CN103115906B - Fluorescence spectrum identification method of Fushunliao schisandra chinensis - Google Patents
Fluorescence spectrum identification method of Fushunliao schisandra chinensis Download PDFInfo
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Abstract
The invention discloses a fluorescence spectrum identification method of Fushunliao schisandra chinensis. The fluorescence spectrum identification method comprises the following steps of: firstly, obtaining a three-dimensional fluorescence spectrum of the Fushunliao schisandra chinensis and deriving the three-dimensional fluorescence spectrum in a data file ASCII (American Standard Code for Information Interchange) format to obtain original data of the three-dimensional fluorescence spectrum; extracting constant wavelength synchronous fluorescence signal data of emission wavelength and excitation wavelength with different wavelength intervals; obtaining a synchronous dynamic spectrum disturbed by different wavelength intervals between the related emission wavelength and excitation wavelength according to the data, so as to establish a two-dimensional related synchronous fluorescence spectrum of the Fushunliao schisandra chinensis; establishing a two-dimensional related synchronous fluorescence spectrum of unknown schisandra chinensis of the source area according to the same method; and comparing the two-dimensional related synchronous fluorescence spectrum with the two-dimensional related synchronous fluorescence spectrum of the Fushunliao schisandra chinensis to judge whether the sample is the Fushunliao schisandra chinensis. According to the method disclosed by the invention, the Fushunliao schisandra chinensis which is used as a geographical indication product and other non-Fushunliao schisandra chinensis can be directly and visually identified to enhance the protection on the Fushunliao schisandra chinensis.
Description
Technical field
The present invention relates to fluorescence spectrum recognition technology, specifically the fluorescence spectrum discrimination method of a kind of Fushun FRUCTUS SCHISANDRAE from Liaoning of China.
Background technology
Chinese medicine Schisandra chinensis is the dry mature fruit of magnoliaceae schisandra.Have convergence astringent or styptic treatment for spontaneous sweating, nourishing generate fluid, the eficac Clinical research of kidney calming finds, the fruit of Chinese magnoliavine can obviously reduce hepatitis serum GPT levels, and has anticancer and activity that is anti-AIDS.Fruit of Chinese magnoliavine plant is divided into Schizandra (namely fructus schisandrae belongs to) and Kadsura 2 genus.The identification of the identification carried out the fruit of Chinese magnoliavine at present mainly north and south fruit of Chinese magnoliavine, spectral discrimination method mainly contains ultraviolet spectrophotometry, second derivative spectra fluorescent spectrometry, infra-red sepectrometry etc.Wherein fluorescence spectroscopy technique has the advantages such as highly sensitive, selectivity is strong, sample size is few, and containing deoxyschizandrin and the anwulignan with fluorescent effect in the fruit of Chinese magnoliavine, becomes the preferred means at present in fruit of Chinese magnoliavine identification.
The existing spectral discrimination method to the fruit of Chinese magnoliavine is mainly for the identification of the north and south fruit of Chinese magnoliavine at present, and with regard to the fruit of Chinese magnoliavine in a certain place of production, its recognition capability is obviously inadequate.Fushun FRUCTUS SCHISANDRAE from Liaoning of China is the authentic medicinal herbs in Liaoning, mainly be distributed in Mountainous Region in East Liaoning, because its skin is thin, meat is thick, grain is large, medicinal ingredient is high, pungent sweet bitter salty bittersweet and famous, have the title of " the distant five tastes ", State General Administration for Quality Supervision implements the protection of national geography famous special product in official approval in 2008 to Fushun FRUCTUS SCHISANDRAE from Liaoning of China, its protection domain is Manchu Autonomous County of Xinbin of Fushun City of Liaoning Province, Manchu Autonomous County of Qingyuan, Fushun County, Shuncheng District, Wanghua District, 7 counties and districts such as economic and technological development zone now have jurisdiction over administrative region, because the fruit of Chinese magnoliavine of Different sources is at the composition of chemical composition, there is some difference in the aspect such as content and drug effect, and the method that there is no at present identifies separately Fushun FRUCTUS SCHISANDRAE from Liaoning of China, and the quality control method lacked Fushun FRUCTUS SCHISANDRAE from Liaoning of China.
