• 제목/요약/키워드: normal and reverse-phase chromatography

검색결과 17건 처리시간 0.021초

Isolation and Identification of Nonpolar Taxane Derivatives from the Plant Cell Culture of Taxus chinensis

  • Gi, Un-Sook;Min, Bumchan;Hong, Seung-Suh;Lee, Hyun-Soo;Kim, Jin-Hyun
    • Journal of Applied Biological Chemistry
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    • 제43권3호
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    • pp.176-179
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    • 2000
  • Nonpolar taxoides extracted from a large-scale cell culture of Taxus chinensis were isolated through the normal and reverse phase column chromatographies, and their compounds were identified via NMR spectroscopy. The complete separation method was systematically established and described. In dichloromethane, dissolved paclitaxel and other taxoids with hexane were precipitated during the purification of paclitaxel from the plant cell culture of T. chinensis through a large-scale process while the relatively nonpolar taxane derivatives remained dissolved in the hexane phase. 13-Deoxy baccatin III (I), baccatin VI (II), taxchinin I (III), $2{\alpha}$, $5{\alpha}$, $10{\beta}$, $14{\beta}$-tetraacetoxy-4(20), 11-taxadiene(IV), 1-deoxy baccatinVI(V), and taxayuntin C (VI) were isolated through column chromatography and identified via NMR spectroscopy. Compounds I and IV were found to the major components, aside from paclitaxel, in the plant cell culture of T. chinensis. The concentrations of I and IV were compared with the that concentration of the paclitaxel in each of plant cell culture. The possible applications of compounds I, II, IV, and V were discussed.

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Purification and Identification of Antioxidant Peptides from Enzymatic Hydrolysate of Spirulina platensis

  • Yu, Jie;Hu, Yuanliang;Xue, Mingxiong;Dun, Yaohao;Li, Shenao;Peng, Nan;Liang, Yunxiang;Zhao, Shumao
    • Journal of Microbiology and Biotechnology
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    • 제26권7호
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    • pp.1216-1223
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    • 2016
  • The aim of this study was to isolate antioxidant peptides from an enzymatic hydrolysate of Spirulina platensis. A novel antioxidant peptide was obtained by ultrafiltration, gel filtration chromatography, and reverse-phase high-performance liquid chromatography, with the 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging assay used to measure the antioxidant activity, and the sequence was determined to be Pro-Asn-Asn (343.15 Da) by electrospray ionization tandem mass spectrometry. This peptide was synthesized to confirm its antioxidant properties, and it exhibited 81.44 ± 0.43% DPPH scavenging activity at 100 μg/ml, which was similar to that of glutathione (82.63 ± 0.56%). Furthermore, the superoxide anion and hydroxyl free-radical scavenging activities and the SOD activity of the peptide were 47.84 ± 0.49%, 54.01 ± 0.82%, and 12.55 ± 0.75%, respectively, at 10 mg/ml. These results indicate that S. platensis is a good source of antioxidant peptides, and that its hydrolysate may have important applications in the pharmaceutical and food industries.

Simultaneous Determination of Vitamin A and E in Infant Formula by HPLC with Photodiode Array Detection

  • Lee, Hong-Min;Kwak, Byung-Man;Ahn, Jang-Hyuk;Jeong, Seung-Hwan;Shim, Sung-Lye;Kim, Kyong-Su;Yoon, Tae-Hyung;Leem, Dong-Gil;Jeong, Ja-Young
    • 한국축산식품학회지
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    • 제31권2호
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    • pp.191-199
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    • 2011
  • The objective of this study was to develop a method to simultaneously quantify vitamins A and E in infant formula. To determine the vitamin A and E content, vitamin A and four different vitamin E isomers (${\alpha}$-, ${\beta}$-, ${\gamma}$-, and ${\delta}$-tocopherol) were separated by high performance liquid chromatography with a photodiode array detector using a Develosil RPAQUEOUS RP-$C_{30}$ column ($4.6{\times}250$ mm, 5 ${\mu}M$). The vitamin A and E contents in the certified reference material determined using this method were within the certified range of standard values. The limits of detection (LODs) and limits of quantitation (LOQs) for vitamin A were 0.02 and 0.06 ${\mu}g/L$, respectively. LODs and LOQs for the vitamin E isomers ranged from 0.20 to 0.55 and from 0.67 to 1.81 ${\mu}g/L$, respectively. Linear analyses indicated that the square of the correlation coefficient for the vitamin A and E isomers was 0.9997-0.9999. The recovery of vitamins ranged from 96.69 to 97.79%. The results demonstrate that this novel method could be used to reliably analyze vitamin A and E content in infant formula.

Streptomyces sp. J46의 세균성구멍병원균 Xanthomonas arboricola pv. pruni에 대한 항균 활성 (Antibacterial Activity of Streptomyces sp. J46 against Bacterial Shot Hole Disease Pathogen Xanthomonas arboricola pv. pruni)

  • 이정은;임다정;김인선
    • 한국환경농학회지
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    • 제40권1호
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    • pp.20-32
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    • 2021
  • BACKGROUND: Bacterial shot hole of stone fruits is a seriuos plant disease caused by Xanthomonas arboricola pv. pruni (Xap). Techniques to control the disease are required. In this study, microorganisms with antibacterial activity were isolated to develop as a microbial agent against the bacterial shot hole. METHODS AND RESULTS: An isolate with the strongest activity among the isolates was identified as Streptomyces avidinii based on 16S rRNA gene sequence analysis and designated Streptomyces sp. J46. J46 showed suppression of bacterial leaf spot with a control value of 90% at 10 times-diluted cell free supernatant. To investigate antibacterial metabolites produced by J46, the supernatant of J46 was extracted with organic solvents, and the extracts were subjected to chromatography works. Antibacterial metabolites were not extractable with organic solvents. Both reverse and normal phase techniques were not successful because the metabolites were extremely water soluble. The antibacterial metabolites were not volatiles but protein compounds based on hydrolysis enzyme treatment. CONCLUSION: Our study suggests that Streptomyces sp. J46 may be a potential as an microbial agent against bacterial shot hole. Further study to identify the metabolites is required in more detail.

