CN108918714A - A kind of detection method of cholesterol level - Google Patents
A kind of detection method of cholesterol level Download PDFInfo
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- CN108918714A CN108918714A CN201810794253.9A CN201810794253A CN108918714A CN 108918714 A CN108918714 A CN 108918714A CN 201810794253 A CN201810794253 A CN 201810794253A CN 108918714 A CN108918714 A CN 108918714A
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- cholesterol
- sample
- detection method
- reference substance
- conical flask
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
Abstract
The invention discloses a kind of detection methods of cholesterol level, include the following steps：S1：Raw material is chosen；S2：Weighing；S3：Bottling charging；S4：Dilution shakes up；S5：Filtering；S6：Sample introduction is analyzed；S7：It calculates.The present invention is by weighing cholesterol reference substance and cholesterol sample as test sample, and the mode detected by using high performance liquid chromatography, manual operation detection error is small, and detection accuracy and detection efficiency are high, and favorable reproducibility, accuracy rate is high, is suitble to be widely popularized.
Description
Technical field
The present invention relates to cholesterol detection technical field, specially a kind of detection method of cholesterol level.
Background technique
Cholesterol is a kind of derivative of cyclopentanoperhy drophenanthrene.Early in 18th century, people have had found that gallbladder is solid from cholelith
The substance of this tool lipid nature is named as cholesterol by alcohol, chemist Ben Xieer in 1816.Cholesterol is widely present in animal
In vivo, especially with the abundantest in brain and nerve fiber, content is also high in kidney, spleen, skin, liver and bile.Its dissolubility and rouge
Fat is similar, not soluble in water, is soluble in ether, chloroform equal solvent.Cholesterol is the indispensable important object of institute of animal tissue cell
Matter, it is not only involved in form cell membrane, and is synthetic bile acid, the raw material of vitamin D and steroid hormone.Cholesterol is through generation
Bile acid, steroid hormone, 7-DHC can also be converted by thanking.
Currently, cholesterol level does not have detection method, only using the height of cholesterol fusing point as cholesterol quality it is good with
Bad, therefore, it is necessary to establish a kind of accurate and effective cholesterol level detection method.
Summary of the invention
The purpose of the present invention is to provide a kind of detection methods of cholesterol level, can accurately measure cholesterol in sample
Content, method is simple, strong operability, favorable reproducibility, and it is the effective ways of cholesterol detection content that accuracy is high, with solution
Certainly the problems mentioned above in the background art.
To achieve the above object, the present invention provides the following technical solutions：A kind of detection method of cholesterol level, including with
Lower step：
S1：Raw material is chosen, and samples cholesterol reference substance and cholesterol sample as detection raw material, equal sampling amount is no less than 4mg, standby
With；
S2：Weighing, weighs cholesterol reference substance and cholesterol sample, and cholesterol reference substance samples 4-6mg, cholesterol sample
4-6mg；
S3：Bottling charging, cholesterol reference substance weighed in S2 and cholesterol sample are respectively put into conical flask, and are added respectively
Enter the highest graduation position that ethyl alcohol is dissolved to conical flask；
S4：Dilution shakes up, and the conical flask bottleneck for completion of bottling in S3 is sealed processing, and rocks conical flask and make internal object
Material dissolution shakes up, and rocking the time is 5-10min, spare；
S5：Filtering, the liquid that completion is rocked in S4 is filtered by filter membrane, and filtrate is collected, spare；
S6：Sample introduction is analyzed, and the filtrate collected in S5 is taken using microsyringe, takes 10-12ul, and the filtrate of acquirement is injected liquid phase
In chromatograph, and record chromatogram；
S7：It calculates, calculates main peak content by external standard method, can be obtained the content of cholesterol.
Preferably, sampling device is weighed using ESJ-A series full-automatic electronic assay balance in the step S2.
Preferably, the volume range of conical flask is 50-60ml in the step S3.
Preferably, ethyl alcohol is dehydrated alcohol in the step S3.
