CN107941741A - The method that methylenum careuleum burst size in Plasma Inactivation bag is detected using near infrared spectrum - Google Patents

Abstract

The invention relates to a near-infrared spectrum detector, comprising a transmission platform, the transmission platform includes a base plate, two vertical plates parallel to each other are vertically arranged on the base plate, and corresponding transmission windows are respectively opened on the two vertical plates. The transmission window is covered with a transparent optical window, and a stage is arranged between the two vertical plates. The object to be tested is located between the two optical windows. held between two optical windows. The present invention also relates to a method for detecting the release amount of methylene blue in a plasma inactivation bag by using near-infrared spectroscopy, comprising: fixing the optical path between two optical windows to 2.0-5.0mm; placing the plasma inactivation bag On the stage, perform near-infrared spectrum detection on multiple plasma standard solutions, and use the PLS method to establish a multivariate analysis model of methylene blue release; perform near-infrared spectrum detection on unknown samples, and use the analysis model to analyze the release of methylene blue. amount released.

Description

A method for detecting the amount of methylene blue released in plasma inactivation bags by near-infrared spectroscopy

technical field

The invention relates to the field of biological detection, in particular to a method for detecting the release amount of methylene blue in a plasma inactivation bag by using near-infrared spectroscopy.

Background technique

In order to prevent virus infection from blood transfusion, the methylene blue light method is widely used to inactivate plasma virus in clinical plasma. This is the only virus inactivation technology approved for clinical use in a single blood component in my country. The inactivation process is carried out in the plasma inactivation bag, and the release of the inactivator methylene blue in the plasma inactivation bag enters with the plasma through gradual dissolution when the plasma flows through the methylene blue storage under sterile conditions Inactivation bag, this process is the release process of methylene blue, and its release amount is a key parameter to determine the degree of virus inactivation. However, the amount of release is related to many factors, including flow rate, ambient temperature, degree of hygroscopicity of methylene blue, and the material and production process of immobilized methylene blue in the storage, etc. If the release amount of methylene blue in the plasma inactivation bag is too high, the residual amount of methylene blue in the plasma will be too large, which will easily lead to cumulative toxicity; if the release amount is too low, the virus inactivation effect cannot be guaranteed. A large number of data show that the virus inactivator methylene blue must be released within a very narrow range, which is 0.9-1.3 μmol/L in my country. In order to detect the release amount as conveniently, quickly and sensitively as possible, a large number of sampling-based detection methods have emerged, including solid phase extraction-spectrophotometry, enhanced fluorescence spectroscopy, HPLC, chemiluminescence, and resonance Rayleigh scattering spectroscopy. Law. Sampling detection requires the use of extraction devices to extract samples from the plasma inactivation bag for testing. Since the extraction operation will cause damage to the plasma inactivation bag, this destructive detection method can easily lead to secondary contamination of plasma during sampling, including bacteria, chemical Pollutants, as well as particulate matter, cause the plasma in the plasma inactivation bag to no longer be used in the human body, resulting in a waste of plasma. It is also based on this that the plasma that has been tested for release can no longer be used clinically. Therefore, it is impossible to carry out such inactivation quality control based on destructive detection for each bag of plasma used in clinical practice. In fact, whether the quality of plasma in the plasma inactivation bag is reliable or not is only a statistical result. The non-destructive detection of the amount of methylene blue released in the inactivation bag is the key issue to realize the quality control of each bag of plasma inactivation.

At present, near-infrared spectroscopy is usually used for non-destructive and rapid detection of samples, which is suitable for quality control in the production process. Among them, the non-destructive testing of samples by diffuse reflectance near-infrared spectroscopy should be the most widely used, but this method has certain limitations: first of all, this method is generally applicable to solid, powder, and paste, but it is difficult to apply to liquid samples. After the light is irradiated into the liquid, only extremely weak diffuse reflection light is produced, and most or even all of it is transmitted light, so it is difficult to quantitatively detect a certain substance in the liquid based on its diffuse reflection near-infrared spectrum.

