CN107238525A - A kind of method of citrinin toxin in extraction wine of rice fermented with red yeast - Google Patents
A kind of method of citrinin toxin in extraction wine of rice fermented with red yeast Download PDFInfo
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- CN107238525A CN107238525A CN201710388758.0A CN201710388758A CN107238525A CN 107238525 A CN107238525 A CN 107238525A CN 201710388758 A CN201710388758 A CN 201710388758A CN 107238525 A CN107238525 A CN 107238525A
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- G—PHYSICS
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
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Abstract
The invention discloses a kind of method for extracting citrinin toxin in wine of rice fermented with red yeast.The method of the present invention comprises the following steps：Wine of rice fermented with red yeast and methanol are mixed, reaction obtains product, that is, obtains citrinin toxin.Tests prove that, the method of the present invention being capable of citrinin toxin in rapid extraction wine of rice fermented with red yeast, and efficiency high, it is time-consuming less, cost is low, the rate of recovery is high, meet Good Laboratory control specification food Physico-chemical tests standard (GB/T 27404), in addition, the extraction of citrinin toxin, is worth with high promotion and application in the also greatly easy red yeast rice of the method and its Related product.
Description
Technical field
The invention belongs to biological technical field, and in particular to a kind of method of citrinin toxin in extraction wine of rice fermented with red yeast.
Background technology
Citrinin is mainly as a kind of mycotoxin produced by Penicillium, aspergillus, monascus.Citrinin distribution is wide
It is general, it is primarily present in the bread baskets such as wheat, corn, oat, and the fabricated product of apple, pear and other fruits.Citrinin has
There is good biocidal property, can suppress bacillus subtilis, staphylococcus aureus, Escherichia coli, streptomyces griseus etc., but due to
Citrinin has very strong renal toxicity without extensive use, and its pollution problem is increasingly paid close attention to by people, and by state
European branch of the natural Mycotoxin identification committee of border school of life and health sciences is classified as one of the toxin that must detect.
China is that red yeast rice is produced and using earliest, most wide country in the world, and wine of rice fermented with red yeast has also had one in China
The history of more than thousand years, its wide in variety, quality better, function are strong, gradually by extensive concern both domestic and external.Citrinin poison in wine of rice fermented with red yeast
Cellulose content also turns into the main contents of China's wine of rice fermented with red yeast export inspection quarantine.Both at home and abroad based on different detection methods to agricultural production
Contained citrinin content of toxins set up different limitation standards in product, food, feed, continuous complete with HPLC technologies
Kind, detection work is increasingly simple, and the pre-treatment of sample is into key problem in technology and difficult point.Separated at present for citrinin toxin pure
Change method is complex, is all generally by concentration, crosses post, extraction and realize that citrinin toxin is separated, finally by TLC or
HPLC carries out qualitative and quantitative detection.
The content of the invention
It is an object of the present invention to provide a kind of method for extracting citrinin toxin in wine of rice fermented with red yeast.
The method of citrinin toxin comprises the following steps in the extraction wine of rice fermented with red yeast that the present invention is provided：Wine of rice fermented with red yeast is mixed with methanol
Even, reaction obtains product, that is, obtains citrinin toxin.
In the above method, the volume ratio of the wine of rice fermented with red yeast and methanol is 1：(0.7-2)；The volume of the wine of rice fermented with red yeast and methanol
Than being specially 1：1.
In the above method, the methanol is hplc grade methanol.
In the above method, the condition of the reaction is room temperature, 200rpm vibrations 30-120min；The duration of oscillation is specific
For 60min.
In the above method, it is described wine of rice fermented with red yeast and methanol solution are mixed before also including the step of wine of rice fermented with red yeast is fully mixed.
In the above method, the step of methods described also includes carrying out HPLC detections to the product.
It is a further object to provide the new application of the above method.
The invention provides application of the above method in detection red yeast rice or its Related product in citrinin content of toxins.
