CN113075190B - Method for rapidly detecting sodium carbonate in raw and fresh milk based on Raman spectrum - Google Patents
Method for rapidly detecting sodium carbonate in raw and fresh milk based on Raman spectrum Download PDFInfo
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Abstract
The invention discloses a method for rapidly detecting sodium carbonate in fresh milk based on Raman spectrum, belonging to food detection technologyDomain. The method comprises collecting fresh milk sample on liquid sample plate, focusing to obtain 50-3500cm ‑1 The Raman spectrum in the range is collected by selecting 6 points at different positions on the measuring sample plate, and the average spectrum is taken as the Raman spectrum of the sample; then carrying out smooth noise reduction pretreatment to obtain a Raman spectrum of the sample; at 1080cm ‑1 And 2900cm ‑1 The characteristic peak at the displacement is used as a basis to qualitatively judge whether sodium carbonate is doped in the fresh milk sample or not, and 2900cm is used as a basis ‑1 And (3) preparing a standard curve according to the peak intensity change of the displacement position, and quantitatively analyzing the sodium carbonate adulterated fresh milk sample. The method disclosed by the invention does not need any sample pretreatment, is simple to operate and short in time consumption, and can be used for realizing on-site and outdoor rapid and high-throughput real-time analysis on the raw milk sample by matching with a handheld Raman analyzer.
Description
Technical Field
The invention relates to a method for rapidly detecting sodium carbonate in fresh milk based on Raman spectrum, and belongs to the technical field of food detection materials.
Background
Milk is rich in proteins, minerals, vitamins and carbohydrates and is known as "white blood". The improvement of the living standard of people in developing countries and their pursuit of health have led to a rapid increase of the global milk consumption, at the same time as the quality and safety of milk is also receiving increasing attention from consumers and producers. At present, the annual production of the fresh milk exceeds 3000 ten thousand tons in China, and in the problems of various safety qualities of the fresh milk, the adulteration has great harm to the physical and mental health of consumers, and the influence on the development of industry is the worst. Common adulterants include nitrogen-containing species adulterants, neutralizing acid adulterants, increasing density adulterants, and extending shelf life adulterants. The neutralization acid adulteration refers to the fraudulent activity that some milk farmers in the market add sodium carbonate to raw fresh milk to neutralize some acidic substances in bad milk in order to avoid the loss caused by the rancidity of the raw fresh milk, so as to improve the economic benefit. Patients with renal insufficiency, or congestive heart failure, may experience discomfort and possibly even serious consequences once they drink such adulterated milk.
The method can not accurately quantify the sodium carbonate in fresh milk by using a common bromothymol blue method, can not meet the requirements of on-site rapid qualitative and quantitative detection, and is inconvenient to be applied to on-site rapid detection in links of production, circulation, sales and the like.
Disclosure of Invention
The technical problems to be solved by the invention are as follows: how to rapidly and qualitatively and quantitatively detect the adulterated sodium carbonate in the fresh milk on site in the links of production, circulation, sales and the like.
In order to solve the problems, the invention provides a method for rapidly detecting the content of sodium carbonate in raw and fresh milk based on Raman spectrum, which comprises the following steps:
step 1: spreading fresh milk sample to be measured on liquid sample plate, selecting 4-8 points at different positions on sample plate, focusing and collecting 50-3500cm -1 Taking the average spectrum of the Raman spectrum at different positions as the Raman spectrum of the sample;
step 2: carrying out smooth noise reduction pretreatment on the Raman spectrum of the sample;
step 3: analysis of pretreated Raman Spectroscopy 1080cm -1 And 2900cm -1 Characteristic peak at displacement of 1080cm -1 If no peak appears at the displacement position, judging that the fresh milk sample to be detected is not doped with sodium carbonate, if 1080cm -1 Peak at displacement and 2900cm -1 And judging that the peak intensity of the displacement is higher than that of the displacement in the normal fresh milk, and judging that the sodium carbonate is doped in the fresh milk sample to be detected.
Preferably, the acquiring conditions of the raman spectrum in the step 1 are as follows: the laser power is 10-50mw, the diameter of the laser beam expander is 5-10mm, and the acquisition time is 3-5 s/time.
Preferably, the temperature of the raw milk sample to be tested in the step 1 is 25+/-5 ℃.
Preferably, the step 3 further comprises quantitatively analyzing the raw milk sample to be detected, which is judged to be adulterated with sodium carbonate.