For the existing near-infrared spectroscopy detection method for rapid quantification of substances in liquids, it is usually necessary to take samples from the container under inspection for detection, which mainly includes two methods. The first one is to place the obtained liquid sample in the cuvette, and then put it into the cuvette rack of the instrument for detection; the second one is to insert the liquid optical fiber probe with a slit into the liquid in the container to be tested, The detection liquid directly flows into the slit of the fiber optic probe as a cuvette to realize the sampling operation, and the fiber optic retransmits the signal to the spectrometer to realize fast detection. No matter which of the two methods is used, it is a destructive detection method, and the defects of this method have been described in detail above. However, using the existing infrared spectrometer to directly detect the plasma bag will encounter great difficulties. First, the plasma inactivation bag contains many air bubbles. If these air bubbles are in the detection area, the measurement results will cause great errors; secondly. , When the plasma inactivation bag is directly detected, because the bag body is soft and its shape is irregular, the optical path of the spectral detection is difficult to fix, and it is difficult to calculate the content of the analyte using the quantitative formula. Of course, the plasma in the inactivation bag contains a large number of biological macromolecules, and the carbon-hydrogen bonds in it will generate a large number of near-infrared spectrum signals, which will also interfere with the detection results and bring certain difficulties to the detection method.

Contents of the invention

In order to solve the above-mentioned technical problems, the purpose of the present invention is to provide a method for detecting the amount of methylene blue released in the plasma inactivation bag by using near-infrared spectroscopy, which overcomes the difficulty of performing near-infrared measurements on irregularly shaped samples in the prior art. Quantitative defects realize the non-destructive, rapid and accurate analysis of the amount of methylene blue released in the plasma inactivation bag.

In one aspect, the present invention provides a near-infrared spectrum detector, including a light source, a monochromator, a detector and a computer processing information system, and also includes a transmission platform, the transmission platform includes a substrate, and the substrate is vertically provided with two The first vertical board and the second vertical board are parallel to each other, and the first vertical board and the second vertical board are respectively provided with corresponding transmission windows, and the transmission windows are covered with transparent optical windows, and the optical windows are located between the two On the opposite side of the vertical plate, there is a stage between the first vertical plate and the second vertical plate, which is used to carry the soft bag containing the object to be tested, and the stage is located below the lower edge of the optical window, The soft bag containing the object to be tested is located between the two optical windows, and at least one vertical plate can move horizontally along the surface of the substrate, so that the soft bag containing the object to be tested is clamped between the two optical windows And the optical path between the two optical windows can be adjusted.

Further, the stage is located at least 2cm below the lower edge of the optical window.

Further, the first vertical plate is fixedly connected with the base plate, the loading table is fixedly connected with the first vertical plate, and the second vertical plate is provided with a strip-shaped hole facing the loading table, and the strip-shaped hole is consistent with the shape of the loading table. Fitting, the stage can be slid into the bar hole.

Furthermore, a guide rail is fixedly connected to the base plate, and the second riser can slide horizontally along the guide rail and can be locked at any position of the guide rail, thereby fixing the distance between the two optical windows (that is, the optical path).

Further, a positioning platform is fixedly connected to the side of the second vertical plate away from the first vertical plate, the second vertical plate is slidably connected to the guide rail through the positioning platform, and the positioning platform is screwed with a screw rod, and the axial direction of the screw rod is the same as that of the second vertical plate. Vertically, a fixed baffle is fixedly connected to the guide rail, the fixed baffle is provided with a through hole, and the screw rod is passed through the through hole.

Further, the material of the optical window is quartz or optical glass.

Further, the transmission window is rectangular.

Further, the area of the optical window is larger than that of the transmission window.