Preferably, filter membrane is 0.45um filter membrane in the step S5.
Preferably, liquid chromatograph minimum peak area is 3000 in the step S6.
Preferably, the formula of the content of calculating cholesterol is in the step S7：
。
Preferably, liquid chromatograph chromatographic condition and system suitability are with octadecylsilane in the step S7
Bonded silica gel is filler；Using methanol as mobile phase, Detection wavelength 210nm, number of theoretical plate should not be low by the calculating of cholesterol peak
In 2000.
Compared with prior art, the beneficial effects of the invention are as follows：The present invention is by weighing cholesterol reference substance and cholesterol
Sample is as test sample, and the mode detected by using high performance liquid chromatography, manual operation detection error is small, detection
Precision and detection efficiency are high, and favorable reproducibility is suitble to be widely popularized.
Specific embodiment
In order to make the objectives, technical solutions, and advantages of the present invention clearer, below in conjunction with specific embodiment, to this
Invention is further elaborated.It should be appreciated that the specific embodiments described herein are merely illustrative of the present invention, not
For limiting the present invention.
Embodiment 1
A kind of detection method of cholesterol level, includes the following steps：
S1：Raw material is chosen, and samples cholesterol reference substance and cholesterol sample as detection raw material, samples 4mg, spare；
S2：Weighing weighs cholesterol reference substance and cholesterol sample in S1, and cholesterol reference substance samples 4mg, and cholesterol sample takes
Sample 4mg；
S3：Bottling charging, cholesterol reference substance weighed in S2 and cholesterol sample are respectively put into conical flask, and second respectively
Alcohol to conical flask highest graduation position；
S4：Dilution shakes up, and the brown conical flask bottleneck for completion of bottling in S3 is sealed processing, and rock conical flask to make inside
Material dissolution shake up, rock the time be 5min, it is spare；
S5：Filtering, the mixed liquor that completion is rocked in S4 is filtered by filter membrane, and filtrate is collected, spare；
S6：Sample introduction is analyzed, and the filtrate collected in S5 is taken using microsyringe, takes 10ul, and the filtrate of acquirement is injected liquid phase color
In spectrometer, and record chromatogram；
S7：It calculates, calculates main peak content by external standard method, can be obtained the content of cholesterol.
Specifically, weighing sampling device using ESJ-A series full-automatic electronic assay balance in the step S2.
Specifically, the volume of brown conical flask is 50ml in the step S3.
Specifically, ethyl alcohol is dehydrated alcohol in the step S3.
Specifically, filter membrane is 0.45um filter membrane in the step S5.
Specifically, liquid chromatograph minimum peak area is 3000 in the step S6.
Specifically, the formula for calculating the content of cholesterol in the step S7 is：
。
Specifically, liquid chromatograph chromatographic condition and system suitability are with octadecylsilane in the step S7
Bonded silica gel is filler；Using methanol as mobile phase, Detection wavelength 210nm, number of theoretical plate is calculated as 2000 by cholesterol peak.
Embodiment 2
A kind of detection method of cholesterol level, includes the following steps：
S1：Raw material is chosen, and samples cholesterol reference substance and cholesterol sample as detection raw material, samples 5mg, spare；
S2：Weighing weighs cholesterol reference substance and cholesterol sample in S2, and cholesterol reference substance samples 5mg, and cholesterol sample takes
Sample 5mg；
S3：Bottling charging, cholesterol reference substance weighed in S2 and cholesterol sample are respectively put into conical flask, and are added respectively
Enter the highest graduation position of ethyl alcohol to conical flask；
S4：Dilution shakes up, and the conical flask bottleneck for completion of bottling in S3 is sealed processing, and rocks conical flask and make internal object
Material dissolution shakes up, and rocking the time is 7.5min, spare；
S5：Filtering, the liquid that completion is rocked in S4 is filtered by filter membrane, and filtrate is collected, spare；
S6：Sample introduction is analyzed, and the filtrate collected in S5 is taken using microsyringe, takes 11ul, and the filtrate of acquirement is injected liquid phase color
In spectrometer, and record chromatogram；
S7：It calculates, calculates main peak content by external standard method, can be obtained the content of cholesterol.
