A quantitative detection method and quantitative detection system for red yeast rice ingredients

Abstract

The invention discloses a quantitative detection method and a quantitative detection system for red yeast rice components, and belongs to the technical field of microbial detection. The invention adopts red yeast rice powder and counting beads to prepare a suspension, and irradiates the suspension in a mixing channel with incident light generated by a light generator, thereby realizing pipeline detection. The forward scattering intensity, the first side scattering intensity, and the second side scattering light are collected to calculate the lateral intensity ratio of the counting beads, and the particle size function is fitted according to the lateral intensity ratio of the counting beads of different wavelengths. The lateral intensity ratio of the component to be measured is calculated according to the scattering data of the component to be measured, and the particle size parameter is predicted based on the particle size function and the lateral intensity ratio of the component to be measured, and the corrected scattering data of the particle size parameter within the particle size gating interval of the component to be measured is extracted, and the concentration of the component to be measured is predicted according to the scattering data of the counting beads and the corrected scattering data, and the particles with a large diameter difference from the component to be measured are excluded according to the particle size parameter, thereby improving the detection accuracy and reducing the measurement error.

Description

Technical Field

[0001] The present invention relates to the technical field of microbial detection, and in particular to a quantitative detection method and a quantitative detection system for red yeast rice components. Background Art

[0002] Red yeast rice is a rice fermentation product prepared by inoculating Monascus fungi on rice and fermenting. The Chinese patent application with publication number CN111387409A discloses a method for processing functional red yeast rice powder, wherein polished rice is added to 2-4 times water and ground into a paddle, boiled, and made into a flowing juice, and polished rice juice is added to a fermentation tank, and monascus is inoculated in the fermentation tank. When the fermentation liquid in the fermentation tank turns red, the fermentation is completed, and the solid is separated and dried to obtain red yeast rice. It can be seen from the processing technology that the main components of red yeast rice include monascus and fermentation products such as red yeast pigment and monacolin K in addition to starch granules and polysaccharide polymers. Red yeast rice has many effective ingredients and is difficult to detect. The Chinese patent application with publication number CN107271582A discloses a method for detecting citrinin toxin in red yeast rice, wherein the red yeast rice sample is mixed with methanol, and the mass volume ratio of the red yeast rice sample to methanol is 1g:5mL, and then the citrinin toxin solution is extracted and citrinin is detected by high performance liquid chromatography. Although this method has a fast detection speed, red yeast rice is a traditional multi-component fermented product, and high performance liquid chromatography is easily interfered by other components. Therefore, it is necessary to provide a rapid and high-precision quantitative detection method for red yeast rice components. Summary of the invention

[0003] In view of the above problems, the present invention provides a quantitative detection method and system for red yeast rice components. The present invention uses incident light generated by a light generator to illuminate the buffer and suspension in the mixing channel, removes impurity signals according to the scattered light intensity at different angles, and obtains the concentration of the component to be tested, thereby improving the detection accuracy.

[0004] The invention objectives of this application can be achieved through the following technical means:

[0005] A method for quantitatively detecting red yeast rice components comprises the following steps:

[0006] Step 1: Prepare a suspension using red yeast rice powder and counting beads, and then prepare a buffer solution;

[0007] Step 2: The first controller sends the buffer into the sheath flow channel, and the second controller sends the suspension into the sample channel, and the buffer and the suspension form a mixing streamline in the mixing channel;

[0008] Step 3: The incident light generated by the light generator illuminates the optical path window of the mixing channel, and the incident light generates forward scattered light, first side scattered light, and second side scattered light after passing through the mixing streamline;

[0009] Step 4: The first detector receives the forward scattered light at a scattering angle θ1, the second detector receives the first side scattered light at a scattering angle θ2, and the third detector receives the second side scattered light at a scattering angle θ3, θ1<θ2 and θ3-θ2=π / 2;

[0010] Step 5: Fix the wavelength of the incident light, the first detector collects the forward scattering intensity I1(t), the second detector collects the first side scattering intensity I2(t), and the third detector collects the second side scattering intensity I3(t), where t is the collection time;

[0011] Step 6: adjusting the flow rate of the first controller or the second controller according to the first side scattering intensity I2(t) and the second side scattering intensity I3(t) within the calibration time T;

[0012] Step 7: I1(t), I2(t), and I3(t) at the same acquisition time t constitute the scattering data, adjust the wavelength of the incident light, generate the scattering data of the counting beads and the scattering data of the components to be measured, and then calculate the lateral intensity ratio of the counting beads, and fit the particle size function according to the lateral intensity ratio of the counting beads at different wavelengths;

[0013] Step 8: Calculate the lateral intensity ratio of the component to be measured according to the scattering data of the component to be measured, and predict the particle size parameters based on the particle size function and the lateral intensity ratio of the component to be measured;

[0014] Step 9: Extract the scattering data of the particle size parameter within the particle size gating interval of the component to be measured. The scattering data is the corrected scattering data. The concentration of the component to be measured is predicted based on the scattering data of the counting beads and the corrected scattering data.