In another aspect, the present invention also provides a method for utilizing near-infrared spectroscopy to detect the release of methylene blue in the plasma inactivation bag, using the above-mentioned near-infrared spectroscopy detector, comprising the following steps:

(1) Adjust the distance between the first vertical plate and the second vertical plate, so that the optical path between the two optical windows is fixed at 2.0-5.0mm; preferably, the optical path is 3.5mm;

(2) Put the plasma inactivation bag on the stage of the near-infrared spectroscopy detector and make it between the two optical windows, and squeeze it slightly above the bag so that the plasma inactivation bag is tightly connected to the The two optical windows are in contact, and the plasma inactivation bag is squeezed to form a regular plane and its surface is pressed into a plane, wherein the plasma inactivation bag contains a plasma standard solution with known methylene blue content;

(3) Under the condition of wave number 4000-10000cm ^-1 , carry out near-infrared spectrum detection to plasma standard solution, repeat test a plurality of samples, according to the near-infrared spectrum detection result of multiple samples, utilize PLS method to establish the methylene blue release amount Multivariate analysis model;

(4) Place the plasma inactivation bag containing methylene blue to be tested on the stage, and squeeze it slightly above the bag so that the plasma inactivation bag is in close contact with the two optical windows, And the plasma inactivation bag is squeezed to form a regular plane;

(5) Near-infrared spectrum detection is carried out under the condition of wave number 4000-10000cm ⁻¹ , and the release amount of methylene blue in the plasma inactivation bag is determined by using the multivariate analysis model of methylene blue release amount.

Further, in step (2), the concentration of methylene blue in the plasma standard solution is 0.312 μmol/L-2.959 μmol/L.

Further, in step (3), the number of plasma standard solutions is 52.

Further, in step (3) and step (5), the near-infrared light scanning resolution of the near-infrared spectroscopy detection is 8 cm ^-1 , the number of scans is 32, and the scan speed is 0.3165.

Further, in step (3) and step (5), the scanning aperture of the near-infrared spectrum detection is 22, and the gain is 1.

Further, in step (3) and step (5), after the near-infrared spectrum is detected, the average absorbance value in the interval of 3999.7-4265.8 cm ^-1 is selected as the drift amount, and the drift is eliminated for the detected near-infrared spectrum.

Further, in step (3) and step (5), after near-infrared spectrum detection, the absorbance in the two intervals of 5350-6600cm ^-1 and 7200-10001cm ^-1 is selected as the information-rich spectrum. Preferably, after near-infrared detection, the absorbance in the two intervals of 5827.9-6464.3 cm ^-1 and 7467.1-9009.9 cm ^-1 is selected as the information-rich spectrum.

Further, in step (3), the number of principal components is selected to be 11, and 11 samples are randomly selected as the prediction set according to the distribution of concentration, and the remaining 41 samples are used as the calibration set, and the release amount of methylene blue is established by the PLS method. multivariate analysis model.

Further, the formula of the multivariate analysis model of methylene blue release is: c=k ₁ A ₁ +k ₂ A ₂ +···+k _n A _n , wherein, c represents the concentration of methylene blue in the sample, A ₁ , A ₂ ···A _n are the absorbance at the wavelengths λ ₁ , λ ₂ ··· _λn corresponding to the information-rich spectral interval selected from the detection spectrum of the sample, k ₁ , k ₂ · ··k _n is the corresponding PLS regression coefficient.

Further, before step (2), it also includes placing an air-filled and unused plasma inactivation bag on the stage of the near-infrared spectrum detector for near-infrared spectrum detection as a spectral background step.

By means of the above solution, the present invention has at least the following advantages:

The structural setting of the transmission platform of the near-infrared spectrum detector of the present invention can realize the fixed optical path during the non-destructive detection of the plasma inactivation bag, and can effectively solve the problem that the air bubbles in the plasma inactivation bag exist in the detection optical path, so that the quantitative results Accurate and reliable. Due to the extrusion of the optical windows, the plasma inactivation bag between the two optical windows is squeezed into a regular plane. The air bubbles in the plasma inactivation bag between them are naturally driven to the plasma inactivation bag outside the extrusion part of the optical window, so that the air bubbles will not appear in the detection area.

