LC-MS (liquid chromatography-mass spectrometry) detection method of 9-propenyladenine

A technology of acryl adenine and a detection method, which is applied in the field of medicine, can solve the problems that the sensitivity cannot meet the detection requirements, and is not effectively controlled, and achieves the effects of high sensitivity, simple method and strong specificity.

Inactive Publication Date: 2018-04-17
ZHENGZHOU TAIFENG PHARMA CO LTD
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] 9-propenyl adenine is a genotoxic impurity brought in by the starting material and produced during the storage of tenofovir disoproxil fumarate. WHO has made strict regulations on its content, requiring that it should not exceed 5 mg · L －1 , so it is necessary to establish a sensitive and specific method to determine its content, but it is not effectively controlled in the current quality standards
Due to the low content of 9-propenyl adenine in the ten...

Method used

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  • LC-MS (liquid chromatography-mass spectrometry) detection method of 9-propenyladenine
  • LC-MS (liquid chromatography-mass spectrometry) detection method of 9-propenyladenine
  • LC-MS (liquid chromatography-mass spectrometry) detection method of 9-propenyladenine

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] 1 Selection of chromatographic conditions

[0027] 1.1 Selection of chromatographic column

[0028] Choose from three octadecylsilane-bonded silica columns. The retention behavior of different chromatographic columns is shown in Table 1:

[0029] Table 1 Retention behavior of 9-propenyl adenine in different chromatographic columns

[0030]

[0031] The results show that XBridge C18 (50mm×2.1mm , 3.5μm) interferes with the detection of genetic impurities in the starting material, and the other two columns can be used for the detection of genetic impurities, but in comparison, the YMC Pack ODS-AQ C18 (150mm×4.6mm, 3μm) chromatographic column has more appropriate retention time and higher sensitivity.

[0032] 1.2 Selection of column temperature

[0033] The commonly used column temperature was investigated, and the results are shown in Table 2:

[0034] Table 2 Effect of different column temperatures

[0035]

[0036] The results showed that the retention time...

Embodiment 2

[0059] 2 Establishment of methodology

[0060] 2.1 Specificity

[0061] Take the blank solvent, the control solution, the test solution and the mixed solution to inject samples respectively, and record the chromatograms. The results showed that the blank solvent, TDF and TDA did not interfere with the detection of 9-propenyladenine.

[0062] 2.2 Investigation of the stability of the sample solution

[0063] Take 1 part of the control solution, measure it at 0-8 hours respectively, record the peak area, and calculate RSD=2.17%, which shows that the control solution has good stability within 6 hours, and the intraday error is small. See Table 7:

[0064] Table 7 Stability of reference substance solution

[0065] .

[0066] 2.3 Limit of quantitation

[0067] Take an appropriate amount of 9-propenyl adenine reference substance, add methanol to make a solution with a certain concentration, and record the chromatogram. The limit of quantification is 5 ng / ml (S / N ≈ 10).

[0...

Embodiment 3

[0093] 3 Determination of 9-propenyl adenine in tenofovir disoproxil fumarate

[0094] (1) Chromatographic conditions

[0095] Chromatographic column: YMC Pack ODS-AQ C18, 150mm×4.6mm, 3μm;

[0096] Column temperature: 30 degrees;

[0097] m / z: 176;

[0098] Flow rate: 1ml / min

[0099] Mobile phase: 0.01mol / L ammonium acetate: acetonitrile=80:20;

[0100] (2) Preparation of the test sample and reference substance solution: take the test sample, accurately weigh it, add methanol to make a 2mg / ml solution, and use it as the test sample solution; take another 9-propenyl adenine reference substance, accurately weigh Weigh it, add methanol to make a 10ng / ml solution as a control solution;

[0101] (3) Determination method: Accurately measure 10 μl of the reference substance solution and the test solution, inject the sample, record the chromatogram, and calculate the peak area according to the external standard method; if the test solution contains 9-propenyl adenine, it should...

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Abstract

The present invention relates to an LC-MS detection method for detecting 9-propenyl adenine. The technical points include the following steps: (1) first determine the chromatographic conditions through the chromatographic column, column temperature and mobile phase; (2) the test sample and the preparation of the reference substance solution; (3) the assay method adopts precise measurement of the reference substance solution and the test solution respectively, injects into the chromatograph, records the chromatogram, and calculates with the peak area according to the external standard method; the test substance does not interfere with 9‑ The detection of propenyl adenine further realizes the present invention. The present invention has the advantages of establishing a LC-MS detection method for determining 9-propenyl adenine in tenofovir disoproxil fumarate. The detection method can accurately detect the content of 9-propenyl adenine; the method has high sensitivity and strong specificity; it can effectively detect the content of 9-propenyl adenine to control the quality of tenofovir disoproxil fumarate.

Description

technical field [0001] The invention belongs to the technical field of medicine, and in particular relates to an LC-MS detection method for detecting 9-propenyl adenine. Background technique [0002] Tenofovir disoproxil fumarate dipivoxil is a new type of nucleotide reverse transcriptase inhibitor developed and marketed by Gilead in the United States. It was approved by the FDA in 2001 for the treatment of HIV-1 infection And chronic hepatitis B, in 2008 FDA approved it for the treatment of HBV infection. Its active ingredient tenofovir is an acyclic 5'-adenosine monophosphate analog, which is phosphorylated by cellular kinases in vivo to generate tenofovir diphosphate, which is incorporated by competing with 5'-deoxyadenosine triphosphate Into the viral DNA chain, causing DNA chain elongation to be blocked and inhibit the replication of the virus. The oral absorption rate of esterified tenofovir is significantly improved, and can increase its uptake by cells. Due to its...

Claims

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Application Information

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IPC IPC(8): G01N30/02
CPCG01N30/02
Inventor 吕艳歌沙薇皇甫光
Owner ZHENGZHOU TAIFENG PHARMA CO LTD
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