Composition of gonadotropic hormone releasing hormone analogue

A gonadotropin and hormone-releasing technology, which is applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of expensive kits, affecting the accuracy of low-concentration quantitative results, and radioactivity

Active Publication Date: 2015-03-25
SHANDONG LUYE PHARMA CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Gonadotropin-releasing hormone analogues (such as triptorelin and goserelin) are currently quantified by radioimmunoassay. Due to the existence of cross-reactivity in immunoassays, the accuracy of low-concentration quantitative results is affected, and they are radioactive. There is potential harm to personnel; and its kit is very expensive, not suitable for clinical and scientific research; the quantitative process of testosterone includes liquid chromatography-mass spectrometry analysis and enzyme-linked immunoassay
Liquid-mass spectrometry analysis uses derivatized sample pretreatment, which is complicated to operate. At the same time, testosterone, as an endogenous compound, makes the analysis more complicated

Method used

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  • Composition of gonadotropic hormone releasing hormone analogue
  • Composition of gonadotropic hormone releasing hormone analogue
  • Composition of gonadotropic hormone releasing hormone analogue

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Example 1 mobile phase is 0.02% propionic acid aqueous solution-methanol detection goserelin and testosterone standard solution

[0025] Standard solution sample preparation

[0026]Precisely weigh the reference substance of goserelin acetate, and prepare a 1.00 mg / mL mother solution with methanol solution for later use. Testosterone stock solution (100 μg / mL) was purchased from Sigma, USA. Using methanol-water-formic acid (60:40:0.08v / v / v) as the diluent, dilute testosterone and goserelin into a standard mixture of the following concentrations, 0.02+0.005, 0.05+0.01, 0.1+0.03, 0.2+0.1 and 0.5+0.3ng / mL.

[0027] Liquid condition chromatographic column: ZORBAX Eclipse plus C 18 (2.1mm x50mm, 1.8μm, Stockport, UK) Mobile phase: A: 0.02% propionic acid aqueous solution, B: methanol; flow rate: 0.4mL / min, column temperature: 40°C, injection volume: 10μl. Agilent1290 high performance liquid chromatography system, including binary infusion pump, automatic sampler, column ...

Embodiment 2

[0033] Example 2 The mobile phase is 0.02% propionic acid aqueous solution-methanol to detect goserelin and testosterone-d 3 Standard solution

[0034] The preparation of standard solution sample is the same as embodiment 1, and testosterone is replaced by testosterone-d during preparation 3 .

[0035] Liquid condition chromatographic column: ZORBAX Eclipse plus C 18 (2.1mm x50mm, 1.8μm, Stockport, UK); mobile phase: A: 0.02% propionic acid aqueous solution, B: methanol; flow rate: 0.4mL / min, column temperature: 40°C, injection volume: 10μl. Agilent1290 high performance liquid chromatography system, including binary infusion pump, automatic sampler, column thermostat. The gradient conditions are as follows:

[0036] Time(min)

A

B

0

90

10

1.8

90

10

2.2

35

65

2.8

30

70

3.8

30

70

4.5

8

92

5.0

90

10

7.0

90

10

[0037] Mass spectrometr...

Embodiment 3

[0040] Example 3 mobile phase is 0.05% propionic acid aqueous solution-methanol detection triptorelin and testosterone standard solution

[0041] The preparation of the standard solution sample was the same as in Example 1, except that goserelin was replaced by triptorelin.

[0042] Liquid conditions Chromatographic column: Eclipse plus RRHD C8 (2.1mm x50mm, 1.8μm, Agilent); mobile phase: A: 0.05% propionic acid aqueous solution, B: methanol; flow rate: 0.4mL / min, column temperature: 40°C, Sample volume: 10 μl. Agilent1290 high performance liquid chromatography system, including binary infusion pump, automatic sampler, column thermostat. The gradient conditions are as follows:

[0043] Time(min)

A

B

0

95

5

1.8

95

5

2.5

40

60

3.2

40

60

5.0

30

70

5.4

30

70

6.2

8

92

6.7

8

92

7.2

95

5

8.0

95

5

[0044] Mass...

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Abstract

The invention relates to a composition of a gonadotropic hormone releasing hormone analogue. A liquid chromatography-mass spectrometry technology is used for determining testosterone and the gonadotropic hormone releasing hormone analogue; furthermore, the invention provides a method for simultaneously and successfully detecting the testosterone and the gonadotropic hormone releasing hormone analogue in a sample; and after the gonadotropic hormone releasing hormone analogue and the composition thereof are used, the content and the change process of the gonadotropic hormone releasing hormone analogue and the testosterone in a body are simultaneously detected.

Description

technical field [0001] The invention relates to a composition of gonadotropin-releasing hormone analogues, which is determined by liquid chromatography and mass spectrometry to simultaneously measure testosterone and gonadotropin-releasing hormone analogues. Background technique [0002] Gonadotropin-releasing hormone (LHRH), a 10-peptide synthesized by the arcuate nucleus of the hypothalamus, has a structure of Pyro-Glu-His-Trp-Ser-Tyr-Gly-Leu-Arg-Pro-Gly-NH 2 , the hypothalamus pulse releases LHRH, which can stimulate the pituitary gland to release gonadotropin-releasing hormone. In 1971, Schall isolated a few milligrams of LHRH from the hypothalamus of 500,000 pigs and sheep, and artificially synthesized and confirmed the structure of LHRH. Therefore, he won the Nobel Prize in Medicine and Physiology Prize. Mammalian LHRH has the same structure, and the 5-6, 6-7, 9-10 amino acid chains of LHRH have poor stability and are easily cleaved by endopeptidases, so the half-life...

Claims

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Application Information

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IPC IPC(8): G01N30/02G01N30/34
Inventor 韩江彬张枢孙考祥刘万卉冷广意王涛
Owner SHANDONG LUYE PHARMA CO LTD
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