• Natural Product Sciences
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• 제24권2호
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• pp.109-114
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• 2018

Artemisia capillaris Thunb. (Compositae) is a native herb of East Asian countries and has used for the treatment of jaundice, high liver fever, and digestive diseases for a long time, as well as being developed as the source of herbal preparations until now. The major components from A. capillaris were chlorogenic acid (1) and its derivatives substituted with caffeoyl moieties, such as 3,5-dicaffeoylquinic acid (2) and 4,5-dicaffeoylquinic acid (3), and coumarins, such as scoparone. In the study, four compounds, chlorogenic acid, 3,5-dicaffeoylquinic acid, 4,5-dicaffeoylquinic acid and scoparone (4) in the 70% ethanolic extract of A. capillaris were simultaneously determined by using HPLC-UVD system. This method was validated with the terms of linearity, precious and accuracy according to ICH guidelines. The developed method was successfully applied for the quantitative analysis of Artemisia genus, A. capillaris, A. iwayomogi, A. princeps, and A. argyi, distributed in Korea.

• 분석과학
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• 제34권4호
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• pp.153-159
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• 2021

In this study, we describe the development of a new high-performance liquid chromatography (HPLC) method for the simultaneous analysis of six bioactive compounds (including gallic acid, epicatechin 3-gallate, quercitrin, afzelin, quercetin, and kaempferol) from Orostachys japonicus. The extraction method was investigated and optimization of the extraction time (min), solvent composition (%), and solvent to material ratio were conducted. As a result, 30 min extraction with 50% methanol and 40:1 mL/g of solvent: material ratio achieved the highest extraction efficiency with a yield of 3.32 mg/g. Furthermore, the developed HPLC method was validated and the correlation coefficient (R) values were within the satisfactory range of 0.9995-0.9999 over the linearity range of 1.53-417 ㎍/mL. The limit of detection and limit of quantification for the six active components were between 0.03-0.08 ㎍/mL and 0.08-0.26 ㎍/mL, respectively. With these newly optimized and developed methods, four batches of O. japonicus were analyzed to confirm the high extraction efficiency of the method and the feasibility of an application.

• 대한수의학회지
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• 제42권2호
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• pp.171-181
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• 2002

Simultaneous multiresidue analysis using liquid chromatography determination for five benzimidazole anthelmintics(thiabendazole, oxibendazole, albendazole, mebendazole and fenbendazole) in bovine muscle, liver and omasum has been described. Blank or benzimidazole-fortified samples(0.5g) were blended with bulk $C_{18}$($40{\mu}m$, 18% load, endcapped, 2g). A column made from the resultant $C_{18}$/animal tissue matrix was first washed with hexane($8m{\ell}$), following which the benzimidazoles were eluted with acetonitrile($8m{\ell}$). Analytes of extracted sample were determined by liquid chromatography with UV detector at 290nm. Correlation coefficients of standard curves for individual benzimidazole isolated from fortified samples, using internal standardization, were linear($0.991{\pm}0.007$ to $0.996{\pm}0.005$) with average relative percentage recoveries from $62.1{\pm}3.8(%)$ to $92.3{\pm}7.5(%)$ for the concentration range($0.2{\sim}6.4{\mu}g/g$), respectively. Recoveries rates of TBZ, MBZ in liver, OBZ, MBZ in muscle and TBZ, MBZ in omasium from fortified benzimidazole were 92.%, 87.3%, 74.5%, 82.7%, 75.2% and 83.5% at condition II, respectively. Condition II showed higher recoveries rates than condition I. These results indicated that the matrix solid phase dispersion(MSPD) methodology is acceptable for the determination of 5 benzimidazole anthelmintics and may also suitable for other matrixes of food animal origin.

• 대한수의학회지
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• 제40권4호
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• pp.741-746
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• 2000

최근 새로 개발중인 항AIDS치료제 KR-V 시리즈를 대상으로 HPLC-UV 검출법을 이용해 랫드 혈장중에서 분석법을 검토하였다. 분석컬럼은 $C_{18}$($5{\mu}m$, $250{\times}2.0mm$ I.D.)을 이용하였으며 이동상은 물과 ACN의 혼합액(40/60, v/v)으로 하였다. 이상의 조건에서 모든 KR-V 물질들은 용출시간 4-12분에 비교적 신속하게 잘 분리되었으며 정량한계는 15-30 ng/ml, 혈장중 회수율은 KR-V 2, 7 및 15를 제외하고는 85%(C.V. <10%) 이상으로 나타내었다. Ester 구조를 포함하고 있는 KR-V 2, 7 및 15는 혈장중에서 극히 불안정하여 유도체 개발의 분자설계상 제외시키는 것이 좋을 것으로 사료되었다. 결론적으로 본 HPLC-UV 검출법온 KR-V 시리즈 물질들의 약물동태연구를 위한 약물 분석법의 유효한 방법으로 검토되었다.

