• 대한예방한의학회지
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• 제15권1호
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• pp.37-47
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• 2011

Objectives : Previous studies have shown that Fructus mume (FM) has anti-platelet effects. The present study was performed to determine the acute oral toxicity and quality control of a crude extract of FM in ICR mice. Methods : We investigated the in vivo single dose acute toxicity of FM 95% ethanol extract. This test was orally administered once by gavage to 20 male and 20 female mice at dose levels of 0 (control group), 1250, 2500 and 5000mg/kg body weight, respectively. Mortalities, clinical findings, autopsy findings and body weight changes were monitored daily for the 14 days following the administration. HPLC analysis was performed for the simultaneous determination of ursolic acid and p-hydroxycinnamic acid in FM. Reverse-phase chromatography using a C18 column and photodiode array detection at 211 nm was used for quantification of the two maker components. The mobile phase for gradient elution consists of water and acetonitrile. Results : We observed survival rates, general toxicity, change of body weight, and autopsy. The mice did not die after single oral administration of maximum dose of FM. Autopsy of animal revealed no abnormal gross finding. Therefore, $LD_{50}$ value of FM for ICR mice was more than 5000mg/kg on oral route. The HPLC analysis showed that ursolic acid and p-hydroxycinnamic acid amounts to 9.75- and 0.12% in the extract with the retention times of 47.99- and 15.38 minutes, respectively. Conclusions : These results suggest that no toxic dose level of FM in mice is considered to be more than 5000mg/kg. Consequently, it was concluded that FM have no effect on acute toxicity and side effect in ICR mice. For the quality control of FM extract, simultaneous determination of ursolic acid and p-hydroxycinnamic acid was established.

• Mass Spectrometry Letters
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• 제8권2호
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• pp.39-43
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• 2017

Meldonium is a drug for treating ischemia by expanding the arteries but it can also enhance the performance of sports players. The World Anti-Doping Agency (WADA) has included it in the list of prohibited substances since 2016. Meldonium is one of the challenging substances for anti-doping testing because it is difficult to recover by general liquid-liquid or solid phase extraction due to its permanent charge and high polarity. Therefore, high-performance liquid chromatography (HPLC) is currently used by injecting a diluted urine sample (known as the "dilute-and-shoot" strategy). There is no loss of target compounds in the extraction/cleanup procedure but its high matrix effect could interfere in their separation or detection from the endogenous urinary compounds. We report a single method using high-resolution mass spectrometry that can be used for both screening and confirmation, which follows the "dilute-and-shoot" strategy. In this method, the endogenous compounds' interfering peaks in the mass spectrum are separated at a high resolution of FWHM 140,000, and the results are suitable for substance detection following the WADA guidelines. The interferences in the obtained mass spectrum of the urine matrix are identified as acetylcholine, lysine, and glutamine by further analysis and database searching. Validation of the method is performed in routine anti-doping testing, and the limit of detection is 50 ng/mL. This method uses simple sample preparation and a general reverse phase HPLC column, and it can be easily applied to other substances.

• 한국식품영양학회지
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• 제30권5호
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• pp.973-982
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• 2017

Gastrodia elata Blume often has been used for the treatment of headaches, convulsions, hypertension, and neurodegenerative diseases. The main active constituents are gastrodin, 4-hydroxybenzyl alcohol, vanillyl alcohol, 4-hydroxybenzylaldehyde and parishin A, B, C and E. Because Gastrodia elata has also unacceptable off-odor (swine barnyard-like) for food, there is a need to reduce it as well as allow for greater utilization as a functional food materials. In this study, a major off-odor producing substance of Gastrodia elata was fractionated by steam distillation and silica gel column chromatography. The substance was identified as p-cresol(4-methyl phenol) by GC-MS analysis and comparison of the retention time with that of an authentic compound in GC. The content of p-cresol in fermented Gastrodia elata was decreased. A fermented sample of Latobacillus sakei for 2 days was reduced to 54.7%, when compared with a unfermented sample. The five parishin derivatives in Gastrodia elata were identified by HPLC-MS analyses, and a comparison of HPLC retention times with those of authentic compounds. When compared with parishin derivatives of an unfermented Gastrodia elata, those of Gastrodia elata fermented by L. sakei, increased to 18.3% for 2 days. Increases of about 14.0~38.4% of the total phenolic compounds and 57.4~77.3% total flavonoids were found in fermented extracts, by 3 lactic acid bacteria strains. They were compared with $97.1{\pm}2.9{\mu}g/g$ and $40.9{\pm}2.0{\mu}g/g$ in the unfermented control, respectively. The extracts of Gastrodia elata Blume that were fermented by lactic acid bacteria had higher DPPH free radical scavenging activity and FRAP reducing power than the unfermented control.

