• Title/Summary/Keyword: HPLC Chromatography

Search Result 2,212, Processing Time 0.025 seconds

Development and Validation of the Determination of Sorafenib in Human Plasma using Tandem Mass Spectrometry Coupled with Liquid Chromatography (고속액체크로마토그래피 텐덤질량분석기법을 이용한 사람 혈장 내 소라페닙 농도분석법의 개발 및 검정)

  • Park, Daejin;Lee, Sunggon;Kim, Woomi
    • Journal of Life Science
    • /
    • v.22 no.11
    • /
    • pp.1456-1462
    • /
    • 2012
  • Sorafenib is a multikinase inhibitor and an oral anticancer drug approved for the treatment of patients with advanced renal cell carcinoma and those with unresectable hepatocellular carcinoma. The purpose of this study was to develop an efficient method of the determination of sorafenib in human plasma using tandem mass spectrometry coupled with liquid chromatography (LC/MS/MS) and validate the method by the guidelines of the Korean Food and Drug Administration (KFDA). Plasma samples ($100{\mu}l$) were added with chlorantraniliprole as an internal standard and then mixed with the 0.1% formic acid-containing extraction solution composed of isopropyl alcohol and ethyl acetate (1:4, v/v). After centrifugation, the supernatant was concentrated at $45^{\circ}C$ under negative pressure and centrifugal force. The residue was reconstituted with a mobile phase and injected into the HPLC instrument using a reverse phase Waters XTerra$^{TM}$ C18 column (particle size $3.5{\mu}m$). Liquid chromatography was carried out within the run time of 5 min using a mobile phase composed of buffer (0.1% formic acid and 10 mM ammonium formate), methanol, and acetonitrile (1:6:3, v/v/v). The analytes were monitored by tandem mass spectrometry in the multiple reaction monitoring method programmed to detect sorafenib at 'm/z 465.2 ${\rightarrow}$ 252.5' and chlorantraniliprole at 'm/z 484.4 ${\rightarrow}$ 286.2' with positive electrospray ionization mode ($ES^+$). The result showed the proper linearity ($r^2$ > 0.99) over the range of 2,000-5,000 ng/ml with good accuracy (90.7-103.9%) and precision (less than 10%). The newly developed method using LC/MS/MS was validated by the guideline of KFDA and identified as more sensitive compared to the previous methods.

Immunostimulating Activity and Characterization of Polysaccharides from Mycelium of Phellinus linteus

  • Lee, Jae Hoon;Soo Muk Cho;Kyung Sik Song;Sang Bae Han;Hwan Mook Kim;Nam Doo Hong;Ick Dong Yoo
    • Journal of Microbiology and Biotechnology
    • /
    • v.6 no.3
    • /
    • pp.213-218
    • /
    • 1996
  • Hot-water extract, Fr. 1, of Phellinus linteus mycelium was fractionated into Fr. 2, 3, 4, and 5 by the difference of solubility in ethanol. The polysaccharide fractions were studied for their immunostimulating activity on in vitro T-independent polyc1onal antibody response to trinitrophenyl-haptened SRBC (sheep red blood cell). The Fr. 4 with the highest immunostimulating activity was subjected to DEAE-cellulose ion exchange chromatography and gave five fractions, 4-I, II, III, IV, and V. The in vitro immunostimulating assay of the five fractions showed that 4-I and 4-III had a similar activity to that of LPS but the other fractions had low activity. By analyses of chemical composition and HPLC, all fractions obtained were found to be heteropolysaccharide-protein complex. The molecular weights ranged from 9, 000 to 15, 000. Sugar analyses showed that glucose, galactose, mannose, arabinose, and xylose were main component. Uronic acid and amino sugar were also detected in the fractions. It should be noted that the molecular weight (15, 000) of 4-III was very small and the structure of 4-III may be different from the known immunostimulating branched $\beta$-(1longrightarrow3)-glucan.