• 한국환경보건학회지
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• 제22권1호
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• pp.36-44
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• 1996

A comparison was made of two detection methods(UV absorbence detection and fluorescence detection with pre-column derivatization, with trifluoroacetic acid) coupled with HPLC for the simultaneous determination of aflatoxin $B_1, B_2, G_1$ and $G_2$. A good separation of the four aflatoxins was achieved on a reversed-phase $C_{18}$ column (30 cm x 3.9 mm) with methanol-acetonitrile-water(20+20+60) for absorbence detection or acetonitrile-water(25+75) for fluorescence detection at the flow rate of 1.0 ml/min. The calibration graphs were linear over the ranges 100 ppb-1 ppm for $B_1/G_1$ and 30~300 ppb for $B_2/G_1$ with absorbence detection, and 1~500 ppb for $B_1/G_1$ and 0.3~150 ppb for $B_2/G_2$ with fluorescence detection. The correlation coefficients were greater than 0.94 and 0.99 for absorbance detection and for fluorescence detection, respectively. The detection limit was 100 ng for $B_1/G_1$ and 30 ng for $B_2/G_2$ with absorbence detection, and 1 ng for $B_1/G_1$ and 0.3 ng for $B_2/G_2$ with fluorescence detection. Recovery rates of aflatoxin $B_1, B_2, G_1$ and $G_2$ added to yeast-extract sucrose broth medium were 66.6%, 59.4%, 67.5% and 59.2%, respectively, for absorbence detection and 82.9%, 71.5%, 80.0% and 69.3%, respectively, for fluorescence detection. The four aflatoxins in culture medium were quantitatively detected by the two methods. The aflatoxins in the rice sample were not detected the absorbence detection method, but were below 10 ppb using the fluorescence detection method. Analysis of aflatoxins by both the absorbence and fluorescence methods coupled with HPLC showed acceptable linearity and good recovery. The absorbence detection was less timeconsuming and safer for treatment. The fluorescence detection was more elective and sensitive though elevated $B_1$ and $G_1$ contents were determined from the TFA-induced conversion of $B_1$ to $B_{2a}$ and $G_1$ to $G_{2a}$.

• 생약학회지
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• 제48권3호
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• pp.232-236
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• 2017

Gastrodia elata has been widely used as a traditional medicinal herb. However, in Korean Pharmacopeia, neither a marker constituent nor an analytical procedure for G. elata has been established. In this study, we suggest gastrodin and gastrodigenin as marker constituents of G. elata, and propose an analytical procedure for simultaneous quantification of these constituents by high-performance liquid chromatography-ultraviolet spectroscopy (HPLC-UV). The analytical method was validated for its linearity, precision, accuracy, and specificity. Based on the validated method, gastrodin and gastrodigenin in 14 commercial G. elata samples were quantified.

• Mycobiology
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• 제41권4호
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• pp.234-242
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• 2013

A total of 62 bacterial isolates were obtained from Gomsohang mud flat, Mohang mud flat, and Jeju Island, Republic of Korea. Among them, the isolate CNU114001 showed significant antagonistic activity against pathogenic fungi by dual culture method. The isolate CNU114001 was identified as Bacillus amyloliquefaciens by morphological observation and molecular data analysis, including 16SrDNA and gyraseA (gyrA) gene sequences. Antifungal substances of the isolate were extracted and purified by silica gel column chromatography, thin layer chromatography, and high performance liquid chromatography. The heat and UV ray stable compound was identified as iturin, a lipopeptide (LP). The isolate CNU114001 showed broad spectrum activity against 12 phytopathogenic fungi by dual culture method. The semi purified compound significantly inhibits the mycelial growth of pathogenic fungi (Alternaria panax, Botrytis cinera, Colletotrichum orbiculare, Penicillium digitatum, Pyricularia grisea and Sclerotinia sclerotiorum) at 200 ppm concentration. Spore germ tube elongation of Botrytis cinerea was inhibited by culture filtrate of the isolate. Crude antifungal substance showed antagonistic activity against cucumber scleotiorum rot in laboratory, and showed antagonistic activity against tomato gray mold, cucumber, and pumpkin powdery mildew in greenhouse condition.

• 한국식품영양과학회지
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• 제45권11호
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• pp.1610-1616
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• 2016

