• 한국수산과학회지
• /
• 제48권3호
• /
• pp.301-307
• /
• 2015

Phlorotannins and marine algal polyphenols, including dieckol, 6,6-bieckol, phloroglucinol, phlorofucofuroeckol-A, and eckol, were isolated from brown seaweeds. These compounds have beneficial bioactivities, and Ecklonia cava has become widely used for the extraction and isolation of phlorotannins. Eckol, in particular, has been to shown to have antioxidant, anti-inflammatory, anticoagulatory, and photoprotective properties. However, due to its low abundance in weaweed, the isolation and purification of eckol are difficult. Its limited availability renders the isolation and purification of eckol labor-intensive processes. Centrifugal partition chromatography (CPC) is an efficient technique for the isolation and purification of eckol. In this study, eckol was isolated from the ethyl acetate fraction of the 70% ethanol extract of E. cava using CPC with a two-phase solvent system of a n-hexane:EtOAc:methanol:water (2:8:3:7, v/v) solution. The purity and anti-inflammatory activity of the isolated eckol were verified by high-performance liquid chromatography and by assaying lipopolysaccharide-induced inflammatory responses in an immortalized murine BV2 microglial cell line, respectively. In conclusion, CPC is a useful technique for simple and efficient isolation of eckol from E. cava.

• 대한수의학회지
• /
• 제37권1호
• /
• pp.195-202
• /
• 1997

A method for the isolation by matrix solid phase dispersion method and liquid chromatographic determination of enrofloxacin and ciprofloxacin in pork muscle tissue is presented. Blank or enrofloxacin and ciprofloxacin spiked samples(0.5g) containing 0.05g oxalic acid were blended with $C_{18}$(octadecylsilyl derivatized silica) packing material. After homogenization, $C_{18}$/muscle tissue matrix was transferred to glass column made from 10ml glass syringe and filter paper, and compressed to 4~4.5ml volume. A column was washed with 8ml of hexane and dried under vacuum. Interfering materials were removed by ethylacetate 8ml and dried, following which enrofloxacin and ciprofloxacin were eluted with 8ml of methanal under gravity. The eluate containing enrofloxacin and ciprofloxacin wase free from interfering compound when analysed by HPLC with UV detection at 278nm. Enrofloxacin and ciprofloxacin showed linear response with UV detector at the range of $0.05{\sim}1.0{\mu}g/ml$ and eluted within 5ml elution volume of methanol from the matrix. Fortified sample containing 0.05g oxalic acid represented more good recoveries than that of control sample. Average percentages of enrofloxacin and ciprofloxacin were $93.30{\pm}4.56%$ and $91.84{\pm}4.17%$, respectively, for the concentration range(0.05, 0.1, 0.25, 0.5 and $0.75{\mu}g/g$). The interassay variability of enrofloxacin was $6.02{\pm}5.33%$ with an intra-assay variability of 4.89% and $6.75{\pm}2.68%$ with 4.54% for ciprofloxacin. Detection limit of enrofloxacin and ciprofloxacin was $0.030{\mu}g/g$ in the spiked sample.

• Bulletin of the Korean Chemical Society
• /
• 제25권10호
• /
• pp.1508-1512
• /
• 2004

A gas chromatography/mass spectrometric (GC/MS) method has been developed for the determination of trace mono-n-butyltin (MBT), di-n-butyltin (DBT), and tri-n-butyltin (TBT) compounds in sediments. Samples were extracted by 10% acetic acid in methanol containing 0.03% tropolone and were then derivatized for GC/MS analysis. Ethylation by sodium tetraethylborate and phenylation by sodium tetraphenylborate were evaluated as a derivatization reaction of the organotins in sample extract. n-Hexane was added into reaction media in the beginning of the reaction for the continuous extraction of derivatized organotins. Ethylation requires less than 2 hours to get proper derivatization yields for MBT, DBT, and TBT altogether and produces relatively low amounts of side reaction products. Compared to ethylation, phenylation requires much longer time but provides relatively lower yield and produces considerable amounts of side reaction products. Therefore, the ethylation reaction was applied for the analysis of organotin compounds in sediment. An isotope dilution mass spectrometric (IDMS) method based on GC/MS has been applied to the accurate determination of DBT compounds in the sediments. The IDMS results from the analyses of sediment samples showed a reasonable repeatability and a good agreement with the values obtained by IDMS based on liquid chromatography/induced coupled plasma/mass spectrometry.

