본 연구는 동백나무 종자에 함유된 생리활성물질인 camelliaside B의 표준정량화를 위해 고성능 액체 크로마토그래피 (HPLC-DAD) 분석법을 확립하고자 실시하였다. 동백나무 종자에 함유된 주요화합물은 MS/MS 분석, UV 흡광도, 화합물의 유지시간 등을 통해 얻은 결과에 바탕을 두어 camelliaside B를 동백나무 종자 추출물의 지표물질로 선정하였다. Camelliaside B의 HPLC 분석법에 대한 특이성, 정확성, 정밀도 및 정량한계 등을 검증하였다. Camelliaside B 분석을 위해 표준화된 HPLC 분석법은 0.99% 이상의 상관계수(R2)로 높은 선형성을 가지는 것을 보여주었다. Camelliaside B의 회수율은 100.42-108.00%였으며, 검출한계 (LOD)와 정량한계 (LOQ)는 각각 0.084㎍/㎖, 0.254㎍/㎖였다. 동백나 무 종자 지표물질 (camelliaside B)의 일중 정밀도는 각각 0.09-0.16 RSD였으며, 일간정밀도는 0.123 RSD였다. 표준화된 분석법으로 지역별로 수집한 동백나무 종자의 camelliaside B의 함량은 0.279-2.05㎎/g 으로 다양하였다. HPLC 분석법으로 확인한 모든 매개변수는 기능성 원료 인정을 위한 제출자료 작성 기준을 충족하였다. 따라서 본 연구에서 확립한 HPLC 분석법은 동백나무 종자 추출물 유래 건강 관련 기능성 제품 또는 의약품 개발의 첫 단계를 제공할 수 있을 것이다.

마에는 다양한 기능성 성분이 함유되어 있어서 식용 및 약용으로 다양하게 이용되고 있다. 특히 allantoin은 대표적인 마의 2차 대사산물로서 의약품 및 기능성 화장품 제조에도 활용되는 고부가가치 산물이다. 본 연구에서는 아미노 결합 정지상 HPLC 컬럼을 이용하여 마의 속과 껍질의 allantoin 함량을 분석하였다. Allantoin 표준용액의 검량 선의 결정계수(R2)값은 0.9999로 높은 직진성을 보였으며, 검출한계(LOD)와 정량한계(LOQ)는 각각 0.0229 mg/mL 및 0.0694 mg/mL로 설정되었다. 마 속과 껍질의 건조중량 기준 allantoin 함량은 각각 3.09±0.02 mg/g, 3.91±0.11 mg/g으로 속보다 껍질에 더 많은 allantoin이 함유되어있음을 확인하였다. 따라서 이러한 결과를 토대로 향후 농업부산물인 마껍질이 새로운 고부가가치 기능성 소재로 활용될 수 있음을 확인하였다.

석유제품 내의 식별제를 정성⋅정량분석하기 위해 HPLC의 적용가능성을 연구하였다. 등유와 자동차용 경유에 함유된 식별제의 정성분석을 위해 HPLC에서 최적의 분석조건(이동상 용매의 비율, 유속 등)을 선정하고, 이를 바탕으로 식별제의 정량분석을 위한 검량곡선을 작성하였다. 특히, 일정 농도 범위에서의 등유는 4.75분에서, 그리고 자동차용 경유는 4.17분의 머무름시간(retention time)을 나타내었고, 등유와 자동차용 경유의 검량곡선 상관계수(R2)가 0.999 이상을 나타내어 정량분석에 적용 가능할 것으로 나타났다. 현행 식별제의 분석방법인 UV/Vis 분광광도계를 이용한 분석결과와 비교분석을 실시하였고, 등유의 경우 약 7 %의 낮은 편차를 보였으며, 자동차용 경유의 경우 약 20 %의 다소 큰 편차를 확인하였다.

To determine sugars content of agricultural products and foods, simultaneous quantitative analysis was carried out on fructose, glucose, sucrose, maltose, and lactose by HPLC-RI. Analysis conditions were set as column ZORBAX carbohydrate (4.6 mm ID×250 mm, 5 μm), the mobile phase of 75% ACN, the column temperature of 35oC, sample injection amount of 10 μL and the flow rate of 1 mL/min. Five standard solutions were isolated without interfering peak within 30 minutes and the calibration curves of standard were confirmed excellent linearity from 0.10% to 1.00% with R2≥0.999. Based on the chromatogram of the standard solution, the limit of quantification (LOQ) and the limit of detection (LOD) values were calculated. The accuracy of the analytical values were highest at 100% water extraction method to the fructose 95.7%, sucrose 98.7%, lactose 102.7% respectively, compared with reference value of a certified reference material (BCR644), by applying the four solvent extraction methods. Using an in-house quality control material (infant formula), repeatability and reproducibility values of this experiment were verified on the basis of AOAC guideline and reference values were set up at 1.17 g/100g of glucose, 0.85 g/100g of maltose, and 45.54 g/100g of lactose. Quality control charts were drawn up and used for sugars analysis of agricultural products.