Summary of the invention
The object of this invention is to provide the fluorescence spectrum discrimination method of a kind of Fushun FRUCTUS SCHISANDRAE from Liaoning of China, from the fruit of Chinese magnoliavine that original producton location is different, the Fushun FRUCTUS SCHISANDRAE from Liaoning of China protected by geography symbol product is identified, strengthen the protection to Fushun FRUCTUS SCHISANDRAE from Liaoning of China.
The object of the present invention is achieved like this: the fluorescence spectrum discrimination method of a kind of Fushun FRUCTUS SCHISANDRAE from Liaoning of China, comprises the steps:
A: set up Fushun FRUCTUS SCHISANDRAE from Liaoning of China two-dimensional correlation synchronous fluorescence collection of illustrative plates,
A-1: get Fushun FRUCTUS SCHISANDRAE from Liaoning of China fruit appropriate, make uniform powder, described powder is mixed with absolute ethyl alcohol in the ratio that mass volume ratio is 1 ︰ 350-400 and carries out ultrasonic process, centrifugal after leaving standstill, get supernatant 0.45 μm of membrane filtration, abandon just filtrate, get subsequent filtrate as analytical solution, income analysis solution carries out 3-D scanning with fluorescence spectrophotometer, obtains triaxial stress condition;
A-2: gained triaxial stress condition is derived with the ASCII fromat of data file, obtain three-dimensional fluorescence spectrum raw data, from obtained three-dimensional fluorescence spectrum raw data, extract the emission wavelength at different wave length interval and the permanent Wavelength synchronous Fluorescent signal data of excitation wavelength, utilize described synchronous fluorescence signal data to obtain the synchronous dynamic spectrum of different wave length interval disturbance between regarding emission wavelength and excitation wavelength;
A-3: by described synchronous dynamic spectrum by 2Dshige software, software dynamic spectrum is set on average, and contour parameter is 8, draws Fushun FRUCTUS SCHISANDRAE from Liaoning of China two-dimensional correlation synchronous fluorescence collection of illustrative plates;
B: the fruit of Chinese magnoliavine two-dimensional correlation synchronous fluorescence collection of illustrative plates setting up original producton location the unknown: the fruit of Chinese magnoliavine two-dimensional correlation synchronous fluorescence collection of illustrative plates of the method establishment original producton location the unknown provided according to described steps A-1, A-2, A-3, the fruit of Chinese magnoliavine two-dimensional correlation synchronous fluorescence collection of illustrative plates of obtained original producton location the unknown is compared with the described Fushun synchronous collection of illustrative plates of FRUCTUS SCHISANDRAE from Liaoning of China two-dimensional correlation, to judge that whether this sample is as Fushun, original producton location FRUCTUS SCHISANDRAE from Liaoning of China.
In described steps A-1, the condition of 3-D scanning is: income analysis solution is placed in 1cm quartz cuvette pond, by following condition: excitation wavelength lambda
ex: 200-700nm, emission wavelength lambda
em: 200-800nm, voltage: 500V, sweep velocity: 12000nm/min, excites and emission scan slit: 5nm, scanning step: 5nm, carries out triaxial stress condition scanning, obtain triaxial stress condition.
The time of ultrasonic process in described steps A-1 is 20-40min.
In described steps A-2, the wavelength interval of emission wavelength and excitation wavelength is followed successively by: 5nm, 10nm, 15nm, 20nm, 25nm, 30nm, 35nm, 40nm, 45nm, 50nm, 55nm, 60nm, 65nm, 70nm, 75nm, 80nm, 85nm, 90nm, 100nm.