Specifically, weighing sampling device using ESJ-A series full-automatic electronic assay balance in the step S2.
Specifically, the volume of brown conical flask is 55ml in the step S3.
Specifically, ethyl alcohol is dehydrated alcohol in the step S3.
Specifically, filter membrane is 0.45um filter membrane in the step S5.
Specifically, liquid chromatograph minimum peak area is 3000 in the step S6.
Specifically, the formula for calculating the content of cholesterol in the step S7 is：
。
Specifically, liquid chromatograph chromatographic condition and system suitability are with octadecylsilane in the step S7
Bonded silica gel is filler；Using methanol as mobile phase, Detection wavelength 210nm, number of theoretical plate is calculated as 2500 by cholesterol peak.
Embodiment 3
A kind of detection method of cholesterol level, includes the following steps：
S1：Raw material is chosen, and samples cholesterol reference substance and cholesterol sample as detection raw material, samples 6mg, spare；
S2：Weighing, weighs cholesterol reference substance and cholesterol sample in S2, and cholesterol reference substance samples 6mg, cholesterol sample
Sample 6mg；
S3：Bottling charging, cholesterol reference substance weighed in S2 and cholesterol sample are respectively put into conical flask, and are added respectively
Enter the highest graduation position of ethyl alcohol to conical flask；
S4：Dilution shakes up, and the conical flask bottleneck for completion of bottling in S3 is sealed processing, and rocks conical flask and make internal object
Material dissolution shakes up, and rocking the time is 10min, spare；
S5：Filtering, the mixed liquor that completion is rocked in S4 is filtered by filter membrane, and filtrate is collected, spare；
S6：Sample introduction is analyzed, and the filtrate collected in S5 is taken using microsyringe, takes 12ul, and the filtrate of acquirement is injected liquid phase color
In spectrometer, and record chromatogram；
S7：It calculates, calculates main peak content by external standard method, can be obtained the content of cholesterol.
Specifically, weighing sampling device using ESJ-A series full-automatic electronic assay balance in the step S2.
Specifically, the volume of brown conical flask is 60ml in the step S3.
Specifically, ethyl alcohol is dehydrated alcohol in the step S3.
Specifically, filter membrane is 0.45um filter membrane in the step S5.
Specifically, liquid chromatograph minimum peak area is 3000 in the step S6.
Specifically, the formula for calculating the content of cholesterol in the step S7 is：
。
Specifically, liquid chromatograph chromatographic condition and system suitability are with octadecylsilane in the step S7
Bonded silica gel is filler；Using methanol as mobile phase, Detection wavelength 210nm, number of theoretical plate is calculated as 3000 by cholesterol peak.
In summary：The present invention by weighing cholesterol reference substance and cholesterol sample as test sample, by using
The mode that high performance liquid chromatography is detected, manual operation detection error is small, and detection accuracy and detection efficiency are high, reproducibility
It is good, it is suitble to be widely popularized.
The foregoing is only a preferred embodiment of the present invention, but scope of protection of the present invention is not limited thereto,
Anyone skilled in the art in the technical scope disclosed by the present invention, according to the technique and scheme of the present invention and its
Inventive concept is subject to equivalent substitution or change, should be covered by the protection scope of the present invention.