• 한국식품영양과학회지
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• 제22권2호
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• pp.132-137
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• 1993

본 연구는 장기간의 에탄올을 섭취시킨 흰쥐에 있어서 최근에 개발된 CL-HPLC를 이용해 측정한 과산화물 및 혈액 생화학치의 변화를 조사하고, 또한 이들 수치에 미치는 아스코르빈산, 알파토코페롤 및 아스코르빈산 알파토코페롤 병합투여의 영향을 알아보기 위하여 실시하였다. 그 결과, 에탄올 투여군은 대조군에 비하여 체중은 감소하였으나, 상대적 간장중량, GPT의 수치가 증가하였으며, 특히 과산화물가가 유의하게(p<0.001) 증가하였다. 반면에 에탄올 투여군에 3주동안 아스코르빈산, 알파토코페롤 및 아스코르빈산.알파토코페롤 병합 투여를 한 결과, 알파토코페롤 투여군 및 아스코르빈산.알파토코페롤 병합투여군에서 상대적 간장중량, 콜레스테롤함량, GPT의 수치 및 과산화물가가 회복되었다. 이 결과는 식이중의 알파토코페롤 및 아스코르빈산.알파토코페롤 투여가 에탄올로 유도된 간장의 과산화지질의 형성을 방지해, 간기능을 회복시키는데 효과적이라는 것을 시사한다 하겠다.

• 대한화장품학회지
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• 제46권1호
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• pp.67-72
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• 2020

본 연구에서는 사람 피부 섬유아세포에서 콜라겐을 파괴하는 matrix metalloproteinase-1 (MMP-1)을 억제하는 새로운 항노화 천연물을 얻고자 하였다. 50 여종의 생약추출물을 스크리닝 한 후 그 중 인동덩굴 추출물을 선정하여 본 연구를 수행하였다. 인동 줄기를 70% 에탄올로 추출한 후 헥산, 에틸아세테이트, 부탄올 차례로 용매 분획을 하였다. 에틸아세테이트층 50 ㎍/mL와 부탄올층 200 ㎍/mL 농도에서 유의적으로 MMP-1 생성을 억제하였다. 이 효능 성분들을 모두 포함하는 분획을 얻기 위해 HP20 레진 컬럼 크로마토그래피를 통해 80% 에탄올 분획을 얻었다(LJ F80). LJ F80과 지표성분 loganin은 농도의존적으로 MMP-1 생성을 억제하였으며 그 활성을 비교하였을 때 같은 농도에서 LJ F80이 더 억제효과가 높게 나타났다. 이 분획에 대한 안정도를 각각 4 ℃, 상온, 40 ℃에 보관하면서 HPLC로 분석하여 16 주간 평가하였다. Loganin을 기준으로 하여 초기값 대비 ± 10%내외의 안정도를 유지하였다. 따라서 인동의 LJ F80은 피부주름개선효과를 나타내는 유효 화장품 원료로 사용될 수 있을 것으로 생각된다.

