Purification and Quantitative Analysis of Destruxins from Meterhizium anisopliae by HPLC

Cheng Ming Liu, Shyuan Shuenn Huang, Yew Min Tzeng

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

Destruxins are metabolic toxins secreted from a fungus, Metarhizium anisopliae, during growth. The structure of destruxins has been identified as a cyclic hexadepsipeptide. Destruxins exhibit insecticidal activities. More than 35 different destruxins have been characterized to have a wide range of activity and specificity. In this report, an extraction solvent, acetonitrile, was found to yield high extraction efficiency. The fermentation broth was mixed with equal volume of acetonitrile and 5% NaCl, which formed two layers. The upper organic solvent layer, containing 80-95% of destruxins, was easily crystallized by lyophilization. The crystal could be redissolved with acetonitrile and separated by semi-preparative HPLC. Each fraction was analyzed and identified with fast atom bombardment mass (FAB) spectrometry. The method for destruxin separation was optimized by adjusting the acetonitrile/water ratio in the gradient elution buffer of the semi-preparative HPLC. Flatter gradient slopes may result in longer retention time and allow for higher quantity of separation. The quantitative analysis of destruxin could be achieved by generating a linear regression curve between concentrations vs. peak areas.

Original languageEnglish
Pages (from-to)1013-1025
Number of pages13
JournalJournal of Liquid Chromatography and Related Technologies
Volume27
Issue number6
DOIs
Publication statusPublished - 2004

Fingerprint

Purification
High Pressure Liquid Chromatography
Chemical analysis
Metarhizium
Fast Atom Bombardment Mass Spectrometry
Freeze Drying
Solvent extraction
Fungi
Linear regression
Fermentation
Organic solvents
Mass spectrometry
Linear Models
Buffers
Atoms
Crystals
acetonitrile
Water
Growth

Keywords

  • Acetonitrile extraction
  • Destruxins
  • Metarhizium anisopliae
  • Semi-preparative HPLC

ASJC Scopus subject areas

  • Analytical Chemistry
  • Clinical Biochemistry

Cite this

Purification and Quantitative Analysis of Destruxins from Meterhizium anisopliae by HPLC. / Liu, Cheng Ming; Huang, Shyuan Shuenn; Tzeng, Yew Min.

In: Journal of Liquid Chromatography and Related Technologies, Vol. 27, No. 6, 2004, p. 1013-1025.

Research output: Contribution to journalArticle

Liu, Cheng Ming ; Huang, Shyuan Shuenn ; Tzeng, Yew Min. / Purification and Quantitative Analysis of Destruxins from Meterhizium anisopliae by HPLC. In: Journal of Liquid Chromatography and Related Technologies. 2004 ; Vol. 27, No. 6. pp. 1013-1025.
@article{eb3df4befed64150b3f8186111f36251,
title = "Purification and Quantitative Analysis of Destruxins from Meterhizium anisopliae by HPLC",
abstract = "Destruxins are metabolic toxins secreted from a fungus, Metarhizium anisopliae, during growth. The structure of destruxins has been identified as a cyclic hexadepsipeptide. Destruxins exhibit insecticidal activities. More than 35 different destruxins have been characterized to have a wide range of activity and specificity. In this report, an extraction solvent, acetonitrile, was found to yield high extraction efficiency. The fermentation broth was mixed with equal volume of acetonitrile and 5{\%} NaCl, which formed two layers. The upper organic solvent layer, containing 80-95{\%} of destruxins, was easily crystallized by lyophilization. The crystal could be redissolved with acetonitrile and separated by semi-preparative HPLC. Each fraction was analyzed and identified with fast atom bombardment mass (FAB) spectrometry. The method for destruxin separation was optimized by adjusting the acetonitrile/water ratio in the gradient elution buffer of the semi-preparative HPLC. Flatter gradient slopes may result in longer retention time and allow for higher quantity of separation. The quantitative analysis of destruxin could be achieved by generating a linear regression curve between concentrations vs. peak areas.",
keywords = "Acetonitrile extraction, Destruxins, Metarhizium anisopliae, Semi-preparative HPLC",
author = "Liu, {Cheng Ming} and Huang, {Shyuan Shuenn} and Tzeng, {Yew Min}",
year = "2004",
doi = "10.1081/JLC-120030175",
language = "English",
volume = "27",
pages = "1013--1025",
journal = "Journal of Liquid Chromatography and Related Technologies",
issn = "1082-6076",
publisher = "Taylor and Francis Ltd.",
number = "6",

}

TY - JOUR

T1 - Purification and Quantitative Analysis of Destruxins from Meterhizium anisopliae by HPLC

AU - Liu, Cheng Ming

AU - Huang, Shyuan Shuenn

AU - Tzeng, Yew Min

PY - 2004

Y1 - 2004

N2 - Destruxins are metabolic toxins secreted from a fungus, Metarhizium anisopliae, during growth. The structure of destruxins has been identified as a cyclic hexadepsipeptide. Destruxins exhibit insecticidal activities. More than 35 different destruxins have been characterized to have a wide range of activity and specificity. In this report, an extraction solvent, acetonitrile, was found to yield high extraction efficiency. The fermentation broth was mixed with equal volume of acetonitrile and 5% NaCl, which formed two layers. The upper organic solvent layer, containing 80-95% of destruxins, was easily crystallized by lyophilization. The crystal could be redissolved with acetonitrile and separated by semi-preparative HPLC. Each fraction was analyzed and identified with fast atom bombardment mass (FAB) spectrometry. The method for destruxin separation was optimized by adjusting the acetonitrile/water ratio in the gradient elution buffer of the semi-preparative HPLC. Flatter gradient slopes may result in longer retention time and allow for higher quantity of separation. The quantitative analysis of destruxin could be achieved by generating a linear regression curve between concentrations vs. peak areas.

AB - Destruxins are metabolic toxins secreted from a fungus, Metarhizium anisopliae, during growth. The structure of destruxins has been identified as a cyclic hexadepsipeptide. Destruxins exhibit insecticidal activities. More than 35 different destruxins have been characterized to have a wide range of activity and specificity. In this report, an extraction solvent, acetonitrile, was found to yield high extraction efficiency. The fermentation broth was mixed with equal volume of acetonitrile and 5% NaCl, which formed two layers. The upper organic solvent layer, containing 80-95% of destruxins, was easily crystallized by lyophilization. The crystal could be redissolved with acetonitrile and separated by semi-preparative HPLC. Each fraction was analyzed and identified with fast atom bombardment mass (FAB) spectrometry. The method for destruxin separation was optimized by adjusting the acetonitrile/water ratio in the gradient elution buffer of the semi-preparative HPLC. Flatter gradient slopes may result in longer retention time and allow for higher quantity of separation. The quantitative analysis of destruxin could be achieved by generating a linear regression curve between concentrations vs. peak areas.

KW - Acetonitrile extraction

KW - Destruxins

KW - Metarhizium anisopliae

KW - Semi-preparative HPLC

UR - http://www.scopus.com/inward/record.url?scp=1842479790&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=1842479790&partnerID=8YFLogxK

U2 - 10.1081/JLC-120030175

DO - 10.1081/JLC-120030175

M3 - Article

AN - SCOPUS:1842479790

VL - 27

SP - 1013

EP - 1025

JO - Journal of Liquid Chromatography and Related Technologies

JF - Journal of Liquid Chromatography and Related Technologies

SN - 1082-6076

IS - 6

ER -