  • PDF

Occurrence of Toxic Alexandrium and Intoxification of Two Mollusk Species by Paralytic Shellfish Poisoning Toxins on the Southeastern Coast of Korea

  • Kim Young-Soo;Lee Jong-Soo;Jang Joon-Ho;Kim Keun-Yong;Kim Chang-Hoon
    • Fisheries and Aquatic Sciences
    • /
    • v.8 no.2
    • /
    • pp.76-82
    • /
    • 2005
  • We analyzed the paralytic shellfish poisoning (PSP) toxins of the toxic marine dinoflagellate Alexandrium tamarense collected from Dadaepo and Gaduck-do in Busan and from Sujeong-ri in Jinhae Bay, Korea, in April 2003. We also analyzed the PSP toxin of mussels (Mytilus galloprovincialis) and oysters (Crassostrea gigas) collected around Busan and Jinhae Bay. PSP toxin analyses were conducted by high performance liquid chromatography (HPLC). Fifteen cultured A. tamarense isolates contained 2.78 to 57.47 fmol/cell, with nearly identical toxin profiles: major components C2, GTX4; minor components C1, GTX1, NEO; and trace components GTX2, GTX3, STX. PSP toxin contents were 0 to $492\;\mu{g}$ STXeq/100 g in mussels and 0 to $48\;\mu{g}$ STXeq/100 g in oysters. Mussels at Gijang and Sujeong-ri contained the most PSP toxin contents ($492\;\mu{g}\;STXeq/100\;g\;and\;252\;\mu{g}\;STXeq/100\;g,\;respectively$), exceeding the quarantine level ($80\;\mu{g}$ STXeq/100 g). Their dominant toxin components were C2, C1, GTX2, and GTX3; the minor components GTX1, GTX4, GTX5, and NEO were sporadically detected. Phytoplankton contained 0.774 fmol/L seawater and 1.228 fmol/L seawater at Gijang and Sujeong-ri in April. At that time, Alexandrium cells were present in the water column at Gijang at 2,577 cells/mL and at Sujeong-ri at 6,750 cells/mL. Overall, we found the high and similar PSP toxin contents in AZexandrium isolates and mussels, and a correlation between occurrence of toxic Alexandrium cells in the water column and mussel intoxification. High densities of toxic Alexandrium cells in the water column immediately preceded shellfish intoxification at Gijang and Sujeong-ri in April.

Analysis of Vitamin $B_{12}$ in the Korean Representative Foods and Dietary Intake Assessment for Koreans

  • Choi, Youn-Ju;Kim, Ji-Yung;Lee, Haeng-Shin;Kim, Cho-Il;Hwang, In-Kyeong;Park, Hye-Kyung;Kim, Tae-Hwa;Oh, Chang-Hwan
    • Food Science and Biotechnology
    • /
    • v.17 no.2
    • /
    • pp.262-266
    • /
    • 2008
  • This study was conducted to provide a vitamin $B_{12}$ database for the representative Korean food items and to assess the dietary intake assessment of vitamin $B_{12}$ for Koreans. The vitamin $B_{12}$ content of 106 foods had been determined by high performance liquid chromatography (HPLC) using column switching method. Rich sources of vitamin $B_{12}$ were meats, milk, and egg ($0.3-3.4\;{\mu}g/100g$). Vegetables and fruits contained vitamin $B_{12}$ below limit of detection (LOD). The major food sources for vitamin $B_{12}$ intake were milk (72.0%), meats (22.3%), egg (3.6%), and fishes & shellfishes (2.1%). Mean vitamin $B_{12}$ intake of Koreans was $3.16\;{\mu}g$/person/day. The proportion of population with intake below estimate average requirement (EAR) and above recommended intake (RI) of vitamin $B_{12}$ was 60.7 and 36.5%, respectively. The vitamin $B_{12}$ intake level of young children with 1-2 years which was 834.6% of RI while the intake level of the older adults 50 years and older was only 70.0% of RI. Also, there were regional differences between urban and rural area. The population with intake below RI was larger than that with intake above RI in Korea.

Determination of Aflatoxin B1 in Rice, Barley, and Feed by Non-instrumental Immunochromatographic Strip-test and High Sensitive ELISA

  • Shim, Won-Bo;Kim, Jung-Sook;Kim, Ji-Young;Choi, Jin-Gil;Je, Jung-Hyun;Kuzmina, Nina Sergeevna;Eremin, Sergei Alexandrovich;Chung, Duck-Hwa
    • Food Science and Biotechnology
    • /
    • v.17 no.3
    • /
    • pp.623-630
    • /
    • 2008
  • A non-instrumental immunochromatographic (ICG) strip-test and direct competitive enzyme-linked immunosorbent assay (DC-ELISA) for aflatoxin B1 (AFB1) determination were developed and optimized. The detection limits of ICG strip-test and DC-ELISA were 0.5 and 0.004 ng/mL, respectively, and these methods possessed a cross-reaction to aflatoxins. The results of spiked samples by both methods were coincided with the amount spiked AFB1 and the comparative analyses of 172 real samples by 2 immunoassays and high performance liquid chromatography (HPLC) showed a good agreement. Especially, the ICG strip-test is easier to perform and quicker, but less sensitivity than DC-ELISA. Both methods could analyze a high sample throughput with short time, but the sample throughput of ICG strip-test was better. Therefore, the ICG strip-test can be used as a simple, easy, non-instrumental, and fast screening technique for AFB1 determination.

