• Title/Summary/Keyword: marker substances

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Effects of Herbal Medicine on Breastfeeding - Analysis of Marker Substances in Saenghwa-tang by HPLC and LC/MS/MS - (산모의 한약 복용이 모유에 미치는 영향 - HPLC와 LC/MS/MS를 이용한 생화탕 지표성분 분석 -)

  • Park, Ka-Young;Lee, Ah-Young;Ban, Ji-Hye;Park, Jung-Kyung;Lee, Eun-Hee
    • The Journal of Korean Obstetrics and Gynecology
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    • v.26 no.4
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    • pp.48-65
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    • 2013
  • Objectives: We took breast milk samples and analyzed them using HPLC and LC/MS/MS, to evaluate the effects of taking Saenghwa-tang during breastfeeding on breast milk. Methods: The study participants were 20 lactating women who admitted in Korean medical postpartum care center. Breast milks were collected from paticipants who have been administrated Saenghwa-tang for more than 3 days. We used HPLC and LC/MS/MS for the determinations of amygdalin, liquiritins, 6-gingerol, decursin and decursinol angelate in Saenghwa-tang. Results: 1. Participants' $Mean{\pm}S.D$ (standard deviation) of age is $31.05{\pm}1.96$, and 15 participants had normal delivery and 5 participants had cesarean delivery. 12 participants were primipara and 8 participants were multipara. $Mean{\pm}S.D$ of lactating date is $9.4{\pm}0.94$. 2. Using HPLC, we learned LOQ level peak that matches the peak retention time of standard components of Saenghwa-tang was not detected from 20 breast milk samples. 3. Using LC/MS/MS, decursin of Angelicae Gigantis Radix was detected from HMSP 02, HMSP 04, HMSP 06, HMSP 11, and the each concentrations are 16, 2, 64, 11 ppb. Liquiritin of Glycyrrhizae Radix was not detected from HMSP 13~HMSP 18. Conclusions: Data obtained by this approach shows that this method is reliable and suitable for determining the safety of taking Saenghwa-tang during breastfeeding.

Affirmative Effect of Hwaweo-jeon (Huayu-jian) in Osteoblast Cells and Tibia Fracture-induced Mice (화어전(化瘀煎)이 조골세포 및 경골골절 유발 생쥐의 골유합에 미치는 영향)

  • Lee, Soo-Hwan;Parichuk, Kira;Cha, Yun-yeop
    • Journal of Korean Medicine Rehabilitation
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    • v.30 no.1
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    • pp.13-29
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    • 2020
  • Objectives This study was performed to decide the bone union effect of Hwaweo-jeon on tibia fractured mice. Methods In this study, laboratory experiments were implemented by the stage of in vitro and in vivo. In in vitro, MC3T3-E1 cells were treated with various concentration of Hwaweo-jeon extract (HWJ). To investigate effect of HWJ for osteoblast, relative mRNA expression of 5 substances (alkaline phosphatase [ALP], runt-related transcription factor 2 [Runx2], osteocalcin [OCN], osterix [OSX] and collagen type II alpha 1 chain [Col2a1]) was used as a marker of osteogenesis. In order to determine HWJ's effect for fracture healing, relative gene expression level of ALP, Runx2, OCN, OSX and Col2a1 were used to find out the influence to osteoblast. Furthermore, receptor activator of nuclear factor kappa-B ligand and osteoprotegerin relative mRNA expression were used to estimate the impact to osteoclast. Also, X-ray was used for the purpose of identifying bone union in tibia-fracture mouse model. Results In in vitro experiment, most part of relative mRNA expression were increased compared to control group. In in vivo and in vitro experiment, HWJ induced osteoblast activitation by verifying relative mRNA expression of 5 substances. And in vivo experiment, we can also identify that HWJ triggered osteoclast activation during early stage of tibia fracture. Furthermore, X-ray pictures show noticeable recovery of tibia fracture. Conclusions HWJ extract promotes bone union by facilitating the osteoblast. But, HWJ may occur liver & kidney toxicity over specific concentration. Therefore, when HWJ is applied to human body, doctors have to follow up the liver function test & renal function test of patient.

