J. P. Jacobsen

Nägele, Horst

doi:10.1007/978-3-476-03835-7

Cited by 2 publications

(2 citation statements)

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“…Twodimensional HILIC coupled with RP chromatography has been proved to be very useful in the separation of Stevia rebaudiana Bertoni extracts [7]. SCX/RPLC shows excellent performance in the 2D preparation of protein digestion [8]. In most of the 2D-HPLC preparation methods, water is usually taken as one of the mobile phase because RPLC is so widely used.…”

Section: Introductionmentioning

confidence: 99%

A novel two‐dimensional preparative chromatography method designed for the separation of traditional animal Tibetan medicine Osteon Myospalacem Baileyi

Jiang

Tao

Wang

et al. 2014

J of Separation Science

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Animal medicine is an important part in traditional Tibetan medicine. However, information about the chemical composition of animal medicine is very limited, and there is a lack of comprehensive chromatographic purification methods. In the present work, animal medicine Osteon Myospalacem Baileyi was taken as an example and a novel two-dimensional preparative chromatographic method was established for the preparation of single compounds with high purity from the extract of Osteon Myospalacem Baileyi. The first-dimension preparation was carried on a DAISO Silica prep column, and ten fractions were obtained from the 112.3 g crude sample within 12 injections. A diol prep column used in nonaqueous mobile phase was selected for the second-dimension preparation. The purity of the compounds isolated from the crude extract was >98%, which indicated that the method built in this work was efficient to manufacture single compounds of high purity from the extract of Osteon Myospalacem Baileyi. Additionally, this method showed great potential in the purification of weakly polar chemicals and it could act as a good example in the purification of other traditional animal medicines.

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Section: Introductionmentioning

confidence: 99%

A novel two‐dimensional preparative chromatography method designed for the separation of traditional animal Tibetan medicine Osteon Myospalacem Baileyi

Jiang

Tao

Wang

et al. 2014

J of Separation Science

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“…In the present study, we are employing a gel-free approach for separation of E. coli cell lysates. Proteome analyses of E. coli have been performed before using 2-D nanoLC using strong cation-exchange and RP-HPLC, both on the digested peptide mixtures, followed by mass spectrometric analysis of the proteins on an ESI-IT instrument [32,33]. In contrast to this approach, we are employing two RP chromatographic separation steps, one normal-flow C4 HPLC on the protein level and one nanoHPLC (C18) on the level of proteolytic peptides, followed by nanoESI FT-ICR MS analysis and identification of E. coli proteins.…”

Section: Introductionmentioning

confidence: 99%

Proteome analysis ofEscherichia coli using high-performance liquid chromatography and Fourier transform ion cyclotron resonance mass spectrometry

Ihling¹,

Sinz²

2005

Proteomics

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The basic problem of complexity poses a significant challenge for proteomic studies. To date two-dimensional gel electrophoresis (2-DE) followed by enzymatic in-gel digestion of the peptides, and subsequent identification by mass spectrometry (MS) is the most commonly used method to analyze complex protein mixtures. However, 2-DE is a slow and labor-intensive technique, which is not able to resolve all proteins of a proteome. To overcome these limitations gel-free approaches are developed based on high performance liquid chromatography (HPLC) and Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR MS). The high resolution and excellent mass accuracy of FT-ICR MS provides a basis for simultaneous analysis of numerous compounds. In the present study, a small protein subfraction of an Escherichia coli cell lysate was prepared by size-exclusion chromatography and proteins were analyzed using C4 reversed phase (RP)-HPLC for pre-separation followed by C18 RP nanoHPLC/nanoESI FT-ICR MS for analysis of the peptide mixtures after tryptic digestion of the protein fractions. We identified 231 proteins and thus demonstrated that a combination of two RP separation steps - one on the protein and one on the peptide level - in combination with high-resolution FT-ICR MS has the potential to become a powerful method for global proteomics studies.

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J. P. Jacobsen

Cited by 2 publications

References 0 publications

A novel two‐dimensional preparative chromatography method designed for the separation of traditional animal Tibetan medicine Osteon Myospalacem Baileyi

A novel two‐dimensional preparative chromatography method designed for the separation of traditional animal Tibetan medicine Osteon Myospalacem Baileyi

Proteome analysis ofEscherichia coli using high-performance liquid chromatography and Fourier transform ion cyclotron resonance mass spectrometry

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