HPLC-MS/MS를 이용하여 벌꿀의 동물용의약품 동시분석을 위한 분석법을 정립하고자 식품공전 중 벌꿀에 규격이 설정된 11종의 동물용의약품을 2개 그룹 Group 1 (streptomycine, dihydrostreptomycine, neomycine)과 Group 2 (oxytetracycline, enrofloxacin, ciprofloxacin, cymiazole, chloramphenicol, amitraz, coumaphos, fluvalinate)으로 나눠서 동시에 분석 할 수 있는 방법을 연구 하였다. 두 그룹모두 RT는 15분 이내였고, 검출한계는 0.0056~0.0643 μg/g, 정량한계는 0.0169~0.1948 μg/g으로 나타났으며 Group 1 (0.05~1.0 μg/g의 농도범위)과 Group 2 (0.01~1.0 μg/mL) 의 검량선을 작성한 결과 각 동물용의약품의 직선상관계수(R2)는 0.9917~0.9987, 0.9923~1.000으로 매우 양호한 상태를 보였고, 최종 농도가 0.25 μg/g, 1.0 μg/g에서의 회수율은 Group 1 65.1~80.6%, Group 2 64.2~90.3%를 나타났다, 또한 area 및 RT 에 대한 inter (n = 3), intra day (n = 6) RSD (%) 분석결과는 area는 10.92%이하, RT은 1.57% 이하로 나타나 양호한 수준을 보여 벌꿀 중 동물용의약품동시분석 방법으로 적합하다고 판단된다.

Lycopene, one of the major carotenoids in human diet, has been suggested as a healthful phytochemical compound. Recently, it is important to quantify lycopene content in the lycopene-containing foods. However, it has been difficult to analyze lycopene content in the lycopene-containing foods using a HPLC because lycopene is not fully extracted during sample pretreatment resulting in less reliable quantitative data. The aim of this study was to investigate the effect of solvent on lycopene extraction from lycopene-containing foods for HPLC quantitative analysis. Two different solvents such as 100% acetone and methanol/acetone (1:1) mixture were used to extract lycopenefrom a commercial lycopene-enriched tomato concentrate (containing approximate 72 mg/g lycopene). All extracted samples were sonicated for 10 min, centrifuged, and filtered before injection into HPLC. The analysis of lycopene content was performed by an Ultimate3000 series HPLC system using a Unisol C18 column on UV detector at 450 nm. When the lycopene-enriched tomato concentrate was extracted using 100% acetone and methanol/acetone mixture for HPLC analysis, the lycopene contents were 75.88 and 39.84 (mg/g), respectively. As a result of quantitative analysis, acetone was able to extract lycopene fully from the lycopene-containing foods. The HPLC method established in this study might be useful to quantify lycopene in the lycopene-containing foods which are manufactured in various typessuchasbeverage, powder, tablet, and etc

We investigated the pharmacokinetics of ferulic acid, a potential antioxidant agent, after intravenous (i.v.) bolus administration in rats. To analyze ferulic acid levels in the plasma, bile, urine and tissue samples, we developed an HPLC-based method which was validated for a pharmacokinetic study by suitable criteria. After i.v. bolus administration of ferulic acid, it rapidly disappeared from blood circulation within 15 min. The mean plasma half-lives at α phase (t1/2α) when administered at doses of 2 and 10 mg/ kg were 1.10 and 1.39 min, respectively. The values of t1/2β at the corresponding doses increased 40% (from 5.02 to 7.01 min) with increasing doses. The total body clearance (CLt) values significantly decreased as the ferulic acid dose increased. On the other hand, steady-state volume of distribution (Vdss) values did not show the significant difference with the increase in dose. Of the various tissues, ferulic acid mainly distributed to the liver and kidney after i.v. bolus administration. The ferulic acid concentrations in various tissues at 2 hr after i.v. bolus administration were below 1.0 μg/g tissue. Ferulic acid was excreted in the bile and urine after i.v. bolus administration at the dose of 10 mg/kg. The cumulative amount of ferulic acid in the bile 2 hr after dosage was comparable with the amount excreted in the urine after 72 hr, indicating that i.v. administered ferulic acid was mainly excreted in the both bile and urine. In conclusion, ferulic acid was rapidly cleared from the circulating blood and transferred to tissues such as the liver and kidney after i.v. bolus administration. Moreover, the majority of ferulic acid appears to be excreted in the bile and urine after i.v. bolus administration.