The method of the invention realizes the non-destructive, fast and accurate analysis of the amount of methylene blue released in the plasma inactivation bag outside the plasma inactivation bag in a non-sampling mode. The correlation coefficients of the model calculated concentration to the real concentration of each sample were 98.16% (calibration set) and 98.93% (prediction set), and the average recoveries of the samples in the high, middle and low concentration groups were 103.6%-110.8%.

The above description is only an overview of the technical solutions of the present invention. In order to understand the technical means of the present invention more clearly and implement them according to the contents of the description, the preferred embodiments of the present invention and accompanying drawings are described in detail below.

Description of drawings

Fig. 1 is a schematic diagram of the three-dimensional structure of the transmission platform of the present invention;

Fig. 2 is a schematic diagram of the front side structure of the transmission platform of the present invention;

Fig. 3 is a schematic diagram of the structure of the left side of the transmission platform of the present invention;

Fig. 4 is a schematic diagram of an enlarged structure of circle A in Fig. 3;

Fig. 5 is a top view structural schematic diagram of the transmission platform of the present invention;

Fig. 6 is the near-infrared spectrogram after baseline drift is eliminated in embodiment 2;

Figure 7 is the residual of the PLS model based on the leave-one-out interactive test;

Fig. 8 is the multivariate analysis model figure of methylene blue release amount;

Explanation of reference signs:

1-second vertical plate; 2-quartz plate; 3-plasma inactivation bag; 4-transmission window; 5-substrate; 6-first vertical plate; 7-stage; 8-positioning platform; 9-screw; 10-guide rail; 11-fixed baffle.

Detailed ways

The specific implementation manners of the present invention will be further described in detail below in conjunction with the accompanying drawings and embodiments. The following examples are used to illustrate the present invention, but are not intended to limit the scope of the present invention.

Example 1

Referring to Fig. 1-5, a kind of near-infrared spectrum detector of the present invention, comprises light source, monochromator, detector and computer processing information system, also comprises a transmission platform, and transmission platform comprises a substrate 5, and flexible connection on the substrate 5 There are two first vertical boards 6 and second vertical boards 1 parallel to each other, and the first vertical boards 6 and second vertical boards 1 are arranged perpendicular to the base plate 5 . Each of the two vertical plates is provided with a corresponding rectangular transmission window 4, and the upper cover of the transmission window 4 is provided with a square transparent quartz plate 2. The quartz plate 2 is located on the opposite side of the two vertical plates, and the area of the quartz plate 2 is larger than that of the transmission window. The area of window 4. The first vertical plate 6 is fixedly connected with the base plate 5, and the first vertical plate 6 is also fixedly connected with a stage 7, the stage 7 is located at least 2 cm below the lower edge of the quartz plate 2, and the stage 7 is used for placing Soft bag for measuring objects. The upper surface of the stage 7 is parallel to the base plate 5, and the second vertical plate 1 is provided with a strip hole facing the stage 7, and the strip hole is adapted to the shape of the stage 7, so that the loading The table 7 is slidably inserted into the bar-shaped hole. Two guide rails 10 are fixedly connected to the base plate 5 , the guide rails 10 are arranged perpendicular to the second riser 1 , and the second riser 1 can slide horizontally along the guide rails 10 . The second riser 1 is fixedly connected with a positioning platform 8 on one side away from the first riser 6, and the positioning platform 8 is threadedly connected with a screw rod 9, and the axial direction of the screw rod 9 is perpendicular to the second riser 1, and the positioning platform 8 and the guide rail 10 Slidingly connected, the guide rail 10 is fixedly connected with a fixed baffle 11, the fixed baffle 11 is provided with a through hole, and the screw rod 9 is penetrated in the through hole.