[0015] In the present invention, the suspension comprises 5.0 g / L red yeast rice powder, 0.1 g / L vitamin C, 0.2 g / L sodium azide, 10 3 The buffer solution consists of 8.0 g / L disodium hydrogen phosphate, 1.44 g / L potassium dihydrogen phosphate, 8.0 g / L sodium chloride, 0.2 g / L potassium chloride and the rest water, and the components to be tested are Monascus spores and hyphae.

[0016] In the present invention, in step 2, the sheath flow channel and the sample channel are coaxial with the mixing channel, the sheath flow channel is located outside the sample channel, the sheath flow channel and the sample channel are connected to the mixing channel through a tapered transition zone, and the cross-sectional area S1 of the sheath flow channel, the cross-sectional area S2 of the sample channel and the cross-sectional area S3 of the mixing channel satisfy: S1=15S2, S3=0.75(S1+S2).

[0017] In the present invention, in step 4, 0<θ1<π / 4<θ2<π / 2 and θ3-θ2=π / 2.

[0018] In the present invention, in step 5, the wavelength of the incident light is 488 nm.

[0019] In the present invention, in step 6, the correlation between the first side scattering intensity I2(t) and the second side scattering intensity I3(t) is calculated. If the correlation is less than the threshold, adjust the flow of the first controller or the second controller, and I2 and I3 are the average values ​​of I2(t) and I3(t) within the calibration time T respectively.

[0020] In the present invention, in step 6, the variance of the first side scattering intensity I2(t) is calculated. If the variance is less than the upper limit of the variance and greater than the lower limit of the variance, the flow of the first controller or the second controller is maintained. If the variance is greater than the upper limit of the variance and Q2≥Q1 / 15, the flow of the first controller is reduced. If the variance is greater than the upper limit of the variance and Q2<Q1 / 15, the flow of the second controller is reduced. If the variance is less than the lower limit of the variance and Q2≥Q1 / 15, the flow of the first controller is increased. If the variance is less than the lower limit of the variance and Q2<Q1 / 15, the flow of the second controller is increased. Q1 is the flow of the first controller and Q2 is the flow of the second controller.

[0021] In the present invention, in step 7, the wavelength of the incident light includes 495 nm, 505 nm, 633 nm, 640 nm, and 650 nm, the scattering data with a wavelength of 495 nm or 650 nm is the scattering data of the counting beads, the scattering data with a wavelength of 505 nm, 633 nm, and 640 nm is the scattering data of the components to be measured, the lateral intensity ratio β of the counting beads is β=(I2(t)-I2') / (I3(t)-I3'), the particle size function is D=aβ+b, I2' and I3' are the first lateral scattering intensity and the second lateral scattering intensity of the organic particles, respectively, a and b are constants, and D is the diameter of the particles in the mixing streamline.

[0022] In the present invention, in step 9, the average number of scattering points N1 of the component to be measured is calculated based on the scattering data of the component to be measured at at least one wavelength, and the average number of scattering points N2 of the counting beads is calculated based on the scattering data of the counting beads at at least one wavelength. The concentration of the component to be measured is C1=C0N1 / N2, and C0 is the concentration of the counting beads corresponding to the selected wavelength.

[0023] A quantitative detection system for realizing the quantitative detection method of red yeast rice components, comprising: a first controller, a second controller, a light generator, a first detector, a second detector, a third detector, a data processing unit, and a data analysis unit.

[0024] The first controller is used to deliver the buffer into the sheath flow channel;

[0025] The second controller is used to deliver the suspension into the sample channel;

[0026] The light generator is used to generate incident light, forward scattered light, first side scattered light and second side scattered light;

[0027] The first detector is used to collect forward scattering intensity;

[0028] The second detector is used to collect the first side scattering intensity;

[0029] The third detector is used to collect the second side scattering intensity;

[0030] The data processing unit is used to calculate the lateral intensity ratio of the counting beads, the lateral intensity ratio of the components to be measured, the particle size parameters, and fit the particle size function;

[0031] The data analysis unit is used to analyze the concentration of the component to be tested.