In above-mentioned application, the red yeast rice or its Related product are wine of rice fermented with red yeast.
Final object of the present invention is to provide a kind of method for detecting citrinin content of toxins in wine of rice fermented with red yeast.
The method of citrinin content of toxins comprises the following steps in the detection wine of rice fermented with red yeast that the present invention is provided：Carry in aforementioned manners
Citrinin toxin in wine of rice fermented with red yeast is taken, citrinin toxin testing sample is obtained, using high performance liquid chromatography to citrinin poison
Plain testing sample is detected, obtains citrinin content of toxins in wine of rice fermented with red yeast.
In the above method or above-mentioned application, it is former that the wine of rice fermented with red yeast does not sell red yeast rice in commercially available wine of rice fermented with red yeast and distillery concretely
Wine.
The present invention is soluble in acetonitrile, methanol, ethyl acetate, the physicochemical property of acetone and other organic solvent using citrinin, sends out
Understand a kind of more easy method for extracting citrinin toxin in wine of rice fermented with red yeast, be conducive to promotion red yeast rice and the hair of red yeast rice the wine industry
Exhibition.Tests prove that, method of the invention can citrinin toxin in rapid extraction wine of rice fermented with red yeast, and efficiency high, it is time-consuming less, cost
The low, rate of recovery it is high (<100ng/mL, rate of recovery scope is in 60-120%；100-1000ng/mL, rate of recovery scope is in 80-
110%) Good Laboratory control specification-food Physico-chemical tests standard (GB/T 27404), is met, in addition, the method is also very big
The extraction of citrinin toxin, is worth with high promotion and application in easy red yeast rice and its Related product.
Brief description of the drawings
Fig. 1 is citrinin content of toxins examination criteria curve spectrum.
Fig. 2 is the HPLC collection of illustrative plates of 2000ng/mL citrinin toxin standard items.
Fig. 3 is wine of rice fermented with red yeast sample A HPLC collection of illustrative plates and the citrinin that 100 μ L various concentrations are added in wine of rice fermented with red yeast sample A
HPLC collection of illustrative plates after toxin standard items.Wherein, Fig. 3 A are wine of rice fermented with red yeast sample A HPLC collection of illustrative plates；Fig. 3 B are in wine of rice fermented with red yeast sample A
The HPLC collection of illustrative plates added after 100 μ L 2000ng/mL citrinin toxin standard items；Fig. 3 C are to add 100 μ in wine of rice fermented with red yeast sample A
HPLC collection of illustrative plates after L 1000ng/mL citrinin toxin standard items；Fig. 3 D are that 100 μ L 500ng/ are added in wine of rice fermented with red yeast sample A
HPLC collection of illustrative plates after mL citrinin toxin standard items.
Embodiment
Experimental method used in following embodiments is conventional method unless otherwise specified.
Material, reagent used etc., unless otherwise specified, are commercially obtained in following embodiments.
Quantitative test in following embodiments, is respectively provided with three repetition experiments, results averaged.
Citrinin standard items in following embodiments are the products in Sigma companies (U.S.).
Hplc grade methanol in following embodiments is the product in Fisher companies (U.S.).
The preparation method of citrinin standard curve is as follows：
With the molten 1mg solids citrinin standard items of 1mL hplc grade methanols, 1000mg/L citrinin standard liquids are configured to, then
2000ng/mL, 1000ng/mL, 500ng/mL, 100ng/mL, 50ng/mL, 10ng/ are configured to respectively with hplc grade methanol solution
ML standard curve working solutions.HPLC is carried out using high performance liquid chromatography to the standard curve working solution of various concentrations quantitatively to examine
Survey, and abscissa made with citrinin concentration, ordinate is made with peak area 0.05min before and after retention time at, according to concentration and
The relation of peak area carries out linear regression, and draws standard curve.Wherein, high performance liquid chromatography condition：Chromatographic column：
Agilent TC-C18,4.6 × 250mm, 5 μm；Mobile phase：Acetonitrile/isopropanol/phosphoric acid (0.08mol/L) (35/10/55, V/
V/V)；Fluorescence detector：Agilent G1321；Detection wavelength：331nm and 500nm；Flow velocity：1.0mL/min；40 DEG C of column temperature；Enter
Sample amount：50μL.