When performing sample detection, such as the sample detection in the plasma inactivation bag 3, first adjust the screw to drive the second vertical plate 1 to move along the guide rail 10, so that it slides to the required distance, and the sliding direction is perpendicular to the plane where the vertical plate is located , and then adjust the screw 9 to fix the second vertical plate 1 at this position, that is, the optical path of the transmission detection is fixed. And during the sliding process of the second vertical plate 1, the stage 7 is slidably inserted into the strip-shaped hole provided thereon to facilitate the adjustment of the optical path and ensure that the quartz plate 2 can sterilize the plasma under different optical paths. Live bag 3 is clamped. Put the plasma inactivation bag 3 on the stage 7, and squeeze it slightly above the bag, so that the plasma inactivation bag is in close contact with the two optical windows, so that the plasma inactivation bag 3 is clamped between the two optical windows. Between 2 pieces of quartz. When the quartz plate 2 squeezes the plasma inactivation bag 3, the plasma inactivation bag 3 between the two quartz plates 2 forms a regular shape, thereby fixing the exact optical path (marking line "5" in Fig. 4 "The distance marked is the detection optical path), and the air bubbles in the plasma inactivation bag 3 migrate to the part of the plasma bag above the upper edge of the quartz sheet 2, so that there are no air bubbles between the quartz sheets 2, preventing it from affecting the detection results .

Example 2

In the following examples, the materials used include: non-inactivated human plasma provided by the Blood Bank of Suzhou Red Cross Center, using the filter in the disposable virus inactivation accessory device to filter out white blood cells to obtain plasma without methylene blue ( i.e. blank plasma). Disposable blood collection and transfusion set containing plasma inactivation bag (transfer bag, specification: 35ml, production batch number: 170428, Sichuan Nangeer Biotechnology Co., Ltd.). Methylene blue hydrochloride (analytically pure, >97% based on dry product content, Sigma-Aldrich Company), the experimental water is triple distilled water. Near-infrared spectrometer (model: NEXUS, American Thermo Company), its transmission platform is modified according to embodiment 1.

The present embodiment is the establishment method of the multivariate analysis model of methylene blue release amount, specifically as follows:

(1) First prepare the methylene blue stock solution: weigh 1.3953 g of methylene blue (analytically pure, with a drying weight loss rate of 10.6%), place it in a 500 mL volumetric flask, and dilute to volume with PBS solution. Accurately measure 1.00ml of methylene blue solution, and then dilute to 1000ml with PBS solution to obtain 7.8μmol/L methylene blue stock solution. Protect from light and keep sealed.

(2) Reconfigure the sample to be tested: firstly, use the methylene blue stock solution and PBS buffer solution (pH=7.4) to prepare primary and secondary dilutions by stepwise dilution method. After that, a total of 52 methylene blue plasma solutions with a concentration of 0.312-2.959 μmol/L were precisely prepared from the stock solution, dilution solution and plasma. Finally, inject the methylene blue plasma solution into each blood collection and transfusion transfer bag with a disposable sterilized syringe to obtain the sample to be tested.

(3) Near-infrared spectroscopy detection of samples

Wipe the outer surface of the plasma inactivation bag to be tested and the quartz plate on the transmission platform with alcohol, and adjust the distance between the first vertical plate 6 and the second vertical plate 1 according to the method in Example 1, so that the distance between the two optical windows is The optical distance between them is fixed at 3.5mm. Afterwards, the plasma inactivation bag was placed on the transmission platform according to the method in Example 1, and the position of the plasma inactivation bag was aligned with the left and right sides as far as possible each time, and the spectrum of the sample was measured. The instrument parameters are: wavenumber 4000-10000cm ^-1 , optical path 3.5mm, resolution 8cm ^-1 , scanning times 32, scanning speed 0.3165, aperture 22, gain 1. Air-filled, unused plasma inactivated bags were used as spectral background before testing samples on the day.

(4) The near-infrared transmission spectrum of the sample in the plasma bag obtained by detection is used as the standard spectrum in the range of 3999.7-4265.8 cm ^-1 to perform baseline drift calibration. Select 5827.9-6464.3cm ^-1 and 7467.1-9009.9cm ^-1 as the rich information interval, take the mean-centered spectrum as the model variable, select the number of principal components as 11, and randomly select 11 samples according to the distribution of concentration as the prediction set. Taking the remaining 41 samples as the calibration set, the multivariate analysis model of methylene blue release was established by PLS method.