The present invention selects the fluorescence data of method to the fruit of Chinese magnoliavine be suitable for process; the fruit of Chinese magnoliavine making the fluorescence pattern of the treated Fushun FRUCTUS SCHISANDRAE from Liaoning of China obtained obviously be different from other places of production (as Jilin, Hubei) through same process the fluorescence pattern that obtains; thus can be directly perceived, vivid make discriminating to as between the Fushun FRUCTUS SCHISANDRAE from Liaoning of China of geography symbol product and other non-Fushun FRUCTUS SCHISANDRAE from Liaoning of China, enhance the protection to Fushun FRUCTUS SCHISANDRAE from Liaoning of China.
Accompanying drawing explanation
Fig. 1 is Fushun FRUCTUS SCHISANDRAE from Liaoning of China triaxial stress condition.
Fig. 2 is Fushun FRUCTUS SCHISANDRAE from Liaoning of China two-dimensional correlation synchronous fluorescence collection of illustrative plates.
Fig. 3 is the fruit of Chinese magnoliavine two-dimensional correlation synchronous fluorescence collection of illustrative plates in Jilin, original producton location.
Fig. 4 is the fruit of Chinese magnoliavine two-dimensional correlation synchronous fluorescence collection of illustrative plates in Hubei, original producton location.
In Fig. 1, EX represents excitation wavelength, and EM represents emission wavelength, and in Fig. 2, Fig. 3, Fig. 4, transverse axis and longitudinal axis numerical value all represent wavelength (unit: nm).
Embodiment
Instrument: Hitachi F-7000 fluorospectrophotometer (Hitachi, Ltd), KQ2200DB type numerical control ultrasonic cleaner (Kunshan Ultrasonic Instruments Co., Ltd.).
Embodiment 1:
A, set up the two-dimensional correlation synchronous fluorescence collection of illustrative plates of Fushun FRUCTUS SCHISANDRAE from Liaoning of China:
Get Fushun FRUCTUS SCHISANDRAE from Liaoning of China fruit sample 10g, make uniform powder, taking 0.05g powder adds in 20ml absolute ethyl alcohol, ultrasonic process 30min, under normal temperature condition leave standstill cooling after centrifugal, get centrifugal after supernatant with 0.45 μm of membrane filtration, discard just filtrate, get subsequent filtrate as analytical solution, scan by the following condition of scanning, obtain the triaxial stress condition shown in Fig. 1:
Excitation wavelength lambda
ex: 200-700nm;
Emission wavelength lambda
em: 200-800nm;
Voltage: 500V;
Sweep velocity (SP): 12000nm/min;
Excite and emission scan slit: 5nm;
Scanning step: 5nm;
Triaxial stress condition with FL Solutions 2.1 for F-7000 workstation with the ASCII(*.TXT of data file) form derive, obtain three-dimensional spectroscopic data, the wavelength interval utilizing Mallab 7.0 software to obtain emission wavelength and excitation wavelength from three-dimensional spectroscopic data is followed successively by 5nm, 10nm, 15nm, 20nm, 25nm, 30nm, 35nm, 40nm, 45nm, 50nm, 55nm, 60nm, 65nm, 70nm, 75nm, 80nm, 85nm, 90nm, the permanent Wavelength synchronous Fluorescent signal data of the different wave length interval disturbance of 100nm, the permanent Wavelength synchronous Fluorescent signal data of this different wave length interval disturbance is utilized to obtain the synchronous dynamic spectrum of different wave length interval disturbance between regarding emission wavelength and excitation wavelength, by synchronous dynamic spectrum by 2Dshige version 1.3 software, software dynamic spectrum is set on average, contour optimum configurations is 8, draw the two-dimensional correlation synchronous fluorescence collection of illustrative plates generating Fushun FRUCTUS SCHISANDRAE from Liaoning of China, as shown in Figure 2.