[0032] The quantitative detection method and quantitative detection system of the red yeast rice component of the present invention have the beneficial effects that: the present invention uses red yeast rice powder and counting beads to prepare a suspension, and the incident light generated by the light generator is used to irradiate the buffer solution and the suspension in the mixing channel, thereby realizing pipeline detection. The forward scattering intensity, the first side scattering intensity, and the second side scattering light are collected to calculate the lateral intensity ratio of the counting beads, and the particle size function is fitted according to the lateral intensity ratio of the counting beads of different wavelengths. The lateral intensity ratio of the component to be measured is calculated according to the scattering data of the component to be measured, and the particle size parameters are predicted based on the particle size function and the lateral intensity ratio of the component to be measured, and the corrected scattering data of the particle size parameters within the particle size gating interval of the component to be measured are extracted, and the concentration of the component to be measured is predicted according to the scattering data of the counting beads and the corrected scattering data, and the particles with a large diameter difference from the component to be measured are excluded according to the particle size parameters, thereby improving the detection accuracy and reducing the measurement error. BRIEF DESCRIPTION OF THE DRAWINGS

[0033] Figure 1 The schematic diagram of the quantitative detection method of red yeast rice components of the present invention;

[0034] Figure 2 The flowchart of the quantitative detection method of red yeast rice components of the present invention;

[0035] Figure 3 Schematic diagram of the sheath flow channel and the sample channel of the present invention;

[0036] Figure 4 A schematic diagram of a detection platform of the present invention;

[0037] Figure 5 The laser light path diagram of the present invention;

[0038] Figure 6 is a scatter diagram of the component to be tested of the present invention;

[0039] Figure 7 is a schematic diagram of the correlation between the first side scattering intensity and the second side scattering intensity of the present invention;

[0040] Figure 8 is a schematic diagram of the scattering of the polysaccharide polymer particles of the present invention;

[0041] Fig. 9 It is a schematic diagram of the particle size function fitted in the present invention;

[0042] Fig.10 The block diagram of the quantitative detection system for realizing the quantitative detection method of red yeast rice components of the present invention.

[0043] Reference numerals in the accompanying drawings: 101 first controller, 102 second controller, 103 power supply, 104 light generator, 105 first detector, 106 second detector, 107 third detector, 108 pressure pump, 109 data processing unit, 201 sheath flow channel, 202 sample channel, 203 mixing channel, 204 tapered transition zone, 301 bottom plate, 302 holding plate, 303 light-transmitting plate, 304 light-blocking plate, 305 first inlet, 306 second inlet, 307 first outlet, 308 light path window, 401 mixing streamline. DETAILED DESCRIPTION

[0044] The technical solutions in the embodiments of the present invention will be described clearly and completely below in conjunction with the accompanying drawings in the embodiments of the present invention. Embodiment 1

[0045] like Figure 1 As shown, the first controller 101 delivers the buffer solution into the sheath flow channel, and the second controller 102 delivers the suspension containing red yeast rice powder and counting beads into the sample channel. The sheath flow channel and the sample channel are connected to the mixing channel through a tapered transition zone. The mixing streamline of the mixing channel is made to flow at a uniform speed by means of a pressure pump 108. The power supply 103 is turned on, and the light generator 104 generates incident light to illuminate the light path window of the mixing channel. After the incident light passes through the mixing streamline, forward scattered light, first side scattered light and second side scattered light are generated and collected by the first detector 105, the second detector 106 and the third detector 107 respectively. The data processing unit 109 calculates the side intensity ratio of the counting beads, the particle size parameters, the side intensity ratio of the component to be measured and the fitted particle size function according to the forward scattering intensity, the first side scattering intensity and the second side scattering intensity. Figures 2 to 9 As shown, the quantitative detection method of red yeast rice components of the present invention includes the following steps.

[0046] Step 1: Prepare a suspension using red yeast rice powder and counting beads, and then prepare a buffer solution. The suspension includes 5.0 g / L red yeast rice powder, 0.1 g / L vitamin C, 0.2 g / L sodium azide, 10 7The counting beads are composed of 8.0 g / L disodium hydrogen phosphate, 1.44 g / L potassium dihydrogen phosphate, 8.0 g / L sodium chloride, 0.2 g / L potassium chloride and the rest of water, and the components to be tested are Monascus spores and hyphae. The counting beads include two particles of different diameters, and the same concentration of fluorescent components FITC (fluorescein isothiocyanate) and APC (Allophycocyanin) are added respectively to scatter incident light with wavelengths of 495 nm and 650 nm.

