CN110699418A - Detection method based on GC-MS sperm counting and application - Google Patents
Detection method based on GC-MS sperm counting and application Download PDFInfo
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Abstract
The invention belongs to the fields of analytical chemistry and clinical medicine, in particular to a detection method based on GC-MS sperm count and an application thereof, comprising the following steps: stage one: research object selection and grouping basis; and a second stage: preparing a sample; and a third stage: sample machine and stage four: data analysis, object of the invention: 1. establishing a detection method of pendimethalin exposure in serum based on GC-MS; 2. pendimethalin exposure is applied as an effect marker for sperm count reduction; 3. the diagnostic kit for developing the pendimethalin exposure detection in the serum can predict the risk of sperm count reduction in a minimally invasive manner in the early stage, is more time-saving and convenient compared with semen collection, can screen the sperm count of male of child bearing age in the early stage, and provides a basis for further deep inspection by a clinician.
Description
Technical Field
The invention belongs to the fields of analytical chemistry and clinical medicine, and particularly relates to a detection method based on GC-MS (gas chromatography-Mass spectrometer) sperm counting and application.
Background
After the scientific and technical research of the national population and family planning committee analyzes the sperms of ten thousand people in total from 1981 to 1996, the semen quality of the male in China is reduced at the speed of 1% per year, and the reduction amplitude is more than 40%. By 2017, the sperm-donation qualified rate of Shanghai sperm bank is only 25%, and the sperm-donation qualified rate is less than 30% by the analysis data of Hubei province human sperm bank from 2011 to 2017. The decline of the sperm donation qualification rate reflects the decline of the sperm level of the whole population to a certain extent. Studies have found that between 1973 and 2011, the number of sperm in men in north america, europe, australia and new zealand has decreased dramatically and fertility has decreased by 50-60%. According to the evaluation data of the world health organization for the last 10 years, compared with the data before thirty years, the number of sperms contained in each milliliter of semen of Chinese men is about 1 hundred million, which is greatly reduced to 2000 to 4000 million at present. Sperm count is closely related to conception and when sperm count is reduced, fertility chances will be reduced. Possible causes of reduced sperm count include: age, smoking or drinking, poor lifestyle, environmental factors, etc., where environmental chemical exposure is more easily overlooked clinically due to insufficient emphasis and limitations of detection techniques.
China is a big country for using pesticides, and the production and use amount are the first world. At present, pendimethalin is the 3 rd herbicide in the world, and becomes the selective herbicide with the largest sales amount by virtue of the broad spectrum, high efficiency and good quality. Pendimethalin is mainly used for weeding in cotton, corn, rice, potato, soybean, peanut, tobacco and vegetable fields. Research has shown that pesticide exposure has adverse effects on male semen quality, which may lead to decreased sperm count, decreased motility, abnormal morphology, etc. However, most studies have evaluated pesticide exposure in the form of questionnaires, and in vivo detection is not performed, and the type and concentration of pesticide exposure cannot be specified. The effect of pendimethalin exposure on male reproductive health is therefore not currently clear.
GC-MS (gas chromatography-mass spectrometer) has been widely used for detecting pesticide residues in the environment and human body due to its advantages of high separation efficiency, simple and convenient quantitative analysis, high sensitivity, strong qualitative ability and the like. The use of GC-MS to analyze the level of pendimethalin exposure in male sera in sperm counting has not received corresponding attention. If the exposure of pendimethalin can be found as an effect marker for sperm count reduction, and a corresponding GC-MS detection method is developed, the stability, accuracy and sensitivity of a detection result are improved, the sperm count of fertile men can be screened and evaluated in an early stage, and the reproductive health level of men in China is further promoted and improved.
Disclosure of Invention
The purpose of the invention is: 1. establishing a detection method of pendimethalin exposure in serum based on GC-MS; 2. pendimethalin exposure is applied as an effect marker for sperm count reduction; 3. a diagnostic kit for the detection of pendimethalin exposure in serum was developed.