섬쑥부쟁이로부터 고요산혈증 개선에 도움을 주는 기능성 식품 개발을 위하여 최적의 에탄올 추출물 탐색과 high performance liquid chromatography-ultraviolet(HPLC-UV) 분석방법에 의한 validation을 실시하였다. 지표성분으로 3,5-dicaffeoylquinic acid(3,5-DCQA)를 선정하여 표준화를 실시하였으며 검출법 확립을 위한 3,5-DCQA 정량분석은 Luna RP-18 칼럼($4.6{\times}250mm$, $5{\mu}m$)을 이용하여 1% 초산용액과 메탄올을 전개용매로 사용하였다. 용출은 1.0 mL/min의 유속으로 기울기 용출(gradient elution) 방법을 이용하였으며, 320 nm 파장에서 검출한 피크 면적을 이용하여 검량곡선을 작성하여 분석하였다. 본 연구에서 확립한 분석법으로 특이성, 직선성, 정밀성, 정확성, 회수율을 검색하였다. 3,5-DCQA의 검량선으로부터 상관계수($R^2$) 0.9999의 우수한 직선성과 intra-day와 inter-day 분석에서 90% 이상의 회수율과 5% 미만의 RSD를 나타내 정밀성과 정확성을 입증하였다. 검출한계는 $2.68{\mu}g/mL$였고 정량한계는 $8.11{\mu}g/mL$로 나타났다. 섬쑥부쟁이 에탄올 추출물(AGE)은 70과 $80^{\circ}C$에서 30, 50, 70, 80% 에탄올로 3, 4, 5, 6시간 동안 각각 추출하였으며, 지표물질의 검량곡선을 활용하여 각각의 AGE로부터 3,5-DCQA의 함량을 분석하였다. 본 시험법으로 분석한 3,5-DCQA의 함량은 $70^{\circ}C$에서 추출한 70% AGE가 $52.59{\pm}3.45mg/Aster$ glehni 100g의 함량을 나타내 가장 우수하게 나타났다. 그러나 섬쑥부쟁이 추출물에 함유된 5-caffeoylquinic acid(5-CQA)의 함량 비교분석은 에탄올 함량이나 추출 시간에 따른 함량 변화가 미미하게 나타났다. 또한, 다양한 AGE에 대하여 XOD 저해 효능을 검색하였을 때, 3,5-DCQA의 함량이 가장 높은 $70^{\circ}C$에서 추출한 70% AGE에서 우수한 효능을 나타내 기능성 원료 표준화를 위한 적합한 분석법임이 검증되었다. 따라서 본 연구를 통하여 확립된 3,5-DCQA의 분석법은 섬쑥부쟁이 에탄올 추출물로부터 개별인정형 건강기능식품 기능성 원료 개발을 위한 유용한 자료로 활용될 것으로 생각한다.

• Safety and Health at Work
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• 제8권2호
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• pp.220-225
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• 2017

Background: Benzene is a known occupational and environmental pollutant. Its urinary metabolite trans, trans-muconic acid (tt-MA) has been introduced by some environmental and occupational health regulatory associations as a biological index for the assessment of benzene exposure; however, recently, doubts have been raised about the specificity of tt-MA for low-level benzene exposures. In the present study, we investigated the association between urinary levels of tt-MA and inhalational exposure to benzene in different exposure groups. Methods: Benzene exposure was assessed by personal air sampling. Collected benzene on charcoal tube was extracted by carbon disulfide and determined by a gas chromatograph (gas chromatography with a flame ionization detector). Urinary tt-MA was extracted by a strong anion-exchange column and determined with high-performance liquid chromatography-UV. Results: Urinary levels of tt-MA in intensive benzene exposure groups (chemical workers and police officers) were significantly higher than other groups (urban and rural residents), but its levels in the last two groups with significant different exposure levels (mean = 0.081 ppm and 0.019 ppm, respectively) showed no significant difference (mean = $388{\mu}g/g$ creatinine and $282{\mu}g/g$, respectively; p < 0.05). Before work shift, urine samples of workers and police officers showed a high amount of tt-MA and its levels in rural residents' samples were not zero. Conclusion: Our results suggest that tt-MA may not be a reliable biomarker for monitoring low-level (below 0.5 ppm) benzene exposures.

• 대한수의학회지
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• 제33권4호
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• pp.605-609
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• 1993

항암치료의 조절인자로서 중요시되는 활성엽산대사물의 장간(腸肝)순환동태를 규명하기 위해 랫드에 $[^3H]pteroylglutamic$ acid(PteGlu)를 정맥주사한 후 고성능 액체 크로마토그래피법(HPLC)을 이용해 담즙중 활성대사물에 관해 조사하였다. HPLC-liquid scintillation counting법의 분석결과, 현저한 다섯개의 방사활성 피크가 출현, 이들은 $H_4PteGlu$, $10-HCO-H_4PteGlu$, $5-CH_3-H_4PteGlu$와 paraaminobenzoyl glutamate(pABG), PteGlu로 각각 동정되었다. 전자 3물질의 활성엽산대사물의 동정은 표준물질과의 비교분석에 의해 HPLC-전기화학검출기를 이용한 retention time profile 및 hydrodynamic, Voltammogram의 일치성 그리고 HPLC-photodiode array분석에 의한 흡광스펙트럼의 일치성에 근거하였다. 후자 그 물질의(pABG 및 PteGlu) 동정은 흡광스펙트럼의 일치성에 근거하였다. 랫드 담즙중$[^3H]PteGlu$의 활성엽산대사물로서 $H_4PteGlu$의 존재가 처음으로 밝혀졌다.