[0047] Step 2: The first controller sends the buffer into the sheath flow channel, and the second controller sends the suspension into the sample channel. The buffer and the suspension form a mixing streamline in the mixing channel. Figure 3 As shown, the sheath flow channel 201 and the sample channel 202 are coaxial with the mixing channel 203, the sheath flow channel 201 is located outside the sample channel 202, and the sheath flow channel 201 and the sample channel 202 are connected to the mixing channel 203 through a tapered transition zone 204. The diameter of the sheath flow channel is 4 times that of the sample channel. The cross-sectional area S1 of the sheath flow channel, the cross-sectional area S2 of the sample channel, and the cross-sectional area S3 of the mixing channel satisfy: S1=15S2, S3=0.75(S1+S2).

[0048] Step 3: The incident light generated by the light generator illuminates the optical path window of the mixing channel. After the incident light passes through the mixing streamline, it generates forward scattered light, first side scattered light, and second side scattered light. Figure 4 As shown, the mixing channel, sheath flow channel and sample channel are placed on a detection platform, and the detection platform includes a bottom plate 301, a holding plate 302, a light-transmitting plate 303, a light-blocking plate 304, a first inlet 305, a second inlet 306, a first outlet 307, and a light path window 308. The incident light enters the mixing channel through the light-transmitting plate and the light path window, and the light-blocking plate prevents ambient light from entering the mixing channel to reduce errors. The buffer enters the sheath flow channel from the first inlet, and the suspension enters the sample channel from the second inlet. The mixed streamline is irradiated by the incident light and flows out from the first outlet.

[0049] Step 4: The first detector receives forward scattered light at a scattering angle θ1, the second detector receives the first side scattered light at a scattering angle θ2, and the third detector receives the second side scattered light at a scattering angle θ3, θ1<θ2 and θ3-θ2=π / 2. 0<θ1<π / 4<θ2<π / 2 and θ3-θ2=π / 2. Figure 5As shown, a first detector 105, a second detector 106, and a third detector 107 are installed at scattering angles θ1, θ2, and θ3 respectively at the same length from the mixing channel and on the same horizontal plane. The incident light generated by the light generator 104 passes through the mixing streamline 401 to generate forward scattered light, first side scattered light, and second side scattered light, which are collected by the first detector, the second detector, and the third detector respectively. The shape of the mixing streamline 401 in the figure is a solid circle.

[0050] Step 5: Fix the wavelength of the incident light, collect the forward scattering intensity I1(t) by the first detector, collect the first side scattering intensity I2(t) by the second detector, and collect the second side scattering intensity I3(t) by the third detector, where t is the collection time. The wavelength of the incident light is 488 nm. Fix the wavelength of the incident light, collect the forward scattering intensity, the first side scattering intensity, and the second side scattering intensity, and adjust the flow rate of the first controller or the second controller to mix the buffer and the suspension evenly to reduce the experimental error.

[0051] Step 6: Adjust the flow of the first controller or the second controller according to the first side scattering intensity I2(t) and the second side scattering intensity I3(t) within the calibration time T. First, calculate the correlation between the first side scattering intensity I2(t) and the second side scattering intensity I3(t). If the correlation is less than a threshold, adjust the flow of the first controller or the second controller. The threshold is 0.8, and whether the first controller or the second controller is adjusted is determined based on the correlation between the first side scattering intensity and the second side scattering intensity. Calculate the variance of the first side scattering intensity I2(t), determine whether the first side scattering intensity is stable based on the variance, and then determine the adjustment direction of the first controller or the second controller. The specific adjustment method is shown in Example 2.

[0052] Step 7: I1(t), I2(t), and I3(t) at the same acquisition time t constitute the scattering data. Adjust the wavelength of the incident light to generate the scattering data of the counting beads and the scattering data of the components to be measured. Then calculate the lateral intensity ratio of the counting beads, and fit the particle size function according to the lateral intensity ratio of the counting beads at different wavelengths. The wavelengths of the incident light include 495 nm, 505 nm, 633 nm, 640 nm, and 650 nm. The scattering data with a wavelength of 495 nm or 650 nm are the scattering data of the counting beads, and the scattering data with a wavelength of 505 nm, 633 nm, and 640 nm are the scattering data of the components to be measured. The lateral intensity ratio of the counting beads β=(I2(t)-I2') / (I3(t)-I3'), the particle size function is D=aβ+b, I2' and I3' are the first lateral scattering intensity and the second lateral scattering intensity of the organic particles, respectively, a and b are constants, and D is the diameter of the particles in the mixing streamline.