In order to solve the technical problem, the detection method based on GC-MS sperm counting and the application thereof comprise the following stages:
stage one: research object selection and grouping basis;
and a second stage: preparing a sample;
the second stage comprises the following steps:
step 1: precipitating serum protein, taking 0.5mL of serum sample, adding 0.5mL of saturated ammonium sulfate, performing vortex oscillation for more than 30s at the solution temperature of 4 ℃, the centrifugal rotation speed of 16000rpm, centrifuging for 15min, taking another EP tube, and transferring the centrifuged supernatant;
step 2: solid phase extraction, firstly, activating a small column, taking the small column, adding 3mL of methanol into the small column, adding 3mL of ultrapure water after the solution is naturally drained off, standing for later use, then loading, transferring all supernatant solution of the EP tube in the step 1 into the activated small column, controlling the flow rate, naturally draining, further removing impurities, adding 1mL of ultrapure water into the small column after the sample is naturally drained off, draining for 15min by using a vacuum pump-20 psi after the sample is naturally drained off, removing residual water phase in the extraction column, finally, collecting the sample, replacing a waste liquid collecting tube by using a 15mL centrifugal tube, adding 4mL of eluent into the small column, adding 4mL of n-hexane after the sample is naturally drained off, pumping for 10min by using a vacuum pump-20 psi, and collecting the object to be detected into the centrifugal tube;
and step 3: concentrating, namely firstly, blowing and concentrating the collected sample dissolved with liquid nitrogen until the nitrogen is blown to be nearly dry, adding 200 mu L of dichloromethane solution after the nitrogen is blown to be nearly dry, carrying out vortex oscillation for 1min, completely transferring the vortexed solution to a brown sample bottle provided with 200 mu L of an inner inserting tube, adding 20 mu L of ethyl acetate solution for redissolution after the liquid is naturally volatilized to be dried, and waiting for detection;
and a third stage: the sample is loaded on the machine, the GC-MS is vacuumized after being started, and the stability is realized, and the loading detection is carried out;
and a fourth stage: and (6) analyzing the data.
Preferably, the kit contains the following reagents:
reagent A: saturated ammonium sulfate solution
And (3) reagent B: chromatographic grade methanol
And (3) reagent C: ultrapure water
And (3) reagent D: eluent
And (3) reagent E: chromatographic grade n-hexane
And (3) reagent F: chromatographic grade dichloromethane
Reagent G: chromatographic grade ethyl acetate
And (3) reagent H: pendimethalin reagent
C18 columella.
Preferably, in stage three, the instrument GC uses high purity helium as a carrier gas at a flow rate of 1 mL/min.
Preferably, the MS mass spectrum end instrument of the stage three instrument is characterized in that the collision gas is high-purity argon, and the flow rate is 1 mL/min;
preferably, the sample injection mode in the third stage is as follows: automatic sample introduction, wherein the sample introduction time is 21 minutes, and the sample introduction needle is used for more than 3 times.
Preferably, the ratio of chromatographic grade dichloromethane/methanol in reagent D is 8: 2.
Preferably, the ratio of reagent H: the concentration of pendimethalin reagent is 0, 0.005, 0.01, 0.05, 0.1, 0.5, 1, 5, 10, 50ug/L respectively.
The invention has the advantages that:
(1) the GC-MS instrument disclosed by the invention is rapid and accurate in analysis, good in stability and higher in clinical diagnosis practical value.
(2) Serum samples of people with reduced sperm count and control people are adopted for verification, and the risk that sperm count is reduced due to pendimethalin exposure is proved.
(3) The exposure detection of pendimethalin in serum provided by the invention can predict the risk of sperm count reduction in a minimally invasive manner at an early stage, is more time-saving and convenient compared with semen collection, can carry out early screening on the sperm count of male of child bearing age, and provides a basis for further deep inspection by a clinician.
Detailed Description
In order to make the technical means, the creation characteristics, the achievement purposes and the effects of the invention easy to understand, the invention is further described with the specific embodiments. Example 1 study selection and grouping basis
The inventor randomly incorporates 38 men with reduced sperm count from a hospital reproductive clinic, matches the control with 38 men according to age, BMI and abstinence time, and takes 76 men as experimental objects for pendimethalin exposure evaluation to collect serum samples meeting the requirements. The method comprises the following specific steps:
first phase associative exploration phase
Study subjects: the inclusion sperm count decreased in 38 men, matched control 38 based on age, BMI, abstinence time, for a total of 76.
Inclusion criteria were:
1. age between 19 and 39 years;
BMI between 17 and 31;
3. abstinence time is 3-7 days;
4. there was no history of other diseases associated with male reproductive disorders.
Grouping basis:
sperm count < 39 x 106/ejaculation, i.e. decreased sperm count.
