A method for detecting adsorption performance of an adsorbent

Abstract

The application provides a method for detecting adsorption performance of an adsorbent, comprising: constructing a first adsorption device, wherein the first adsorption device comprises a cation resin adsorber and a neutral macroporous resin adsorber connected in series; performing equal-ratio reduction processing on the first adsorption device according to a preset ratio value to obtain a second adsorption device; obtaining simulated plasma, wherein the simulated plasma contains a preset concentration of a to-be-detected product, and the to-be-detected product comprises bilirubin and / or bile acid; injecting the simulated plasma into the second adsorption device for adsorption processing according to preset circulation parameters to obtain a first concentration of the to-be-detected product in the simulated plasma; determining a first simulated adsorption rate of the to-be-detected product according to the preset concentration of the to-be-detected product and the first concentration of the to-be-detected product; obtaining a calibration factor, and calibrating the first simulated adsorption rate of the to-be-detected product according to the calibration factor to determine an adsorption rate of the to-be-detected product. The application can pre-evaluate the effect of the adsorbent in the DPMAS on removing bilirubin and bile acid.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of detection, in particular to a method for detecting adsorption performance of an adsorbent. BACKGROUND

[0002] Bilirubin is a product of heme metabolism degradation, and is an endogenous toxin. Under normal physiological conditions, serum proteins bind with bilirubin, helping bilirubin to be transferred to the liver and then excreted. When liver function is abnormal, bilirubin metabolism is blocked, causing bilirubin to accumulate in the blood. High concentration of bilirubin in the blood can cause obstructive jaundice, and even acute renal failure. The level of bilirubin in the body is generally positively correlated with the degree of liver cell lesion. Similarly, bile acid is also positively correlated with the degree of liver cell lesion.

[0003] For patients with severe hepatitis or liver failure, they have basically lost the ability to metabolize bilirubin and bile acid. Generally, blood purification is used to remove pathogenic substances. Blood perfusion and plasma adsorption can effectively remove high concentrations of bilirubin and bile acid in the body of patients. Preparing an adsorbent with good blood compatibility, strong adsorption performance and high specificity is the key to the development of blood perfusion and plasma adsorption technology. At present, the double plasma molecular adsorption system (DPMAS) is a broad-spectrum resin adsorbent that can adsorb middle and large molecular toxins based on plasma bilirubin adsorption therapy. After blood is drawn from the patient's body, it is separated into plasma and blood cells by a plasma separator. The blood cells are returned to the body, and the plasma is sequentially returned to the body after flowing through an anion exchange resin and a neutral macroporous adsorbent column. DPMAS combines an anion resin adsorber and a neutral macroporous adsorbent adsorber. The two adsorbers are used in combination to achieve synergistic effect. Not only can they remove inflammatory mediators, but also can remove bilirubin and bile acid in plasma, improve the success rate of treatment, and improve the prognosis of patients.

[0004] Pre-evaluation of the effect of the adsorbent in DPMAS on the removal of bilirubin and bile acid can provide reference value for the later application of DPMAS. However, at present, there is no method in the prior art for pre-evaluating the effect of the adsorbent in DPMAS on the removal of bilirubin and bile acid. SUMMARY

[0005] The present application aims to solve the problem that there is no method in the prior art for pre-evaluating the effect of the adsorbent in DPMAS on the removal of bilirubin and bile acid.

[0006] To solve the above problems, the present application provides a method for detecting adsorption performance of an adsorbent, comprising the following steps:

[0007] The first adsorption device is constructed, and the first adsorption device comprises an anion resin adsorber and a neutral macroporous resin adsorber connected in series.

[0008] According to the preset proportion value, the first adsorption device is processed by equal proportion reduction to obtain a second adsorption device.

[0009] Analog plasma is obtained, and the analog plasma contains a preset concentration of a to-be-tested product, and the to-be-tested product comprises bilirubin and / or bile acid.

[0010] According to the preset cycle parameter, the analog plasma is injected into the second adsorption device for adsorption treatment to obtain a first concentration of the to-be-tested product in the analog plasma.

[0011] According to the preset concentration of the to-be-tested product and the first concentration of the to-be-tested product, a first simulated adsorption rate of the to-be-tested product is determined.

