CN109612957A - A kind of method of quick detection lycopene - Google Patents
A kind of method of quick detection lycopene Download PDFInfo
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- CN109612957A CN109612957A CN201910133660.XA CN201910133660A CN109612957A CN 109612957 A CN109612957 A CN 109612957A CN 201910133660 A CN201910133660 A CN 201910133660A CN 109612957 A CN109612957 A CN 109612957A
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- lycopene
- detection
- ultrasonic
- chloroform
- extraction
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
- G01N21/31—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/286—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q involving mechanical work, e.g. chopping, disintegrating, compacting, homogenising
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/34—Purifying; Cleaning
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/38—Diluting, dispersing or mixing samples
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/38—Diluting, dispersing or mixing samples
- G01N2001/386—Other diluting or mixing processes
Abstract
The invention discloses a kind of methods of quickly detection lycopene, include the following steps: step 1, lycopene is extracted from raw material；Pure Lycopene is dissolved in chloroform by step 2, and is configured to the detection liquid of multiple gradient concentrations；Step 3 takes detection liquid to be put into NanoDrop 2000C ultramicrospectrophotometer, detects the absorption peak of 510-520nm wavelength, draws standard curve；Step 4, the content of establishing criteria curve conversion lycopene；It only needs few sample size that can be accurately detected lycopene with this method, improves the efficiency and quality of detection.
Description
Technical field
The present invention relates to detection field, especially a kind of method for detecting lycopene.
Background technique
Existing lycopene detection method is usually to use visible point of high performance liquid chromatography (HPLC), Conventional UV-
Light photometry (UV-VIS).It needs a large amount of mobile phases, complicated for operation, chromatographic column to be easy blocking in HPLC method detection process to seep
One leakage, detection lycopene sample generally require 30min, thus it is cumbersome, time-consuming, low efficiency.The detection of UV-VIS method
Required sample size is also very big, detected value is inaccurate；Market, which needs to find a kind of method and can lead to too small amount of sample, quickly to be detected
Lycopene out, the present invention solve such problems.
Summary of the invention
To solve the deficiencies in the prior art, the purpose of the present invention is to provide a kind of method of quickly detection lycopene,
It only needs few sample size that can be accurately detected lycopene, improves the efficiency and quality of detection.
In order to achieve the above objectives, the present invention adopts the following technical scheme that:
A kind of method of quick detection lycopene, includes the following steps:
Step 1 extracts lycopene from raw material；
Pure Lycopene is dissolved in chloroform by step 2, and is configured to the detection liquid of multiple gradient concentrations；
Step 3 takes detection liquid to be put into NanoDrop 2000C ultramicrospectrophotometer, detection 510-520nm wavelength
Absorption peak draws standard curve；
Step 4, the content of establishing criteria curve conversion lycopene.
A kind of method of quickly detection lycopene above-mentioned,
Step 1 extracts lycopene from tomato；
Tomato cleans stalk, in refiner broken wall after chopping；
Chloroform is added in strained tomatoes, mixing is sufficiently stirred, ultrasonic 25-35min, shaking table are protected from light in ultrasonic tank
170-190r/min revolving speed concussion extraction extraction 5-7h；
Extract liquor filters removal impurity through 0.40-0.50um filter.
A kind of method of quickly detection lycopene above-mentioned,
Step 1, from the mould middle extraction lycopene of three spore cloth Laplaces；
Chloroform is added in trispore Bruce mould body after fermented and cultured；
Mixing is sufficiently stirred, ultrasonic 25-35min, the concussion leaching of shaking table 170-190r/min revolving speed are protected from light in ultrasonic tank
Mention extraction 5-7h；
Extract liquor filters removal impurity through 0.40-0.50um filter.
A kind of method of quickly detection lycopene above-mentioned,
Step 1 extracts lycopene from rhodotorula glutinis；
By the rhodotorula glutinis thallus after fermented and cultured in Ultrasonic cell smash after broken wall, chloroform is added；
Mixing is sufficiently stirred, ultrasonic 25-35min, the concussion leaching of shaking table 170-190r/min revolving speed are protected from light in ultrasonic tank
Mention extraction 5-7h；
Extract liquor filters removal impurity through 0.40-0.50um filter.
A kind of method of quickly detection lycopene above-mentioned,
Pure Lycopene is dissolved in chloroform by step 2, and be configured to final concentration of 1,1.5,2,2.5,5,
10, the detection liquid of 20,40,80,100 μ g/mL.