The standard curve of drafting is as shown in Figure 1.It is according to the calibration curve equation that standard curve is obtained：Y=
0.15375654x-1.8281142 (y is peak area, and x is citrinin content of toxins), R2=0.99987.Fig. 2 is 2000ng/mL
The HPLC collection of illustrative plates of citrinin toxin standard items.
The detection method of the citrinin toxin standard solution rate of recovery is as follows：
Accurately measured after 1.9mL hplc grade methanols, be placed in 10mL centrifuge tubes with pipettor, be separately added into the dense of 100 μ L
Spend for 2000ng/mL, 1000ng/mL, 500ng/mL citrinin toxin standard items, turn upside down for several times to after well mixed, turn
Into 2mL micro-samplings bottle, carry out HPLC and quantitatively detect, data processing is carried out with quantified by external standard method.Every kind of citrinin toxin mark
Quasi- addition standard items are repeated 3 times, and are averaged.
As a result show：Theoretical addition citrinin content of toxins is respectively 200ng/mL, 100ng/mL, 50ng/ in 1mL samples
ML, actually detected average value (being the actually detected values of the addition citrinin toxin standard items in following embodiments) is respectively
177.30ng/mL, 94.23ng/mL, 59.01ng/mL, the rate of recovery are respectively 88.65%, 94.23%, 118.01%.
The method of citrinin toxin and the detection of citrinin content of toxins in embodiment 1, a kind of extraction wine of rice fermented with red yeast
Citrinin Toxic extraction and citrinin poison in original wine of rice fermented with red yeast are carried out to wine of rice fermented with red yeast sample A (commercially available wine of rice fermented with red yeast) respectively
Citrinin Toxic extraction in plain standard addition sample, then carries out HPLC and quantitatively detects.Comprise the following steps that：
1st, citrinin Toxic extraction in original wine of rice fermented with red yeast
The wine of rice fermented with red yeast sample A that 1mL has fully been mixed is accurately measured with pipettor, is placed in 10mL centrifuge tubes, 1mL colors are added
Spectrum level methanol, room temperature is placed on shaking table, and 200rpm is acutely shaken after 60min, is gone in 2mL micro-samplings bottle, using efficient
Liquid chromatography (HPLC) carries out efficient liquid phase chromatographic analysis to sample liquid, and data processing is carried out with quantified by external standard method.Wine of rice fermented with red yeast
Sample A is repeated 3 times, and is averaged.
High performance liquid chromatography condition：Chromatographic column：Agilent TC-C18,4.6 × 250mm, 5 μm；Mobile phase：Acetonitrile/
Isopropanol/phosphoric acid (0.08mol/L) (35/10/55, V/V/V)；Fluorescence detector：Agilent G1321；Detection wavelength：331nm
And 500nm；Flow velocity：1.0mL/min；40 DEG C of column temperature；Sample size：50μL.
The eluent at the peak consistent with citrinin toxin standard items retention time (retention time about 0.05min) is collected, is obtained
To the citrinin content of toxins in original wine of rice fermented with red yeast sample A.Fig. 3 A are wine of rice fermented with red yeast sample A HPLC collection of illustrative plates.
