.028-$0.192{\mu}g/mL$ and 0.093-$0.540{\mu}g/mL$, respectively. The RSD (%) of the intra and inter day validations were 0.03-0.84% and 0.05 -1.00%, respectively. Recovery was 96.14-01.90% and RSD (%) was less than 1.5%. Conclusions : The established simultaneous analysis methods will help management to improve the quality of Yijin-tang.

HPLC-PDA를 이용한 제빵 제품의 글루텐 함량 분석법 검증

나예림,허성원,박성훈 한국산업식품공학회 2023 산업 식품공학 Vol.27 No.3

A rapid analytical method was developed and optimized to determine gluten content in bread. Existing gluten quantification methods were inappropriate for bread with high gluten content because they were optimized to analyze shallow gluten content. To overcome this problem, the first method of quantifying the gluten content in bread was developed by modifying the gluten analysis method in cereal grains. Heat-stable gliadin was selectively quantified for gluten quantification using high-performance liquid chromatography (HPLC), and gliadin peaks were separated using an Agilent SB-C8 column. The specificity, linearity, accuracy, precision, limit of detection (LOD), and limit of quantification (LOQ) were measured for validation. The calibration curve of gliadin had high linearity (R2 = 0.9996), and LOD and LOQ were 0.03 and 0.10 g/100 g, respectively. The relative standard deviation (RSD) values of intra- and inter-day precision were less than 2.49% and 1.54%, respectively. Recovery ranged from 90.73% to 93.87%, with RSD values less than 2.06%. These results indicate that the HPLC method for quantifying gluten in bakery products is efficient, reliable, and reproducible.

HPLC-PDA를 이용한 보중익기탕 중 Liquiritin, Nodakenin, Hesperidin 및 Glycyrrhizin의 동시분석

서창섭,김정훈,신현규 대한약학회 2013 약학회지 Vol.57 No.3

Bojungikgi-tang has been widely used for enhancement of physical fitness in Korea. The convenient, simple, and accurate high-performance liquid chromatography (HPLC) method was established for simultaneous determination of four marker compounds, liquiritin, nodakenin, hesperidin, and glycyrrhizin in Bojungikgi-tang (Buzhongyiqi-tang in Chinese, Hochuekkito in Japanese), a traditional Korean herbal prescription. The column for optimizing HPLC separation was used a Gemini C18 column at column oven temperature of 40 o C with 1.0% (v/v) aqueous acetic acid (A) and 1.0% (v/v) acetic acid in acetonitirle (B) by gradient flow. The flow rate was 1.0 ml/min and the detector was a photodiode array (PDA) set at 254 nm, 280 nm, and 335 nm. Calibration curves of four components were acquired with r 2 values ≥0.9999. The recoveries were found to range 92.11~105.68% with relative standard deviations (RSDs, %) value less than 2.50%. The RSD values of intra- and inter-day precision were 0.07~2.50% and 0.16~1.99%, respectively. The contents of liquiritin, nodakenin, hesperidin and glycyrrhizin in Bojungikgi-tang were 3.85~3.92 mg/g, 2.27~2.32 mg/g, 4.14~4.19 mg/g, and 3.39~3.45 mg/g, respec- tively. The established simultaneous analysis method will be effective for quality control of Bojungikgi-tang.

HPLC-PDA를 이용한 오약순기산 중 6종 성분의 동시분석

서창섭,김정훈,신현규,Seo, Chang-Seob,Kim, Jung-Hoon,Shin, Hyeun-Kyoo 대한한의학방제학회 2012 大韓韓醫學方劑學會誌 Vol.20 No.2