In order to know the characteristic roles of salivary protein complex (SPC) the gel-filtration chromatography was performed using the unstimulated and the stimulated whole saliva separately. The first and second dominant SPC peaks were fractionated and analyzed by immunoprecipitation HPLC (IP-HPLC) using antibodies against the essential salivary proteins including α-amylase, mucin-1, proline rich proteins (PRPs), histatin, cystatin, LL-37, lysozyme, lactoferrin, -defensin-1, -2, -3, IgA, transglutaminase 4 (TGase 4), mucocidin, α1-antitrypsin, cathepsin G. In the gel-filtration chromatography the stimulated whole saliva showed much reduced amount of SPCs than the unstimulated whole saliva, but the proportional patterns of both whole saliva were almost similar each other. Through IP-HPLC analysis both of the first and second dominant SPCs were variably positive for the essential salivary proteins, however, α-amylase, mucin-1, PRPs, lysozyme, and cathepsin G were predominant in the first dominant SPC, while cystatin, lactoferrin, β-defensin-1, -2,-3, IgA, mucocidin, TGase 4, and α1-antitrypsin were predominant in the second dominant SPC. And more, the α1-antitrypsin and cathepsin G which were mostly derived from gingival crevicular fluid were also consistently found in the SPCs. These data may suggest that the first dominant SPC, rich in α-amylase, mucin-1, PRPs, lysozyme, and cathepsin G, may play a role in food digestion, protein degradation, and mucosa lubrication, while the second dominant SPC, rich in cystatin, lactoferrin, β-defensin-1, -2, -3, mucocidin, IgA, TGase 4, and α1-antitrypsin, may play a role in the mucosa protection and antimicrobial defense.

본 연구는 식품산업에서 큰 비중을 차지하고 있는 음료류에 대한 인공감미료 (삭카린나트륨, 아스파탐, 아세설팜 칼륨, 수크랄로스, 싸이클라메이트)의 함유현황 파악을 위한 동시분석법을 확립하였다. 희석 및 여과의 간편하고 효율적인 전처리 후 HPLC/MS/MS를 이용하여 5종 인공감 미료 동시분석 조건을 검토하였다. 컬럼은 C18(2.1 mm × 150 mm, 3.5 um), 이동상은 2% methanol (1 mM ammonium acetate)과 95% methanol (1 mM ammonium acetate)을 사용하여 농도구배 조건으로 각 성분을 분리하였으며 ESI/ SRM 방식으로 정량 분석하였다. 0.1~5.0 mg/L의 농도범 위에서 각 인공감미료 검량선은 1에 가까운 높은 직선성을 나타냈다. 검출한계, 정량한계는 각각 0.001~0.022 mg/ L, 0.004~0.073 mg/L로 저농도의 감미료 분석이 가능함을 확인하였다. 회수율은 92.76~113.50%, 정밀도는 10.91% 이하로 양호하였다. 이상의 결과로 본 분석법은 음료 중 인공감미료 분석에 적합하다고 판단되었다. 확립된 분석법으로 시중 유통 중인 음료 102건 분석한 결과 42건에 서 아스파탐, 아세설팜칼륨, 수크랄로스가 검출되었으며 삭카린나트륨과 싸이클라메이트는 검출되지 않았다. 검출된 인공감미료는 표시사항과 일치하였으며 사용대상과 사용량의 기준에 적합하였다.

하동지역에서 생산되는 잎 녹차(우전, 세작, 중작, 대작)에 함유되어 있는 catechin류, alkaloid류 및 theanine를 HPLC를 이용하여 분석을 함과 동시에 녹차 추출물을 이용하여 총 페놀 물질과 항산화능을 측정하였다. Catechin류와 alkaloid류, theanine, 총 페놀 화합물의 함량은 물 추출물 보다 80% 알코올 추출물에서 더 높았다. 총 catechin과 alkaloid의 함량은 80% 에탄올로 추출한 우전 (172.33 mg/g, 30.80 mg/g)에서 가장 높았다. Theanine의 함량도 80% 에탄올 추출물에서 높았고 55.36에서 37.48 mg/g의 범위였다. 녹차의 총페놀 화합물은 우전에서 가장 높았고, DPPH법, FTC법 및 TBA법을 이용한 항산화 활성 측정에서도 우전에서 높은 결과를 나타내었다.