• 한국식품위생안전성학회지
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• 제36권5호
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• pp.376-381
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• 2021

Red beet roots (Beta vulgaris L.)는 천연색소로 붉은 색계열의 betacyanins은 75-95%의 betanine와 이성질체인 isobetanine 15-45%으로 존재한다. 본 연구는 비트레드를 사용한 식품에 대해 HPLC-DAD를 이용하여 지표성분인 betanine 및 isobetanine에 대해 분석법을 확립하였으며, 유효성 검증을 위해 직선성, 검출한계, 정량한계, 정확성, 정밀성, 측정불확도를 측정하였다. 캔디류, 빙과류, 코코아 가공품의 matrix에 적용하여 matrix matched calibration법을 사용하였으며 R²이 0.9998 이상으로 높은 직선성을 보였다. 검출한계와 정량한계는 각각 0.16-0.32 mg/L, 0.48-0.97 mg/L으로 확인되었다. 분석법의 정확성 및 정밀성을 검증하기 위해 intra-day 및 inter-day 반복 실험 결과, 회수율은 96.0-103.1 %, 100.0-102.2 %이였으며, RSD는 0.5-3.3 %, 0.9-3.8 %로 산출되었다. 측정불확도는 매트릭스 및 측정 농도에 따라 1.71-12.43%로 평가되었다. 또한, 확립된 분석법의 적용성 검토를 위해서 비트레드 색소를 사용한 가공식품 26종을 분석한 결과, betanine과 isobetanine을 정량 할 수 있었다 (8.4-3,823.4 mg/kg).

• 생명과학회지
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• 제29권4호
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• pp.455-460
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• 2019

이전 연구에서 우리는 Lactobacillus plantarum ilchiwhangchil 1785와 Lactobacillus plantarum ilchiwhangchil 2020 균주로 발효시킨 황칠나무 잎 가지 추출물의 효과적인 항균 활성을 확인하였다. 본 연구에서는 황칠나무 잎 가지 추출물을 L. plantarum ilchiwhangchil 1785와 L. plantarum ilchiwhangchil 2020 균주로 발효시킨 발효액이 인간모유두세포에 미치는 육모효과를 알아보았다. 황칠나무 잎 가지 발효 추출물의 농도가 1, $10{\mu}g/ml$ 범위에서 인간모유두세포는 세포사멸 증가 없이 유의적으로 세포 성장이 농도의존적으로 증가하였다. 또한 황칠나무 잎 가지 발효물로 처리한 세포가 대조군보다 세포성장이 활발함을 이중염색법으로 확인하였다. 더욱이 황칠나무 잎 가지 발효 추출물 처리한 세포는 처리하지 않은 세포에 비해 각각 18.84%, 23.31% 세포 이동성이 증가하였다. 고성능액체크로마토그래피(HPLC)를 이용하여 육모효과의 활성 물질을 분석하였다. 모발성장효과가 알려진 ${\beta}$-sitosterol 함량이 황칠나무 잎 가지 추출물에 비해 황칠나무 잎 가지 발효 과정에서 약 10배 증가되는 것을 확인하였다. 결과를 종합하면, 우리는 본 연구에서 황칠나무 잎 가지 추출물 발효액이 모발 성장을 촉진시킴을 확인하였다.

• 대한본초학회지
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• 제32권4호
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• pp.25-32
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• 2017

Objectives : Chungsimyeonja-eum (CSYJE; Qingxinlianzi-yin in Chinese; Seishinrenshi-in in Japanese), a traditional herbal medicine, has been used for treating mouth dryness, hyperuresis. This study was designed to determine preservation period of CSYJE. We investigated the stability and biological activity of CSYJE depending on the preservation temperature and periods. Methods : CSYJE decoction was preserved for 0-6 or 12 months at room temperature (RT, $23{\pm}1^{\circ}C$) or refrigeration ($4^{\circ}C$). To evaluate the stability of CSYJE decoction, pH and dissolved solids content were measured. High-performance liquid chromatography (HPLC) analysis was performed to determine marker compounds-liquiritin apioside, liquiritin, baicalin, wogonoside, and glycyrrhizin-in CSYJE. To determine anti-inflammatory effect of CSYJE, prostaglandin $E_2$ ($PGE_2$) was measured in LPS-stimulated RAW 264.7 macrophages. Results : Both at RT and $4^{\circ}C$, pH was decreased depending on the preservation periods. There was no changes in dissolved solids content depending on the preservation temperature and periods. Both at RT and $4^{\circ}C$, the contents of liquiritin apioside and liquiritin were slightly increased at 1 month of storage. The level of baicalin was decreased time-dependently and the disappearance rate at RT is larger than at $4^{\circ}C$. CSYJE inhibited $PGE_2$ production in LPS-stimulated RAW 264.7 cells and maintained inhibitory effect by 12 months both at RT and $4^{\circ}C$. Conclusions : Based on the disappearance rate of the baicalin in CSYJE, the preservation period is recommended within 8 months for RT and 12 months for $4^{\circ}C$.

