Method for detecting a radical compound derived from a lipid and a compound derived from the radical compound

Patent metadata

Field	Value
Publication number	US-2018328951-A1
Application number	US-201815965096-A
Country	US
Kind code	A1
Filing date	Apr 27, 2018
Priority date	Apr 28, 2017
Publication date	Nov 15, 2018
Grant date	—

Abstract

[Means for solution] A fluorescent nitroxide (NBD-Pen) is made to act to scavenge a lipid radical or a fragment radical thereof, and fluorescent detection liquid chromatography (LC/FL) and mass spectrometry (MS) are combined to identify lipid-derived radicals contained in a lipid extract.

First claim

1 . A method for detecting a radical compound derived from a lipid present in a sample, the method comprising:
a step of obtaining a nitroxide derivative mixture by adding a nitroxide derivative selected from the following Compounds (A), (B) and (C):

or a mixture thereof to the sample; and
a step of analyzing its structure by analyzing the obtained nitroxide mixture with a fluorescence detection liquid chromatography, fractionating adducts in which a radical compound derived from a lipid is attached to the nitroxide derivative, and mass analyzing the respective fractions.

2 . The method of claim 1 , wherein the sample is a lipid extract obtained by extracting lipids from a liver excised from a biological object.

3 . The method of claim 1 , wherein the nitroxide derivative or mixture thereof are Compound (C).

4 . The method of claim 1 , further comprising a step of inducing radicals by adding to a sample a radical inducing agent which is any one of lipoxygenase, 2,2′-azobis(2-methylpropionamidine) dihydrochloride, 2,2′-azobis(4-methoxy-2,4′-dimethylvaleronitrile, wherein into the radical induced sample, the nitroxide derivative or a mixture thereof is added.

5 . A compound selected from a group consisting of the following Compounds (A′), (B′) and (C′):

6 . The compound of claim 5 , wherein R is selected from a group consisting of CH 3 , C 2 H 5 , C 3 H 7 , C 4 H 9 , C 6 H 13 , C 7 H 13 , C 7 H 13 O, C 8 H 15 O, C 9 H 15 , C 9 H 15 O 2 , C 9 H 13 O 3 , C 10 H 17 O, C 11 H 19 O 2 , and C 12 H 21 O 2 and, wherein LA represents linoleic acid.

7 . The compound of claim 5 , wherein R is selected from a group consisting of AA-O 2 , CH 3 , C 2 H 5 , C 3 H 7 , C 4 H 9 , C 6 H 13 , C 6 H 9 O 3 , C 7 H 13 , C 7 H 13 O, C 8 H 13 O 2 , C 8 H 15 O, C 9 H 17 , C 9 H 15 O 2 , C 9 H 13 O 3 , C 10 H 17 O, C 11 H 19 , C 11 H 17 O, C 11 H 19 O, C 11 H 19 O 2 , C 12 H 19 O, C 12 H 19 O 2 , C 13 H 19 O 2 , C 14 H 21 O, C 14 H 23 O, and C 15 H 23 O, and wherein AA represents arachidonic acid.

8 . The compound of claim 5 , wherein R is selected from a group consisting of ALA-O 2 , CH 3 , C 5 H 9 O, C 6 H 9 O, C 6 H 11 O, C 7 H 11 O, C 8 H 13 , C 8 H 11 O, C 8 H 11 O 3 , C 8 H 13 O, C 9 H 13 O, C 9 H 13 O 2 , C 10 H 15 O, C 11 H 17 , C 11 H 15 O, C 11 H 17 O, C 11 H 15 O 3 , C 15 H 21 O 2 , and C 15 H 21 O 3 and, wherein ALA represents α-linolenic acid.

9 . The compound of claim 5 , wherein R is selected from a group consisting of EPA-O 2 , CH 3 , C 5 H 9 O, C 6 H 9 O, C 6 H 11 O, C 7 H 11 O, C 8 H 13 , C 8 H 11 O, C 8 H 11 O 3 , C 8 H 13 O, C 9 H 13 O, C 9 H 13 O 2 , C 10 H 15 O, C 11 H 17 , C 11 H 15 O, C 11 H 17 O, C 11 H 15 O 3 , C 12 H 17 O, C 12 H 17 O 2 , C 14 H 21 O, C 14 H 19 O, C 15 H 21 O, C 15 H 21 O 2 , C 15 H 21 O 3 , C 17 H 23 O, and C 18 H 25 O and, wherein EPA represents eicosapentaenoic acid.

10 . The compound of claim 5 , wherein R is selected from a group consisting of DHA-O 2 , CH 3 , C 5 H 9 O, C 6 H 9 O, C 6 H 11 O, C 7 H 11 O, C 8 H 13 , C 8 H 11 O, C 8 H 11 O 3 , C 8 H 13 O, C 9 H 13 O, C 9 H 13 O 2 , C 10 H 15 O, C 11 H 17 , C 11 H 15 O, C 11 H 17 O, C 11 H 15 O 3 , C 12 H 17 O, C 12 H 17 O 2 , C 14 H 21 O, C 14 H 19 O, C 15 H 21 O, C 15 H 21 O 2 , C 15 H 21 O 3 , C 17 H 23 O, C 18 H 25 O, and C 21 H 31 O 4 and, wherein DHA represents docosahexaenoic acid.

11 . The compound of claim 6 , which is Compound (C′):

Assignees

• Yamada Ken Ichi
• Fuso Pharmaceutical Ind

Inventors

• Yamada Ken-Ichi
• Matsuoka Yuta
• Yamamoto Keiichi

Classifications

• G01N33/92 Primary

  involving lipids, e.g. cholesterol {, lipoproteins, or their receptors (steroid hormones G01N33/743)} · CPC title

• G01N33/582

  with fluorescent label · CPC title

• C07D271/12

  condensed with carbocyclic rings or ring systems · CPC title

• C11C1/002

  Sources of fatty acids, e.g. natural glycerides, characterised by the nature, the quantities or the distribution of said acids · CPC title

• C11B1/10

  by extracting · CPC title