[0012] A calibration factor is obtained, and the first simulated adsorption rate of the to-be-tested product is calibrated according to the calibration factor to determine the adsorption rate of the to-be-tested product.

[0013] The method for detecting the adsorption performance of the adsorbent provided by the application constructs two sets of adsorption devices, the first adsorption device can be a DPMAS equal proportion device, and the anion resin adsorber and the neutral macroporous resin adsorber in the second adsorption device are obtained by equal proportion reduction of the anion resin adsorber and the neutral macroporous resin adsorber in the first adsorption device, respectively, wherein the second adsorption device can measure the adsorption rate of the adsorbent for adsorbing the to-be-tested product (bilirubin and / or bile acid) under simulated conditions, and the calibration factor can be obtained through the first adsorption device, the second adsorption device and different plasma, the relationship between the first simulated adsorption rate and the adsorption rate of the to-be-tested product is established according to the calibration factor, and the adsorption rate of the to-be-tested product can be determined according to the first simulated adsorption rate and the calibration factor, so that the adsorption rate of the to-be-tested product can be determined more conveniently and accurately, and the adsorption rate of the adsorbent for adsorbing bilirubin and / or bile acid measured finally can reflect the adsorption rate under real conditions, so that the effect of the adsorbent in the DPMAS for removing bilirubin and / or bile acid can be evaluated more truly and accurately; in addition, when the concentration of the to-be-tested product in the plasma is within the preset concentration range in the subsequent detection, the calibration factor obtained is applicable, and it is not necessary to determine the calibration factor again, so that the detection method is simpler and faster, the consumption of plasma is reduced, and the use of device materials is reduced by using the second adsorption device reduced by equal proportion for adsorption treatment, which is beneficial to saving the detection cost. BRIEF DESCRIPTION OF DRAWINGS

[0014] Figure 1 It is a structural schematic diagram of the first adsorption device provided in the embodiments of the application.

[0015] Figure 2 This is a schematic flowchart of the method for detecting the adsorption performance of an adsorbent provided in an embodiment of the present invention. Detailed Implementation

[0016] This invention provides a method for detecting the adsorption performance of an adsorbent, which can detect the adsorption rate of bilirubin and / or bile acids in DPMAS, thereby enabling a pre-evaluation of the effect of the adsorbent in DPMAS in removing bilirubin and bile acids, and providing reference value for the later application of DPMAS.

[0017] To make the above-mentioned objects, features and advantages of the present invention more apparent and understandable, specific embodiments of the present invention will be described in detail below with reference to the accompanying drawings.

[0018] It should be noted that, unless otherwise specified, the embodiments and features described in the present invention can be combined with each other.

[0019] Furthermore, the terms "comprising," "including," "containing," and "having" are non-restrictive and can refer to the addition of other steps and components that do not affect the results. Unless otherwise specified, all materials, equipment, and reagents are commercially available.

[0020] Furthermore, although the present invention describes each step in the preparation process in the form of steps S100, S200, S300, etc., this description is only for ease of understanding. The form of steps S100, S200, S300, etc. does not indicate a limitation on the order of the steps.

[0021] Figure 1 This is a simplified structural diagram of the first adsorption device in this embodiment. (Combined with...) Figure 1 As shown, the first adsorption device includes a blood pool 1, a blood input pipeline 2, a blood pump 3, an anion exchange resin adsorber 4, a connecting pipe 5, a neutral macroporous resin adsorber 6, and a blood return pipeline 7. One end of the blood input pipeline 2 is connected to the output end of the blood pool 1, and the other end is connected to the input end of the anion exchange resin adsorber 4. The blood pump 3 is mounted on the first blood input pipeline 2 and is located between the blood pool 1 and the anion exchange resin adsorber 4. The anion exchange resin adsorber 4 and the neutral macroporous resin adsorber 6 are connected in series. Furthermore, the connecting pipe 5 connects to the anion exchange resin adsorber 4 and the neutral macroporous resin adsorber 6. One end of the blood return pipeline 7 is connected to the output end of the neutral macroporous resin adsorber 6, and the other end of the blood return pipeline 7 is connected to the input end of the blood pool 1. Thus, the blood in the blood pool 1 passes through the anion exchange resin adsorber 4 and the neutral macroporous resin adsorber 6 in sequence. The bilirubin and bile acids in the blood are absorbed by the adsorbents in the two adsorbers, thereby achieving the purpose of purifying the blood. Then, the purified blood is returned to the blood pool 1 to form a circulation loop.