[0053] Step 8: Calculate the lateral intensity ratio of the component to be measured based on the scattering data of the component to be measured, and predict the particle size parameters based on the particle size function and the lateral intensity ratio of the component to be measured. Substitute the lateral intensity ratio of the component to be measured into the particle size function to obtain the particle size parameters. For specific methods, refer to Example 3. If the particle size of the component to be measured is within a specific range, determine whether the particle corresponding to the collection time t is the component to be measured based on the particle size parameters.

[0054] Step 9: Extract the scattering data of the particle size parameter within the particle size gate interval of the component to be measured, which is the corrected scattering data, and predict the concentration of the component to be measured based on the scattering data of the counting beads and the corrected scattering data. Calculate the average number of scattering points N1 of the component to be measured based on the scattering data of the component to be measured at at least one wavelength, and calculate the average number of scattering points N2 of the counting beads based on the scattering data of the counting beads at at least one wavelength. The concentration of the component to be measured is C1=C0N1 / N2, and C0 is the concentration of the counting beads corresponding to the selected wavelength.

[0055] Red yeast rice has a complex composition, and the particle size parameters generated by the scattering data of proteins, cell walls, etc. are different from those of the components to be measured. The present invention calculates the particle size parameters based on the particle size function and the lateral intensity ratio of the components to be measured. The particle size parameters express the diameter of the particles corresponding to the scattering data of the components to be measured at different sampling times.

[0056] The particle size gating interval of the component to be tested is set according to the diameter of the Monascus spores and hyphae, usually (2μm, 11μm). The scattering data of the component to be tested whose particle size parameter is within the particle size gating interval of the component to be tested is extracted, and the scattering data of the component to be tested is the corrected scattering data. The corrected scattering data eliminates particles with a large difference in diameter from the component to be tested, thereby improving the detection accuracy.

[0057] The concentration of the component to be measured is predicted based on the scattering data of the counting beads and the corrected scattering data. When the incident light irradiates the counting beads or the component to be measured in the mixing streamline, scattering data of corresponding intensity is generated. The number of times the scattering data is excited is related to the concentration. The scattering data is plotted into a scattering diagram (i.e., FSC vs. SSC). Figure 6 As shown in the figure, the horizontal axis is the forward scattering intensity, the vertical axis is the first side scattering intensity, and the dots are excitation events (scattering data is excited). Under the same experimental conditions, the ratio of the counting bead concentration C0 to the concentration of the component to be tested C1 is equal to the ratio of the number of counting bead excitations N2 to the number of excitations N1 of the component to be tested. That is, the concentration of the component to be tested is C1=C0N1 / N2.

[0058] In this embodiment, the average number of scattering points N1 of the component to be measured is calculated based on the scattering data of the component to be measured at one wavelength, and the average number of scattering points N2 of the counting beads is calculated based on the scattering data of the counting beads at one wavelength. In another embodiment, different numbers of scattering points are obtained based on the scattering data of different wavelengths, and then the average number of scattering points is obtained by averaging.

[0059] Furthermore, for the same concentration of the tested component, the scattering diagram is concentrated in a stable area. In the actual test process, some proteins, starch particles, etc. will produce similar scattering data. The present invention can also set other gates to exclude interfering data. For example Figure 6 As shown, a rectangular or elliptical gate is defined to segment the scattering pattern around a core group of particles (event group), excluding particles outside the gate, ensuring that the gate contains the particles of interest as accurately as possible. Embodiment 2

[0060] This embodiment further discloses a method for adjusting the flow rate of the first controller or the second controller in step 6. The components of the red yeast rice suspension are complex. In order to control the separation degree of the components of the red yeast rice suspension and ensure that the components to be measured accurately scatter the incident light, it is necessary to optimize the flow rates of the sheath flow and the sample flow in combination with the specific red yeast rice.

[0061] Step 601: Calculate the correlation between the first side scattering intensity I2(t) and the second side scattering intensity I3(t). The scattering intensity changes with time. In this embodiment, the Pearson correlation coefficient is used. , I2 and I3 are the means of I2(t) and I3(t) within the calibration time T, respectively.