A kind of method of quickly detection lycopene above-mentioned,
Step 3 takes detection liquid to be put into NanoDrop 2000C ultramicrospectrophotometer, detects the absorption of 515nm wavelength
Peak value draws standard curve.
A kind of method of quickly detection lycopene above-mentioned,
Step 4 takes the 2 μ L of lycopene extracted in raw material, is put into NanoDrop 2000C ultramicrospectrophotometer,
Detect the absorption peak of 510-520nm wavelength wavelength, establishing criteria curve converts the content of lycopene, and detection knot is obtained in 2s
Fruit.
The invention has the beneficial effects that:
The present invention detects lycopene using the ultraviolet-visible spectrophotometer module of NanoDrop 2000/2000c
Content, sample size needed for more accurate compared to the testing result of other detection methods, repeated higher, detection seldom only need 2 μ L i.e.
Testing result can be obtained in 2s；
It is 515nm by experimental selection best detection wavelength, improves detection accuracy, avoid interfering.
Detailed description of the invention
Fig. 1 is that the Pure Lycopene of present invention experiment one absorbs peak figure；
Fig. 2 is that the beta carotene standard items of present invention experiment one absorb peak figure；
Fig. 3 is lycopene content testing result comparison diagram of three kinds of detection methods to same sample.
Specific embodiment
Specific introduce is made to the present invention below in conjunction with the drawings and specific embodiments.
A kind of method of quick detection lycopene, includes the following steps:
Step 1 extracts lycopene from raw material；
It is as follows that △ extracts the step of lycopene from tomato:
Tomato cleans stalk, in refiner broken wall after chopping；Strained tomatoes 5g is taken, chloroform 5mL is added, is sufficiently stirred mixed
It is even, ultrasonic 25-35min, shaking table 170-190r/min revolving speed concussion extraction extraction 5-7h are protected from light in ultrasonic tank；Extract liquor warp
0.40-0.50um filter filtering removal impurity.As a preference, ultrasonic 30min is protected from light in ultrasonic tank, shaking table 180r/
Min revolving speed concussion extraction extraction 6h.Extract liquor filters removal impurity through 0.45um filter.
The step of △ mould from three spore cloth Laplaces middle extraction lycopene, is as follows:
The trispore Bruce mould body 5g after fermented and cultured is taken, chloroform 5mL is added；Mixing, Yu Chao is sufficiently stirred
Ultrasonic 25-35min, shaking table 170-190r/min revolving speed concussion extraction extraction 5-7h are protected from light in sound sink；Extract liquor is through 0.40-
0.50um filter filtering removal impurity.As a preference, being protected from light ultrasonic 30min in ultrasonic tank, shaking table 180r/min turns
Speed concussion extraction extraction 6h, extract liquor filter removal impurity through 0.45um filter.
It is as follows that △ extracts the step of lycopene from rhodotorula glutinis:
Rhodotorula glutinis thallus 5g after weighing fermented and cultured, in Ultrasonic cell smash after broken wall, broken wall condition is
Chloroform 5mL is added in 80W, ultrasonic 1min, interval 1min；Mixing is sufficiently stirred, ultrasonic 25- is protected from light in ultrasonic tank
35min, shaking table 170-190r/min revolving speed concussion extraction extraction 5-7h；Extract liquor filters off removal of impurities through 0.40-0.50um filter
Matter.As a preference, ultrasonic 30min is protected from light in ultrasonic tank, and shaking table 180r/min revolving speed concussion extraction extraction 6h, extraction
Liquid filters removal impurity through 0.45um filter.
Pure Lycopene is dissolved in chloroform by step 2, and be configured to final concentration of 1,1.5,2,2.5,5,
10, the detection liquid of 20,40,80,100 μ g/mL.
Step 3 takes detection liquid to be put into NanoDrop 2000C ultramicrospectrophotometer, detection 510-520nm wavelength
Absorption peak draws standard curve；
Step 4 takes the 2 μ L of lycopene extracted in raw material, is put into NanoDrop 2000C ultramicrospectrophotometer,
Detect the absorption peak of 510-520nm wavelength, the content of establishing criteria curve conversion lycopene: mKind(μg/gThallus)=AAbsorption peak
Testing result is obtained in × 0.0241-0.0484,2s.