Example 2GC-MS detection of pendimethalin exposure correlation analysis with sperm count
Sample pretreatment
1.1 precipitation of serum proteins
Taking 0.5mL serum sample, adding 0.5mL reagent A, vortexing and shaking for more than 30s, centrifuging at 16000rpm for 15min at 4 ℃, taking another EP tube, and transferring the centrifuged supernatant.
1.2 solid phase extraction
1.2.1 pillar activation
And (3) taking a small column, adding 3mL of reagent B into the small column, and after the small column is drained naturally, adding 3mL of reagent C into the small column for later use after the small column is drained naturally.
1.2.2 Loading
And (3) completely transferring the supernatant solution of the EP tube in the step 1.1 to an activated small column, controlling the flow rate, and naturally draining.
1.2.3 removing impurities
After the sample naturally flows out, 1mL of reagent C is added into the small column, and after the sample naturally flows out, the sample is pumped by a vacuum pump at-20 psi for 15min to remove the residual water phase in the extraction column as far as possible.
1.2.4 sample Collection
And replacing the waste liquid collecting pipe with a 15mL centrifuge tube, adding 4mL of the reagent D into the small column, adding 4mL of the reagent E after the natural flow-out, pumping for 10min by using a vacuum pump at-20 psi after the natural flow-out, and collecting the object to be detected into the centrifuge tube.
1.3 concentration
1.3.1 the collected sample is concentrated by blowing the solution into liquid nitrogen, and the nitrogen is blown to near dryness.
1.3.2 after nitrogen blow-dry, 200. mu.L of reagent F was added and vortexed for 1 min.
1.3.3 vortex and transfer all to a brown sample bottle with 200. mu.L of internal cannula, and allow the liquid to evaporate naturally.
1.3.4 adding 20 microliter of reagent G for redissolving and testing.
2. Sample machine
The concrete steps and parameters are as follows:
after the GC-MS is started, vacuumizing is performed and the like is stable, TSQSeries dashboard software is started, and the experiment can be started after the instrument parameters and the tuning parameters pass through the back.
Instrument GC acquisition parameters: high purity helium was used as a carrier gas at a flow rate of 1 mL/min. A Saimerfi TR-Pest II column (30m 0.25mm 0.25 μm) was used.
GC set parameters
c. Instrument MS Mass Spectrometry end instrument setup parameters: the collision gas is high-purity argon, and the flow rate is 1 mL/min.
MS recommended parameters
And (3) sample introduction mode: automatic sample introduction, wherein the sample introduction time is 21 minutes, and the sample introduction needle is used for more than 3 times.
Data processing: data were collected online using tracefinder3.3 software (seemefly). The sample name, location number, sample type, etc. are input.
And leading the peak area, the peak height, the peak-out time value and the like out to an excel table.
3. Data analysis
Welch' st-test was used to compare the level of pendimethalin exposure in healthy controls and in the reduced sperm count group; linear regression was used to analyze the correlation of pendimethalin content in serum with sperm count. Correlation of pendimethalin content in serum with reduced sperm count was analyzed using Logistic regression. All data were analyzed statistically after lg transformation.
4. Difference in pendimethalin exposure in serum samples from sperm count-reduced groups and control groups
The pendh' st-test found that the level of pendimethalin exposed water was higher in the group with decreased sperm count than in the control group. Linear regression found that sperm counts decreased progressively with increasing serum pendimethalin concentrations (β ═ 0.22, 95% CI: -0.4, -0.04). Logistic regression found that the risk of decreased sperm count increased with increasing concentration of pendimethalin (OR ═ 2.58, 95% CI:1.13, 5.89).
EXAMPLE 3 preparation of a kit for detecting Male serum pendimethalin concentration and early screening Male sperm count
The higher level of pendimethalin concentration in the serum of the sperm count reduction group is determined through GC-MS detection, the pendimethalin concentration in the serum can be used as an effect marker of sperm count reduction, and the detection of pendimethalin exposure can be used for early screening of semen quality. The kit comprises a batch of reagents and consumables for detecting pendimethalin in serum, wherein the reagents comprise a reagent (saturated ammonium sulfate solution) for precipitating sample protein, a reagent (chromatographic grade methanol, ultrapure water, eluent, chromatographic grade n-hexane) for solid phase extraction and a reagent (chromatographic grade dichloromethane and chromatographic grade ethyl acetate) for a sample re-dissolving machine. Other examples include pendimethalin standard curve concentration gradient, a C18 column for standard curve drawing for quantitative analysis and solid phase extraction. The kit has the value that the pendimethalin content in the serum can be detected only by 500 mul of male serum, the risk of occurrence of sperm count reduction is evaluated, the kit can be used for early screening of the semen quality, and dynamic monitoring and observation of the treatment effect are easy to carry out.