Preferably, the statorThe quantitative analysis is as follows: manufacture 2900cm -1 Standard curve of peak intensity and sodium carbonate mass fraction at displacement position, measuring 2900cm of fresh milk sample -1 And comparing the peak intensity at the displacement position with the prepared standard curve to determine the content of sodium carbonate in the fresh milk sample to be detected.
Preferably, the method for manufacturing the standard curve comprises the following steps: preparing 6-10 adulterated fresh milk samples containing sodium carbonate with different mass fractions, obtaining Raman spectrum of the adulterated fresh milk samples, taking 2900cm -1 The peak intensity at the displacement and the mass fraction of sodium carbonate form a standard curve, and the correlation equation is as follows: y=424.3833x+3.253.
Preferably, the number of the prepared adulterated fresh milk samples is 7, and the mass fraction of sodium carbonate in the 7 adulterated fresh milk samples is 0.02%, 0.03%, 0.04%, 0.06%, 0.07%, 0.08% and 0.10% in sequence.
Compared with the prior art, the invention has the beneficial effects that:
1. the method for rapidly detecting the sodium carbonate content in the raw and fresh milk does not need complex sample pretreatment steps, has simple and rapid determination process, and has good practical application value.
2. The detection method can simultaneously perform qualitative and quantitative analysis on sodium carbonate in fresh milk, can rapidly detect the content of adulterated sodium carbonate, is convenient and rapid, and is suitable for any occasion.
Drawings
FIG. 1 is a Raman spectrum of undoped raw fresh milk;
FIG. 2 is a Raman spectrum contrast graph of undoped raw fresh milk (a), 1% sodium carbonate doped raw fresh milk (b), sodium carbonate powder (c);
FIG. 3 is a graph of Raman spectrum contrast of samples of adulterated raw fresh milk containing 0.02% (a), 0.03% (b), 0.04% (c), 0.06% (d), 0.07% (e), 0.08% (f), and 0.10% (g) sodium carbonate by mass;
FIG. 4 is a mass fraction of sodium carbonate and 2990cm -1 Linear relationship of peak height at displacement.
Detailed Description
In order to make the invention more comprehensible, preferred embodiments accompanied with figures are described in detail below.
Example 1
Raman spectroscopic detection of undoped raw fresh milk:
detecting milk sample with DXR laser micro Raman spectrometer, collecting proper amount of undoped fresh milk on liquid sample plate, focusing, and collecting 50-3500cm -1 The Raman spectrum in the range is collected for 5 s/time, the laser is a 532nm DPSS laser, the power is 10mw, the diameter of the laser beam expander is 5mm, 6 points at different positions on the measuring sample plate are selected for collection, the obtained average spectrum is subjected to pretreatment by adopting Adjacent-Averaging after the fluorescent background is subtracted, and a Raman spectrum of the undoped fresh milk is obtained, as shown in a in fig. 1 and 2.
The most intense raman peak of raw milk is probably due to three major components in milk: fat, protein and lactose. The average fat content in milk is about 3.9%, the average protein content is about 3.25%, and the average lactose content is about 4.6%, the three components remaining dissolved or suspended in water. The salt, vitamins and other ingredients account for about 87.3% of the total milk ingredients.
As shown in FIG. 1, at 2900cm -1 The nearby peak may be attributed to 2894cm -1 (C-H asymmetric stretching vibration) and 2854cm -1 The superposition of (C-H symmetrical stretching vibration) is classified into characteristic peaks of lactose and lactic acid molecules. Spectrum at 1656cm -1 There is a peak in the vicinity (c=o stretch), which is related to COOH groups of proteins and lactic acid, and is also the c=c stretch pattern of unsaturated fatty acids. At 1446cm -1 Where the and CH is observed 2 The peak intensity associated with the shear vibration was high at 1300cm -1 Here, CH associated with lipid molecules was observed 2 Torsional vibration. Finally, 1076-850cm -1 The spectral regions are attributed to CO, CH stretch and COC deformation modes of lactose molecules. 1000cm -1 The following areas present different vibration pattern combinations in the milk sample.