Experiment one, the choice experiment of absorption peak:
Lycopene, beta carotene standard items are dissolved in chloroform, 2 μ L is taken to be detected, using chloroform as
Blank control obtains peak figure.As Fig. 1 Pure Lycopene absorbs peak figure: there are three absorption peak, corresponding wavelength for lycopene
Respectively 456nm, 484nm, 515nm.As Fig. 2 beta carotene standard items absorb peak figure: beta carotene there are three absorption peak,
Wavelength is respectively 442nm, 461nm, 489nm.In order to avoid the interference of beta carotene, 515nm is chosen as lycopene
Detection wavelength.Step as one embodiment, step 3 take detection liquid to be put into NanoDrop 2000C ultramicron spectrophotometric
Meter detects the absorption peak of 515nm wavelength, draws standard curve.
Experiment two, the precise verification experiment of three kinds of method detection lycopenes:
Pass through the three mould extraction pigments of spore cloth Laplace；
Trispore Bruce mould body 5g after weighing fermented and cultured is added chloroform 5mL, is protected from light in ultrasonic tank super
Sound 30min, shaking table 180r/min revolving speed concussion extraction extraction 6h, extract liquor filter removal impurity through 0.45um filter, are examined
Survey liquid.
Test experience 2-1: Conventional UV-visible spectrophotometry (UV-VIS) detects lycopene content；
Pure Lycopene is dissolved in chloroform, and be configured to final concentration of 1,1.5,2,2.5,5,10,20,40,
80, the detection liquid of 100 μ g/mL；1mL detection liquid is taken to detect the absorption peak of 502nm wavelength in ultraviolet-visible spectrophotometer,
Draw standard curve；Resulting detection liquid 1mL is taken to detect in ultraviolet-visible spectrophotometer, establishing criteria curve conversion tomato
Lycopene content.
Test experience 2-2: high performance liquid chromatography (HPLC) detects lycopene content；
Pure Lycopene is dissolved in chloroform, and be configured to final concentration of 1,1.5,2,2.5,5,10,20,40,
80, then the detection liquid of 100 μ g/mL takes 10 μ L to carry out high performance liquid chromatography (HPLC) detection after 0.22 μm of filter filters.
Chromatographic column is Diamonsil C18 column (250mm × 4.6mm), and column temperature is set as 30 DEG C, and mobile phase A is acetonitrile: water=9: 1
(v/v), Mobile phase B is ethyl acetate, and eluent gradient is arranged are as follows: 0-15min, 100%-40% mobile phase A, 0%-60%
Mobile phase B；16min-30min, 40% mobile phase A, 60% Mobile phase B；31min-35min, 100% Mobile phase B, 0% flowing
Phase A, flow velocity are set as 1mL/min.The absorption peak of Detection wavelength 502nm draws standard curve.Resulting extract liquor is further
It is filtered through 0.22 μm of filter, 10 μ L is taken to carry out high performance liquid chromatography (HPLC) detection, establishing criteria curve conversion lycopene
Content.
Test experience 2-3: the invention patent method detects lycopene content；
Pure Lycopene is dissolved in chloroform, and be configured to final concentration of 1,1.5,2,2.5,5,10,20,
40, the detection liquid of 80,100 μ g/mL；It takes detection liquid to be put into NanoDrop 2000C ultramicrospectrophotometer, detects 510-
The absorption peak of 520nm wavelength draws standard curve；
It takes the resulting detection liquid of 2 μ L to be put into NanoDrop 2000C ultramicrospectrophotometer, detects the suction of 515nm wavelength
Peak value is received, testing result is obtained in 2s, the content of establishing criteria curve conversion lycopene:
mKind(μg/gThallus)=AAbsorption peak×0.0241-0.0484。
Experimental result:
Three kinds of methods repeat to detect the lycopene content of same sample three times, as shown in figure 3,
The results show that Conventional UV-visible spectrophotometry (UV-VIS) numerical value is relatively low and deviation is larger, the invention patent
Method (Nano) testing result and high performance liquid chromatography (HPLC) result approach and deviation is smaller.
Interpretation of result:
The results show that the invention patent method testing result is more accurate, repeatability is higher, it is few to detect required sample size.
The present invention detects lycopene using the ultraviolet-visible spectrophotometer module of NanoDrop 2000/2000c
Content, it is more accurate compared to the testing result of other detection methods, repeatability is higher, detect needed for sample size it is few, it is only necessary to 2 μ L are
Testing result can be obtained in 2s；It is 515nm by experimental selection best detection wavelength, improves detection accuracy, avoid interfering.