The specific kit comprises the following components:
reagent A: saturated ammonium sulfate solution
And (3) reagent B: chromatographic grade methanol
And (3) reagent C: ultrapure water
And (3) reagent D: eluent (chromatographic grade dichloromethane/methanol ratio 8:2)
And (3) reagent E: chromatographic grade n-hexane
And (3) reagent F: chromatographic grade dichloromethane
Reagent G: chromatographic grade ethyl acetate
And (3) reagent H: the concentration of pendimethalin is 0, 0.005, 0.01, 0.05, 0.1, 0.5, 1, 5, 10, 50ug/L
C18 columella (200mg/3ml, ProEIut).
Claims (7)
1. The detection method based on GC-MS sperm counting and the application thereof are characterized in that: the method comprises the following steps:
stage one: research object selection and grouping basis;
and a second stage: preparing a sample;
the second stage comprises the following steps:
step 1: precipitating serum protein, taking 0.5mL of serum sample, adding 0.5mL of saturated ammonium sulfate, performing vortex oscillation for more than 30s at the solution temperature of 4 ℃, the centrifugal rotation speed of 16000rpm, centrifuging for 15min, taking another EP tube, and transferring the centrifuged supernatant;
step 2: solid phase extraction, firstly, activating a small column, taking the small column, adding 3mL of methanol into the small column, adding 3mL of ultrapure water after the solution is naturally drained off, standing for later use, then loading, transferring all supernatant solution of the EP tube in the step 1 into the activated small column, controlling the flow rate, naturally draining, further removing impurities, adding 1mL of ultrapure water into the small column after the sample is naturally drained off, draining for 15min by using a vacuum pump-20 psi after the sample is naturally drained off, removing residual water phase in the extraction column, finally, collecting the sample, replacing a waste liquid collecting tube by using a 15mL centrifugal tube, adding 4mL of eluent into the small column, adding 4mL of n-hexane after the sample is naturally drained off, pumping for 10min by using a vacuum pump-20 psi, and collecting the object to be detected into the centrifugal tube;
and step 3: concentrating, namely firstly, blowing and concentrating the collected sample dissolved with liquid nitrogen until the nitrogen is blown to be nearly dry, adding 200 mu L of dichloromethane solution after the nitrogen is blown to be nearly dry, carrying out vortex oscillation for 1min, completely transferring the vortexed solution to a brown sample bottle provided with 200 mu L of an inner inserting tube, adding 20 mu L of ethyl acetate solution for redissolution after the liquid is naturally volatilized to be dried, and waiting for detection;
and a third stage: the sample is loaded on the machine, the GC-MS is vacuumized after being started, and the stability is realized, and the loading detection is carried out;
and a fourth stage: and (6) analyzing the data.
2. The GC-MS sperm count-based detection method and use of claim 1, characterized in that: the kit contains the following reagents:
reagent A: saturated ammonium sulfate solution
And (3) reagent B: chromatographic grade methanol
And (3) reagent C: ultrapure water
And (3) reagent D: eluent
And (3) reagent E: chromatographic grade n-hexane
And (3) reagent F: chromatographic grade dichloromethane
Reagent G: chromatographic grade ethyl acetate
And (3) reagent H: pendimethalin reagent
C18 columella.
3. The GC-MS sperm count-based detection method and use of claim 1, characterized in that: and in the third stage, the instrument GC uses high-purity helium gas as carrier gas, and the flow rate is 1 mL/min.
4. The GC-MS sperm count-based detection method and use of claim 1, characterized in that: and a step three instrument MS mass spectrum end instrument, wherein collision gas is high-purity argon, and the flow rate is 1 mL/min.
5. The GC-MS sperm count-based detection method and use of claim 1, characterized in that: stage three sample introduction mode: automatic sample introduction, wherein the sample introduction time is 21 minutes, and the sample introduction needle is used for more than 3 times.
6. The GC-MS sperm count based detection method and use of claim 2, characterized in that: the ratio of chromatographic grade dichloromethane/methanol in reagent D was 8: 2.
7. The GC-MS sperm count based detection method and use of claim 2, characterized in that: and (3) reagent H: the concentration of pendimethalin reagent is 0, 0.005, 0.01, 0.05, 0.1, 0.5, 1, 5, 10, 50ug/L respectively.