• 대한본초학회지
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• 제28권6호
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• pp.47-51
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• 2013

Objectives : The Illicii Veri Fructus was not only traditional medicine but also food in Asia. The aim of this study was selection of optimum solvent in the fruit of Illicii Veri Fructus because an appropriate solvent affect a medicinal effect. Methods : Illicii Veri Fructus was carried out ultrasonic-assisted extraction as various solvents. Two main compounds, p-anisaldehyde and anethole, were successfully analyzed by high performance liquid chromatography-photodiode array detector (HPLC-PDA) and carried out method validation according to ICH guideline. The optimum solvent selected by comparing with yields of two main ingredients. Results : The p-anisaldehyde and anethole were detected at approximately 8.0 min and 19.8 min, respectively. It was all below 5.0% that RSD of retention time and peak area for two main peaks. Calibration curves of two compounds were good linearity as $R^2$ >0.9999. All of the precisions and accuracy were good intra-day and inter-day as below 5.0% RSD. Limited of detection (LOD) of p-anisaldehyde and anethole were analyzed as $0.134{\mu}g/mL$ and $4.286{\mu}g$, respectively. Limited of quantification (LOQ) of two compounds were $0.407{\mu}g$ and $12.989{\mu}g$, respectively. As a result of this study, p-anisladehyde was detected as 0.209 ~ 0.467%, however anethole was not detected in the distilled water. Conclusions : Anethole was main component as 5.329 ~ 6.815% except for water extraction. Methanol extraction among various solvents was detected the highest contents of p-anisaldehyde and anethole as 0.467(${\pm}0.008$)% and 6.815(${\pm}0.220$)%, respectively.

• 한국자연치유학회지
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• 제12권1호
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• pp.7-15
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• 2023

배경: 야생 망초(줄기와 잎)의 성분을 분석한 결과에서 생리활성 성분의 함유가 확인되어 화합물의 규명이 필요하다. 목적: 망초에는 항산화성 물질이 함유되었다는 선행연구 결과가 있어서 화합물의 분자적 구조를 연구하는 것이 목적이었다. 방법: 망초(줄기와 잎) 초음파 분쇄물을 1차로 90% 메탄올로, 다음에 유기 용매로 추출하였다. 다음에 HPLC, LC/MS 크로마토그래피 등으로 분획하였고, 분핵물을 NMR 분광기로 정밀 분석하여 분자의 구조를 동정하였다. 결과: 망초 100 g을 초음파기로 파쇄하여 90% methanol로 추출하고, 감압 농축하여 조 추출물 11.96 g을 얻었다. 조추출물을 유기용매로 재추출하여 n-hexane으로 123.8 mg, dichloromethane으로 448.2 mg, ethyl acetate(EA)로는 1047.7 mg, butanol로는 2563.8 mg, water로는 7.04 g을 얻었다. EA 추출물을 LC-MS 크로마토그래피로 분획하고, 정밀 분획하여 F1~F20의 20개를 얻었다. F15 분획을 다시 분획하여 9개를 얻었다. F17 분획을 재분획하여 10개의 분획을 얻었다. F15-7 분획물을 LC-MS 분석과 NMR 분광기로 분석한 결과는 이 화합물의 구조가 3,5-di-caffeoylquinic acid로 확인되었다. F17-4와 F17-5의 구조를 조사한 결과는 Quercetin-3-o-β-galactose로 동정 되었다. F17-10의 구조를 확인한 결과는 1,3,4-tri-caffeoylquinic acid로 확인되었다. 결론: 본 연구에서 망초 성분에도 항산화성 물질이 있었으며, 이의 활용성을 기대하며, 식물에는 항산화성 물질인 phenol 성분을 다양하게 함유하였다.