Feasibility Study on RI Biochip Application to Detection of Risk Factors of Atherosclerosis (RI검출 바이오칩의 혈관계 질환 발생 위험인자 검지에 대한 타당성 연구)

  • Ko, Kyong-Cheol;Choi, Mi Hee;Park, Sang Hyun;Cho, Kyung-Hyun;Lee, Ki-Teak
    • Journal of Radiation Industry
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    • v.3 no.1
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    • pp.25-29
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    • 2009
  • Microarrays can be used to screen thousands of binding events in a parallel and high throughput fashion and are of major importance in disease diagnosis and drug discovery. The use of radioisotope is conventionally regarded as one of the most sensitive detection methods. Atherosclerosis is a common disorder affecting arterial blood vessels. It happens when fat, cholesterol, and other substances made in the arterial blood vessels form a hard substances called plaque. Lipoprotein-associated phospholipase $A_2$ ($Lp-PLA_2$), a phospholipase $A_2$ enzyme, is used as a marker for cardiac disease. The detection of $Lp-PLA_2$ was accomplished by using radioactive [$^3H-acetyl$] PAF as a substrate and a feasibility study on RI biochip application to detection of $Lp-PLA_2$, a risk factors of atherosclerosis was performed. Inhibitive activity of a native plant extract was also determined by using the RI biochip. It was found to be applicable to a high-throughput screening of inhibitors for developing atherosclerosis therapeutic agents.

A Rapid Method for Estimating the Levels of Urinary Thiobarbituric Acid Reactive Substances for Environmental Epidemiologic Survey

  • Kil, Han-Na;Eom, Sang-Yong;Park, Jung-Duck;Kawamoto, Toshihiro;Kim, Yong-Dae;Kim, Heon
    • Toxicological Research
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    • v.30 no.1
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    • pp.7-11
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    • 2014
  • Malondialdehyde (MDA), used as an oxidative stress marker, is commonly assayed by measuring the thiobarbituric acid reactive substances (TBARS) using HPLC, as an indicator of the MDA concentration. Since the HPLC method, though highly specific, is time-consuming and expensive, usually it is not suitable for the rapid test in large-scale environmental epidemiologic surveys. The purpose of this study is to develop a simple and rapid method for estimating TBARS levels by using a multiple regression equation that includes TBARS levels measured with a microplate reader as an independent variable. Twelve hour urine samples were obtained from 715 subjects. The concentration of TBARS was measured at three different wavelengths (fluorescence: ${\lambda}-_{ex}$ 530 nm and ${\lambda}-_{em}$ 550 nm; ${\lambda}-_{ex}$ 515 nm and ${\lambda}-_{em}$ 553 nm; and absorbance: 532 nm) using microplate reader as well as HPLC. 500 samples were used to develop a regression equation, and the remaining 215 samples were used to evaluate the validity of the regression analysis. The induced multiple regression equation is as follows: TBARS level (${\mu}M$) = -0.282 + 1.830 ${\times}$ (TBARS level measured with a microplate reader at the fluorescence wavelengths ${\lambda}-_{ex}$ 530 nm and ${\lambda}-_{em}$ 550 nm, ${\mu}M$) -0.685 ${\times}$ (TBARS level measured with a microplate reader at the fluorescence wavelengths ${\lambda}-_{ex}$ 515 nm and ${\lambda}-_{em}$ 553 nm, ${\mu}M$) + 0.035 ${\times}$ (TBARS level measured with a microplate reader at the absorbance wavelength 532 nm, ${\mu}M$). The estimated TBARS levels showed a better correlation with, and are closer to, the corresponding TBARS levels measured by HPLC compared to the values obtained by the microplate method. The TBARS estimation method reported here is simple and rapid, and that is generally in concordance with HPLC measurements. This method might be a useful tool for monitoring of urinary TBARS level in environmental epidemiologic surveys with large sample sizes.

Protective Effects of In Vitro Gastrointestinal Digests of Abalone (Haliotis discus hannai) Intestines against Oxidative Stress in RAW264.7 Macrophage Cells

  • Nguyen, Phuong-Hong;Kim, Sun-Ae;Choi, Il-Whan;Jung, Won-Kyo
    • Fisheries and Aquatic Sciences
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    • v.13 no.3
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    • pp.216-223
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    • 2010
  • Abalone (Haliotis discus hannai), mostly distributed and maricultured in southwestern coastal areas of South Korea, is recognized as an economically important species in the fishery industry. Abalone intestines are one of the by-products of abalone processing. To investigate abalone intestines as bioactive substances, abalone intestine gastrointestinal digests (AIGIDs) of various molecular weights (MWs) were prepared using in vitro gastrointestinal digestion and an ultrafiltration system, and tested for inhibitory effects against reactive oxygen species (ROS) and oxidative stress in macrophage cells treated with hydrogen peroxide ($H_2O_2$). In our results, among AIGIDs, AIGID-III (MW=5-10 kDa) showed potent inhibitory activities for lipid peroxidation and free radicals. Additionally, the results clearly indicated that AIGID-III treatment could prevent cytotoxic damage of macrophages by $H_2O_2$-induced oxidative stress due to its potent scavenging ability against cellular ROS. These results suggest that AIGIDs may have protective and therapeutic potential for oxidative stress syndromes and immune diseases through ROS inhibition in macrophage cells.