2nd, citrinin Toxic extraction in citrinin toxin standard addition sample
The wine of rice fermented with red yeast sample A that 1mL has fully been mixed is accurately measured with pipettor, is placed in 10mL centrifuge tubes, is separately added into
100 μ L concentration is 2000ng/mL, and 1000ng/mL, 500ng/mL citrinin toxin standard items turn upside down for several times to mixing
After uniform, 0.9mL hplc grade methanols are added, room temperature is placed on shaking table, and 200rpm is acutely shaken after 60min, goes to 2mL micro-
Measure in sample injection bottle.Efficient liquid phase chromatographic analysis is carried out to sample liquid using high performance liquid chromatography (HPLC) respectively, with external standard method
Data processing is quantitatively carried out, the citrinin content of toxins in citrinin toxin standard addition sample is respectively obtained, then calculate respectively
The rate of recovery.Every kind of citrinin toxin standard addition wine of rice fermented with red yeast sample is repeated 3 times, and is averaged.
Testing result is as shown in table 1.Wherein, actual content (ng/mL) is citrinin content of toxins in wine of rice fermented with red yeast sample
Actual average detected value；Theoretical content (ng/mL) be wine of rice fermented with red yeast sample in be separately added into 0,2000ng/mL, 1000ng/mL,
Theoretical content after 500ng/mL citrinin toxin standard items；The rate of recovery (%) is actual content/theoretical content * 100%.
When spiked levels are 0, citrinin content of toxins is that 30.90ng/mL (is the tangerine in original wine of rice fermented with red yeast sample A
Mycin toxin average content)；When spiked levels are 2000ng/mL, citrinin content of toxins is 216.27ng/mL, and theory contains
Measure as 230.90ng/mL (200.00ng/mL+30.90ng/mL), the rate of recovery is 93.66% (216.27ng/mL/230.90ng/
mL).When spiked levels are 1000ng/mL, citrinin content of toxins is 127.77ng/mL, and theoretical content is 130.90ng/mL
(100.00ng/mL+30.90ng/mL), the rate of recovery is 97.61% (127.77ng/mL/130.90ng/mL).Work as spiked levels
During for 500ng/mL, citrinin content of toxins is 79.22ng/mL, and theoretical content is 80.90ng/mL (50.00ng/mL+
30.90ng/mL), the rate of recovery is 97.92% (97.92ng/mL/80.90ng/mL).Above recovery of standard addition meets laboratory
Quality control specifications-food Physico-chemical tests standard (GB/T 27404) (spiked levels<100ng/mL, rate of recovery scope is in 60-
120%；100-1000ng/mL, rate of recovery scope is in 80-110%), illustrating the method for the citrinin toxin that the present invention is extracted is
Feasible, and this method is easier.
Fig. 3 B are the HPLC collection of illustrative plates added in wine of rice fermented with red yeast sample A after 100 μ L 2000ng/mL citrinin toxin standard items；
Fig. 3 C are the HPLC collection of illustrative plates added in wine of rice fermented with red yeast sample A after 100 μ L 1000ng/mL citrinin toxin standard items；Fig. 3 D be
The HPLC collection of illustrative plates added in wine of rice fermented with red yeast sample A after 100 μ L 500ng/mL citrinin toxin standard items.
Citrinin content of toxins is detected in the wine of rice fermented with red yeast sample A of table 1
| Spiked levels (ng/mL) | Actual content (ng/mL) | Theoretical content (ng/mL) | The rate of recovery (%) |
| 0 | 30.90 | 30.90 | |
| 2000 | 216.27 | 230.90 | 93.66 |
| 1000 | 127.77 | 130.90 | 97.61 |
| 500 | 79.22 | 80.90 | 97.92 |
The method of citrinin toxin and the detection of citrinin content of toxins in embodiment 2, a kind of extraction wine of rice fermented with red yeast
Citrinin Toxic extraction and citrinin poison in original wine of rice fermented with red yeast are carried out to wine of rice fermented with red yeast sample B (commercially available wine of rice fermented with red yeast) respectively
Citrinin Toxic extraction in plain standard addition sample, then carries out HPLC and quantitatively detects.Comprise the following steps that：
1st, citrinin Toxic extraction in original wine of rice fermented with red yeast
The wine of rice fermented with red yeast sample B that 1mL has fully been mixed is accurately measured with pipettor, is placed in 10mL centrifuge tubes, 1mL colors are added
Spectrum level methanol, room temperature is placed on shaking table, and 200rpm is acutely shaken after 60min, is gone in 2mL micro-samplings bottle, using efficient
Liquid chromatography (HPLC) carries out efficient liquid phase chromatographic analysis to sample liquid, and data processing is carried out with quantified by external standard method.Wine of rice fermented with red yeast
Sample B is repeated 3 times, and is averaged.
