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Method for measuring trace triptorelin

A triptorelin and trace technology, applied in the field of trace triptorelin determination, can solve problems such as injury to testing personnel, unsuitability for clinical and scientific research, and expensive kits

Active Publication Date: 2013-06-19
诺桥制药(成都)有限责任公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The extremely low clinical dosage of triptorelin may cause extremely strong clinical adverse reactions. In these cases, it is urgent to establish a simple, specific and highly sensitive detection technology for trace triptorelin. At present, plasma and The detection of trace amounts of triptorelin in biological samples such as excreta is determined by radioimmunoassay. Self-degradation products can also react with immune reagents, increasing the error of the results, and it is radioactive, which may potentially harm the testing personnel; and its kit is very expensive and must be equipped with advanced equipment, which is not suitable for clinical and scientific research

Method used

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  • Method for measuring trace triptorelin
  • Method for measuring trace triptorelin
  • Method for measuring trace triptorelin

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0013] Example 1 The mobile phase is 0.02% acetic acid aqueous solution-methanol to detect triptorelin

[0014] Preparation of Standard Series Solution Samples Accurately weigh 25 mg of triptorelin standard, put it in a 25 ml volumetric flask, add methanol: water: formic acid (80: 20: 0.04) solution to dissolve, and dilute to the mark to prepare 1 mg / ml stock solution. Take an appropriate amount of triptorelin stock solution and dilute with methanol: water: formic acid (60:40:0.08) to 0.005, 0.01, 0.03, 0.1, 0.3, 1.0, 3.0, 10.0 ng / ml.

[0015] LC Conditions Column: Venusil MP-C18 column (2.1 mm x 50 mm, 3 μm, Agela). Mobile phase: A: 0.02% acetic acid aqueous solution, B: methanol; flow rate: 0.6mL / 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:

[0016] Time(min)

A

B

...

Embodiment 2

[0020] Example 2 The mobile phase is 0.02% acetic acid aqueous solution-methanol to detect triptorelin

[0021] The preparation of standard series solution sample is the same as embodiment 1

[0022] LC Conditions Column: Venusil MP-C18 column (2.1 mm x 50 mm, 3 μm, Agela). Mobile phase: A: 0.02% acetic acid aqueous solution, B: methanol; flow rate: 0.6mL / 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:

[0023] Time(min)

A

B

0

90

10

1

90

10

2.3

40

60

2.7

40

60

2.71

2

98

4

2

98

4.01

90

10

5.5

90

10

[0024] Mass spectrometry conditions: QTRAP5500 mass spectrometer, equipped with ion spray ionization source, ...

Embodiment 3

[0027] Example 3 The mobile phase is 0.05% acetic acid aqueous solution-methanol detection triptorelin

[0028] The preparation of standard series solution sample is the same as embodiment 1

[0029] LC Conditions Column: Venusil MP-C18 column (2.1 mm x 50 mm, 3 μm, Agela). Mobile phase: A: 0.05% acetic acid aqueous solution, B: methanol; flow rate: 0.4 mL / min, column temperature: 30°C, injection volume: 10 μl. Agilent 1290 high performance liquid chromatography system, including binary infusion pump, automatic sampler, column thermostat. The gradient conditions are as follows:

[0030] Time(min)

A

B

0

90

10

1

90

10

4.0

50

50

5.0

50

50

5.1

5

95

6.5

5

95

7.0

90

10

9.0

90

10

[0031] Mass spectrometry conditions: QTRAP5500 mass spectrometer, equipped with ion spray ionization sourc...

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Abstract

The invention relates to a method for measuring trace triptorelin. The trace triptorelin in a sample is measured by using a liquid chromatography and mass spectrometry technology. The method is simple in operations and accurate in results, and has good specificity and high sensitivity. The invention also relates to applications of the measurement method for the qualitatively or quantitatively measuring the content of the triptorelin in a biological sample and for pharmacokinetic studies of triptorelin preparations.

Description

technical field [0001] The invention relates to a method for determining trace triptorelin by using liquid chromatography and mass spectrometry. Background technique [0002] Triptorelin is a synthetic decapeptide that is an analog of gonadotropin-releasing hormone (GnRH). The improvement of its structure is to replace the sixth left-handed amino acid (glycine) in the natural molecular structure with D-tryptophan. Triptorelin has the same effect as GnRH, but its plasma half-life is prolonged and its affinity for GnRH receptors is stronger, so triptorelin becomes a strong agonist of GnRH receptors. After triptorelin injection, it initially stimulates the pituitary gland to secrete gonadotropins (Gn), namely luteinizing hormone (LH) and follicle stimulating hormone (FSH). When the pituitary gland enters a refractory period after prolonged stimulation, the release of gonadotropins decreases, thereby reducing the sex steroids (testosterone or estrogen) to castrate levels. The...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/88G01N30/06
Inventor 韩江彬沙春洁宋涛王涛孙考祥刘万卉
Owner 诺桥制药(成都)有限责任公司
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