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An immunoassay method and kit to leucomalachite green and malachite green

A technology for leuco malachite green and malachite green, which is applied in the field of immunoassay methods and kits for detecting or measuring leuco malachite green and malachite green, and can solve the problems of time-consuming experiment errors and the like

Inactive Publication Date: 2008-03-26
RANDOX LAB LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This makes the immunoassay more time consuming and more prone to experimental error than the present invention due to the use of additional steps

Method used

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  • An immunoassay method and kit to leucomalachite green and malachite green
  • An immunoassay method and kit to leucomalachite green and malachite green
  • An immunoassay method and kit to leucomalachite green and malachite green

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Experimental program
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Effect test

Embodiment 1

[0055] Embodiment 1: Preparation p-[3-(ethyl carboxyl) propoxy] benzaldehyde (2)

[0056] To a suspension of 4-hydroxybenzaldehyde, 1, (20 g, 164 mMol) and potassium carbonate (68 g, 492 mMol) in acetonitrile (300 ml) was added ethyl 4-bromobutyrate (35.2 ml, 246 mMol). The mixture was then heated to reflux overnight. After cooling to room temperature, the solid was filtered off and the solvent was removed in vacuo. Water (200ml) was then added to the residue and extracted with ethyl acetate (3x200ml). The combined organic phases were washed with brine (1x200ml), dried over sodium sulfate, filtered and evaporated to dryness. The resulting residue was purified by flash chromatography on silica gel using 30% ethyl acetate in hexane as eluent to give 2 (27 g, 69.8%) as a colorless oil.

[0057] IR spectrum (film): 1733.7, 1692.5, 1601.2, 1577.9, 1509.8, 1257.7, 1161.1cm -1 .

Embodiment 2

[0058] Embodiment 2: Preparation of [3-(ethyl carboxyl) propoxy] leuco malachite green (3)

[0059] To the mixture of p-[3-(ethylcarboxy)propoxy]benzaldehyde 2 (10g, 42.4mMol) and N,N-dimethylaniline (12.38g, 106mMol), concentrated sulfuric acid (2ml) was added, and the mixture Heat at 120°C for 48 hours. The solution was then cooled to room temperature, water (200ml) was added, the mixture was neutralized with solid sodium carbonate, extracted with ethyl acetate (3x100ml), washed with brine (1x100ml), dried over sodium sulfate, filtered and concentrated to dryness in vacuo. The resulting crude product was purified by flash chromatography on silica gel using 25% ethyl acetate in hexanes as eluent to give ester 3 (6.9 g, 35%) as an orange oil.

[0060] IR spectrum (film): 1735.3, 1612.2, 1518.3, 1242.6cm -1 .

Embodiment 3

[0061] Example 3: Preparation of p-(3-carboxypropoxy) leuco malachite green (hapten A)

[0062] To a solution of ester, 3, (8.4g, 18.75mMol) dissolved in a mixture of methanol and THF (2:1, 240ml) was added sodium hydroxide (2N, 40ml). The mixture was stirred at room temperature for 16 hours. The solvent was removed in vacuo and water (100ml) was added. The solution was neutralized to pH 7 by adding HCl (2N) and the resulting precipitate was collected by filtration and dried in a desiccator over phosphorus pentoxide. Hapten A was obtained as a blue / green solid (3.2 g, 40%).

[0063] NMR 13 C, solvent δ-MeOH (Figure 5): 178.8, 156.9, 148.9, 137.7, 133.6, 130.6, 129.9, 114.1, 112.9, 66.6, 54.2, 40.9, 30.9, 24.6.

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Abstract

The invention provides haptens, immunogens comprising such haptens coupled to an antigenicity-conferring carrier material, conjugates comprising such haptens bonded to a labelling agent as well as, antibodies raised against such immunogens and capable of binding with leucomalachite green and malachite green.

Description

field of invention [0001] The present invention relates to novel immunogens, antibodies and conjugates for use in immunoassays for the detection and / or determination of leucomalachite Green and Malachite Green. The present invention also relates to an immunoassay method and a kit for detecting or measuring leuco malachite green and malachite green. "Detecting" refers to the definitive analysis of the presence or absence of leuco malachite green and / or malachite green in a sample. "Determining" refers to the quantitative analysis of the amount of leuco malachite green and / or malachite green in a sample. Background of the invention [0002] Malachite green is a topical insect repellent, fungicide and fungicide used in fish farming. However, it is a potential carcinogen, mutagen and teratogen. Illegal and public safety use in edible fish such as trout and eel has been recognized since 1933. After absorption, malachite green is rapidly reduced to its chromophore-free metabol...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/53G01N33/566
CPCG01N33/84G01N33/53
Inventor 埃尔奥亚特·本奇克罗伯特·I·麦康奈尔斯蒂芬·P·菲茨杰拉德安德鲁·P·劳里
Owner RANDOX LAB LTD
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