The Effective Preparation of Flavonoids from Scutellaria baicalensis GEORGI by Diaion HP-20 Resin

  • Yu, Young-Beob
    • Korean Journal of Plant Resources
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    • v.27 no.6
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    • pp.635-641
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    • 2014
  • Scutellaria baicalensis $G_{EORGI}$ (Scutellariae Radix) has been used to clear heat and to dry dampness in the stomach or intestines, which manifests as diarrhea or dysenteric disorder. In this study, we investigated the effective preparation of active components in Scutellariae Radix using the methods of solvent extraction and absorption fractionation for the development of new functional food or pharmaceuticals. The marker substances, baicalin, baicalein, wogonoside, and wogonin were directly isolated from the Scutellariae Radix. There chemical structures were elucidated by spectroscopic analysis. The Scutellariae Radix was extracted with hot water. To enhance yield of flavonoids in Scutellariae Radix, the hot water extract was dissolved in ethanol with concentration dependent manner. The precipitates were separated using centrifugal techniques at 10,000 rpm. Supernatant liquid was applied to the HPLC for quantification of major compounds. Separately, the hot water extract was absorbed on Diaion HP-20 resin. And then, the absorbed fraction was eluted with methanol for HPLC. The contents of baicalin, baicalein, wogonoside and wogonin in different treatment methods were analyzed by HPLC. Total amount of four major components were 16.9% in 50% ethanol extract, 21.7% in 70% ethanol extract, 20.5% in 90% ethanol extract, and 39.3% in absorbed fraction of Diaion HP-20 resin. In these results, we found that resin absorption method is suitable for the extraction of enriched flavonoids from Scutellariae Radix.

Genetic Diversity and Metabolite Analysis of Gastrodia elata by Inter-Simple Sequence Repeats (ISSR) Markers (ISSR 표지에 의한 천마의 유전 다양성분석 및 기능성 물질분석)

  • Kim, Hyun Tae;Kim, Ji Ah;Park, Eung Jun
    • Korean Journal of Medicinal Crop Science
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    • v.20 no.6
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    • pp.440-446
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    • 2012
  • Gastrodia elata, an achlorophyllous orchid plant, is rare medicinal plant. We investigated the genetic diversity in G. elata from 4 locations by using Inter-Simple Sequence Repeats (ISSR) markers. Shannon's information Index (S.I.) indicating genetic diversity ranged from 0.255 (Pocheon) to 0.322 (Muju) with the mean of 0.29. The level of genetic diversity was lower than other plant and most genetic diversity was allocated among individuals within populations (26.81%). The UPGMA dendrogram based on genetic distance failed in showing decisive geographic relationship. In the case of gastrodin (GA), the major components in G. elata, Sangju was highest. The ergothionine (ERG) was detected a lot of contents in Muju and Pocheon. In conclusion, our results is very important information for explaining relationship of genetic variation and functional substances without the effects of environment factors and developing genetic marker by ISSR in G. elata, which may be responsible for the development of breeds with a lot of functional substance in G. elata.

Acute Oral Toxicity Study of Standardized Herbal Preparations(Gami-Samhwang-San, SH-21-B) in Rats (HPLC로 표준화한 가미삼황산(加味三黃散) 분획물(SH-21-B)의 랫드에 대한 단회경구투여독성시험)

  • Yu, Young-Beob;Kim, Seon-Hyeong;Yoon, Yoo-Sik
    • Toxicological Research
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    • v.21 no.3
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    • pp.255-261
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    • 2005
  • Gami-Samhwang-San, a herbal prescription for obesity treatment, is composed of seven crude herbs such as Rehmanniae Radix Preparata, Ephedrae Herba, Scutellariae Radix, Acori Gramineri Rhizoma, Polygalae Radix, Typhae Pollen, Armeniacae Semen, Menthae Herba. In this study, marker substances in n-butanol fraction (SH-21-B) from Gami-Samhwang-San were analyzed by high performance liquid chromatography (HPLC) and acute toxicity of standardized SH-21-B was evaluated by good laboratory practices (GLP) guideline of Korea Food and Drug Administration. Therefore we confirmed that there were baicalin of 15.92%, amygdalin of 6.57% and ephedrine of 2.49% in SH-21-B. SH-21-B was administered in rats at dose of 0 mg/kg, 2,000 mg/kg, and 5,000mg/kg. Clinical signs of both sexes of rats were observed daily for 14 days after single oral administration. Two female rats one administered at 2,000 mg/kg and the other administered at 5,000 mg/kg, died, but no dead animal was observed among male rats. Therefore $LD_{50}$ in the female rat is observed to be 8,710 mg/kg, and MLD (Minimun Lethal Dose) of the male rat is observed to be more than 5,000 mg/kg.