[0022] The adsorbent in the anion resin adsorber 4 is an anion exchange resin, which contains functional groups (for example, quaternary amine groups) with positive charges, while the bilirubin and bile acid molecules have carboxyl anions with negative charges, and the two are combined through ionic bonds, so as to achieve the purpose of removing bilirubin and bile acid. The anion resin adsorber 4 can adopt a bilirubin adsorber. Specifically, the anion resin adsorber 4 can adopt a BS bilirubin adsorber produced by Jianfan Biotechnology Group Co., Ltd., such as a BS330 plasma bilirubin adsorber. Such adsorbers have BS ion exchange resin, a specific adsorbent for bilirubin, inside, which specifically adsorbs bilirubin, bile acid and other toxins by electrostatic force and lipophilic binding.

[0023] The adsorbent in the neutral macroporous resin adsorber 6 is a neutral macroporous adsorption resin, which is polymerized by styrene and divinylbenzene, and mainly removes bilirubin, bile acid and the like through three-dimensional molecular sieves and intermolecular van der Waals forces. The neutral macroporous resin adsorber 6 can adopt a hemoperfusion device. Specifically, the neutral macroporous resin adsorber 6 can adopt a HA hemoperfusion device produced by Jianfan Biotechnology Group Co., Ltd., such as a HA330-II disposable hemoperfusion device. Such hemoperfusion devices have HA neutral macroporous resin, a broad-spectrum adsorbent, inside, which has a macroporous structure and a large surface area, and adsorbs middle-molecular-weight toxins by van der Waals forces and skeleton molecular sieves.

[0024] It should be noted that the first adsorption device in the embodiment is a simplified device of DPMAS, but its working principle is the same as that of DPMAS. The structure diagram and working principle of DPMAS are prior art, and will not be described here.

[0025] In combination with Figure 2 As shown in the figure, the embodiment of the present application provides a method for detecting the adsorption performance of an adsorbent, which comprises the following steps:

[0026] Step S100, a first adsorption device is constructed, the first adsorption device comprising an anion resin adsorber and a neutral macroporous resin adsorber connected in series, and the first adsorption device is processed by equal ratio reduction according to a preset ratio value to obtain a second adsorption device.

[0027] The types of the adsorbents in the anion resin adsorber and the neutral macroporous resin adsorber in the first adsorption device are the same as those in the anion resin adsorber and the neutral macroporous resin adsorber in the second adsorption device, but the anion resin adsorber and the neutral macroporous resin adsorber in the second adsorption device are obtained by reducing the anion resin adsorber and the neutral macroporous resin adsorber in the first adsorption device by n times in proportion, and correspondingly, the amounts of the adsorbents in the anion resin adsorber and the neutral macroporous resin adsorber in the second adsorption device are also reduced by n times in proportion. Specifically, the 3D printing technology can be used to reduce the components in the first adsorption device by n times in proportion to obtain the second adsorption device.

[0028] In the embodiment, the preset ratio value is n, and 15≤n≤25, so that the detection accuracy can be improved while reducing the amount of blood.

[0029] In the embodiment, two sets of adsorption devices are constructed. The first adsorption device can be a proportional device of DPMAS, and the plasma separator can be arranged in the first adsorption device according to the DPMAS to obtain a true and accurate adsorption rate. The second adsorption device is a proportional reduction device of the first adsorption device. The second adsorption device can measure the adsorption rate of the adsorbent adsorbing bilirubin and / or bile acid under simulated conditions, and the first adsorption device can be used to calibrate the adsorption rate under the simulated conditions, so that the finally measured adsorption rate of the adsorbent adsorbing bilirubin and / or bile acid can more reflect the adsorption rate under the real conditions, thereby the effect of the adsorbent in the DPMAS on removing bilirubin and / or bile acid can be more truly and accurately evaluated. Moreover, the second adsorption device with proportional reduction can also reduce the amount of plasma, which is beneficial to saving the detection cost.

[0030] In step S200, the simulated plasma containing a preset concentration of the to-be-measured product is obtained. The to-be-measured product includes bilirubin and / or bile acid. According to preset circulation parameters, the simulated plasma is injected into the second adsorption device for adsorption treatment, and the first concentration of the to-be-measured product in the simulated plasma is obtained.