The basic principles, main features and advantages of the invention have been shown and described above.The technical staff of the industry should
Understand, the above embodiments do not limit the invention in any form, all obtained by the way of equivalent substitution or equivalent transformation
Technical solution is fallen within the scope of protection of the present invention.
Claims (7)
1. a kind of method of quickly detection lycopene, which comprises the steps of:
Step 1 extracts lycopene from raw material；
Pure Lycopene is dissolved in chloroform by step 2, and is configured to the detection liquid of multiple gradient concentrations；
Step 3 takes detection liquid to be put into NanoDrop 2000C ultramicrospectrophotometer, detects the absorption of 510-520nm wavelength
Peak value draws standard curve；
Step 4, the content of establishing criteria curve conversion lycopene.
2. a kind of method of quickly detection lycopene according to claim 1, which is characterized in that
Step 1 extracts lycopene from tomato；
Tomato cleans stalk, in refiner broken wall after chopping；
Chloroform is added in strained tomatoes, mixing is sufficiently stirred, ultrasonic 25-35min, shaking table 170- are protected from light in ultrasonic tank
190r/min revolving speed concussion extraction extraction 5-7h；
Extract liquor filters removal impurity through 0.40-0.50um filter.
3. a kind of method of quickly detection lycopene according to claim 1, which is characterized in that
Step 1, from the mould middle extraction lycopene of three spore cloth Laplaces；
Chloroform is added in trispore Bruce mould body after fermented and cultured；
Mixing is sufficiently stirred, ultrasonic 25-35min, shaking table 170-190r/min revolving speed concussion extraction extraction are protected from light in ultrasonic tank
Take 5-7h；
Extract liquor filters removal impurity through 0.40-0.50um filter.
4. a kind of method of quickly detection lycopene according to claim 1, which is characterized in that
Step 1 extracts lycopene from rhodotorula glutinis；
By the rhodotorula glutinis thallus after fermented and cultured in Ultrasonic cell smash after broken wall, chloroform is added；
Mixing is sufficiently stirred, ultrasonic 25-35min, shaking table 170-190r/min revolving speed concussion extraction extraction are protected from light in ultrasonic tank
Take 5-7h；
Extract liquor filters removal impurity through 0.40-0.50um filter.
5. a kind of method of quickly detection lycopene according to claim 1, which is characterized in that
Pure Lycopene is dissolved in chloroform by step 2, and be configured to final concentration of 1,1.5,2,2.5,5,10,20,
40, the detection liquid of 80,100 μ g/mL.
6. a kind of method of quickly detection lycopene according to claim 1, which is characterized in that
Step 3 takes detection liquid to be put into NanoDrop 2000C ultramicrospectrophotometer, detects the absorption peak of 515nm wavelength
Value draws standard curve.
7. a kind of method of quickly detection lycopene according to claim 1, which is characterized in that
Step 4 takes the 2 μ L of lycopene extracted in raw material, is put into NanoDrop 2000C ultramicrospectrophotometer, detection
The absorption peak of 510-520nm wavelength wavelength, establishing criteria curve convert the content of lycopene, obtain testing result in 2s.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1928098A (en) * | 2005-09-06 | 2007-03-14 | 童志清 | Method for purifying lycopene |
| CN106174165A (en) * | 2016-06-27 | 2016-12-07 | 徐州工程学院 | A kind of short flash extracting method of lycopene |
| CN107119098A (en) * | 2017-05-11 | 2017-09-01 | 天津北洋百川生物技术有限公司 | Addition growth factor produces bata-carotene and detection method during the fermentation |
-
2019
- 2019-02-22 CN CN201910133660.XA patent/CN109612957A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1928098A (en) * | 2005-09-06 | 2007-03-14 | 童志清 | Method for purifying lycopene |
| CN106174165A (en) * | 2016-06-27 | 2016-12-07 | 徐州工程学院 | A kind of short flash extracting method of lycopene |
| CN107119098A (en) * | 2017-05-11 | 2017-09-01 | 天津北洋百川生物技术有限公司 | Addition growth factor produces bata-carotene and detection method during the fermentation |
Non-Patent Citations (8)
| Title |
|---|
| 何晗 等: "超声波提取番茄红素新工艺研究", 《食品科技》 * |
| 侯纯明 等: "紫外分离测定番茄红素", 《粮食与食品工业》 * |
| 姚佳，蒲彪: "番茄红素提取工艺及检测方法的研究进展", 《食品与发酵科技》 * |
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| 王海兵等: "红酵母番茄红素提取工艺优化", 《食品科学》 * |
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Application publication date: 20190412 |