• Title/Summary/Keyword: HPLC resolution

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Enantiomer Separation of α-Amino Acid Esters as Nitrobenzoxadiazole Derivatives Using Chiral Columns (키랄 컬럼을 사용한 아미노산 에스테르의 니트로벤조옥사디아졸 유도체의 광학분리)

  • Yun, Won Nam;Kim, Ji Yeon;Lee, Wonjae
    • KSBB Journal
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    • v.28 no.6
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    • pp.423-427
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    • 2013
  • A new convenient derivatization method of ${\alpha}$-amino acid esters as nitrobenzoxadiazole (NBD) derivatives for chiral resolution was introduced and the enantiomer separation of ${\alpha}$-amino acid esters as NBD derivatives was performed by normal HPLC using chiral columns based on polysaccharide derivatives. The NBD derivatives were readily prepared by stirring NBD-Cl and ${\alpha}$-amino acid methyl ester HCl with $NaHCO_3$ in ethanol. The performance of Chiralpak IA was superior to the other chiral stationary phases for enantiomer resolution of NBD derivatives of several ${\alpha}$-amino acid methyl esters. Owing to fluorescence detection as well as strong UV absorption, it is expected that the convenient analytical method developed in this study will be very useful for enantiomer separation of ${\alpha}$-amino acid esters as NBD derivatives on polysaccharide-derived chiral columns.

Studies on the Separation of Immunoglobulin and Immunological Response from Korean Native Cattle (한우초유중 Ig의 분리 ${\cdot}$ 정제 및 면역 반응에 관한 연구)

  • Baick, Seung-Cheon;Kim, Yong-Hwi;Shin, Je-Ho;Yu, Je-Hyeon
    • Journal of Dairy Science and Biotechnology
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    • v.15 no.1
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    • pp.1-9
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    • 1997
  • This study was conducted to efficiently separate the Ig from Korean native cattle colostrum and to utilize them as an immunogen for the production of antibodies aginst rabbit. The results obtained were as follows : 1. About 84% of Ig G could be separated from Korean native cattle colostrum by·gel filtration using Superose 12 column on HPLC. The separation profile of Korean native cattle colostral immunoglobulin was similar that of Holstein colostral Ig. 2. Separation of Korean native cattle colostral Ig by anion exchange chromatography using Mono Q column on HPLC was poor resolution chromatographic pattern. 3. Hi-Trap Protein G column showed better results than the Protein A Sepharose CL-4B column in the Ig G binding capacity from Korean native cattle colostral Ig. 4. Protein G Sepharose Fast Flow system resulted in higher Ig g binding capacity as the industrial size scale-up approach. 5. Sufficient titer reaction of antibody to Korean native cattle colostral Ig G was confirmed by ELISA.

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Determination of Isomaltooligosaccharides in Yoghurts by Using HPLC-ELSD (HPLC-ELSD를 이용한 발효유 제품 중의 Isomaltooligosaccharides 분석법 개발)

  • Ko, Jinhyouk;Lee, Moon-Seok;Kwak, Byung-Man;Ahn, Jang-Hyuk;Park, Jong-Su;Kwon, Joong-Ho
    • Food Science of Animal Resources
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    • v.33 no.3
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    • pp.417-424
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    • 2013
  • A rapid and simple analytical method for the determination of 9 isomaltooligosaccharides (IMO) species in yoghurts was developed using dispersive solid phase extraction (dSPE) clean-up technic and high performance liquid chromatography with evaporative light-scattering detector (HPLC-ELSD). In this study, 9 IMO were extracted from samples simply with chemical reagent using ISO22662 IDF198 method and additional dSPE clean-up. The optimum instrument conditions for the determination were used carbohydrate ES $5{\mu}$ column with gradient elution of water and acetonitrile and ELS detector. The linearity of this method was expressed as the correlation coefficient ($r^2$), the results of IMO 9 species were shown in 0.9999. LOD and LOQ were respectively 7.9-22.1 mg/kg, 25.9-72.8 mg/kg. The accuracy of intra- and inter-day measurements were in the range from $84.3{\pm}4.5$ to $104.9{\pm}6.5%$, and the preceision of the intra- and inter-day measurements were in the range from 0.8 to 7.7%. The recoveries were from $84.3{\pm}4.5$ to $104.9{\pm}6.5%$. The determination results of IMO 9 species for the 9 yoghurts circulated in the market were in the range from $0.317{\pm}0.007$ to $1.624{\pm}0.050$ g/100 g. The newly developed method is appropriate for the determination of IMO in yoghurts, is a rapid and simple method with excellent resolution in compared with previous method.

Facile Synthesis of 2',5'-Dideoxy-, 2',3'-Dideoxy- and 3'-Deoxy-1, N6-ethenoadenosine Nucleosides

  • Chae, Whi-Gun
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.4 no.1
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    • pp.17-20
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    • 1999
  • Facile synthetic methods of 2',5'-dideoxy-, 2',3'-dideoxy- and 3'-deoxy-1, N6-ethenoadenosine nucleosides by either an enzymatic dideoxyribosyl transfer reaction or a simple chemical reaction were proposed. The synthesis products were isolated and purified by preparative HPLC and their structures were confirmed by 1H NMR (500 MHz) and FAB-MS including high resolution mass measurement. These modified nucleoside analogs have not been reported yet. Therefore, these modified nucleoside analogs are of potential value to be studied further for biological activity such as anticancer or antiviral.