[0031] In this embodiment, simulated plasma can be prepared by means of the following method: Prepare a sodium carbonate solution with a concentration of 0.01 mol / L to 0.25 mol / L, and let the sodium carbonate solution stand at room temperature; prepare a bilirubin solution and a bile acid solution, wherein the concentration of the bilirubin solution is 100 μmol / L to 1000 μmol / L, and the concentration of the bile acid solution is 100 μmol / L to 1000 μmol / L, and let the bilirubin solution and bile acid solution stand at room temperature; take plasma from a healthy human body, add the bilirubin solution and bile acid solution to the plasma, and mix thoroughly by shaking to obtain simulated plasma. That is, the preset concentration of bilirubin in the simulated plasma is 100 μmol / L to 1000 μmol / L, and the preset concentration of bile acid is 100 μmol / L to 1000 μmol / L.

[0032] When preparing bilirubin solutions, dimethyl sulfoxide (DMSO) and sodium carbonate solution need to be added; similarly, DMSO needs to be added when preparing bile acid solutions to increase their solubility. Simulated plasma should be prepared just before use to avoid denaturation or contamination from storage over time, which could affect the accuracy of test results. After preparation, both bilirubin and simulated plasma should be wrapped in aluminum foil and stored in a cool, dark place to prevent structural instability and deformation of bilirubin, which could also affect the accuracy of test results.

[0033] In this embodiment, the preset circulation parameters include a first preset circulation parameter and a second preset circulation parameter. Both the first and second preset circulation parameters include plasma flow rate, plasma volume, and circulation time. The plasma flow rate and plasma volume in the first preset circulation parameter are n times the plasma flow rate and plasma volume in the second preset circulation parameter. The circulation time in the first preset circulation parameter is the same as the circulation time in the second preset circulation parameter. The first preset circulation parameter can be set according to the circulation parameters of DPMAS, and the first preset circulation parameter can be consistent with the circulation parameters of DPMAS. The second preset circulation parameter can be determined according to the first preset circulation parameter and the preset ratio value n.

[0034] In some optional embodiments, the plasma flow rate v1 in the first preset circulation parameters is 20ml / L to 50ml / L, the plasma volume V1 is 3600ml to 5400ml, and the circulation time t1 is 120min to 180min. Then, the plasma flow rate v2 = v1 / n, the plasma volume V2 = V1 / n, and the circulation time t2 = t1 in the second preset circulation parameters.

[0035] In this embodiment, the simulated adsorption rate of the sample to be tested in the first adsorption device is determined according to the first preset circulation parameter, the simulated adsorption rate of the sample to be tested in the second adsorption device is determined according to the second preset circulation parameter, and the preset circulation parameters suitable for the volumes of the two devices are set according to the volumes of the two adsorption devices, so that the effect of the adsorbent in the DPMAS on the removal of bilirubin and bile acids can be accurately evaluated.

[0036] It should be noted that, before the plasma (including simulated plasma, ex vivo plasma and plasma) is injected into the first adsorption device and the second adsorption device in this embodiment, a preflush (such as sodium chloride injection) is used to preflush the first adsorption device and the second adsorption device. After the preflush is completed, the preflush in the first adsorption device and the second adsorption device is emptied, wherein the volume of the preflush in the second adsorption device is 1 / n of the volume of the preflush in the first adsorption device, the preflush time of the second adsorption device is the same as the preflush time of the first adsorption device, and the flow rate of the preflush in the second adsorption device is 1 / n of the flow rate of the preflush in the first adsorption device. Thus, the accuracy of the detection result can be improved.

[0037] After the simulated plasma is prepared and the preset circulation parameters are set, the simulated plasma is injected into the second adsorption device according to the second preset circulation parameter, and the simulated plasma sequentially passes through the anion resin adsorber and the neutral macroporous resin adsorber. The bilirubin and bile acids in the simulated plasma are absorbed by the adsorbents in the two adsorbers, and the first concentration of the sample to be tested in the simulated plasma after the adsorption treatment by the second adsorption device is detected.

[0038] In step S300, the first simulated adsorption rate of the sample to be tested is determined according to the preset concentration of the sample to be tested and the first concentration of the sample to be tested.

