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Kit for detecting concentration of oleic acid in serum or plasma, and preparation method and application of kit

A kit and the technology of oleic acid, which are applied in the field of kits and preparations for detecting the concentration of oleic acid in serum or plasma, can solve the problems of low accuracy and complicated methods, and achieve reliable diagnosis and remarkable technical progress

Inactive Publication Date: 2019-02-01
SHANGHAI EAST HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] Aiming at the above-mentioned technical problems in the prior art, the present invention provides a kit and method for detecting OA concentration in serum or plasma using ultra-high performance liquid chromatography tandem mass spectrometry (UHPLC-MS / MS) technology. A kit and method for detecting the concentration of OA in serum or plasma need to solve the technical problems that the method for detecting the concentration of OA in serum or plasma is complicated and has low accuracy in the prior art

Method used

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  • Kit for detecting concentration of oleic acid in serum or plasma, and preparation method and application of kit
  • Kit for detecting concentration of oleic acid in serum or plasma, and preparation method and application of kit
  • Kit for detecting concentration of oleic acid in serum or plasma, and preparation method and application of kit

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Embodiment 1

[0030] The establishment of the ultra high performance liquid chromatography tandem mass spectrometry (UHPLC-MS / MS) detection method of embodiment 1 serum or plasma oleic acid (OA) content

[0031] 1. Instruments: high performance liquid chromatography Agilent 1290 UPLC (Agilent, USA), tandem triple quadrupole mass spectrometer AB API 5000 (AB sciex, USA), METTLER TOLEDO balance (Max=220g, d=0.1mg) (METTLER , Germany), SB-C18 rapid separation high-throughput narrow-bore column (2.1mm*100mm*1.8μm) (Agilent, USA), Eppendorf 5424R low-temperature high-speed centrifuge (Eppendorf, Germany).

[0032] 2. Commercial reagents: standard oleic acid was purchased from sigma company with a purity of 98.0%; oleic acid isotope marker OA-[ 13 C 5 ] was purchased from IsoSciences, with an isotope concentration of 1030±1 μg / mL; methanol (LC / / MS / MS grade), acetonitrile (LC / / MS / MS grade), and formic acid (LC / / MS / MS grade) were purchased from American Fisher Corporation. The test water was pre...

Embodiment 2O

[0080] Example 2 OA detection clinical preliminary application

[0081] 1. Research object

[0082]Serum samples were collected at Shanghai Dongfang Hospital.

[0083] NAFLD group: The subjects were 100 patients diagnosed with NAFLD in the Fatty Liver Disease Clinic of the Department of Traditional Chinese Medicine between December 2016 and February 2017, including 42 males and 58 females, with an average age of 55.6 years.

[0084] Diagnostic criteria:

[0085] (1) No history of drinking or alcohol consumption equivalent to less than 140g / week (female <70g / week);

[0086] (2) Viral hepatitis, drug-induced liver disease, total parenteral nutrition, hepatolenticular degeneration, autoimmune liver disease and other specific diseases that can cause fatty liver are excluded;

[0087] (3) Liver ultrasonography meets the diagnostic criteria for diffuse fatty liver;

[0088] (4) Serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), and γ-glutamyltransferase (GGT...

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Abstract

The invention provides a kit for detecting concentration of oleic acid in serum or plasma. The kit comprises a mobile phase A, a mobile phase B, a standard substance stock solution, an isotope internal standard substance stock solution, a sample extracting solution and a diluents, wherein mobile phase A is ultrapure water, and the mobile phase B is a solution formed by methanol and acetonitrile according to the volume ratio of 1 to 1; the standard substance stock solution is 20 mg / mL of oleic acid solution; the isotope internal standard substance stock solution is 20mg / mL of oleic acid-[13C5]solution; the sample extracting solution is a solution formed by methanol and acetonitrile according to the equal volume ratio of 1 to 1; and the diluents is bovine serum albumin which does not contain free fatty acids, wherein the concentration of the bovine serum albumin is 50 mg / mL. The invention further provides a preparation method of the kit, and a method for detecting the concentration of oleic acid by adopting the kit. The kit is an effective tool for quick, stable, reliable diagnosis, treatment and curative effect monitoring of diseases related to abnormal oleic acid metabolism.

Description

Technical field: [0001] The invention belongs to the field of biological detection, and relates to a detection kit, in particular to a kit for detecting the concentration of oleic acid (Oleic acid, OA) in serum or plasma, and a preparation method and application thereof. Background technique: [0002] Oleic acid (OA) is a monounsaturated Omega-9 fatty acid with molecular formula C18H34O2. Oleic acid in human blood is divided into exogenous oleic acid and endogenous oleic acid. Exogenous oleic acid comes from dietary intake, and endogenous oleic acid is passed through stearoyl-coenzyme Adesaturase (stearoyl-coenzyme Adesaturase, SCD) catalyzes the production of saturated fatty acids. Oleic acid is the main component of free fatty acid (FFA). When glycogen is depleted, adipose tissue will decompose neutral fat into free fatty acid for energy use. Under normal circumstances, FFA is the three major energy substances, an important part of sugar, protein and fat metabolism, and ...

Claims

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Application Information

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IPC IPC(8): G01N30/02G01N30/06
CPCG01N30/02G01N30/06G01N2030/027
Inventor 范列英金中淦宗明
Owner SHANGHAI EAST HOSPITAL
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