[0062] Step 602: Determine whether the flow needs to be adjusted based on the correlation. If the correlation is less than the threshold, adjust the flow of the first controller or the second controller. The threshold can be set to 0.85. Figure 7 When the components are uniform and stable, the first side scattering intensity and the second side scattering intensity show a strong positive correlation. The ideal state in the figure is Pea=1. If the correlation decreases, the mixing streamline may produce turbulence, the stability of the suspension may deteriorate, and the detection accuracy may be affected. The inlet flow needs to be optimized. If the correlation coefficient is greater than or equal to the threshold, no adjustment is required.

[0063] Step 603: Adjust the flow rate of the first controller or the second controller according to the variance of the first side scattering intensity I2(t) and the current flow rate of the first controller or the second controller. .

[0064] Specifically, if the variance is less than the upper limit of the variance and greater than the lower limit of the variance, maintain the flow rate of the first controller or the second controller. If the variance is greater than the upper limit of the variance and Q2≥Q1 / 15, reduce the flow rate of the first controller, Q1=Q1-ΔQ, ΔQ=(Q2-Q1 / 15) / M, M is the number of adjustment steps, usually 2 to 20, to ensure the convergence of the adjustment. If the variance is greater than the upper limit of the variance and Q2<Q1 / 15, reduce the flow rate of the second controller, Q2=Q2-ΔQ. If the variance is less than the lower limit of the variance and Q2≥Q1 / 15, increase the flow rate of the first controller, Q1=Q1+ΔQ. If the variance is less than the lower limit of the variance and Q2<Q1 / 15, increase the flow rate of the second controller, Q2=Q2+ΔQ. Q1 is the flow rate of the first controller, Q2 is the flow rate of the second controller, and ΔQ is the flow rate increment of the first controller and the second controller. The values ​​of the upper limit of the variance and the lower limit of the variance are related to the power of the incident light and the scattering intensity in each direction. Usually the first side scattering intensity is around 2000Lx, and the upper limit of the variance can be 40000Lx. 2 , the lower limit of variance can be 900Lx 2 It should be noted that, if the variance of the first side scattering intensity I2(t) does not converge when adjusting the flow rate of the first controller or the second controller, resulting in failure to adjust it within the interval, the task is terminated and the suspension is prepared again. Embodiment 3

[0065] This embodiment further discloses a method for calculating the particle size parameters in step 7 and step 8. Red yeast rice is rich in polysaccharides, which form polysaccharide polymers in the suspension, and the particle size of the polysaccharide polymers is similar to the particle size of Monascus spores. The present invention excludes the signal interference of organic particles including polysaccharide polymers when calculating the particle size parameters, which can further improve the detection accuracy.

[0066] First, calculate the lateral intensity ratio β of the counting beads, β=(I2(t)-I2') / (I3(t)-I3'), where I2' and I3' are the first lateral scattered light intensity and the second lateral scattered light intensity of the organic particles, respectively. Figure 8 , the polysaccharide polymer is fitted into a solid sphere, and a three-dimensional coordinate system is established, with the center of the sphere as the coordinate origin, the direction of the incident light as the X-axis, the direction of the horizontal plane perpendicular to the X-axis as the Y-axis, and the direction perpendicular to the XOY plane as the Z-axis. The polysaccharide polymer scatters the incident light in three directions of the three-dimensional coordinate system, generating forward scattering intensity I1', first side scattering intensity I2', and second side scattering intensity I3'. The first side scattering intensity of organic particles I2'=λ 2 i(θ2)I0 / (8π 2 r1 2 ), the second side scattered light intensity of organic particles I3'=λ 2 i(θ3)I0 / (8π 2 r22 ), λ is the wavelength of the incident light, I0 is the intensity of the incident light, i(θ2) is the scattering value of the intensity distribution function at θ2, i(θ3) is the scattering value of the intensity distribution function at θ3, r1 is the distance between the second detector and the mixing streamline, and r2 is the distance between the third detector and the mixing streamline.

[0067] Then, the particle size function is fitted according to the lateral intensity ratio of the counting beads at different wavelengths. Fig. 9 , the solute particle size of the Monascus suspension is related to the lateral intensity ratio of the counting beads, and the particle size parameters can be predicted based on the lateral intensity ratio of the counting beads. The particle size function is a linear function, that is, D=aβ+b, where a and b are constants. In this embodiment, the average value of the first lateral scattered light intensity and the second lateral scattered light intensity can be obtained based on the scattering data of a wavelength of 495 nm or 650 nm, and then the average value of the lateral intensity ratio of the counting beads is calculated. The diameter of the counting beads and the average value of the lateral intensity ratio are substituted into the particle size function to calculate the constants a and b. In another embodiment, the lateral intensity ratio of the counting beads is calculated based on the first lateral scattered light intensity and the second lateral scattered light intensity at each sampling moment, the lateral intensity ratio of the T group of counting beads and the corresponding counting beads are fitted into a linear function, and then the constants a and b are determined based on the slope and intercept of the linear function.