• 한국식품과학회지
• /
• 제37권4호
• /
• pp.537-541
• /
• 2005

곡류, 서류, 두류 및 그 가공품 총 200건을 대상으로 benzo(a)pyrene등 8개 PAHs 화합물에 대하여 분석하였다. 시료 중 PAHs 화합물을 n-hexane으로 추출한 후 Sep-Pak Florisil cartridge로 정제한 후 HPLC-fluorescence로 분석하였다. 분석한 대상식품 중 PAHs 평균 함량은 각각 benzo(a)anthracene $0.13{\mu}g/kg$, chrysene $0.08{\mu}g/kg$, benzo(b)fluoranthene $0.06{\mu}g/kg$, benzo(k)fluoranthene $0.03{\mu}g/kg$, benzo(a)pyrene $0.08{\mu}g/kg$, dibenzo(a,h)anthracene $0.15{\mu}g/kg$, benzo(g,h,i)perylene $0.45{\mu}g/kg$, indeno(1,2,3-c,d)pyrene $0.14{\mu}g/kg$이었다.

• 분석과학
• /
• 제22권1호
• /
• pp.109-117
• /
• 2009

조리 어패류 중 8종의 PAHs 실태파악을 위하여 어류 168건, 패류 40건을 분석하였다. 시료를 알칼리분해하여 n-hexane으로 추출하고 세척한 후 Sep-Pak Florisil Cartridge로 정제하여 HPLC/FLD로 정량 분석하였다. 각각의 PAHs에 대한 회수율은 약 88~112%였다. 조리어류에서 개별 PAH 평균 농도는 benzo(a)anthracene 불검출, chrysene 불검출, benzo(b)fluoranthene $0.0009{\mu}g/kg$, benzo(k)fluoranthene 불검출, benzo(a)pyrene $0.01{\mu}g/kg$, dibenzo(a,h)anthracene 불검출, benzo(g,h,i)perylene 불검출, indeno(1,2,3-c,d) pyrene 불검출이었다. 조리패류에서 개별 PAH 평균 농도는 benzo(a)anthracene $1.84{\mu}g/kg$, chrysene $3.51{\mu}g/kg$, benzo(b)fluoranthene $0.81{\mu}g/kg$, benzo(k)fluoranthene $0.38{\mu}g/kg$, benzo(a)pyrene $0.39{\mu}g/kg$, dibenzo(a,h)anthracene $0.04{\mu}g/kg$, benzo(g,h,i)perylene $0.20{\mu}g/kg$, indeno(1,2,3-c,d)pyrene 불검출이었다.

• 분석과학
• /
• 제26권1호
• /
• pp.99-105
• /
• 2013

염모제품 중 유효성분의 품질검사는 식품의약품안전청의 "의약외품에 관한 기준 및 시험방법"에 따라 제품에 표기된 산화염료들을 박층크로마토그래프법(TLC방법)으로 확인시험을 하도록 되어있다. 그러나 TLC방법은 원료분량이 미량이거나, $R_f$ 값이 비슷한 성분들이 존재하면 확인시험이 어려운 문제점이 있다. 본 연구에서는 미량성분 검출이 용이하며 분석시간이 짧다고 보고된 UPLC를 이용하기 위한 시료 전처리 및 분석조건을 탐색하였다. 유효성분들을 분석할 수 있는 검출한계는 6.7-77.9 ${\mu}g/L$, 정량한계는 22.3-259.7 ${\mu}g/L$ 이었으며, 회수율은 ${\alpha}$-naphthol를 제외하고는 96.2-101.5%로 양호하였다. 유효성분 추출하기 위한 시료 전처리에는 헥산-증류수를 사용하였다. 시료 전처리 시 복잡한 추출과정을 거치지 않고, UPLC방법은 빠르고 정확하게 염모제품에 함유되어 있는 유효성분들을 동시에 분석 할 수 있었다.