Levels of organochlorine pesticides and PCB congeners in Korean human tissues

  • Yoo, Young-Chan;Lee, Sang-Ki;Yang, Ja-Youl;Kim, Ki-Wook;Lee, Soo-Yeun;Oh, Seung-Min;Chung, Kyu-Hyuck
    • Proceedings of the PSK Conference
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    • 2002.10a
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    • pp.283.2-283.2
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    • 2002
  • Organochlorine pesticides and polychlorinated biphenyls (PCBs) have been used intensively in agriculture and industry for a long time. They belong to a group of contaminants whose occurrence in the environment is a serious concern to environmental chemists and toxicologists due to their resistance to degradation in the environment as well as their potential toxicity. Also. the lipophilic characteristics of these substances are responsible for their ability to bioaccumulate in tissues and organs rich in lipids of men and animals through food chain. Therefore, the measure of the levels of organochlorine pesticides and PCBs in human tissues are good markers in detemining the extent to exposure and evaluating the hazards. This study was preformed to compare concentrations of organochlorine pesticides(${\alpha}-BHC, {\beta}-BHC, {\gamma}-BHC, {\delta}-BHC$, p.p'-DDT,p.p'-DDD,p.p.'-DDE. endrin. dieldrin. aldrin) and seven marker PCBs(PCB nos. 28. 52. 101. 118. 138. 153. 180) in liver. kidney cortex, lung blood and adipose tissue collected at autopsies of 10men and 10 women using gas chromatography equipped with electron capture detector to express the data on a lipid adjusted basis. From the results, the significant differences in the levels of organochlorines of PCBs between sexes, districts where they had lived and ages were also investigated.

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Effects of Extracts of Monascus-fermented Angelica gigas Nakai in High-fat Diet-induced Obesity in Rats (고지방 식이 유도 비만 흰쥐에서 홍국발효 참당귀 추출물의 항비만 효과)

  • Kim, Young Wan;Kim, Tae Hoon;Sim, So Yeon;Ahn, Hee Young;Park, Kyu Rim;Kim, Jung Wook;Cho, Young Su
    • Journal of Life Science
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    • v.28 no.1
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    • pp.58-67
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    • 2018
  • This study aimed to identify the effects of extracts of fermented Angelica gigas Nakai (A. gigas) with a Monascus purpureus strain on anti-obesity in a high-fat diet (HFD)-induced obesity rat model. Male Sprague-Dawley rats were randomly divided into seven dietary groups (n=8 per group), as follows: a normal diet group (N) and six HFD groups (C: control, HFD and no treatment; AG: HFD +10% A. gigas extracts; FAG2.5: HFD +2.5% fermented A. gigas extracts; FAG5: HFD +5% fermented A. gigas extracts; FAG10: HFD +10% fermented A. gigas extracts; GC: HFD + Garcinia cambogia extracts). FAG-fed rats exhibited effectively lowered rates of increasing body weight and visceral fat accumulation in the HFD-induced obesity model. The activities of several hepatic marker enzymes, including aspartate aminotransferase (AST), alanine aminotransferase (ALT), lactate dehydrogenase (LDH), and alkaline phosphatase (ALP), were increased with HFD-induced obesity, but levels of these marker enzymes were significantly decreased in FAG-fed rats. The consumption of FAG reduced triglyceride concentrations in serum to normal levels. FAG-fed rats showed effectively increased leptin concentrations in the HFD-induced obesity model. HFD ingestion induced a significant increase in the thiobarbituric acid reactive substances (TBARS) levels, which was decreased in FAG-fed rats. Hematoxylin and eosin staining and Oil Red O staining of the liver showed that the lipid deposits were decreased via FAG feeding. Moreover, hematoxylin and eosin staining of epididymal adipose adipose tissue showed that the adipocyte were decreased by feeding FAG.