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Detection method based on GC-MS (gas chromatography-mass spectrometry) sperm counting and application

A GC-MS and sperm counting technology, applied in the field of analytical chemistry and clinical medicine, to achieve rapid and accurate analysis, good stability, and high practical value for clinical diagnosis

Active Publication Date: 2020-01-17
ZHEJIANG CELLPRO BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Use of GC-MS to analyze pendimethalin exposure in male serum for sperm counts has not received commensurate attention

Method used

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  • Detection method based on GC-MS (gas chromatography-mass spectrometry) sperm counting and application
  • Detection method based on GC-MS (gas chromatography-mass spectrometry) sperm counting and application
  • Detection method based on GC-MS (gas chromatography-mass spectrometry) sperm counting and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] In order to make the technical means, creative features, goals and effects achieved by the present invention easy to understand, the present invention will be further described below in conjunction with specific embodiments. Embodiment 1 Research Object Selection and Grouping Basis

[0035] The inventors randomly recruited 38 males with reduced sperm count from the reproductive clinic of the hospital, and matched 38 males according to age, BMI, and abstinence time. A total of 76 males were used as experimental subjects for pendimethalin exposure assessment, and serum samples meeting the requirements were collected. details as follows:

[0036] The first stage of association exploration

[0037] Research objects: 38 men with reduced sperm count were included, and 38 matched controls were matched according to age, BMI, and abstinence time, a total of 76 people.

[0038] Inclusion criteria:

[0039] 1. Between 19 and 39 years old;

[0040] 2. BMI between 17 and 31;

...

Embodiment 2

[0045] Example 2 Correlation analysis between GC-MS detection of pendimethalin exposure and sperm count

[0046] Sample pretreatment

[0047] 1.1 Precipitation of serum proteins

[0048] Take 0.5mL serum sample, add 0.5mL reagent A, vortex for more than 30s, centrifuge at 16000rpm for 15min at 4°C, take another EP tube, and transfer the centrifuged supernatant.

[0049] 1.2 Solid phase extraction

[0050] 1.2.1 Cartridge Activation

[0051] Take a small column, add 3mL of reagent B to the small column, after the natural flow is exhausted, add 3mL of reagent C, wait for the natural flow to be exhausted, and set aside.

[0052] 1.2.2 Sample loading

[0053] Transfer all the supernatant solution of the EP tube in the above step 1.1 to the activated small column, control the flow rate, and let it drain naturally.

[0054] 1.2.3 Impurity removal

[0055] After the sample drains off naturally, add 1mL of reagent C to the small column. After draining off naturally, use a vacuum...

Embodiment 3

[0080] Example 3 Production of kits for detecting male serum pendimethalin concentration and early screening male sperm count

[0081] The concentration of pendimethalin in the serum of the reduced sperm count group was determined to have a higher level by GC-MS detection. The concentration of pendimethalin in the serum can be used as a marker of the effect of sperm count reduction. The detection of pendimethalin exposure can be used for semen Early screening of quality. This kit contains a batch of reagents and consumables for the detection of pendimethalin in serum, including reagents for precipitation of sample proteins (saturated ammonium sulfate solution), reagents for solid phase extraction (chromatographic grade methanol, ultra-pure water, eluent, chromatographic grade n-hexane) and reagents for sample reconstitution (chromatographic grade dichloromethane and chromatographic grade ethyl acetate). Others include pendimethalin standard curve concentration gradient, a C18...

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Abstract

The invention belongs to the field of analytical chemistry and clinical medicine, and is particularly relates to a detection method based on GC-MS (gas chromatography-mass spectrometry) sperm countingand application; the detection method comprise the steps: stage one, selecting research objects and providing grouping basis; stage two, preparing a sample; stage three, loading the sample; and stagefour, carrying out data analysis. The objects of the invention are to: establish the GC-MS based detection method for pendimethalin exposure in serum; apply pendimethalin exposure as an effect markerof sperm count reduction; and develop a diagnostic kit for pendimethalin exposure detection in serum. The detection of pendimethalin exposure in serum provided by the invention helps predict the riskof reduced sperm count in an early stage through a minimally invasive mode; compared with semen collection, the pendimethalin exposure detection is more time-saving and handier, is suitable for earlyscreening for the sperm count of a male of childbearing age, and can provide basis for further in-depth examination for clinicians.

Description

technical field [0001] The invention belongs to the field of analytical chemistry and clinical medicine, in particular to a detection method and application based on GC-MS sperm counting. Background technique [0002] After analyzing the sperm of more than 10,000 people from 1981 to 1996, the Institute of Science and Technology of the National Population and Family Planning Commission found that the semen quality of Chinese men is declining at a rate of 1% per year, and the rate of decline is more than 40%. As of 2017, the qualified rate of sperm donation in the Shanghai sperm bank was only 25%, while the analysis data of the human sperm bank in Hubei Province from 2011 to 2017 also found that the qualified rate of sperm donation was less than 30%. The decline in the qualified rate of sperm donation reflects to a certain extent that the sperm level of the overall population is declining. Studies have found that between 1973 and 2011, men in North America, Europe, Australia ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/02G01N30/02
CPCG01N33/5005G01N30/02Y02A90/10
Inventor 曾桥胡西陵陈继勇
Owner ZHEJIANG CELLPRO BIOTECH
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