[0039] After the simulated plasma is treated by the second adsorption device, the first concentration of the sample to be tested is obtained, and the first simulated adsorption rate of the sample to be tested can be calculated according to the following formula:

[0040] The first simulated adsorption rate η1 =

(C1-C2) / C1

[0041] Wherein, C1 is the preset concentration of the sample to be tested, that is, the concentration of bilirubin and / or bile acids in the simulated plasma before adsorption;

[0042] C2 is the first concentration of the sample to be tested, that is, the concentration of bilirubin and / or bile acids in the simulated plasma after the adsorption treatment by the second adsorption device.

[0043] In step S400, a calibration factor is obtained, the first simulated adsorption rate of the sample to be tested is calibrated according to the calibration factor, and the adsorption rate of the sample to be tested is determined.

[0044] Since the first simulated adsorption rate is obtained by using the simulated blood plasma and using the second adsorption device that is proportionally reduced, the first simulated adsorption rate cannot truly and accurately reflect the adsorption rate of the adsorbent in the DPMAS to adsorb bilirubin and bile acids, and therefore it is necessary to calibrate the first simulated adsorption rate of the test sample according to the calibration factor to obtain the adsorption rate of the test sample, so that the finally obtained adsorption rate of the test sample can more reflect the adsorption rate under the real condition, thereby the effect of the adsorbent in the DPMAS to remove bilirubin and / or bile acids can be more truly and accurately evaluated.

[0045] The calibration factor includes a first calibration factor F1, a second calibration factor F2 and a third calibration factor F3, wherein the first calibration factor F1 represents the matrix effect of using the simulated blood plasma to represent the liver disease blood plasma, the second calibration factor F2 represents the kinetic effect of using the proportionally reduced second adsorption device to represent the actual perfusion, and the third calibration factor F3 represents the influence of individual differences and toxin production and redistribution and the like.

[0046] Under the condition that the concentration of the test sample is the same and the experimental conditions are the same, the first calibration factor F1 is determined by the dynamic cycle adsorption difference of the first adsorption device in different matrices. Different matrices refer to injecting simulated blood plasma and ex vivo blood plasma into the first adsorption device for adsorption treatment, respectively. The ex vivo blood plasma represents the blood plasma taken from the body of a patient with liver disease to the outside of the body, and the ex vivo blood plasma contains the test sample. In order to ensure the accuracy of the first calibration factor F1, the preset concentration of the test sample in the simulated blood plasma is the same as the concentration of the test sample in the ex vivo blood plasma, that is, the concentration of bilirubin and / or bile acids in the simulated blood plasma can be prepared according to the concentration of bilirubin and / or bile acids in the ex vivo blood plasma.

[0047] Specifically, determining the first calibration factor includes:

[0048] According to the first preset cycle parameter, the simulated blood plasma is injected into the first adsorption device for adsorption treatment to obtain a second concentration of the test sample in the simulated blood plasma, and according to the preset concentration of the test sample and the second concentration of the test sample, a second simulated adsorption rate η2 of the test sample is determined.

[0049] Before the adsorption treatment, a third concentration of the test sample in the ex vivo blood plasma is obtained, the ex vivo blood plasma is injected into the first adsorption device for adsorption treatment according to the first preset cycle parameter, a fourth concentration of the test sample in the ex vivo blood plasma is obtained, and according to the third concentration of the test sample and the fourth concentration of the test sample, a third simulated adsorption rate η3 of the test sample is determined.

[0050] The first calibration factor F1 is calculated according to the following formula: the first calibration factor F1 = η2 / η3.

[0051] In the embodiment, when the first calibration factor F1 is determined, the second simulation adsorption rate η2 of the analyte and the third simulation adsorption rate η3 of the analyte are determined, and the experimental conditions are the same except that the matrix injected into the first adsorption device is different, and the preset concentration of the analyte in the simulation plasma and the third concentration of the analyte in the ex vivo plasma are the same.

[0052] According to the simulation plasma, the first adsorption device and the second adsorption device, the second calibration factor F2 is determined, and the second calibration factor F2 is determined by the difference in dynamic cycle adsorption of the first adsorption device and the second adsorption device under the condition that the concentration of the analyte in the simulation plasma is the same and the experimental conditions are the same.