For the low wavenumber region of the near-infrared spectrum, there will usually be a spectral baseline. It should be a horizontal line, and the spectra of samples of different concentrations in this interval should completely overlap, have no statistical difference, and have an absorbance mean of 0. In Fig. 6, the spectral baseline of each sample is in the interval of 3999.7-4265.8 cm ^-1 . However, in the original spectrum, the baselines of samples with different concentrations drifted by about 7, and this concentration-independent drift will affect the accuracy of methylene blue release. The average absorbance value of the detected spectrum of each sample in the range of 3999.7-4265.8 cm ^-1 is used as the drift amount, and the drift is eliminated to obtain the drift-eliminated spectra of all samples.

From the spectrum after baseline drift elimination (Figure 6), it can be found that the plasma sample of methylene blue has two obvious absorption peaks in the range of 5350-6600cm ^-1 and 7200-10001cm ^-1 in the entire spectral range, which are the absorption peak spectral regions . For the former, it consists of two high and low shoulder peaks. In the low wave number region of 5350-5828cm ^-1 , the spectrum not only has low absorption, but is basically a horizontal line without peaks, and there are wavy burrs. This type of low signal-to-noise ratio The spectral signal will affect the precision of the quantitative analysis model. The spectrum in the high wavenumber range of 6464-6600cm ^-1 is the end spectrum of the absorption peak, with low absorption and weak signal, which also shows a low signal-to-noise ratio. Therefore, the spectrum at 5827.9-6464.3 cm ^-1 in the absorption peak spectral region is selected as the signal-rich spectrum. Similarly, in the 7200-10001cm ^-1 absorption peak spectral region, select the 7467.1-9009.9cm ^-1 spectrum as the rich signal spectrum.

Using the spectra of 5827.9-6464.3cm ^-1 and 7467.1-9009.9cm ^-1 as the rich information spectrum, using the mean-centered spectrum as the model variable, and using the calibration set samples according to the leave-one-out interactive test, calculate the scores of different principal components When the PLS model predicts the residual of methylene blue concentration (Fig. 7). We can find that as the number of principal components increases, the predicted concentration residuals tend to decrease gradually, indicating that the accuracy of the model gradually increases. When the number of principal components is greater than or equal to 11, the degree of reduction in the prediction residual decreases significantly, indicating that the continued increase in the number of principal components has a low contribution to the accuracy of the model. Of course, too large a number of principal components will also lead to overfitting of the model, so 11 is selected as the number of principal components.

Taking the information-rich spectra of the selected two intervals as the identification variable of the release of methylene blue, based on the previously determined principal component numbers, centering the spectral mean, and using the PLS method to establish a multivariate analysis model for the release of methylene blue, in which methylene blue For the high and low distribution of blue concentration, 11 samples are randomly selected as the prediction set, and the remaining 41 samples are the prediction set. The results are shown in Figure 8. ▲ in the figure represents the calibration set sample, and ● represents the prediction set sample. It can be found that in the multivariate analysis model, the calibration concentration and the real concentration of the calibration sample can correspond to each other well, and they are all distributed near the real concentration (diagonal line) in the entire detection concentration interval, and the correlation coefficient Rcorr=98.16 %. Using the established model to predict the release of methylene blue in the independent prediction set samples (i.e. sample detection), it is not difficult to find that the predicted concentrations of these samples still show a good correspondence, and they can also be distributed in Near the real concentration, the correlation coefficient Rpred is as high as 98.93%. This shows that the model established by the present invention has good accuracy.

In order to illustrate the accuracy of the model, the samples of high, middle and low concentration groups were designed and the recovery rate test was carried out. The results are shown in Table 1. It can be seen from Table 1 that no matter the concentration is high or low, each concentration group has a good recovery rate, the recovery rate is 103.6%-110.8%, and there is no correlation between the concentration level and randomness, which shows that the established non-destructive testing method is accurate and reliable. Of course, the lowest recovery rate is as low as 96%, and the highest is as high as 119%, which is an acceptable accuracy for a non-destructive and rapid detection method, especially for the detection of biological samples.