Claims (8)
1. a kind of detection method of cholesterol level, it is characterised in that：Include the following steps：
S1：Raw material is chosen, and samples cholesterol reference substance and cholesterol sample as detection raw material, equal sampling amount is no less than 4mg, standby
With；
S2：Weighing, cholesterol reference substance and cholesterol sample, cholesterol reference substance sample 4-6mg, cholesterol sample 4-
6mg；
S3：Bottling charging, cholesterol reference substance weighed in S2 and cholesterol sample are respectively put into conical flask, and add ethyl alcohol
To the highest graduation position of conical flask；
S4：Dilution shakes up, and the conical flask bottleneck for completion of bottling in S3 is sealed processing, and rocks conical flask and make internal object
Material dissolution shakes up, and rocking the time is 5-10min, spare；
S5：Filtering, the liquid that completion is rocked in S4 is filtered by filter membrane, and filtrate is collected, spare；
S6：Sample introduction is analyzed, and the filtrate collected in S5 is taken using microsyringe, takes 10-12ul, and the filtrate of acquirement is injected liquid phase
In chromatograph, and record chromatogram；
S7：It calculates, calculates main peak content by external standard method, can be obtained the content of cholesterol.
2. a kind of detection method of cholesterol level according to claim 1, it is characterised in that：Title in the step S2
Take sampling device using ESJ-A series full-automatic electronic assay balance.
3. a kind of detection method of cholesterol level according to claim 1, it is characterised in that：Taper in the step S3
The volume range of bottle is 50-60ml.
4. a kind of detection method of cholesterol level according to claim 1, it is characterised in that：Ethyl alcohol in the step S3
For dehydrated alcohol.
5. a kind of detection method of cholesterol level according to claim 1, it is characterised in that：Filter membrane in the step S5
For 0.45um filter membrane.
6. a kind of detection method of cholesterol level according to claim 1, it is characterised in that：Liquid phase in the step S7
Chromatograph minimum peak area is 3000.
7. a kind of detection method of cholesterol level according to claim 1, it is characterised in that：It is calculated in the step S7
The formula of the content of cholesterol is：
。
8. a kind of detection method of bilirubin according to claim 1, it is characterised in that：Liquid chromatogram in the step S7
Instrument chromatographic condition and system suitability are using octadecylsilane chemically bonded silica as filler；Using methanol as mobile phase, inspection
Survey wavelength is 210nm, and number of theoretical plate is calculated by cholesterol peak should be not less than 2000.
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| CN201810794253.9A CN108918714A (en) | 2018-07-19 | 2018-07-19 | A kind of detection method of cholesterol level |
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| CN201810794253.9A CN108918714A (en) | 2018-07-19 | 2018-07-19 | A kind of detection method of cholesterol level |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114689759A (en) * | 2022-04-15 | 2022-07-01 | 安徽丰本生物科技有限公司 | Method for measuring content of cholesterol in fish oil |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102507791A (en) * | 2011-11-17 | 2012-06-20 | 吉林大学 | Method for measuring cholesterol content in birds' egg and product thereof |
| CN102731557A (en) * | 2011-04-02 | 2012-10-17 | 北京市理化分析测试中心 | Steroid substance derivation method and its analysis detection method |
| CN106198798A (en) * | 2016-07-06 | 2016-12-07 | 汤臣倍健股份有限公司 | A kind of detection method of cholesterol level |
-
2018
- 2018-07-19 CN CN201810794253.9A patent/CN108918714A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102731557A (en) * | 2011-04-02 | 2012-10-17 | 北京市理化分析测试中心 | Steroid substance derivation method and its analysis detection method |
| CN102507791A (en) * | 2011-11-17 | 2012-06-20 | 吉林大学 | Method for measuring cholesterol content in birds' egg and product thereof |
| CN106198798A (en) * | 2016-07-06 | 2016-12-07 | 汤臣倍健股份有限公司 | A kind of detection method of cholesterol level |
Non-Patent Citations (3)
| Title |
|---|
| 廖连英 等: "食用植物油中胆固醇含量的快速测定", 《化学分析计量》 * |
| 李杰 等: "高效液相色谱法快速测定黑甜玉米油中的胆固醇", 《食品科学》 * |
| 陈永伟 等: "油脂中胆固醇含量的快速测定法", 《理化检验-化学分册》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114689759A (en) * | 2022-04-15 | 2022-07-01 | 安徽丰本生物科技有限公司 | Method for measuring content of cholesterol in fish oil |
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Application publication date: 20181130 |
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