• 한국식품위생안전성학회지
• /
• 제23권1호
• /
• pp.1-5
• /
• 2008

미강 추출 상업용 유통 감마오리자놀의 콜레스테롤 자동산화에 의한 C-7 산화 콜레스테롤 유도체 생성 저해 효과가 수용성 모델 시스템을 이용하여 검토되었다. C-7 콜레스테롤 산화 유도체 (C-7 oxidized cholesterol derivatives: C-7 OCDs) 생성을 위해 콜레스테롤과 감마오리자놀이 분산된 수용성 모델시스템은 구리이온을 촉매로 pH 5.5와 $80^{\circ}C$의 가혹 조건에서 20시간 동안 반응되었다. 산화 유도 기간에 따른 C-7콜레스테롤 산화 유도체 (7-ketocholesterol, 7{\alpha}$-hydroxy-cholesterol과 7b-hydroxycholesterol)의 생성 정도와 감마오리자놀 및 콜레스테롤 변화 추이 정도가 핵산과 에틸아세테이트를 이용한 용매 추출법과 고속 액체크로마토그래프 (high-performance liquid chromatography) 테크닉을 이용 정량적으로 분석되었다. 분석 결과 콜레스테롤 산화 유도 기간에 따른 7-ketocholesterol 생성비율은 7-hydroxycholesterol 이성체 (${\alpha}$-형:${\beta}$-형) 대비 약 2:1의 비율로 생성되었으며, 7-hyoxycholesterol 이성체에 있어서는 ${\alpha}$-형 대비 ${\beta}$-형의 생성 정도가 약 1:2의 비율로 나타났고, 총 C-7 산화콜레스테롤의 생성은 상대적인 고농도(300 ppm) 감마오리자놀 처리 모델 시스템에서 효과적으로 저해되었다.

• Journal of Pharmaceutical Investigation
• /
• 제35권4호
• /
• pp.265-271
• /
• 2005

A rapid, selective and sensitive reversed-phase HPLC method for the determination of etodolac in human serum was developed, validated, and applied to the pharmacokinetic study of etodolac. Etodolac and internal standard, ibuprofen were extracted from human serum by liquid-liquid extraction with hexane/isopropanol (95:5, v/v) and analyzed on a Luna C18(2) column with the mobile phase of 1% aqueous acetic acid-acetonitrile (4:6, v/v). Detection wavelength of 227 nm and flow rate of 1.0 mL/min were fixed for the study. The assay robustness for the changes of mobile phase pH, organic solvent content, and flow rate was confirmed by $3^3$ factorial design using a fixed etodolac concentration $(1\;{\mu}g/mL)$ with respect to its peak area and retention time. And also, the ruggedness of this method was investigated at three different laboratories using same quality control (QC) samples. This method showed linear response over the concentration range of $0.05-40\;{\mu}g/mL$ with correlation coefficients greater than 0.999. The lower limit of quantification using 0.5 mL of serum was 0.05 ${\mu}g/mL$, which was sensitive enough for pharmacokinetic studies. The overall accuracy of the quality control samples ranged from 92.00 to 110.00% for etodolac with overall precision (% C.V.) being 1.08-10.11%. The percent recovery for human serum was in the range of 76.73-115.30%. Stability studies showed that etodolac was stable during storage, or during the assay procedure in human serum. The peak area and retention time of etodolac were not significantly affected by the changes of mobile phase pH, organic solvent content, and flow rate under the conditions studied. This method showed good ruggedness (within 15% C.V.) and was successfully used for the analysis of etodolac in human serum samples for the pharmacokinetic studies of orally administered Lodin XL tablet (400 mg as etodolac) at three different laboratories, demonstrating the suitability of the method.