Table 1 Accuracy of non-invasive detection of methylene blue release in plasma by near-infrared spectroscopy

The above results show that the present invention uses the good penetrability of near-infrared light to establish a non-destructive detection model and method for the release of plasma virus inactivator methylene blue in blood collection and transfusion transfer bags by using near-infrared transmission spectroscopy. The near-infrared transmission spectrum of the sample in the plasma bag obtained by non-destructive testing outside the plasma bag was used as the identification variable, and the baseline drift calibration was performed based on the spectrum in the interval 3999.7-4265.8cm- ¹ , and 5827.9-6464.3cm ^-1 and 7467.1- The spectrum in the 9009.9cm ^-1 interval is an information-rich spectrum, the mean-centered spectrum is used as the model variable, the number of principal components is selected as 11, 11 samples are randomly selected as the prediction set, and the remaining 41 samples are used as the correction set, and the PLS Methods A multivariate analysis model for methylene blue release was established. The formula for this model is as follows:

c=k ₁ A ₁ +k ₂ A ₂ +···+k _n A _n , where c represents the concentration of methylene blue in the sample, A ₁ , A ₂ ···A _n are the detected The wavelengths corresponding to the selected information-rich spectral intervals in the spectrum are the absorbance at λ ₁ , λ ₂ ···λ _n , and k ₁ , k ₂ ···k _n are the corresponding PLS regression coefficients.

The correlation coefficients of the model-calculated concentration to each sample concentration were 98.16% (calibration set) and 98.93% (prediction set), and the average recoveries of samples in the high, middle and low concentration groups were 103.6%-110.8%. This shows that the near-infrared transmission spectroscopy method established by the present invention can realize the non-destructive, rapid and accurate analysis of the release amount of methylene blue in the collection and transportation transfer bag outside the plasma bag. It indicates that under the condition of no secondary pollution, the monitoring of the quality of plasma virus inactivation in the collection, transportation and transfer bag can be realized.

Example 3

The present embodiment provides a method for detecting the release of methylene blue in plasma using the multivariate analysis model of the release of methylene blue established above, specifically as follows:

Wipe the outer surface of the plasma inactivation bag to be tested and the quartz plate on the transmission platform with alcohol, adjust the distance between the first vertical plate 6 and the second vertical plate 1 according to the method in Example 1, so that the distance between the two optical windows The optical distance between them is fixed at 3.5mm. Then put the plasma inactivation bag with unknown methylene blue content (that is, the plasma inactivation bag that released methylene blue) on the stage, and align the left and right sides of the plasma inactivation bag each time as much as possible, and measure the sample spectrum. The instrument parameters are: wavenumber 4000-10000cm ^-1 , optical path 3.5mm, resolution 8cm ^-1 , scanning times 32, scanning speed 0.3165, aperture 22, gain 1. Air-filled, unused plasma inactivated bags were used as spectral background before testing samples on the day. Then the measured near-infrared transmission spectrum is calibrated with the spectrum in the range of 3999.7-4265.8 cm ^-1 as the standard for baseline drift. Select 5827.9-6464.3cm ^-1 and 7467.1-9009.9cm ^-1 as the rich information interval, and substitute the absorbance at each wavelength in this interval into the formula of the model established in Example 2 to calculate the methylene blue in the plasma inactivation bag content.

The above is only a preferred embodiment of the present invention, and is not intended to limit the present invention. It should be pointed out that for those of ordinary skill in the art, some improvements can be made without departing from the technical principle of the present invention. and modifications, these improvements and modifications should also be considered as the protection scope of the present invention.