B, set up the fruit of Chinese magnoliavine two-dimensional correlation synchronous fluorescence collection of illustrative plates that original producton location is Jilin, Hubei:
According to the test method given by steps A and condition, draw the synchronous collection of illustrative plates of two-dimensional correlation of the fruit of Chinese magnoliavine of each 3 batches that original producton location is Jilin, Hubei respectively, the synchronous collection of illustrative plates of the two-dimensional correlation of 3 batches in the same place of production is almost identical, Fig. 3 represents that original producton location is the synchronous collection of illustrative plates of two-dimensional correlation of 1 batch of Jilin, and Fig. 4 represents that original producton location is the synchronous collection of illustrative plates of two-dimensional correlation of 1 batch of Hubei.
On the basis of embodiment 1, carry out following 3 tests respectively, to prove confidence level and the stability of embodiment 1 result.
(1) precision test: the sample METHOD FOR CONTINUOUS DETERMINATION in Example 13 times, obtain 3 Fushun FRUCTUS SCHISANDRAE from Liaoning of China two-dimensional correlation synchronous fluorescence collection of illustrative plates of same batch, calculate position and the fluorescence intensity of fluorescence peak in collection of illustrative plates, result shows its coefficient of variation < 3%, the two-dimensional correlation synchronous fluorescence collection of illustrative plates made is consistent, proves that instrument precision is good.
(2) stability test: the analytical solution in Example 1, measure when 0.5h, 1h, 2h, 3h, 5h, 10h respectively, obtain the two-dimensional correlation synchronous fluorescence collection of illustrative plates of 6 Fushun FRUCTUS SCHISANDRAE from Liaoning of China of same batch, calculate position and the fluorescence intensity of fluorescence peak in collection of illustrative plates, result shows its coefficient of variation < 3%, the two-dimensional correlation synchronous fluorescence collection of illustrative plates made is consistent, shows that analytical solution is stable in 10h.
(3) reappearance test: the 5 increment product getting different batches, the method step provided according to steps A measures, obtain the two-dimensional correlation synchronous fluorescence collection of illustrative plates of 5 Fushun FRUCTUS SCHISANDRAE from Liaoning of China of different batches, calculate position and the fluorescence intensity of fluorescence peak in collection of illustrative plates, result shows coefficient of variation < 3%, the two-dimensional correlation synchronous fluorescence collection of illustrative plates made is consistent, shows that the present invention has good reappearance.
Interpretation of result: from obtained three two-dimensional correlation synchronous fluorescence collection of illustrative plates (Fig. 2, Fig. 3, Fig. 4), Fushun FRUCTUS SCHISANDRAE from Liaoning of China and the self correlated peak in the fruit of Chinese magnoliavine in other places of production (Jilin, Hubei) and intersect the going out peak number, position, positive and negatively all there is significant difference of peak, its difference is fairly obvious and directly perceived, thus can pass through the fruit of Chinese magnoliavine in directly perceived, the vivid discriminating Fushun the Liao Dynasty place of production of two-dimensional correlation fluorescence pattern.
Method provided by the present invention is the fluorescence spectrum discrimination method for Fushun FRUCTUS SCHISANDRAE from Liaoning of China, and in addition, the method also provides scientific basis and the technical support of identification and verification to the geography symbol product of other kinds.