[0053] Specifically, determining the second calibration factor includes:

[0054] According to the second preset cycle parameter, the simulation plasma is injected into the second adsorption device for adsorption treatment to obtain the fifth concentration of the analyte in the simulation plasma, and the fourth simulation adsorption rate η4 of the analyte is determined according to the preset concentration of the analyte and the fifth concentration of the analyte.

[0055] According to the first preset cycle parameter, the simulation plasma is injected into the first adsorption device for adsorption treatment to obtain the sixth concentration of the analyte in the simulation plasma, and the fifth simulation adsorption rate η5 of the analyte is determined according to the preset concentration of the analyte and the sixth concentration of the analyte.

[0056] The second calibration factor F2 is calculated according to the following formula: the second calibration factor F2 = η4 / η5.

[0057] In the embodiment, when the second calibration factor F2 is determined, the fourth simulation adsorption rate η4 of the analyte and the fifth simulation adsorption rate η5 of the analyte are determined, and the experimental conditions are the same except that the first adsorption device and the second adsorption device are different, and the first preset cycle parameter and the second preset cycle parameter are different.

[0058] In the case that the concentration of the analyte is the same and the experimental conditions are the same, the third calibration factor F3 is determined by the difference in dynamic cycle adsorption of the first adsorption system under simulation conditions and real conditions. The simulation conditions refer to that the first adsorption device is injected with simulation plasma for adsorption treatment under laboratory conditions, and the real conditions refer to that the first adsorption device is used to adsorb the plasma of a patient with liver disease in actual application. The plasma of the patient with liver disease (referred to as plasma) contains the analyte, and in order to ensure the accuracy of the third calibration factor F3, the preset concentration of the analyte in the simulation plasma and the concentration of the analyte in the plasma are the same, that is, the concentration of bilirubin and / or bile acid in the simulation plasma can be prepared according to the concentration of bilirubin and / or bile acid in the plasma.

[0059] Specifically, determining the third calibration factor includes:

[0060] According to the first preset cycle parameter, the simulated plasma is injected into the first adsorption device for adsorption treatment to obtain a seventh concentration of the to-be-tested product in the simulated plasma, and a sixth simulated adsorption rate η6 of the to-be-tested product is determined according to the preset concentration of the to-be-tested product in the simulated plasma and the seventh concentration of the to-be-tested product.

[0061] Before the adsorption treatment, an eighth concentration of the to-be-tested product in the plasma is obtained, the plasma is injected into the first adsorption device for adsorption treatment according to the first preset cycle parameter to obtain a ninth concentration of the to-be-tested product in the plasma, and a seventh simulated adsorption rate η7 of the to-be-tested product is determined according to the eighth concentration of the to-be-tested product and the ninth concentration of the to-be-tested product.

[0062] The third calibration factor F3 is calculated according to the following formula: the third calibration factor F3 = η6 / η7.

[0063] In the embodiment, when determining the third calibration factor F3, the sixth simulated adsorption rate η6 of the to-be-tested product and the seventh simulated adsorption rate η7 of the to-be-tested product are determined, and the experimental conditions are the same except that the use scenarios of the first adsorption device are different, and the preset concentration of the to-be-tested product in the simulated plasma and the eighth concentration of the to-be-tested product in the plasma are the same.

[0064] It should be noted that when determining the third calibration factor F3, the plasma of the liver disease patient is subjected to adsorption treatment by the first adsorption device in actual application, so as to avoid that the adsorption performance of the adsorbent in the DPMAS under the actual use state cannot be truly and accurately reflected by injecting the simulated plasma into the first adsorption device for adsorption treatment under laboratory conditions. Therefore, the plasma of the liver disease patient is subjected to adsorption treatment by the first adsorption device in actual application, so as to calibrate the adsorption rate under simulated conditions, thereby facilitating the true and accurate evaluation of the effect of the adsorbent in the DPMAS on removing bilirubin and / or bile acid.

[0065] In the embodiment, the preset concentration of the to-be-tested product in the simulated plasma can be adjusted according to the concentration of the to-be-tested product in the ex vivo plasma or the concentration of the to-be-tested product in the plasma, for example, the simulated plasma can be diluted to keep the preset concentration of the to-be-tested product in the simulated plasma consistent with the concentration of the to-be-tested product in the ex vivo plasma, and keep the preset concentration of the to-be-tested product in the simulated plasma consistent with the concentration of the to-be-tested product in the plasma.