Example 2
Raman spectroscopic detection of sodium carbonate powder:
detecting sodium carbonate powder by using a DXR laser micro-Raman spectrometer, placing a proper amount of sodium carbonate powder on a solid sample plate, focusing, and obtaining 50-3500cm -1 The Raman spectrum in the range is collected for 5 s/time, the laser is a 532nm DPSS laser, the power is 10mw, the diameter of the laser beam expander is 5mm, the measuring sample plate is collected at 6 points at different positions, the obtained average spectrum is used for preprocessing the spectrum by adopting Adjacent-Averaging after the fluorescent background is subtracted from the obtained average spectrum, and the Raman spectrum diagram of sodium carbonate powder is obtained, wherein the Raman spectrum diagram of sodium carbonate is shown as c in fig. 2, and the concentration of sodium carbonate is 1080cm -1 There is a strong absorption peak, which is attributed to CO 3 2- CO stretching vibration in ion, i.e. 1080cm -1 Is a characteristic peak of sodium carbonate.
Example 3
Qualitative analysis of sodium carbonate adulterated raw fresh milk by Raman spectrum:
adding sodium carbonate into fresh milk, mixing to obtain 1.00% sodium carbonate sample, detecting with DXR laser micro Raman spectrometer, collecting proper amount of fresh milk on liquid sample plate, focusing, and obtaining 50-3500cm -1 The Raman spectrum in the range is collected for 5 s/time, the laser is a 532nm DPSS laser, the power is 10mw, the diameter of the laser beam expander is 5mm, 6 points at different positions on a measuring plate are selected to be collected, the obtained average spectrum is used for preprocessing the spectrum by adopting Adjacent-Averaging after fluorescent background is subtracted, and a Raman spectrum of undoped fresh milk is obtained, as shown in b in figure 2, the spectrum c is found to be 1080cm by comparing a (Raman spectrum of undoped fresh milk), b (Raman spectrum of fresh milk doped with 1% sodium carbonate) and c (Raman spectrum of sodium carbonate powder) in figure 2 -1 There appears a distinct characteristic peak with a peak intensity significantly higher than that of the peak at the point in b, but almost no 1080cm is observed in a -1 The peak intensity at the displacement indicates 1080cm -1 The characteristic signal peak of sodium carbonate is shown, and the Raman signal from sodium carbonate is detected in 1% sodium carbonate adulterated fresh milk; at the same time, a spectrum b at 2900cm relative to a spectrum a was also observed by comparison -1 The signal is enhanced, whereas the Raman spectrum c of the sodium carbonate powder does not appear obvious hereThe signal, the characteristic peak is C-H vibration signal, can be attributed to carbonic acid generated by chemical reaction of lactic acid and sodium carbonate in fresh milk, so that Raman spectrum is 1080cm -1 And 2900cm -1 The characteristic peak at the position can be used as the basis for qualitatively detecting whether sodium carbonate is doped in fresh milk, but is 2900cm -1 The change of the peak intensity of the characteristic peak can be used as the basis for quantitatively detecting the adulterated sodium carbonate in the fresh milk.
Example 4
And (3) manufacturing a standard curve:
adding sodium carbonate with different mass into raw and fresh milk, and uniformly mixing to obtain 7 adulterated raw and fresh milk samples with different sodium carbonate concentrations, wherein the mass fraction of the sodium carbonate is 0.02%, 0.04%, 0.06%, 0.07%, 0.08%, 0.09% and 0.10% in sequence. The DXR laser microscopic Raman spectrometer is used for detecting 7 adulterated fresh milk samples, the detection method is the same as that of the embodiment 3, 7 samples are obtained by detection, and the 7 samples are corresponding to 2900cm in the Raman spectrum as shown in the figure 3 -1 The peak height at shift is plotted against the mass fraction of sodium carbonate concentration to yield a standard curve, as shown in fig. 4. As can be seen from FIG. 4, 2900cm -1 The peak height (Y) at the displacement is linearly related to the mass fraction (X) of sodium carbonate in the raw milk, and the correlation equation is as follows: y=424.3833x+3.253, correlation coefficient R 2 0.989.
Example 5
Qualitative and quantitative detection of adulterated fresh milk to be detected:
to verify the accuracy of the qualitative and quantitative analysis in example 4 and 5, 20 blind samples (numbered 1-20) were prepared in the laboratory as samples to be tested, and the 20 samples to be tested were detected by using a DXR laser micro-Raman spectrometer, respectively, in the same manner as in example 3, to obtain 20 Raman spectra of the blind samples, if the Raman spectra were 1080cm -1 If no peak appears, judging that the fresh milk is not added with sodium carbonate, if 1080cm -1 Peak at displacement and 2900cm -1 The peak intensity of the displacement is higher than that of the displacement in the normal fresh milk, so that the sample is proved to be the fresh milk added with sodium carbonate; then the sample to be tested is measured at 2900cm -1 Displacement positionAnd (3) comparing the peak intensity of the sample with the standard curve obtained in the example 4 to determine the sodium carbonate content in the sample to be detected. In the Raman spectrum of 20 blind samples, no. 5 and No. 18 are 1080cm -1 Characteristic peaks and 2900cm were observed at the displacement -1 The signal peak at the displacement is enhanced, the two samples are proved to be fresh milk samples doped with sodium carbonate, and the No. 5 and No. 18 samples are respectively arranged at 2900cm -1 The signal intensity at the displacement is compared with a standard curve to obtain the detection results of 20 blind samples, wherein the sodium carbonate content of the sample No. 5 is 0.06%, the sodium carbonate content of the sample No. 18 is 0.1%, and the detection results are shown in Table 1.