Claims (3)
1. a fluorescence spectrum discrimination method for Fushun FRUCTUS SCHISANDRAE from Liaoning of China, is characterized in that, comprise the steps:
A: set up Fushun FRUCTUS SCHISANDRAE from Liaoning of China two-dimensional correlation synchronous fluorescence collection of illustrative plates;
A-1: get Fushun FRUCTUS SCHISANDRAE from Liaoning of China fruit appropriate, make uniform powder, described powder is mixed with absolute ethyl alcohol in the ratio that mass volume ratio is 1 ︰ 350-400 and carries out ultrasonic process, centrifugal after leaving standstill, get supernatant 0.45 μm of membrane filtration, abandon just filtrate, get subsequent filtrate as analytical solution, income analysis solution carries out 3-D scanning with fluorescence spectrophotometer, obtains triaxial stress condition;
A-2: gained triaxial stress condition is derived with the ASCII form of data file, obtain three-dimensional fluorescence spectrum raw data, from obtained three-dimensional fluorescence spectrum raw data, extract the emission wavelength at different wave length interval and the permanent Wavelength synchronous Fluorescent signal data of excitation wavelength, utilize described synchronous fluorescence signal data to obtain the synchronous dynamic spectrum of different wave length interval disturbance between regarding emission wavelength and excitation wavelength; The wavelength interval of described emission wavelength and excitation wavelength is followed successively by: 5nm, 10nm, 15nm, 20nm, 25nm, 30nm, 35nm, 40nm, 45nm, 50nm, 55nm, 60nm, 65nm, 70nm, 75nm, 80nm, 85nm, 90nm, 100nm;
A-3: by described synchronous dynamic spectrum by 2Dshige software, software dynamic spectrum is set on average, and contour parameter is 8, draws Fushun FRUCTUS SCHISANDRAE from Liaoning of China two-dimensional correlation synchronous fluorescence collection of illustrative plates;
B: the fruit of Chinese magnoliavine two-dimensional correlation synchronous fluorescence collection of illustrative plates setting up original producton location the unknown: the fruit of Chinese magnoliavine two-dimensional correlation synchronous fluorescence collection of illustrative plates of the method establishment original producton location the unknown provided according to described steps A-1, A-2, A-3, the fruit of Chinese magnoliavine two-dimensional correlation synchronous fluorescence collection of illustrative plates of obtained original producton location the unknown and described Fushun FRUCTUS SCHISANDRAE from Liaoning of China two-dimensional correlation synchronous fluorescence collection of illustrative plates are compared, to judge that whether this sample is as Fushun, original producton location FRUCTUS SCHISANDRAE from Liaoning of China.
2. according to the fluorescence spectrum discrimination method of the Fushun FRUCTUS SCHISANDRAE from Liaoning of China described in claim 1, it is characterized in that, in described steps A-1, the condition of 3-D scanning is: income analysis solution is placed in 1cm quartz cuvette pond, by following condition: excitation wavelength lambda ex: 200-700nm, and emission wavelength lambda em: 200-800nm, voltage: 500V, sweep velocity: 12000nm/min, excites and emission scan slit: 5nm, scanning step: 5nm, carry out triaxial stress condition scanning, obtain triaxial stress condition.
3. according to the fluorescence spectrum discrimination method of the Fushun FRUCTUS SCHISANDRAE from Liaoning of China described in claim 1, it is characterized in that, the time of ultrasonic process in described steps A-1 is 20-40min.
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| CN104062274A (en) * | 2014-04-29 | 2014-09-24 | 江西农业大学 | Genetic optimization algorithm-based synchronous fluorescence spectrum characteristic wavelength selection method |
| CN105092549B (en) * | 2015-07-28 | 2017-11-17 | 江苏大学 | A kind of extracting method of fluorescence spectrum information |
| CN105334197A (en) * | 2015-11-14 | 2016-02-17 | 常州大学 | Method for analyzing disinfection by-product on basis of fluorescence spectrum technology |
| CN106092990A (en) * | 2016-06-16 | 2016-11-09 | 宁夏林业研究院股份有限公司 | A kind of three-dimensional fluorescence spectrum discrimination method of lycium barbarum |
| CN107192700A (en) * | 2017-07-25 | 2017-09-22 | 潍坊学院 | A kind of multiple light courcess rapid fluorescence spectrum detection device controlled for traditional Chinese medicine quality and method |
| CN108519361B (en) * | 2018-04-11 | 2020-07-03 | 南京市产品质量监督检验院 | Method for identifying tree species of dalbergia odorifera and dalbergia odorifera by fluorescence spectrum |
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