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.

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 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.

Triacylglycerols of the seeds of Ginkgo biloba have been resolved by high-performace liquid chromatography(HPLC in the silver-ion and reverse-phase modes. The fatty acids were identified by a combination of capillary gas chromatography and gas-chromatography /mass spectrometry as the methyl and /or picolinyl ester. The main components are C18:2Ω6(39.0mol%), C18:1Ω7(asclepic acid 21.5mol%), and C18:1Ω9(oleic acid, 13.8mol%). Considerable amounts of unusual acid such as C20:3δ5,11,14 (5.7mol%), C18:2δ5,9(2.8mol%), and C18:3δ5,9,12(1.6mol%), were checked. In addition, an anteiso-branched fatty acid, 14-methylhexadecanoic acid, was also present as a minor component(0.9 mol%). The triacylglycerols were separated into 17 fractions by reverse-phase HPLC, and the fractionation was achieved according to the partition numnber(PN) in which a δ5-non methylene interrupted double bond(5-NMDB) showed different behaviour from a methylene interrupted double bond in a molecule with a given cahinlength. Silver-ion HPLC exhibited excellent resolution in which fractions(23 fractions) were resolved on the basis of the number and configuration of double bonds. In this instance, the strength of interaction of a δ5-NMDB system with silver ions seemed to be weaker than a methylene interrupted double bond system. The principal triacylglycerol species are as follows ; (C18:2Ω6)2/C18:1Ω7, C18:1Ω9/C18:1Ω7/C18:2Ω6, (C18:1Ω7)2/C18:2Ω6, C16:1Ω7/C18:1Ω9/C20:3δ5,11,14, C16:1Ω7/C18:1Ω7/C20:3δ5,11,14, C18:1Ω9/C18:1Ω7/C18:2Ω6, C18:1Ω9/C18:2δ5,5/C20:3δ5,11,14, (C18:1Ω7)2/C18:2Ω6 and (C18:1Ω9)2/C18:2Ω6, while simple triacylglycerols without C18:2Ω6)3 were not present. Stereospecific analysis showed that fatty acids with δ5-NMDB system and saturated chains were predominantly located at the site of sn-3 carbon of glycerol backbones. It is evident that there is asymmetry in the distribution of fatty acids in the TG molecules of Ginkgo nut oils.

Background : Alpinia Officinarum Rhizome (高良薑) is recorded as a roots of Alpinia officinarum Hance (Zingiberaceae) in Korea Pharmacopoeia and there is no established confirmation test and method for quantitative determination using marker compound. Using the six marker compounds, pinocembrin, (5S)-5-hydroxy-7- (4-hydroxy-3-methoxyphenyl) -1-phenyl-3-heptanone, pinocembrin, galangin, kaemferide, galangin 3-methyl ether, A simultaneous analysis method was developed to utilize the quality control. Methods and Results : In this study, YMC-pack column (ODS C18, 250 × 4.6 ㎜, 5 ㎛) from YMC was used and the temperature was set at 30℃. A soultion of 0.1% formic acid was using water (mobile phase A) and acetonitrile (mobile B), respectively, and the mobile phase B was set to 15 - 35% for 0 - 15 minutes and 35 - 40% for 15 - 60 minutes. And 270 ㎚ was set as the detection wavelength. The order of the components is pinocembrin (1, Rt: 23.2 min), (5S)-5-hydroxy-7-(4-hydroxy-3-methoxyphenyl)-1-phenyl-3-heptanone (2, Rt: 37.8 min), pinocembrin (3, Rt: 44.9 min), galangin (4, Rt: 46.7 min), kaemferide (5, Rt: 48.6 min), galangin 3-methyl ether (6, Rt: 52.8 min) and Based on the retention time, ultraviolet spectrum and MS value, it was confirmed by comparison with standard compounds. Conclusion : In this study, we developed the simultaneous analysis method by six marker components of the Alpina officinarum Hance extract to develop a quality test method to scientifically identify Alpinia officinarum Hance.