Method for measuring concentrations of four arsenic compounds in granulocytes by using HPLC (High Performance Liquid Chromatography)-ICP (Inductively Coupled Plasma)-MS method and application

A technology of HPLC-ICP-MS and arsenic compounds, which is applied in the field of HPLC-ICP-MS method to determine the concentration of four kinds of arsenic compounds in granulocytes, can solve the problems of inaccurate determination and achieve individualized medication and targeted therapy, Reduce toxicity, quantify accurate effects

Active Publication Date: 2019-12-31
HARBIN MEDICAL UNIVERSITY
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  • Claims
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Problems solved by technology

[0004] In order to solve the problem that existing methods cannot accurately measure As in granulocytes III 、As V 、DMA V and MMA V For the problem of the concentration o...

Method used

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  • Method for measuring concentrations of four arsenic compounds in granulocytes by using HPLC (High Performance Liquid Chromatography)-ICP (Inductively Coupled Plasma)-MS method and application
  • Method for measuring concentrations of four arsenic compounds in granulocytes by using HPLC (High Performance Liquid Chromatography)-ICP (Inductively Coupled Plasma)-MS method and application
  • Method for measuring concentrations of four arsenic compounds in granulocytes by using HPLC (High Performance Liquid Chromatography)-ICP (Inductively Coupled Plasma)-MS method and application

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Experimental program
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Embodiment 1

[0046] Instruments and reagents used in this embodiment:

[0047] Instruments: HPLC-ICP-MS NexI0N 350D (Perkin Elmer Instruments Co., Ltd.), Ultrapure Water Instrument (Thermofisher Technology Co., Ltd.), Ultrasonic Cleaner (Branson, USA) One-third precision electronic balance (Sartorius company), multifunctional low-temperature high-speed centrifuge (Eppendorf company), vortex mixer (Branson, Germany); the flow cytometer model is BD FACSAria IIu.

[0048] Reagents: arsenite standard solution GBW08666 (National Academy of Metrology), arsenate standard solution GBW08667 (National Academy of Metrology), monomethyl arsenic standard solution GBW08668 (National Academy of Metrology), dimethyl arsenic standard solution GBW08669 (National Academy of Metrology) , anhydrous sodium dihydrogen phosphate, potassium nitrate, anhydrous sodium acetate, and disodium edetate (99.00%, superior grade) were purchased from Sinopharm Chemical Reagent Co., Ltd., erythrocyte lysate-FACS lysingsolutio...

Embodiment 2

[0064] The HPLC-ICP-MS method provided in this embodiment measures the method for the concentration of four kinds of arsenic compounds in granulocytes, comprising the following steps:

[0065] Step 1. Determine the detection conditions of HPLC-ICP-MS:

[0066] HPLC conditions: anion exchange column: Hamilton PRP-X100, 150mm×4.1mm, 5μm; flow rate: 0.9mL / min; mobile phase: 3mmol / L anhydrous sodium dihydrogen phosphate, 13mmol / L anhydrous sodium acetate , a mixed solution composed of 0.2 mmol / L disodium edetate and 4 mmol / L potassium nitrate; the injection volume is 50 μL;

[0067] Inductively coupled plasma mass spectrometry conditions: detection mode: DRC plus oxygen mode, nebulizer gas flow: 0.9L / min, auxiliary gas flow: 1.2L / min, plasma gas flow: 18L / min, inductively coupled plasma RF power: 1600W, oxygen Flow rate: 0.6ml / min;

[0068] Step 2, prepare standard solutions of four kinds of arsenic compounds:

[0069] As was prepared at a concentration of 1000ng / mL respectivel...

Embodiment 3

[0083] The HPLC-ICP-MS method provided in this embodiment measures the method for the concentration of four kinds of arsenic compounds in granulocytes, comprising the following steps:

[0084] Step 1. Determine the detection conditions of HPLC-ICP-MS:

[0085] HPLC conditions: anion exchange column: Hamilton PRP-X100, 150mm×4.1mm, 5μm; flow rate: 0.9mL / min; mobile phase: 3mmol / L anhydrous sodium dihydrogen phosphate, 13mmol / L anhydrous sodium acetate , a mixed solution composed of 0.2 mmol / L disodium edetate and 4 mmol / L potassium nitrate; the injection volume is 50 μL;

[0086] Inductively coupled plasma mass spectrometry conditions: detection mode: DRC plus oxygen mode, nebulizer gas flow: 0.9L / min, auxiliary gas flow: 1.2L / min, plasma gas flow: 18L / min, inductively coupled plasma RF power: 1600W, oxygen Flow rate: 0.6ml / min;

[0087] Step 2, prepare standard solutions of four kinds of arsenic compounds:

[0088] As was prepared at a concentration of 1000ng / mL respectivel...

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Abstract

The invention relates to a method for measuring the concentrations of four arsenic compounds in granulocytes by using an HPLC (High Performance Liquid Chromatography)-ICP (Inductively Coupled Plasma)-MS method and an application, and belongs to the technical field of blood sample detection. In order to solve the problem that the existing method cannot accurately measure the concentrations of fourdifferent arsenic compounds AsIII, AsV, DAV and MMVA in granulocytes, the invention provides a method for measuring the concentrations of four arsenic compounds in granulocytes by using the HPLC-ICP-MS method. The method comprises the steps of determining a detection condition, preparing a standard solution, drawing a standard curve, collecting and processing a granulocyte sample, performing sample detection and data processing. According to the method for measuring the concentrations of four arsenic compounds in granulocytes by using the HPLC-ICP-MS method and the application, each arsenic compound in granulocytes can be effectively extracted and accurately quantified. When the method is applied to the detection of the concentrations of four arsenic compounds in peripheral blood granulocytes of a patient suffering from APL (Acute Promyelocytic Leukemia, the metabolism and distribution characteristics of each arsenic form can be clarified, a technical basis is provided for revealing ofthe action mechanism of arsenic acid and reduction of toxic reactions, and when the method is used for assisting clinics, personalized medicine and targeted therapy are implemented.

Description

technical field [0001] The invention belongs to the technical field of blood sample detection, and in particular relates to a method and an application of an HPLC-ICP-MS method for measuring the concentration of four arsenic compounds in granulocytes. Background technique [0002] Arsenous Acid Injection-As 2 o 3 It is a drug independently developed by my country in 1971 for the treatment of acute promyelocytic leukemia-APL. The application of this drug can significantly improve the cure rate of APL patients. After arsenous acid enters the human body, trivalent inorganic arsenic-As Ⅲ form exists. Production of pentavalent monomethylarsenic acid-MMA by hepatic metabolism Ⅴ and pentavalent dimethyl arsenic acid-DMA Ⅴ , through a redox reaction to generate pentavalent inorganic arsenic-As Ⅴ . Studies have found that the therapeutic effect of arsenous acid is closely related to the PML / RARA fusion protein. Arsenous acid or its metabolites can enter the cell through the c...

Claims

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Application Information

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IPC IPC(8): G01N30/02G01N30/06G01N30/72G01N30/86
CPCG01N30/02G01N30/06G01N30/72G01N30/86G01N2030/047
Inventor 海鑫于凯江王新羽郭美华
Owner HARBIN MEDICAL UNIVERSITY
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