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Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1166598A (en) * | 1996-02-27 | 1997-12-03 | A·阿米拉夫 | Method and device for introduction of sample into gas chromatograph |
| ES2225604T3 (en) * | 2000-08-25 | 2005-03-16 | Basf Aktiengesellschaft | CONTINUOUS PROCEDURE FOR THE PREPARATION OF A STABLE CONCENTRATED HERBICIAN WATER COMPOSITION FOR SUSPENSIONS. |
| EP2339352A1 (en) * | 2009-12-28 | 2011-06-29 | BIOCRATES Life Sciences AG | Use of endogenous metabolites for early diagnosing sepsis |
| CN103293266A (en) * | 2013-06-08 | 2013-09-11 | 国家烟草质量监督检验中心 | GC-MS/MS (gas chromatography-tandem mass spectrometry) method for analyzing dinitroaniline pesticide residue in tobacco and tobacco product |
| CN103529155A (en) * | 2013-09-27 | 2014-01-22 | 中国检验检疫科学研究院 | Method for measuring content of wood preservatives of wooden products for children |
| CN106596741A (en) * | 2015-10-16 | 2017-04-26 | 江苏维赛科技生物发展有限公司 | Method for detecting basic orange content of food by high performance liquid chromatography |
| WO2018057647A1 (en) * | 2016-09-23 | 2018-03-29 | Alveo Technologies, Inc. | Methods and compositions for detecting analytes |
| WO2018092170A1 (en) * | 2016-11-15 | 2018-05-24 | Lta-Biotech S.R.L. | Method for in vitro diagnosis of male infertility |
| CN110704810A (en) * | 2019-11-14 | 2020-01-17 | 浙江星博生物科技股份有限公司 | Detection method and application based on ICP-MS (inductively coupled plasma-mass spectrometry) semen quality |
-
2019
- 2019-11-14 CN CN201911110723.6A patent/CN110699418B/en active Active
Patent Citations (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1166598A (en) * | 1996-02-27 | 1997-12-03 | A·阿米拉夫 | Method and device for introduction of sample into gas chromatograph |
| ES2225604T3 (en) * | 2000-08-25 | 2005-03-16 | Basf Aktiengesellschaft | CONTINUOUS PROCEDURE FOR THE PREPARATION OF A STABLE CONCENTRATED HERBICIAN WATER COMPOSITION FOR SUSPENSIONS. |
| EP2339352A1 (en) * | 2009-12-28 | 2011-06-29 | BIOCRATES Life Sciences AG | Use of endogenous metabolites for early diagnosing sepsis |
| CN103293266A (en) * | 2013-06-08 | 2013-09-11 | 国家烟草质量监督检验中心 | GC-MS/MS (gas chromatography-tandem mass spectrometry) method for analyzing dinitroaniline pesticide residue in tobacco and tobacco product |
| CN103529155A (en) * | 2013-09-27 | 2014-01-22 | 中国检验检疫科学研究院 | Method for measuring content of wood preservatives of wooden products for children |
| CN106596741A (en) * | 2015-10-16 | 2017-04-26 | 江苏维赛科技生物发展有限公司 | Method for detecting basic orange content of food by high performance liquid chromatography |
| WO2018057647A1 (en) * | 2016-09-23 | 2018-03-29 | Alveo Technologies, Inc. | Methods and compositions for detecting analytes |
| CN109996888A (en) * | 2016-09-23 | 2019-07-09 | 阿尔韦奥科技公司 | For testing and analyzing the method and composition of object |
| WO2018092170A1 (en) * | 2016-11-15 | 2018-05-24 | Lta-Biotech S.R.L. | Method for in vitro diagnosis of male infertility |
| CN110704810A (en) * | 2019-11-14 | 2020-01-17 | 浙江星博生物科技股份有限公司 | Detection method and application based on ICP-MS (inductively coupled plasma-mass spectrometry) semen quality |
Non-Patent Citations (3)
| Title |
|---|
| JIYEON HAM等: "Pendimethalin induces apoptosis in testicular cells via hampering ER-mitochondrial function and autophagy", 《ENVIRONMENTAL POLLUTION》 * |
| 张芬 等: "常见除草剂的睾丸毒性作用及其机制研究进展", 《职业与健康》 * |
| 黄婷 等: "农药除草剂雄性生殖毒性研究进展", 《实用预防医学》 * |
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