In vitro Antioxidative Activities and Phenolic Composition of Hot Water Extract from Different Parts of Cudrania tricuspidata

  • Jeong , Chang-Ho;Choi, Gwi-Nam;Kim, Ji-Hye;Kwak, Ji-Hyun;Heo, Ho-Jin;Shim, Ki-Hwan;Cho, Bok-Rai;Bae, Young-Il;Choi, Jine-Shang
    • Preventive Nutrition and Food Science
    • /
    • v.14 no.4
    • /
    • pp.283-289
    • /
    • 2009
  • We evaluated total phenolics and antioxidative activities of water extracts from different parts of Cudrania tricuspidata (specifically, the leaves, stems, roots, and fruits). The antioxidative activities of these samples were determined using five methods, including 1,1-diphenyl-2-picrylhydrazyl (DPPH), 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid (ABTS) radical scavenging, reducing power, ferric reducing ability of plasma (FRAP), and $\beta$-carotene/linoleic acid system. The water extract of leaves exhibited the higher DPPH, ABTS radical scavenging activities, reducing power, and FRAP than water extract of stem, roots, and fruits. Inhibition values on linoleic oxidation of water extracts from leaves, stems, roots, and fruits were calculated as 45.98%, 33.03%, 39.73%, and 25.48% at 10 mg/mL, respectively. The water extract of C. tricuspidata leaves had the highest amount of toal phenolics (73.60$\pm$0.28 mg/g). High-performance liquid chromatography (HPLC) analysis showed that quercetin is the predominant phenolic compound in water extract of leaves. Thus, our study verified that the water extract of leaves has strong antioxidant activities which are correlated with its high level of phenolic compounds, particularly quercetin. This water extract of C. tricuspidata leaves can be used as an effective and safe source of antioxidants.

Effect of Do-In (Prunus persica L. BATSCH) Water Extract (PPE) on Concentration of Extracellular Acetylcholine in the Rat Hippocampus

  • Gong Dae-Jong;Kim Geun-Woo;Koo Byung-Soo
    • The Journal of Korean Medicine
    • /
    • v.25 no.4
    • /
    • pp.1-7
    • /
    • 2004
  • Objective : This study was designed to examine the effects of orally administered Prunus persica water extract (PPE), which is used as herbal medicine, for treatment of Yu Xue (stasis of blood) and tacrine on the basal concentration of extracellular acetylcholine in the hippocampus of rats. Methods: To investigate the effects of PPE and tacrine on concentration of extracellular acetylcholine in the hippocampus of rats, the microdialysis technique, under the same experimental conditions, was used. And we used male Wistar rats which were 7 weeks of age and 210-290 g. PPE was extracted with boiling water, and the rats were anesthetized with pentobarbital Na. Their skulls were exposed and a hole was drilled for implantation of a microdialysis probe. In order to increase the recovery of acetylcholine, a probe with a long membrane was used. One day after surgery, the microdialysis probe was perfused with Ringer's solution at a flow rate of 1.5 l/min. The acetylcholine concentration in dialysis samples was measured by high-performance liquid chromatography (HPLC) with electrochemical detection. AChE activity was measured using the radiometric method, as described by Sherman. Results : The comparative effects of PPE and tacrine on hippocampal extracellular acetylcholine concentration was that these cholinesterase inhibitors produced dose-dependent increases in the extracellular acetylcholine concentration. And the effect of PPE and tacrine on rat brain AChE activity was that PPE produced maximal inhibition at 1 h after administration, when AChE activity was 44% of the intact level. AChE activity gradually recovered thereafter, and reached 78% of the intact level at 12 h after administration. Conclusion : In this study, PPE has a potent activity and a long-lasting effect on the central cholinergic system, in terms of the basal concentration of extracellular acetylcholine in the hippocampus and the AChE activity in the brain of rats. And oral administration of PPE increased dose-dependently the basal concentration of extracellular acetylcholine in the hippocampus of rats. PPE may be one of the more useful cholinesterase inhibitors for the treatment of Alzheimer's disease.