[0066] After the first calibration factor F1, the second calibration factor F2 and the third calibration factor F3 are determined, a calibration factor F is calculated according to the following formula: the calibration factor F = F1 × F2 × F3.

[0067] The adsorption rate of the to-be-tested product is calculated according to the following formula:

[0068] The adsorption rate of the to-be-tested product = the first simulated adsorption rate η1 / calibration factor F.

[0069] The method for detecting the adsorption performance of the adsorbent provided in the embodiment is applicable to the calibration factor F obtained when the concentration of the to-be-detected product in the plasma (including the simulated plasma, the ex vivo plasma, and the blood plasma) is within the preset concentration range, and the calibration factor F obtained can be directly used without the need for re-determination, thereby reducing the consumption of the plasma. After the concentration of the to-be-detected product (bilirubin and / or bile acid) in the blood of a patient with liver disease is determined, the simulated plasma containing the to-be-detected product at the same concentration is prepared, and the simulated plasma is injected into the second adsorption device for adsorption treatment, so as to obtain the concentration of the to-be-detected product in the simulated plasma after the adsorption treatment. According to the concentration of the to-be-detected product in the simulated plasma before the adsorption treatment and the concentration of the to-be-detected product in the simulated plasma after the adsorption treatment, the first simulated adsorption rate of the to-be-detected product can be determined. According to the first simulated adsorption rate and the calibration factor, the adsorption rate of the to-be-detected product can be determined, and the relationship between the first simulated adsorption rate and the adsorption rate of the to-be-detected product is established. The adsorption rate of the to-be-detected product can be more conveniently and accurately determined, so as to predict the effect of the adsorbent in the DPMAS on the removal of bilirubin and bile acid, thereby providing a reference for the later application of the DPMAS. In addition, the method for detecting the adsorption performance of the adsorbent provided in the embodiment adopts the second adsorption device with a proportional reduction for adsorption treatment, which can not only reduce the consumption of the plasma, but also reduce the use of device consumables, thereby being beneficial to saving the detection cost.

[0070] In order to further illustrate the present application, the present application will be further illustrated in combination with specific examples. The experimental methods used in the examples in the present application are all conventional methods unless otherwise specified. The materials and reagents used in the examples in the present application are all commercially available unless otherwise specified.

[0071] Example 1

[0072] The embodiment provides a method for detecting the adsorption performance of an adsorbent, comprising the following steps:

[0073] (1) Constructing an adsorption device: a first adsorption device with the same proportion as the DPMAS is arranged, the anion resin adsorber and the neutral macroporous resin adsorber in the first adsorption device are the same as those in the DPMAS, the anion resin adsorber and the neutral macroporous resin adsorber in the first adsorption device are connected in series, the anion resin adsorber and the neutral macroporous resin adsorber in the first adsorption device are both reduced by 20 times in proportion by using the 3D printing technology, two micro adsorbers are obtained, adsorbents are respectively filled in the two micro adsorbers, the types of the adsorbents are the same as those in the anion resin adsorber and the neutral macroporous resin adsorber, and the amounts of the adsorbents in the two micro adsorbers are respectively 1 / 20 of the amounts of the adsorbents in the anion resin adsorber and the neutral macroporous resin adsorber, and the two micro adsorbers are connected in series by using a disposable infusion tube, so as to obtain a second adsorption device.

[0074] (2) Preparation of simulated plasma:

[0075] Preparation of sodium carbonate solution: 0.53 g of anhydrous sodium carbonate was accurately weighed into a 100 mL volumetric flask, dissolved with water for injection and diluted to the mark, shaken uniformly, and a 0.05 mol / L sodium carbonate solution was prepared, which was placed at room temperature and ready for use;

[0076] Preparation of bilirubin (M = 584.7) solution: Under light-proof conditions, about 120 mg of bilirubin standard was accurately weighed, 6 mL of DMSO (dimethyl sulfoxide) was added, shaken to mix, then 12 mL of 0.05 mol / L sodium carbonate aqueous solution was added, shaken to dissolve completely, and placed at room temperature, wrapped with tin foil to avoid light and ready for use;

[0077] Preparation of bile acid (M = 408.6) solution: About 60 mg of cholic acid standard was accurately weighed, 5 mL of DMSO was added, shaken to dissolve completely, and placed at room temperature and ready for use;

[0078] Preparation of simulated plasma solution: 540 mL of healthy human plasma was taken, and the above prepared bilirubin solution and cholic acid solution were added, wrapped with tin foil to avoid light, and shaken at 80 rpm / min in a constant temperature shaker for 20 min, mixed uniformly, and a simulated plasma with a total bilirubin concentration of about 350 μmol / L and a total bile acid concentration of 250 μmol / L was obtained, which was prepared and used immediately.

