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Simple and efficient method for detecting concentration of venlafaxine and active metabolite O-desmethylvenlafaxine in human blood serum

A technology for desvenlafaxine and human detection, applied in the field of medicine, can solve the problems of high cost of solid-phase extraction cartridge, shortened life of chromatographic column, reduced sensitivity, etc., and achieves optimized blood sample processing process, easy operation and low cost Effect

Inactive Publication Date: 2018-12-11
SHANGHAI MENTAL HEALTH CENT (SHANGHAI PSYCHOLOGICAL COUNSELLING TRAINING CENT)
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Precipitated protein is a fast but rough purification technology, which can only remove about 90-95% of the protein, while the high salt in the matrix and other endogenous interference substances such as phospholipids are not removed at all, and the processed blood sample is easy to clog Particle size packing chromatographic column shortens the life of the chromatographic column, and leads to sample dilution, resulting in a decrease in sensitivity, and the cost of the solid phase extraction column is high

Method used

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  • Simple and efficient method for detecting concentration of venlafaxine and active metabolite O-desmethylvenlafaxine in human blood serum
  • Simple and efficient method for detecting concentration of venlafaxine and active metabolite O-desmethylvenlafaxine in human blood serum
  • Simple and efficient method for detecting concentration of venlafaxine and active metabolite O-desmethylvenlafaxine in human blood serum

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Preparation of solution

[0039] 0.05mol / L sodium dihydrogen phosphate solution: weigh NaH 2 PO 4 ·H 2 O 3.44893g was dissolved in water and diluted to 500mL, and 10μL of triethylamine was added, and the pH was adjusted to 2.90 with phosphoric acid solution.

[0040]Venlafaxine hydrochloride standard stock solution: Accurately weigh 2.33 mg of venlafaxine hydrochloride and put it in a 10 mL brown volumetric flask, dissolve it with 50% methanol-water and dilute to the mark, shake well, and use it as a stock solution (206 μg / mL) , Store in a 4°C refrigerator for later use.

[0041] O-desvenlafaxine standard stock solution: Accurately weigh 2.12mg of O-desvenlafaxine, put it in a 10mL brown volumetric flask, dissolve and dilute to the mark with 50% methanol-water, shake well, and use it as a reserve The solution (212 μg / mL) was stored in a 4°C refrigerator for later use.

[0042] Internal standard solution: Accurately weigh 2.33mg of mexiletine hydrochloride, put it i...

Embodiment 2

[0045] Plasma sample processing

[0046] Take 500 μL of plasma sample into a 5 mL stoppered centrifuge tube, add 30 μL of 50 μg / mL internal standard solution, vortex at 1500 r / min for 3 min, add 3 mL of ether, vortex at 1500 r / min for 3 min, then centrifuge at 3000 r / min for 15 min for extraction. For extraction, take the upper organic layer and dry it under nitrogen flow at 40°C, add 100 μL of mobile phase to reconstitute, and vortex for 10 seconds to obtain the sample solution for plasma samples, inject 10 μL by HPLC, and the HPLC conditions For: chromatographic column Agilent Eclipse XDB-C18 (5μm, 4.6×150mm), mobile phase is sodium dihydrogen phosphate (0.05mol / L)-acetonitrile (72:28), flow rate 0.05mL / min, column temperature 30°C, fluorescence The excitation wavelength is 276nm, and the emission wavelength is 598nm.

Embodiment 3

[0048] Choice of Chromatographic Conditions

[0049] Condition 1: Chromatographic column Agilent Eclipse XDB-C18 (5μm, 4.6×150mm), the mobile phase is sodium dihydrogen phosphate (0.05mol / L)-acetonitrile (75:25), the flow rate is 0.5mL / min, the column temperature is 30°C, The fluorescence excitation wavelength is 276nm, the emission wavelength is 598nm, and the injection volume is 10μL; see Figure 7 A

[0050] Condition 2: Chromatographic column Agilent Eclipse XDB-C18 (5μm, 4.6×150mm), the mobile phase is sodium dihydrogen phosphate (0.05mol / L)-acetonitrile (72:28), the flow rate is 0.5mL / min, the column temperature is 30°C, The fluorescence excitation wavelength is 276nm, the emission wavelength is 598nm, and the injection volume is 10μL; see Figure 7 b.

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Abstract

The invention provides a simple and efficient method for detecting the concentration of venlafaxine and an active metabolite O-desmethylvenlafaxine in human blood serum and relates to the technical field of medicines. The simple and efficient method is finished through the following steps: (1) treating a blood serum sample; (2) preparing a solution; and (3) selecting chromatographic conditions. According to the simple and efficient method provided by the invention, a blood sample treatment process is optimized and the simple and efficient method is simple to operate and low in cost; a fluorescence method has strong specificity and high sensitivity; and a capability of accurately determining the venlafaxine and the active metabolite O-desmethylvenlafaxine is greatly improved and the reliability of the determination method is improved.

Description

technical field [0001] The invention relates to the technical field of medicine, in particular to a simple and efficient method for detecting the concentration of venlafaxine and its active metabolite O-desvenlafaxine in human plasma. Background technique [0002] Depressive disorders are a group of common mood disorders characterized by significant and persistent depression and other symptoms, which have the characteristics of high prevalence, high relapse rate, and high disability rate. According to the statistics of the World Health Organization in 2012, depressive disorder has become the fourth largest disease in the world, and it is estimated that by 2020 it may become the second largest human disease after ischemic heart disease. Venlafaxine is the first dual 5-hydroxytryptamine (5-HT) and norepinephrine (NE) reuptake inhibitors (SNRIs), which is different from other antidepressants with unique chemical structure and neuropharmacological activity new antidepressants. ...

Claims

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Application Information

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IPC IPC(8): G01N30/06G01N30/02
CPCG01N30/02G01N30/06G01N2030/027G01N2030/062
Inventor 毛叶萌申书娟张晨
Owner SHANGHAI MENTAL HEALTH CENT (SHANGHAI PSYCHOLOGICAL COUNSELLING TRAINING CENT)
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