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Method for detecting ulipristal acetate and metabolites thereof in blood plasma by LC-MS (liquid chromatography-mass spectrometry) method

A technology of uliplast acetate and LC-MS, which is applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problem of large amount of plasma consumption, long detection time, and inability to detect the active metabolite monodemethyl-unixilast acetate special problems, to avoid interference

Active Publication Date: 2022-03-04
四川尚锐分析检测有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, this method consumes a large amount of plasma, and can only detect uliplast acetate, but cannot detect its active metabolite monodesmethyl- uliplast acetate at the same time, and the detection time is longer

Method used

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  • Method for detecting ulipristal acetate and metabolites thereof in blood plasma by LC-MS (liquid chromatography-mass spectrometry) method
  • Method for detecting ulipristal acetate and metabolites thereof in blood plasma by LC-MS (liquid chromatography-mass spectrometry) method
  • Method for detecting ulipristal acetate and metabolites thereof in blood plasma by LC-MS (liquid chromatography-mass spectrometry) method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Example 1 Liquid Chromatography-Tandem Mass Spectrometry for the Detection of Uliprist Acetate and Uliprist Monodesmethyl Acetate in Plasma

[0045] 1. Pretreatment of plasma samples

[0046] Take 0.1mL plasma sample, add 20μL internal standard working solution (100ng / mL) and mix for 2min, add 3ml extraction solvent dichloromethane / isopropanol (9 / 1v / v), shake for 10min, centrifuge (4000rpm 5min), Take 2.4mL of the supernatant and add it to a 5ml EP tube, evaporate to dryness under nitrogen at 45°C, and redissolve with 50μL of acetonitrile / water (1:1v / v) mixed solvent. The reconstituted solution was transferred to a 96-well plate for LC-MS / MS analysis. see results figure 1 , figure 2 and image 3 , representing the typical mass spectra of Ulipristal Acetate, Monodesmethyl-Ulipristal Acetate and Ulipristal Acetate-d6, respectively.

[0047] 2. Preparation of standard series samples and quality control samples

[0048] (1) Preparation of uliplast acetate stock solut...

Embodiment 2

[0063] Embodiment 2 methodological verification

[0064] 1. Selective

[0065] Take 0.1 mL (n=6) of blank plasma from different sources, process the samples with the method in Example 1 respectively, take (without internal standard) samples for mass spectrometry analysis, and obtain the chromatogram of the blank plasma samples. Get different sources of blank plasma (n=6) to prepare respectively 0.1mL of standard curve minimum point concentration plasma sample, process sample with the method of embodiment 1, carry out mass spectrometry, standard curve minimum point concentration plasma sample carries out mass spectrometry analysis, evaluation method selective.

[0066] The results showed that endogenous substances did not interfere with the determination of uliplast acetate, monodesmethyl-uliplast acetate and uliplast acetate-d6. For a typical blank plasma sample chromatogram and a typical LLOQ chromatogram see Figure 4 , Figure 5 and Image 6 .

[0067] 2. Standard cur...

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Abstract

The invention discloses a method for detecting ulipristal acetate and an active metabolite mono-demethylation-ulipristal acetate in blood plasma by adopting an LC-MS (Liquid Chromatography-Mass Spectrometry) method. The method comprises the following steps: extracting a blood plasma sample by using dichloromethane / isopropanol, and performing LC-MS / MS detection. The method can be used for detecting ulipristal acetate, can also be used for detecting an active metabolite monodemethylation-ulipristal acetate, and has the characteristics of rapidness, high sensitivity and capability of simultaneously monitoring two components by using a small amount of plasma; the method can be applied to research on pharmacokinetics and bioequivalence of ulipristal acetate and active metabolite monodemethylation-ulipristal acetate.

Description

technical field [0001] The invention relates to the technical field of drug analysis, in particular to a liquid chromatography-tandem mass spectrometry method for detecting uliplast acetate and an active metabolite monodemethyl- uliplast acetate in plasma. Background technique [0002] Ulipristal acetate (UPA) (11β)-17α-(acetyloxy)-11-[4-(dimethylamino)phenyl]-19-norpregna-4,9-di En-3,20-dione, a selective progesterone receptor modulator, has a steroidal structure. The drug is approved as emergency contraception in the United States and other countries. The same is true in Europe and has been approved for the treatment of uterine fibroids, and the drug has entered the clinic for the treatment of ovarian and breast cancer. With many potential applications of UPA currently being evaluated, it is imperative to develop a well-validated analytical method with high applicability. [0003] Prior art Journal of Chromatography B, 1059 (2017) 43-48) discloses a method for detecting...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02G01N30/06G01N30/72G01N30/86
CPCG01N30/02G01N30/06G01N30/72G01N30/8679
Inventor 李志娜黄静彭显峰黄小兰曾思瑶杨婷秀
Owner 四川尚锐分析检测有限公司
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