High performance liquid chromatography condition：Chromatographic column：Agilent TC-C18,4.6 × 250mm, 5 μm；Mobile phase：Acetonitrile/
Isopropanol/phosphoric acid (0.08mol/L) (35/10/55, V/V/V)；Fluorescence detector：Agilent G1321；Detection wavelength：331nm
And 500nm；Flow velocity：1.0mL/min；40 DEG C of column temperature；Sample size：50μL.
The eluent at the peak consistent with citrinin toxin standard items retention time (retention time about 0.05min) is collected, is obtained
To the citrinin content of toxins in original wine of rice fermented with red yeast sample B.
2nd, citrinin Toxic extraction in citrinin toxin standard addition sample
The wine of rice fermented with red yeast sample B that 1mL has fully been mixed is accurately measured with pipettor, is placed in 10mL centrifuge tubes, is separately added into
100 μ L concentration is 2000ng/mL, and 1000ng/mL, 500ng/mL citrinin toxin standard items turn upside down for several times to mixing
After uniform, 0.9mL hplc grade methanols are added, room temperature is placed on shaking table, and 200rpm is acutely shaken after 60min, goes to 2mL micro-
Measure in sample injection bottle.Efficient liquid phase chromatographic analysis is carried out to sample liquid using high performance liquid chromatography (HPLC) respectively, with external standard method
Data processing is quantitatively carried out, the citrinin content of toxins in citrinin toxin standard addition sample is respectively obtained, then calculate respectively
Recovery of standard addition.Every kind of citrinin toxin standard addition wine of rice fermented with red yeast sample is repeated 3 times, and is averaged.
Testing result is as shown in table 2.When spiked levels are 0, citrinin content of toxins is that 30.39ng/mL is (as original
Citrinin toxin average content in wine of rice fermented with red yeast sample B)；When spiked levels are 2000ng/mL, citrinin content of toxins is
210.73ng/mL, theoretical content is 230.39ng/mL (200.00ng/mL+30.39ng/mL), and the rate of recovery is 91.47%
(210.73ng/mL/230.39ng/mL).When spiked levels are 1000ng/mL, citrinin content of toxins is 119.87ng/
ML, theoretical content is 130.39ng/mL (100.00ng/mL+30.39ng/mL), and the rate of recovery is 91.93% (119.87ng/mL/
130.39ng/mL).When spiked levels are 500ng/mL, citrinin content of toxins is 75.44ng/mL, and theoretical content is
80.39ng/mL (50.00ng/mL+30.39ng/mL), the rate of recovery is 93.84% (75.44ng/mL/80.39ng/mL).More than
Recovery of standard addition meets Good Laboratory control specification-food Physico-chemical tests standard (GB/T 27404) (spiked levels<
100ng/mL, rate of recovery scope is in 60-120%；100-1000ng/mL, rate of recovery scope is in 80-110%), illustrate the present invention
The method of the citrinin toxin of extraction is feasible, and this method is easier.