Claims (10)

1. a kind of near-infrared spectrometer, including light source, monochromator, detector and computer treatmenting information system, its feature It is：The infrared spectrometer further includes a transmission platform, and the transmission platform includes a substrate, is equipped with vertically on the substrate Corresponding transmission is respectively offered on two pieces of starting staves being parallel to each other and the second riser, the starting stave and the second riser Window, the transmissive window is covered with transparent optics window, and the optics window is located on the opposite side of two pieces of risers, described An objective table is equipped between starting stave and the second riser, the soft bag of determinand is filled for carrying, the objective table is located at light The lower section of window lower edge is learned, for the soft bag for filling determinand between two optics windows, at least one riser can be along institute The surface for stating substrate moves horizontally, and the soft bag for filling determinand is clamped between two optics windows and two optical windows Light path between piece can adjust.

2. near-infrared spectrometer according to claim 1, it is characterised in that：The starting stave is consolidated with the substrate Fixed connection, the objective table are fixedly connected with the starting stave, and objective table described in face is offered on second riser Bar hole, the bar hole are adapted with the shape of the objective table, and the objective table can be slidably inserted into the bar hole.

3. near-infrared spectrometer according to claim 1, it is characterised in that：It is fixedly connected with and leads on the substrate Rail, second riser can slide along guide rail level and can be locked in any position of guide rail.

4. near-infrared spectrometer according to claim 1, it is characterised in that：The material of the optics window is quartz Or optical glass.

A kind of 5. method that methylenum careuleum burst size in Plasma Inactivation bag is detected using near infrared spectrum, it is characterised in that using power Profit requires the near-infrared spectrometer any one of 1-4, comprises the following steps：

(1) the distance between described starting stave and the second riser are adjusted, is fixed as the light path between two optics windows 2.0-5.0mm；

(2) by Plasma Inactivation bag be placed on the objective table of the near-infrared spectrometer and be located at two optics windows it Between, Plasma Inactivation bag is closely contacted with two optics windows, contain wherein filling known methylenum careuleum in the Plasma Inactivation bag The plasma standard solution of amount；

(3) in wave number 4000-10000cm^-1Under conditions of near infrared spectrum detection is carried out to plasma standard solution, retest is more A sample, according to the near infrared spectrum testing result of multiple samples, the multi-variables analysis of methylenum careuleum burst size is established using PLS methods Model；

(4) the Plasma Inactivation bag to be measured containing methylenum careuleum is placed on the objective table, make Plasma Inactivation bag closely with two Optics window contacts；

(5) in wave number 4000-10000cm^-1Under conditions of carry out near infrared spectrum detection, utilize the more of the methylenum careuleum burst size Variable analysis model determines the burst size of methylenum careuleum in Plasma Inactivation bag.

6. according to claim 5 using the method for methylenum careuleum burst size near infrared spectrum detection Plasma Inactivation bag, it is special Sign is：In step (2), the concentration of methylenum careuleum is 0.312 μm of ol/L-2.959 μm of ol/L in the plasma standard solution.

7. according to claim 5 using the method for methylenum careuleum burst size near infrared spectrum detection Plasma Inactivation bag, it is special Sign is：In step (3) and step (5), the computerized near infrared scan resolution ratio of the near infrared spectrum detection is 8cm^-1, scanning Number is 32 times, sweep speed 0.3165.

8. according to claim 5 using the method for methylenum careuleum burst size near infrared spectrum detection Plasma Inactivation bag, it is special Sign is：In step (3), after near infrared spectrum detection, 3999.7-4265.8cm is selected^-1The absorbance average conduct in section Drift value, drift elimination is carried out to infrared spectrum testing result.

9. according to claim 5 using the method for methylenum careuleum burst size near infrared spectrum detection Plasma Inactivation bag, it is special Sign is：In step (3) and step (5), after near infrared spectrum detection, 5350-6600cm is selected^-1And 7200-10001cm^-1 The absorbance in two sections is as rich informative spectral.

10. according to claim 5 using the method for methylenum careuleum burst size near infrared spectrum detection Plasma Inactivation bag, it is special Sign is：Before step (2), the near infrared light will be placed on filled with air and original Plasma Inactivation bag by further including The step of spectral background being used as on the objective table of spectrometer after progress near infrared spectrum detection.