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Studies on the Development of the Resources of Korean Crude Drugs (II) -Amygdalin Contents in the Seeds of Some Prunus spp. in Korea- (한국산(韓國産) 생약자원(生藥資源) 개발연구(開發硏究) (제2보)(第2報) -Prunus tomentosa의 약효성분에 관하여-)

  • Choi, Eung-Chil;Han, Dae-Suk;Kim, Young-Ho;Hong, In-Hyuk
    • Korean Journal of Pharmacognosy
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    • v.14 no.2
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    • pp.64-66
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    • 1983
  • HPLC is a useful method for identifying amygdalin which has been the subject of considering controversy for cancer remedy. The resolution of amygalin was satisfactory and the separation of amygdalin was accomplished using carbohydrate analysis column, mobile phase of acetonitrile: $H_2O$ (90:10), and UV detector. Quantitative analysis showed that amygdalin contents of Prunus tomentosa, P. serrulata, and P. ishidoyana (Amygdalaceae) were 2.18, 2.41, and 2.26%, respectively. It was suggested that the seed of Prunus species was worth useful resource of amygdalin.

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HPLC를 이용한 ketoprofen racemate의 분리

  • Yun, Tae-Ho;Kim, Hyeong-Won;Kim, In-Ho
    • 한국생물공학회:학술대회논문집
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    • 2001.11a
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    • pp.693-696
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    • 2001
  • Ketoprofen racemate is very useful pharmaceutical intermediate, but the hardness of separation to each enantiomer is most a barrier to apply pharmaceutical industry. To overcome this difficulty, we would like to separate S-ketoprofen from ketoprofen racemate. To design suitable mobile phase, hexane and t-BME(tri-buthyl-methyle-ether) were used to elute peaks of ketoprofen racemate separately. When the ratio of hexane/t-BME/acetic acid was 60/40/0.1(%v/v), each component was separated successfully. In order to separate large amount of S-ketoprofen, sample concentration was increased from 100ppm to 2000ppm. In this case, resolution was decreased due to the overlapping of peaks(1.65${\rightarrow}$0.94).

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Hydroxylation of Resveratrol with DoxA In Vitro: An Enzyme with the Potential for the Bioconversion of a Bioactive Stilbene

  • Rimal, Hemraj;Yu, Sang-Cheol;Lee, Joo-Ho;Yamaguchi, Tokutaro;Oh, Tae-Jin
    • Journal of Microbiology and Biotechnology
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    • v.28 no.4
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    • pp.561-565
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    • 2018
  • The late-stage doxorubicin biosynthesis pathway acting enzyme (DoxA) from Streptomyces peucetius CYP129A2 exhibited substrate promiscuity towards the stilbene group of compounds such as resveratrol. DoxA along with two accessory enzymes ferrdoxin reductase and ferredoxin from spinach hydroxylated resveratrol at the 3'-position in vitro to produce piceatannol. The product was identified by HPLC-PDA and high-resolution HR-qTOF-ESI/MS analyses in positive mode. The ESI/MS fragments resembled the hydroxylated product of resveratrol.

Chromatographic chiral resolution of several racemic drugs containing primary amino moiety on a chiral stationary phase

  • Lee, Won-Jae;Baek, Chae-Sun;Jin, Jing-Yu
    • Proceedings of the PSK Conference
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    • 2003.04a
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    • pp.278.3-279
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    • 2003
  • A chiral stationary phase (CSP) prepared by bonding (18-crown-6)-2,3,11,12-tetracarboxylic acid (18-C-6-TA) to aminopropyl silica gel by HPLC was used in resolving several racemic drugs containing primary amino moiety. Most compounds used in this study were resolved on the CSP using 80% methanol in water (V/V) containing 10mM sulfuric acid as a mobile phase. These results on the CSP were compared to those on the similar CSP derived from 18-C-6-TA of the same chiral selector by different connecting method.

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High Temperature Size Exclusion Chromatography for High Throughput Analysis

  • Chang, Tai-Hyun;Park, Soo-Jin;Cho, Hee-Sook;Kim, Young-Tak;Ihm, Kyu-Hyun
    • Proceedings of the Polymer Society of Korea Conference
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    • 2006.10a
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    • pp.202-202
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    • 2006
  • With a modern size exclusion chromatography (SEC) column, molecular weight analysis of a polymer sample takes about 10 min. However, it is desirable to reduce the analysis time further, in particular, for high throughput measurements required in combinatorial analyses or 2D-HPLC analyses. We implemented the high temperature SEC for the purpose. By inserting a narrow bore tubing between the column and the detector, a sufficient backpressure can be maintained to prevent the mobile phase from boiling and the effluent is cooled down enough when it reaches the detector. Therefore, a normal SEC detector can be used without any modification. The SEC resolution is greatly improved at the elevated temperature at high flow rate which allows high speed operation.

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Metabolomics Approach for Classification of Medicinal Plants

  • Lee, Dong-Ho
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2010.05a
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    • pp.5-5
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    • 2010
  • Selection of specific medicinal sources as well as bioactive compounds is important for the preparation of medicine and related products with good quality. It is necessary to pay close attention for choosing correct medicinal sources, particularly in case of medicinal plants, because of their diversity, which can affect the quality and efficacy of medicine. Discrimination of plants based on morphological or genetic characteristics has been used as a conventional classification method of pharmaceutical sources so far; however, more need demands more general methods for accurate quality assessment of medicinal plants. In this study, ultra performance liquid chromatography/quadrupole time-of-flight mass spectrometry (UPLC/Q-TOF MS) technique applied to this metabolic profiling is a powerful tool due to its higher sensitivity, resolution, and speed compared to conventional HPLC technique. The metabolite profiling of several medicinal plants including Panax ginseng was carried out using UPLC/Q-TOF MS and total metabolites were then subsequently applied to various statistical tools to compare the patterns. The developed metabolomics tool with UPLC/Q-TOF MS successfully identified and classified the samples tested according to their origins.

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