Citrinin content of toxins is detected in the wine of rice fermented with red yeast sample B of table 2
| Spiked levels (ng/mL) | Actual content (ng/mL) | Theoretical content (ng/mL) | The rate of recovery (%) |
| 0 | 30.39 | 30.39 | |
| 2000 | 210.73 | 230.39 | 91.47 |
| 1000 | 119.87 | 130.39 | 91.93 |
| 500 | 75.44 | 80.39 | 93.84 |
The method of citrinin toxin and the detection of citrinin content of toxins in embodiment 3, a kind of extraction wine of rice fermented with red yeast
Citrinin Toxic extraction and citrinin poison in original wine of rice fermented with red yeast are carried out to wine of rice fermented with red yeast sample C (commercially available wine of rice fermented with red yeast) respectively
Citrinin Toxic extraction in plain standard addition sample, then carries out HPLC and quantitatively detects.Comprise the following steps that：
1st, citrinin Toxic extraction in original wine of rice fermented with red yeast
The wine of rice fermented with red yeast sample C that 1mL has fully been mixed is accurately measured with pipettor, is placed in 10mL centrifuge tubes, 1mL colors are added
Spectrum level methanol, room temperature is placed on shaking table, and 200rpm is acutely shaken after 60min, is gone in 2mL micro-samplings bottle, using efficient
Liquid chromatography (HPLC) carries out efficient liquid phase chromatographic analysis to sample liquid, and data processing is carried out with quantified by external standard method.Wine of rice fermented with red yeast
Sample C is repeated 3 times, and is averaged.
High performance liquid chromatography condition：Chromatographic column：Agilent TC-C18,4.6 × 250mm, 5 μm；Mobile phase：Acetonitrile/
Isopropanol/phosphoric acid (0.08mol/L) (35/10/55, V/V/V)；Fluorescence detector：Agilent G1321；Detection wavelength：331nm
And 500nm；Flow velocity：1.0mL/min；40 DEG C of column temperature；Sample size：50μL.
The eluent at the peak consistent with citrinin toxin standard items retention time (retention time about 0.05min) is collected, is obtained
To the citrinin content of toxins in original wine of rice fermented with red yeast sample C.
2nd, citrinin Toxic extraction in citrinin toxin standard addition sample
The wine of rice fermented with red yeast sample C that 1mL has fully been mixed is accurately measured with pipettor, is placed in 10mL centrifuge tubes, is separately added into
100 μ L concentration is 2000ng/mL, and 1000ng/mL, 500ng/mL citrinin toxin standard items turn upside down for several times to mixing
After uniform, 0.9mL hplc grade methanols are added, room temperature is placed on shaking table, and 200rpm is acutely shaken after 60min, goes to 2mL micro-
Measure in sample injection bottle.Efficient liquid phase chromatographic analysis is carried out to sample liquid using high performance liquid chromatography (HPLC) respectively, with external standard method
Data processing is quantitatively carried out, the citrinin content of toxins in citrinin toxin standard addition sample is respectively obtained, then calculate respectively
Recovery of standard addition.Every kind of citrinin toxin standard addition wine of rice fermented with red yeast sample is repeated 3 times, and is averaged.
Testing result is as shown in table 3.When spiked levels are 0, citrinin content of toxins is that 28.74ng/mL is (as original
Citrinin toxin average content in wine of rice fermented with red yeast sample C)；When spiked levels are 2000ng/mL, citrinin content of toxins is
208.56ng/mL, theoretical content is 228.74ng/mL (200.00ng/mL+28.74ng/mL), and the rate of recovery is 91.18%
(208.56ng/mL/228.74ng/mL).When spiked levels are 1000ng/mL, citrinin content of toxins is 118.25ng/
ML, theoretical content is 128.74ng/mL (100.00ng/mL+28.74ng/mL), and the rate of recovery is 94.99% (118.25ng/mL/
128.74ng/mL).When spiked levels are 500ng/mL, citrinin content of toxins is 73.54ng/mL, and theoretical content is
78.74ng/mL (50.00ng/mL+28.74ng/mL), the rate of recovery is 93.40% (73.54ng/mL/78.74ng/mL).More than
Recovery of standard addition meets Good Laboratory control specification-food Physico-chemical tests standard (GB/T 27404) (spiked levels<
100ng/mL, rate of recovery scope is in 60-120%；100-1000ng/mL, rate of recovery scope is in 80-110%), illustrate the present invention
The method of the citrinin toxin of extraction is feasible, and this method is easier.