• Journal of Pharmaceutical Investigation
• /
• 제36권1호
• /
• pp.23-29
• /
• 2006

A rapid, selective and sensitive reversed-phase HPLC method for the determination of promethazine in human serum was developed, validated, and applied to the pharmacokinetic study of promethazine. Promethazine and internal standard, chlorpromazine, were extracted from human serum by liquid-liquid extraction with n-hexane containing 0.8% isopropanol and analyzed on a Capcell Pak CN column with the mobile phase of acetonitrile-0.2 M potassium dihydrogen phosphate (42:58, v/v, adjusted to pH 6.0 with 1 M NaOH). Detection wavelength of 251 nm and flow rate of 0.9 mL/min were fixed for the study. The assay robustness for the changes of mobile phase pH, organic solvent content, and flow rate was confirmed by $3^{3}$ factorial design using a fixed promethazine concentration (10 ng/mL) with respect to its peak area and retention time. In addition, the ruggedness of this method was investigated at three different laboratories using same quality control (QC) samples. This method showed linear response over the concentration range of 1-40 ng/mL with correlation coefficients greater than 0.999. The lower limit of quantification using 1 mL of serum was 1 ng/mL, which was sensitive enough for pharmacokinetic studies. The overall accuracy of the quality control samples ranged from 96.15 to 105.40% for promethazine with overall precision (% C.V.) being 6.70-11.22%. The relative mean recovery of promethazine for human serum was 63.54%. Stability (freeze-thaw and short-term) studies showed that promethazine was stable during storage, or during the assay procedure in human serum. However, the storage at $-80^{\circ}C$ for 4 weeks showed that promethazine was not stable. Extracted serum sample and stock solution were not allowed to stand at ambient temperature for 12 hr prior to injection. The peak area and retention time of promethazine were not significantly affected by the changes of mobile phase pH, organic solvent content, and flow rate under the conditions studied. This method showed good ruggedness (within 15% C.V.) and was successfully used for the analysis of promethazine in human serum samples for the pharmacokinetic studies of orally administered Himazin tablet (25 mg as promethazine hydrochloride) at three different laboratories, demonstrating the suitability of the method.

• 한국미생물·생명공학회지
• /
• 제41권3호
• /
• pp.272-277
• /
• 2013

본 연구에서는 gas chromatography/mass spectrometry (GC/MS)와 gas chromatography (GC)를 이용하여 주목 식물세포 Taxus chinensis 유래 타르 성분을 최초로 동정/정량하였다. 또한 식물세포배양으로부터 항암물질 paclitaxel 정제를 위한 전처리 과정에서 이들 타르 성분들의 제거 양상을 확인하였다. GC/MS 분석을 통하여 체류시간을 비교한 결과, 5종류의 타르성분이 체류시간 6.374, 8.208, 15.209, 20.045, 24.474분에서 각각 2-picoline, o-xylene, 2,5-xylenol, 1-methylnaphthalene, acenaphthene이 동정되었다. 또한 표준물질을 이용한 spike testing을 수행한 결과 동일 물질임을 재확인 하였다. GC 분석을 통하여 동정된 5종류 타르성분을 정량한 결과, 메탄올 추출물에 총 0.6805 wt% (2-picoline: 0.2512 wt%, 2,5-xylenol: 0.1586 wt%, acenaphthene: 0.1240 wt%, 1-methylnaphthalene: 0.0942 wt%, o-xylene: 0.0525 wt%) 타르 성분이 존재하였다. 액-액 추출을 수행한 결과, 메탄올 추출물 시료 내 총 타르 성분의 양 대비 42%의 타르 성분이 제거됨을 확인할 수 있었다. 1-Methylnaphthalene, acenaphthene, 2,5-xylenol, 2-picoline의 경우에는 각각 75.90, 59.92, 35.94, 29.74%로 높은 제거율을 보인 반면 oxylene의 경우에는 10.86%의 제거율로 상대적으로 적게 제거됨을 알 수 있었다. 흡착제 처리 후 2-picoline과 o-xylene의 양도 상당히 줄었지만 이들 2 종류의 타르 성분을 제외한 나머지 세 종류의 타르 성분(2,5-xylenol, 1-methylnaphthalene, acenaphthene)은 완전히 제거됨을 확인할 수 있었다. 흡착제 처리 공정에서 제거되지 않은 두 종류의 타르 성분(2-picoline, o-xylene)은 헥산 침전 공정에 의해 완전히 제거 가능하였다.