  • PDF

Residues of New Fungicide, Isopyrazam on Cucumber and Oriental Melon (Pyrazolecarboxamide계 살균제 Isopyrazam의 오이 및 참외 중 잔류특성)

  • Han, Ye-Hoon;Lee, Cheol-Yong;Park, Kwee-Doo;Park, Kwang-Wook;Lee, Kyu-Seung
    • The Korean Journal of Pesticide Science
    • /
    • v.17 no.1
    • /
    • pp.13-19
    • /
    • 2013
  • Isopyrazam, a new pyrazolecarboxamide fungicide developed by Syngenta, was highly active against foliar powdery mildew pathogens on cucumber, oriental melon and various vegetables. Following foliar applications on cucumber and oriental melon, crop residues were determined using high performance liquid chromatography. For all studies, limit of quantification was 0.02 mg/kg and minimum detection level was 2.0 ng and recoveries were 83.0-88.0% on cucumber, 92.4-104.5% on oriental melon. Isopyrazam was detected 0.07-0.72 mg/kg on cucumber and < 0.02-0.68 mg/kg on oriental melon, respectively. The TMDI (Theoretical Maximum Dailly Intake) of isopyrazam on cucumber and oriental melon was estimated to less than 1.765% of ADI.

Isolation and Identification of Cytotoxic and Biological Active Toxin from the Puffer Fish Arothron stellatus

  • Veeruraj, Anguchamy;Pugazhvendan, Sampath Renuga;Ajithkumar, Thipramalai Thankappan;Arumugam, Muthuvel
    • Toxicological Research
    • /
    • v.32 no.3
    • /
    • pp.215-223
    • /
    • 2016
  • This study is to investigate the biological, biochemical and cytotoxic effects of puffer fish (Arothron stellatus) toxin extracts under in-vitro condition. Extracted toxins from various organs of puffer fish were purified by using active charcoal column, and Bio-gel-P2 column chromatography. The lethality of toxin was tested in crabs, which consists of neurotoxic compounds. The degree of the brine shrimp lethality assay was found directly proportional to the concentration of the toxin extracts, which was well supported by hemolytic assay. The experimental results suggested that the gonad was found higher toxins than the liver and muscles. The mortality rate of brine shrimp nauplii was increased with the raise of concentrations of toxin level. Among the different doses and time dependent cytotoxic effect of human cervical carcinoma (HeLa) cells were showed $4.0{\mu}g/mL$ of toxin, which was effectively inhibited cancer cell proliferation. HPLC and TLC analysis was revealed that the A. stellatus toxin contains tetrodotoxin (TTX), related compounds 4-epi TTX and anhydro-TTX. The present results suggested that the A. stellatus contain TTX as a major and anh-TTX as a minor toxin. It could be the potential candidate in the field of anticancer drug discovery against human cervical cancer cells. The present data is confirming that the puffer fish toxin as an interesting source of novel bioactive natural compounds with potent applications in pharmacology.

Re-evaluation of physicochemical and NMR data of triol ginsenosides Re, Rf, Rg2, and 20-gluco-Rf from Panax ginseng roots

  • Cho, Jin-Gyeong;In, Seo-Ji;Jung, Ye-Jin;Cha, Byeong-Ju;Lee, Dae-Young;Kim, Yong-Bum;Yeom, Myeonghun;Baek, Nam-In
    • Journal of Ginseng Research
    • /
    • v.38 no.2
    • /
    • pp.116-122
    • /
    • 2014
  • Ginseng roots were extracted with aqueous methanol, and extracts were suspended in water and extracted successively with ethyl acetate and n-butanol. Column chromatography using the n-butanol fraction yielded four purified triol ginseng saponins: the ginsenosides Re, Rf, Rg2, and 20-gluco-Rf. The physicochemical, spectroscopic, and chromatographic characteristics of the ginsenosides were measured and compared with reports from the literature. For spectroscopic analysis, two-dimensional nuclear magnetic resonance (NMR) methods such as $^1H$-$^1H$ correlation spectroscopy, nuclear Overhauser effect spectroscopy, heteronuclear single quantum correlation, and heteronuclear multiple bond connectivity were employed to identify exact peak assignments. Some peak assignments for previously published $^1H$-and $^{13}C$-NMR spectra were found to be inaccurate. This study reports the complete NMR assignment of 20-gluco-Rf for the first time.