Citrinin content of toxins is detected in the wine of rice fermented with red yeast sample C of table 3
| Spiked levels (ng/mL) | Actual content (ng/mL) | Theoretical content (ng/mL) | The rate of recovery (%) |
| 0 | 28.74 | 28.74 | |
| 2000 | 208.56 | 228.74 | 91.18 |
| 1000 | 118.25 | 128.74 | 91.85 |
| 500 | 73.54 | 78.74 | 93.40 |
The method of citrinin toxin and the detection of citrinin content of toxins in embodiment 4, a kind of extraction wine of rice fermented with red yeast
Citrinin Toxic extraction in original wine of rice fermented with red yeast is carried out to wine of rice fermented with red yeast sample D (not selling red yeast rice former wine in distillery) respectively
With citrinin Toxic extraction in citrinin toxin standard addition sample, then carry out HPLC and quantitatively detect.Comprise the following steps that：
1st, citrinin Toxic extraction in original wine of rice fermented with red yeast
The wine of rice fermented with red yeast sample D that 1mL has fully been mixed is accurately measured with pipettor, is placed in 10mL centrifuge tubes, 1mL colors are added
Spectrum level methanol, room temperature is placed on shaking table, and 200rpm is acutely shaken after 60min, is gone in 2mL micro-samplings bottle, using efficient
Liquid chromatography (HPLC) carries out efficient liquid phase chromatographic analysis to sample liquid, and data processing is carried out with quantified by external standard method.Wine of rice fermented with red yeast
Sample D is repeated 3 times, and is averaged.
High performance liquid chromatography condition：Chromatographic column：Agilent TC-C18,4.6 × 250mm, 5 μm；Mobile phase：Acetonitrile/
Isopropanol/phosphoric acid (0.08mol/L) (35/10/55, V/V/V)；Fluorescence detector：Agilent G1321；Detection wavelength：331nm
And 500nm；Flow velocity：1.0mL/min；40 DEG C of column temperature；Sample size：50μL.
The eluent at the peak consistent with citrinin toxin standard items retention time (retention time about 0.05min) is collected, is obtained
To the citrinin content of toxins in original wine of rice fermented with red yeast sample D.
2nd, citrinin Toxic extraction in citrinin toxin standard addition sample
The wine of rice fermented with red yeast sample D that 1mL has fully been mixed is accurately measured with pipettor, is placed in 10mL centrifuge tubes, is separately added into
100 μ L concentration is 2000ng/mL, and 1000ng/mL, 500ng/mL citrinin toxin standard items turn upside down for several times to mixing
After uniform, 0.9mL hplc grade methanols are added, room temperature is placed on shaking table, and 200rpm is acutely shaken after 60min, goes to 2mL micro-
Measure in sample injection bottle.Efficient liquid phase chromatographic analysis is carried out to sample liquid using high performance liquid chromatography (HPLC) respectively, with external standard method
Data processing is quantitatively carried out, the citrinin content of toxins in citrinin toxin standard addition sample is respectively obtained, then calculate respectively
Recovery of standard addition.Every kind of citrinin toxin standard addition wine of rice fermented with red yeast sample is repeated 3 times, and is averaged.
Testing result is as shown in table 4.When spiked levels are 0, citrinin content of toxins is that 74.39ng/mL is (as original
Citrinin toxin average content in wine of rice fermented with red yeast sample D)；When spiked levels are 2000ng/mL, citrinin content of toxins is
260.94ng/mL, theoretical content is 274.39ng/mL (200.00ng/mL+74.39ng/mL), and the rate of recovery is 95.10%
(260.94ng/mL/274.39ng/mL).When spiked levels are 1000ng/mL, citrinin content of toxins is 166.60ng/
ML, theoretical content is 174.39ng/mL (100.00ng/mL+74.39ng/mL), and the rate of recovery is 95.53% (166.60ng/mL/
174.39ng/mL).When spiked levels are 500ng/mL, citrinin content of toxins is 121.46ng/mL, and theoretical content is
124.39ng/mL (50.00ng/mL+74.39ng/mL), the rate of recovery is 97.64% (121.46ng/mL/124.39ng/mL).
