Method for detecting prostaglandin substances in biological sample

A technology for prostaglandins and biological samples, applied in the field of quantitative analysis of a series of prostaglandins in biological samples, can solve problems such as difficulty in carrying out research work, and achieve the effects of reducing the amount of biological samples, simplifying sample processing steps, and shortening retention time.

Active Publication Date: 2021-09-07
北京现代药物代谢研究院
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

This harsh requirement for operation and equipment makes it difficult for many laboratories to carry out relevant research work

Method used

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  • Method for detecting prostaglandin substances in biological sample
  • Method for detecting prostaglandin substances in biological sample
  • Method for detecting prostaglandin substances in biological sample

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Add 1mL of acetonitrile / water mixture into the anti-adsorption EP tube, and add 1mg / mL PGE in sequence 1 Acetonitrile solution 20 μL, 1 μL / mL DIPEA acetonitrile solution 50 μL, 1 mg / mL HATU acetonitrile solution 50 μL, shake at room temperature for 10 minutes; add 1 mg / mL 5-(aminomethyl)naphthalene-2-sulfonic acid aqueous solution 50 μL, shake at room temperature Shake for 20 minutes.

[0035] After the reaction was completed, 50 μL of the reaction mixture was taken out from the reaction solution and added to 4950 μL of acetonitrile water 1 / 1 (v / v) to dilute into the test solution. Mass spectrometry not found [PGE 1 -H] - signal (m / z 353.23Da), and the corresponding PGE 1 The signal of benzenesulfonic acid derivative is obvious (m / z 572.27Da), indicating that PGE 1 Fully derivatized.

[0036] Triple TOF TM 6600 + Type tandem mass spectrometer, equipped with ESI ionization source; negative ion detection; ion injection voltage -5000V; temperature 500°C; source gas...

Embodiment 2

[0042] According to the derivatization scheme described in Example 1, the equivalent amount of limatoprost was converted into 4-(aminomethyl)benzenesulfonic acid derivatives; and the mass spectrometry conditions were optimized.

[0043] Column: ACQUITY TM BEH C 18 Column, 100mm×2.1mm I.D., 1.7μm particle size; mobile phase: aqueous phase: 5% acetonitrile + 95% water (containing 1mMol ammonium acetate + 0.02% acetic acid) pH 5, organic phase: acetonitrile; gradient elution; column The temperature is 40°C; the flow rate is 0.3mL / min; the injection volume is 20μL. The gradient elution in the chromatographic conditions, the program is shown in Table 2, wherein A is 5% acetonitrile + 95% water (containing 1mMol ammonium acetate + 0.02% acetic acid), and B is acetonitrile.

[0044] Table 2 Gradient elution program

[0045]

[0046] The mass spectrometry conditions are: Triple Quad TM Model 6500 tandem mass spectrometer, equipped with ESI ionization source, SelexION differen...

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Abstract

The invention discloses a method for detecting a prostaglandin substance in a biological sample, which comprises the following steps of: equivalently converting a prostaglandin substance to be detected in the biological sample into an aminomethyl aromatic sulfonic acid derivative by a chemical derivatization method, and carrying out liquid chromatography-mass spectrometry online detection by taking aromatic sulfonic acid derivative negative ions [M-H] <-> as precursor ions during detection. The [M-H] <-> has a higher mass spectral response, a shorter reverse phase chromatography retention time than the underived prostaglandin, and meanwhile, the cracking conversion efficiency of specific product ions [M-H2O-C15Residue]<-> can be improved during gas phase collision cracking of the mass spectrum Compared with an ion pair selected for mass spectrum multi-reaction monitoring at present, the high-specificity ion is used for quantifying the prostaglandin substances, and the signal intensity can be improved by two times. According to the detection method, sample treatment steps can be simplified, the use amount of biological samples can be reduced, or the requirements on instruments can be relaxed on the basis of ensuring the signal-to-noise ratio, and the requirements of accurately analyzing a series of prostaglandin substances in the biological samples in scientific research and production activities can be met.

Description

technical field [0001] The invention belongs to the technical field of drug analysis and research, and relates to a method for quantitatively analyzing a series of prostaglandin substances in biological samples. Background technique [0002] Prostaglandins are physiologically active unsaturated fatty acids with 20 carbon atoms produced by enzymatic metabolism of arachidonic acid in organisms. Such as figure 1 As shown, the series of prostaglandins have a common structure such as an aliphatic five-membered ring and two side chains (the A chain has 7 carbon atoms and the head end is a carboxyl group; the B chain has 8 carbon atoms and contains an allyl alcohol residue). [0003] Prostaglandins are released outside the cell after they are synthesized. Each prostaglandin has its own specific receptors. Prostaglandins combined with their specific receptors can mediate cell proliferation, differentiation, apoptosis, platelet aggregation, etc., and participate in Inflammation and...

Claims

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Application Information

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IPC IPC(8): G01N30/02G01N30/06G01N30/34G01N30/86
CPCG01N30/02G01N30/06G01N30/34G01N30/8675G01N2030/045G01N2030/062G01N2030/067
Inventor 顾景凯孙冬孟祥骏宋诗文
Owner 北京现代药物代谢研究院
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