[0068] Finally, the scattering data with wavelengths of 488nm, 500nm, and 505nm are the scattering data of the component to be measured. The lateral intensity ratio of the component to be measured is calculated according to the scattering data of the component to be measured, and the particle size parameters are calculated based on the particle size function and the lateral intensity ratio of the component to be measured.

[0069] Furthermore, the intensity distribution function i(θ) is related to the scattering angle. The Mie scattering theory can be used to predict the scattering field of spherical particles to electromagnetic waves, and then predict the scattering value of the intensity distribution function. i(θ)=|S(m,θ,ε)| 2 = , a n and b n is the Mie scattering coefficient, which is a function related to the particle refractive index m and the dimensionless parameter ε, δ n and is a function related to the scattering angle θ. In red yeast rice, the diameters of the polysaccharide polymer and the Monascus are in the micrometer range, which is much larger than the wavelength of the incident light. Therefore, in a simpler embodiment, the intensity distribution function can be simplified to i(θ)=sin 4 (θ / 2). Embodiment 4

[0070] like Fig.10As shown, a quantitative detection system for realizing the quantitative detection method of the red yeast rice component includes: a first controller, a second controller, a light generator, a first detector, a second detector, a third detector, a data processing unit, a data analysis unit, and a data display unit. The first controller is used to send a buffer into the sheath flow channel to adjust the flow rate of the buffer. The second controller is used to send a suspension into the sample channel to adjust the flow rate of the suspension. The light generator is used to generate incident light, and the incident light generates forward scattered light, a first side scattered light, and a second side scattered light after passing through a mixing streamline. The first detector is used to receive the forward scattered light and collect the forward scattered intensity. The second detector is used to receive the first side scattered light and collect the first side scattered intensity. The third detector is used to receive the second side scattered light and collect the second side scattered intensity. The data processing unit is used to generate scattering data of counting beads and scattering data of the component to be measured according to the forward scattering intensity, the first side scattering intensity and the second side scattering intensity, calculate the lateral intensity ratio of the counting beads according to the scattering data of the counting beads, fit the particle size function according to the lateral intensity ratio of different wavelengths of the counting beads, calculate the lateral intensity ratio of the component to be measured according to the scattering data of the component to be measured, and predict the particle size parameters based on the particle size function and the lateral intensity ratio of the component to be measured. The data analysis unit is used to extract the scattering data of the particle size parameter within the particle size gating interval of the component to be measured to generate corrected scattering data, and predict the concentration of the component to be measured according to the scattering data of the counting beads and the corrected scattering data. The data display unit is used to display the concentration of the component to be measured.

[0071] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions and improvements made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims

