Method of detecting pneumocandin compounds

A technology for echinocandin and sample detection, which is applied in the direction of measuring devices, biological testing, material inspection products, etc., can solve the problems of limiting the usefulness of analysis purposes, low stability, and low solubility of echinocandin

Inactive Publication Date: 2012-05-30
克塞里尔制药公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the disadvantage of this method is the low solubility and slightly lower stability of echinocandins in the loading solution
In addition, this mobile phase is not well suited for mass spectrometry, limiting the method's usefulness for analytical purposes

Method used

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  • Method of detecting pneumocandin compounds
  • Method of detecting pneumocandin compounds
  • Method of detecting pneumocandin compounds

Examples

Experimental program
Comparison scheme
Effect test

Embodiment I

[0046] In this experiment, an Agilent 1200 HPLC system coupled to an Agilent 6520 quadrupole time-of-flight (Q-TOF) mass spectrometer was used. The Agilent 1200HPLC system is composed of a binary pump, a degasser, a constant temperature autosampler and a constant temperature column chamber (the temperature is set at 25°C). A Supelco Ascentis Express Hydrophilic Interaction Chromatography (HILIC) 15 cm x 4.6 mm, 2.7 micron column was used. The mobile phase consisted of 15% v / v 0.1% w / w ammonium acetate (pH=4.5) and 85% v / v acetonitrile. The flow rate was 1 ml / min. The parameters of the MS ion source are as follows: the nebulizer pressure is 50 psig, the drying gas flow rate is 10 L / min, the drying gas temperature is 350° C., and the capillary outlet voltage is 250 volts. LC-MS / MS analysis was performed in negative ion mode with deprotonation. Separation of echinocandin B at m / z 1063 in quadrupole (Q) 0 or Echinocandin C 0 . The separated pseudomolecular ions were then fra...

Embodiment II

[0054] In this experiment, a Thermo Fisher Surveyor HPLC system coupled to a Thermo Fisher LXQ linear ion trap mass spectrometer was used. The Surveyor HPLC system is composed of a quaternary pump, a degasser, a constant temperature autosampler and a constant temperature column chamber (the temperature is set at 40°C). A Supelco Ascentis Si Hydrophilic Interaction Chromatography 15 cm x 2.1 mm, 5 micron column was used. The mobile phase consisted of 13% v / v 0.1% w / w ammonium acetate (pH=4.5) and 87% v / v acetonitrile. The flow rate was 0.2 ml / min. The parameters of the MS ion source are as follows: sheath gas 35 (arbitrary units), auxiliary gas 15 (arbitrary units), capillary column temperature 350 °C, spray voltage 5 kV, LC-MS / MS in negative ion mode and deprotonation change.

[0055] echinocandin B 0 or Echinocandin C 0 was isolated (at m / z 1063) and fragmented in the ion trap (with a collision energy of 13). The ion trap was set to scan between m / z 290 to 1100 (Figures...

Embodiment III

[0063]In this experiment, an Agilent 1200 HPLC system coupled to an Agilent 6410 triple quadrupole (QQQ) mass spectrometer was used. The Agilent 1200HPLC system is composed of a binary pump, a degasser, a constant temperature autosampler, and a constant temperature column chamber (the temperature is set at 25°C). A Supelco Ascentis Express Hydrophilic Interaction Chromatography 15 cm x 4.6 mm, 2.7 micron column was used. The mobile phase consisted of 15% v / v 0.1% w / w ammonium acetate (pH=4.5) and 85% v / v acetonitrile. The flow rate was 1 ml / min. The parameters of the MS ion source are as follows: the nebulizer pressure is 50 psig, the drying gas flow rate is 10 L / min, the drying gas temperature is 325° C., and the capillary outlet voltage is 4000 volts. LC-MS / MS was performed in negative ion mode and deprotonation was performed. Separation of echinocandin B at m / z 1063 in the first quadrupole (Q) 0 or Echinocandin C 0 . The separated pseudomolecular ions are then fragmen...

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Abstract

The present invention concerns a method of detecting the antifungal cyclic hexapeptides Pneumocandin B0 and/or Pneumocandin C0 specific fragment is/are detected using MS in negative mode.

Description

technical field [0001] The present invention relates to methods for detecting echinocandin compounds. Background technique [0002] Pneumocandins are antifungal cyclic hexapeptides with lipid side chains (see Schwarts et al., 1992, Journal of Antibiotics, Vol. 45, No. 12, pp. 1853-1866; Masurekar et al. , 1992, Journal of Antibiotics, Vol. 45, No. 12, pp. 1867-1874; Hensens et al., 1992, Journal of Antibiotics, Vol. 45, No. 12, pp. 1875-1885; Schmatz et al. , 1992, Journal of Antibiotics, Vol. 45, No. 12, pp. 1886-1891; and Adefarati et al., 1992, Journal of Antibiotics, Vol. 45, No. 12, pp. 1953-1957; and the United States Patent US 5,021,341). [0003] The antifungal activity of echinocandins is related to the inhibition of 1,3β-glucan biosynthesis. The action of 1,3β-glucan synthase, a multisubunit enzyme, is involved in fungal cell wall construction, septum deposition, and ascospore wall assembly. The catalytic subunit of this enzyme complex has been identified in mo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68
CPCG01N33/6848G01N27/62G01N33/53
Inventor 安德斯·布伦斯维克马丁·曼森
Owner 克塞里尔制药公司
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