국상(Kuksang) 뽕잎(Morus alba L.)으로부터 α-amylase와 α-glucosidase 저해 물질 분리 및 동정 (Isolation and Identification of Inhibitory Compounds from Morus alba cv. Kuksang on α-amylase and α-glucosidase)

  • 최무영;조영제
    • 생명과학회지
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    • 제25권8호
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    • pp.870-879
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    • 2015
  • 109종의 뽕잎 종류 중 국상 품종 뽕잎으로부터 α-amylase와 α-glucosidase 저해작용을 가지는 페놀성화합물을 분리하여 저해활성을 검토하였다. 국상 뽕잎의 물추출물에서 α-amylase와 α-glucosidase 저해 활성이 각각 93.8%와 48.7%를 나타내었다. 국상뽕잎 추출물의 페놀화합물의 함량은 물 추출물에서 9.7±0.2 mg/g, 60% ethanol 추출물에서 14.3±0.2 mg/g으로 나타났으며, 국상뽕잎 물 추출물 200 μg/ml의 농도에서 α-amylase와 α-glucosidase 저해활성이 각각 100%와 82.6%로 나타났다. 국상뽕잎으로부터 α-amylase와 α-glucosidase 저해활성을 가지는 천연물을 분리하기 위하여 국상뽕잎 물추출물을 Sephadex LH-20과 MCI-gel을 이용하여 normal phase type인 EtOH→H2O와 reverse phase type인 H2O→MeOH로 유기용매의 농도를 증가시키며 gradient로 용출하여 정제하였다. α-Amylase와 α-glucosidase 저해활성을 가지는 정제물은 FAB-MS, NMR과 IR spectrum을 구조 분석한 결과 quercetin으로 동정되었다.

Isolation, Purification, and Identification of Taxol and Related Taxanes from Taxol-Producing Fungus Aspergillus niger subsp. taxi

  • Li, Dan;Fu, Dongwei;Zhang, Yue;Ma, Xueling;Gao, Liguo;Wang, Xiaohua;Zhou, Dongpo;Zhao, Kai
    • Journal of Microbiology and Biotechnology
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    • 제27권8호
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    • pp.1379-1385
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    • 2017
  • The content of taxol in the bark of yews is very low, and this is not affordable from the environmental point of view. Thus, it is a necessity to look for alternative sources of taxol production to solve its supply. Currently, a large portion of the taxol in the market comes from chemical semi-synthesis, but the semi-synthetic precursors such as baccatin III and 10-deacetyl-baccatin III are extracted from needles and twigs of yew trees. Taxol-producing fungi as a renewable resource is a very promising way to increase the scale of taxol production. Our group has obtained a taxol-producing endophytic fungus, Aspergillus niger subsp. taxi HD86-9, to examine if A. niger can produce the taxanes. Six compounds from the fermentation broth of strain HD86-9 were isolated and identified by $^1H$ NMR, $^{13}C$ NMR, and ESI-MS. The results showed that the six compounds included four taxane diterpenoids (taxol, cephalomannine, baccatin III, and 10-deacetyl-baccatin III) and two non-taxane compounds (${\beta}-sitosterol$ and flavonoid isovitexin). The study verified that the taxanes can be produced by the A. niger, which is very important to taxol production via chemical semi-synthesis. Additionally, the finding is potentially very significant to solve the taxol semi-synthetic precursors extracted from needles and twigs of yew trees, and the precursor production can be easily increased through the culture condition optimization, genetic breeding, and metabolic engineering of the A. niger.

HPLC/DAD/ESI-MS를 이용한 혈장시료 중 갑상선 호르몬 분석 (Determination of thyroid hormones in plasma samples by high performance liquid chromatograph/diode array detector/electrospray ionization mass spectrometer)

  • 곽선영;문명희;표희수
    • 분석과학
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    • 제20권5호
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    • pp.424-433
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    • 2007
  • 혈장시료 중의 갑상선 호르몬을 고체상 추출법을 이용하여 추출한 후 HPLC/DAD/ESI-MS (high-performance liquid chromatograph/diode array detector/electrospray ionization-mass spectrometer)를 사용하여 분석하였다. 7종의 thyroid hormones의 HPLC 분리조건은 Hypersil ODS 컬럼(4.6 mm I.D., 250 mm length, particle size $5{\mu}m$)을 사용하고 ammonium formate buffer와 아세토나이트릴을 이동상으로 하여 기울기 용리한 결과 완전 분리가 가능하였으며, UV spectra 및 질량스펙트럼을 확인할 수 있었다. 고체상추출법에 의한 전처리 최적 조건을 조사한 결과 시료를 pH 3으로 한 후 C18 고체상을 사용하여 4 mL의 메탄올로 용리할 경우 회수율이 74.5-115.7%로 나타났다. HPLC/ESI-MS를 이용하여 20-500 ng/mL범위에서 검량선을 작성한 결과 $r^2$값은 0.9939-0.9978으로 나타났으며 검출한계는 20-50 ng/mL (38.1-162.8 pmol/mL)로 계산되었다. 정상인의 혈장에서 thyroxine의 정량이 가능하였으며 그 결과 50.98-112.97 ng/mL로 검출되었다.