1. A method for quantitatively detecting red yeast rice components, characterized in that: The following steps are involved: Step 1: Prepare a suspension using red yeast rice powder and counting beads, and then prepare a buffer solution; Step 2: The first controller sends the buffer into the sheath flow channel, and the second controller sends the suspension into the sample channel, and the buffer and the suspension form a mixing streamline in the mixing channel; Step 3: The incident light generated by the light generator illuminates the optical path window of the mixing channel, and the incident light generates forward scattered light, first side scattered light, and second side scattered light after passing through the mixing streamline; Step 4: The first detector receives the forward scattered light at a scattering angle θ1, the second detector receives the first side scattered light at a scattering angle θ2, and the third detector receives the second side scattered light at a scattering angle θ3, θ1<θ2 and θ3-θ2=π / 2; Step 5: Fix the wavelength of the incident light, the first detector collects the forward scattering intensity I1(t), the second detector collects the first side scattering intensity I2(t), and the third detector collects the second side scattering intensity I3(t), where t is the collection time; Step 6: adjusting the flow rate of the first controller or the second controller according to the first side scattering intensity I2(t) and the second side scattering intensity I3(t) within the calibration time T; Step 7: I1(t), I2(t), and I3(t) at the same acquisition time t constitute the scattering data, adjust the wavelength of the incident light, generate the scattering data of the counting beads and the scattering data of the components to be measured, and then calculate the lateral intensity ratio of the counting beads, and fit the particle size function according to the lateral intensity ratio of the counting beads at different wavelengths; Step 8: Calculate the lateral intensity ratio of the component to be measured according to the scattering data of the component to be measured, and predict the particle size parameters based on the particle size function and the lateral intensity ratio of the component to be measured; Step 9: Extract the scattering data of the particle size parameter within the particle size gate interval of the component to be measured, the scattering data is the corrected scattering data, and predict the concentration of the component to be measured based on the scattering data of the counting beads and the corrected scattering data, wherein: In step 6, the correlation between the first side scattering intensity I2(t) and the second side scattering intensity I3(t) is calculated. If the correlation is less than the threshold, adjust the flow of the first controller or the second controller. I2 and I3 are the average values ​​of I2(t) and I3(t) within the calibration time T. Calculate the variance of the first side scattering intensity I2(t). If the variance is less than the upper limit of the variance and greater than the lower limit of the variance, maintain the flow of the first controller or the second controller. If the variance is greater than the upper limit of the variance and Q2≥Q1 / 15, reduce the flow of the first controller. If the variance is greater than the upper limit of the variance and Q2<Q1 / 15, reduce the flow of the second controller. If the variance is less than the lower limit of the variance and Q2≥Q1 / 15, increase the flow of the first controller. If the variance is less than the lower limit of the variance and Q2<Q1 / 15, increase the flow of the second controller. Q1 is the flow of the first controller, and Q2 is the flow of the second controller. In step 7, the wavelength of the incident light includes 495 nm, 505 nm, 633 nm, 640 nm, and 650 nm. The scattering data with a wavelength of 495 nm or 650 nm is the scattering data of the counting beads. The scattering data with a wavelength of 505 nm, 633 nm, and 640 nm is the scattering data of the components to be measured. The lateral intensity ratio β of the counting beads is β=(I2(t)-I2') / (I3(t)-I3'). The particle size function is D=aβ+b. I2' and I3' are the first lateral scattering intensity and the second lateral scattering intensity of the organic particles, respectively. a and b are constants. D is the diameter of the particles in the mixing streamline. In step 9, the average number of scattering points N1 of the component to be measured is calculated based on the scattering data of the component to be measured at at least one wavelength, and the average number of scattering points N2 of the counting beads is calculated based on the scattering data of the counting beads at at least one wavelength. The concentration of the component to be measured is C1=C0N1 / N2, and C0 is the concentration of the counting beads corresponding to the selected wavelength.

2. The method for quantitatively detecting red yeast rice components according to claim 1, characterized in that: The suspension includes 5.0 g / L red yeast rice powder, 0.1 g / L vitamin C, 0.2 g / L sodium azide, 10 3 The buffer solution consists of 8.0 g / L disodium hydrogen phosphate, 1.44 g / L potassium dihydrogen phosphate, 8.0 g / L sodium chloride, 0.2 g / L potassium chloride and the rest water, and the components to be tested are Monascus spores and hyphae.

3. The method for quantitative detection of red yeast rice components according to claim 1, characterized in that: In step 2, the sheath flow channel and the sample channel are coaxial with the mixing channel, the sheath flow channel is located outside the sample channel, the sheath flow channel and the sample channel are connected to the mixing channel through a tapered transition zone, and the cross-sectional area S1 of the sheath flow channel, the cross-sectional area S2 of the sample channel, and the cross-sectional area S3 of the mixing channel satisfy: S1=15S2, S3=0.75(S1+S2).

4. The method for quantitative detection of red yeast rice components according to claim 1, characterized in that: In step 4, 0<θ1<π / 4<θ2<π / 2 and θ3-θ2=π / 2.

5. The method for quantitative detection of red yeast rice components according to claim 1, characterized in that: In step 5, the wavelength of the incident light is 488 nm.

6. A quantitative detection system for realizing the quantitative detection method of red yeast rice components according to claim 1, characterized in that: include: a first controller, a second controller, a light generator, a first detector, a second detector, a third detector, a data processing unit, and a data analysis unit, The first controller is used to deliver the buffer into the sheath flow channel; The second controller is used to deliver the suspension into the sample channel; The light generator is used to generate incident light, forward scattered light, first side scattered light and second side scattered light; The first detector is used to collect forward scattering intensity; The second detector is used to collect the first side scattering intensity; The third detector is used to collect the second side scattering intensity; The data processing unit is used to calculate the lateral intensity ratio of the counting beads, the lateral intensity ratio of the components to be measured, the particle size parameters, and fit the particle size function; The data analysis unit is used to analyze the concentration of the component to be tested.

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