Objectives : Oyaksungi-san(Wuyaoshunqisan) has been used for treatment of stroke and rheumatoid arthritis in Korea. In this study, a simple and accurate high-performance liquid chromatography(HPLC) method was established for simultaneous determination of six main components, liquiritin, ferulic acid, naringin, hesperidin, neohesperidin, and glycyrrhizin in Oyaksungi-san, a traditional Korean herbal prescription. Methods : The analytical column for separation of six constituents was used a Gemini $C_{18}$ column maintained at $40^{\circ}C$. The mobile phase consisted of two solvent systems, 1.0% (v/v) acetic acid in $H_2O$ (A) and 1.0% (v/v) acetic acid in acetonitrile (B) by gradient flow. The flow rate was 1.0 mL/min and the detector was a photodiode array (PDA) set at 254 nm for glycyrrhizin, 280 nm for liquiritin, naringin, hesperidin, and neohesperidin, and 320 nm for ferulic acid. Results : Calibration curves were acquired with $r^2$ values ${\geq}0.9998$. The results of recovery test were 91.58%-105.90% with a relative standard deviations (RSDs, %) value less than 2.0%. The values of RSD for intra- and inter-day precision were 0.03%-1.72% and 0.03%-1.63%, respectively. The contents of the six compounds in Oyaksungi-san were 0.33-9.30 mg/g. Conclusions : The newly established HPLC method will be helpful to improve quality control of Oyaksungi-san.

HPLC-PDA를 이용한 닭고기 중 Nitroxoline 분석법 개발

조윤제 ( Yoon Jae Cho ),채영식 ( Young Sik Chae ),김재은 ( Jae Eun Kim ),김재영 ( Jae Young Kim ),강일현 ( Ri Hyun Kang ),이상목 ( Sang Mok Lee ),도정아 ( Jung Ah Do ),오재호 ( Jae Ho Oh ),장문익 ( Moon Ik Chang ),홍진환 ( Jin Hwa 한국환경농학회 2013 한국환경농학회지 Vol.32 No.1

Research Article : 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 (r2) > 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.

HPLC-PDA에 의한 사군자탕 중 Liquiritin과 Glycyrrhizin의 동시분석

신현규,서창섭,김정훈 한국생약학회 2011 생약학회지 Vol.42 No.3

A high-performance liquid chromatography (HPLC) method was developed for quantitative analysis of liquiritin and glycyrrhizin in Sagunja-tang (SGT, Sijunzi-tang in Chinese), a traditional Korean medicine. HPLC analysis was performed using a Gemini C18 column operating at 40℃, and photodiode array (PDA) detection at 254 nm and 280 nm for quantification of the two components in SGT. The mobile phase using a gradient flow consisted of two solvent systems. Solvent A was 1.0%(v/v) aqueous acetic acid and solvent B was acetonitrile with 1.0% (v/v) acetic acid. Calibration curves were acquired with r^2values > 0.9998, and the relative standard deviations (RSDs, %) for intra- and inter-day precision were not exceed 4.0%. The recovery of each component was in the range of 91.85 - 108.62%, with a RSD less than 4.0%. The contents of the two components in SGT were 7.94 - 13.83 mg/g.

GC/MS and HPLC/PDA characterization of essential oils and phenolic compounds from the aerial parts of common rue (Ruta graveolens)

Chang-Dae Lee,Hak-Dong Lee,Yunji Lee,Hwan Myung Lee,Sanghyun Lee 한국응용생명화학회 2023 Journal of Applied Biological Chemistry (J. Appl. Vol.66 No.-

Two different extraction methods were used to evaluate the medical value of common rue, Ruta graveolens L. (RGL). The results of our 2,2-diphenyl-1-picrylhydrazyl and 2,2'-azino-bis-3- ethylbenzothiazoline-6-sulphonic acid assays indicated that the antioxidant activity of RGL essential oil extract obtained through steam distillation was very low, whereas ethanol (EtOH) extracts of RGL showed higher antioxidant activity. RGL essential oil was extracted by steam distillation and characterized by GC/MS analysis. Furthermore, EtOH extracts of RGL were obtained under reflux and analyzed by HPLC/PDA. The GC/MS results indicated that the ketone compounds 2-undecanol acetate, nonyl cyclopropanecarboxylate, and 2-nonanone accounted for more than 70% of the composition of RGL essential oil. The HPLC/ PDA analyses indicated that the RGL extracts were rich in phenolic compounds such as protocatechuic acid, rutin, psoralen, xanthotoxin, and bergapten, among which rutin was the most abundant. Collectively, our results demonstrated that RGL contains high levels of phenolic compounds and could thus be commercialized as a valuable plant-derived antioxidant.