Comparative example 1
The bromothymol blue assay detects sodium carbonate content in the 20 blind samples of example 5:
placing 5mL of sample into a test tube, keeping the test tube at an inclined position, then carefully adding 5 drops of 0.04wt% bromothymol blue ethanol solution along the tube wall, slowly inclining and rotating the test tube for 2-3 weeks to enable the reagent and fresh milk to be in contact with each other but not to be mixed with each other, then standing the test tube vertically for 2min, observing the color of a ring layer indicator at the junction of two liquid levels, if the ring layer is yellow, indicating that the fresh milk does not contain sodium carbonate, if the ring layer is green or blue, indicating that the fresh milk contains 0.05-1wt% sodium carbonate; under this test, the color of the ring layer indicator of 20 samples is shown in Table 1, and the results show that the 5 and 18 samples of 20 samples are sodium carbonate-spiked raw milk, and the results are the same as those of example 5.
Table 1 results of the tests of example 5 and comparative example
While the above embodiments have been described in detail with reference to the present invention, it should be understood that they have been presented by way of example only, and not by way of limitation, and that various modifications and additions may be made by those skilled in the art without departing from the scope of the invention.
Claims (4)
1. The method for rapidly detecting sodium carbonate in raw and fresh milk based on Raman spectrum is characterized by comprising the following steps of:
step 1: spreading fresh milk sample to be measured on liquid sample plate, selecting 4-8 points at different positions on sample plate, focusing and collecting 50-3500cm -1 Taking the average spectrum of the Raman spectrum at different positions as the Raman spectrum of the sample;
step 2: carrying out smooth noise reduction pretreatment on the Raman spectrum of the sample;
step 3: analysis of pretreated Raman Spectroscopy 1080cm -1 And 2900cm -1 Characteristic peak at displacement of 1080cm -1 If no peak appears at the displacement position, judging that the fresh milk sample to be detected is not doped with sodium carbonate, if 1080cm -1 Peak at displacement and 2900cm -1 The peak intensity of the displacement is higher than that of the displacement in the normal fresh milk, and the sample is judged to be adulterated with sodium carbonate;
the step 3 is followed by quantitative analysis of the fresh milk sample to be detected, which is judged to be adulterated with sodium carbonate, wherein the quantitative analysis is as follows: manufacture 2900cm -1 Standard curve of peak intensity and sodium carbonate mass fraction at displacement position, measuring 2900cm of fresh milk sample -1 Comparing the peak intensity at the displacement position with the prepared standard curve to determine the content of sodium carbonate in the fresh milk sample to be detected;
the manufacturing method of the standard curve comprises the following steps: preparing 6-10 adulterated fresh milk samples containing sodium carbonate with different mass fractions, obtaining Raman spectrum of the adulterated fresh milk samples, taking 2900cm -1 The peak intensity at the displacement and the mass fraction of sodium carbonate form a standard curve, and the correlation equation is as follows: y=424.3833x+3.253.
2. The method for rapidly detecting sodium carbonate in raw milk based on raman spectrum according to claim 1, wherein the acquisition conditions of raman spectrum in step 1 are as follows: the laser power is 10-50mw, the diameter of the laser beam expander is 5-10mm, and the acquisition time is 3-5 s/time.
3. The method for rapidly detecting sodium carbonate in raw milk based on Raman spectrum according to claim 1, wherein the temperature of the raw milk sample to be detected in the step 1 is 25+/-5 ℃.
4. The method for rapidly detecting sodium carbonate in raw milk based on Raman spectrum according to claim 1, wherein the number of the prepared adulterated raw milk samples is 7, and the mass fraction of sodium carbonate in the 7 adulterated raw milk samples is 0.02%, 0.03%, 0.04%, 0.06%, 0.07%, 0.08% and 0.10% in sequence.
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