Above recovery of standard addition meets Good Laboratory control specification-food Physico-chemical tests standard (GB/T 27404) (spiked levels<
100ng/mL, rate of recovery scope is in 60-120%；100-1000ng/mL, rate of recovery scope is in 80-110%), illustrate the present invention
The method of the citrinin toxin of extraction is feasible, and this method is easier.
Citrinin content of toxins is detected in the wine of rice fermented with red yeast sample D of table 4
| Spiked levels (ng/mL) | Actual content (ng/mL) | Theoretical content (ng/mL) | The rate of recovery (%) |
| 0 | 74.39 | 74.39 | |
| 2000 | 260.94 | 274.39 | 95.10 |
| 1000 | 166.60 | 174.39 | 95.53 |
| 500 | 121.46 | 124.39 | 97.64 |
Claims (10)
1. a kind of method for extracting citrinin toxin in wine of rice fermented with red yeast, comprises the following steps：Wine of rice fermented with red yeast and methanol are mixed, reaction,
Product is obtained, that is, obtains citrinin toxin.
2. according to the method described in claim 1, it is characterised in that：The volume ratio of the wine of rice fermented with red yeast and methanol is 1：(0.7-2).
3. method according to claim 1 or 2, it is characterised in that：The volume ratio of the wine of rice fermented with red yeast and methanol is 1：1.
4. according to any described methods of claim 1-3, it is characterised in that：The methanol is hplc grade methanol.
5. according to any described methods of claim 1-4, it is characterised in that：The condition of the reaction is room temperature, 200rpm shakes
Swing 30-120min.
6. according to any described methods of claim 1-5, it is characterised in that：The condition of the reaction is room temperature, 200rpm shakes
Swing 60min.
7. according to any described methods of claim 1-6, it is characterised in that：It is described that wine of rice fermented with red yeast and methanol solution are mixed into preceding go back
The step of including wine of rice fermented with red yeast is fully mixed.
8. any described method answering in citrinin content of toxins in detection red yeast rice or its Related product in claim 1-7
With.
9. application according to claim 8, it is characterised in that：The red yeast rice or its Related product are wine of rice fermented with red yeast.
10. a kind of method for detecting citrinin content of toxins in wine of rice fermented with red yeast, comprises the following steps：With any in claim 1-7
Described method extracts citrinin toxin in wine of rice fermented with red yeast, citrinin toxin testing sample is obtained, using high performance liquid chromatography pair
The citrinin toxin testing sample is detected, obtains citrinin content of toxins in wine of rice fermented with red yeast.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111088171A (en) * | 2020-01-14 | 2020-05-01 | 北京工商大学 | Fermentation method capable of bidirectionally adjusting synthesis amount of monascus pigment in monascus purpureus |
-
2017
- 2017-05-27 CN CN201710388758.0A patent/CN107238525A/en active Pending
Non-Patent Citations (1)
| Title |
|---|
| 熊勇华 等: "橙色红曲菌液体发酵产桔霉素特性的研究", 《食品科学》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111088171A (en) * | 2020-01-14 | 2020-05-01 | 北京工商大学 | Fermentation method capable of bidirectionally adjusting synthesis amount of monascus pigment in monascus purpureus |
| CN111088171B (en) * | 2020-01-14 | 2021-03-16 | 北京工商大学 | Fermentation method capable of bidirectionally adjusting synthesis amount of monascus pigment in monascus purpureus |
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Application publication date: 20171010 |