[0079] (3) Set the preset circulation parameters:

[0080] Set the circulation process parameters, start the laboratory circulation experiment, and the circulation process parameters are as follows:

[0081] The first preset circulation parameters are: ① plasma flow rate v: 30 mL / min; ② plasma volume V: 3600 mL; ③ circulation time t: 120 min;

[0082] The second preset circulation parameters are: ① plasma flow rate: 1.5 mL / min; ② plasma volume: 180 mL; ③ circulation time: 120 min.

[0083] (4) Determine the first simulated adsorption rate: according to the first preset circulation parameters, the simulated plasma was injected into the second adsorption device for adsorption treatment, the concentrations of bilirubin and bile acid in the simulated plasma after adsorption treatment were obtained, and the first simulated adsorption rate of bilirubin and the first simulated adsorption rate of bile acid were determined according to the concentrations of bilirubin and bile acid in the simulated plasma before adsorption treatment and the concentrations of bilirubin and bile acid in the simulated plasma after adsorption treatment;

[0084] (5) determining the adsorption rate of bilirubin and bile acid: determining the calibration factor, calibrating the first simulated adsorption rate of bilirubin according to the calibration factor to obtain the adsorption rate of bilirubin, and calibrating the first simulated adsorption rate of bile acid according to the calibration factor to obtain the adsorption rate of bile acid.

[0085] Example 2

[0086] The present embodiment provides a method for determining a first calibration factor, comprising the following steps:

[0087] (1) determining the first calibration factor of bilirubin: taking the ex vivo plasma with total bilirubin concentrations of 350 μmol / L, 200 μmol / L and 400 μmol / L, respectively, according to the first preset cycle parameters in Example 1, the three kinds of ex vivo plasma with different bilirubin concentrations are injected into the first adsorption device for adsorption treatment, and according to the concentration of bilirubin in the ex vivo plasma before adsorption treatment and the concentration of bilirubin in the ex vivo plasma after adsorption treatment, the third simulated adsorption rate η3 of bilirubin is determined.

[0088] The simulated plasma prepared in Example 1 is respectively prepared into simulated plasma with total bilirubin concentrations of 350 μmol / L, 200 μmol / L and 400 μmol / L, respectively, according to the first preset cycle parameters in Example 1, the three kinds of simulated plasma with different bilirubin concentrations are injected into the first adsorption device for adsorption treatment, and according to the concentration of bilirubin in the simulated plasma before adsorption treatment and the concentration of bilirubin in the simulated plasma after adsorption treatment, the second simulated adsorption rate η2 of bilirubin is determined, and the first calibration factor F1 is calculated according to the following formula: F1 = η2 / η3.

[0089] Repeat the experiment several times to obtain the final F1, and the adsorption rate results of the adsorbent and the first calibration factor F1 results under different concentrations of bilirubin are shown in Table 1.

[0090] Table 1: First calibration factor results of bilirubin

[0091]

[0092]

[0093] Finally, the first calibration factor F1 of bilirubin is obtained as F1 = (1.3 + 1.26 + 1.28) / 3 = 1.28.

[0094] (2) The first calibration factor of bile acid is determined by the same method as determining the first calibration factor of bilirubin in step (1), and the final F1 is obtained by repeating the experiment several times, and the adsorption rate results of the adsorbent and the first calibration factor F1 results under different concentrations of bile acid are shown in Table 2.

[0095] Table 2 First calibration factor results table of bile acids

[0096]

[0097] The first calibration factor F1 of bile acids is finally obtained as F1 = (1.02 + 1.03 + 1.03) / 3 = 1.03.

[0098] Example 3

[0099] The present example provides a method for determining a second calibration factor, comprising the following steps:

[0100] (1) Determining the second calibration factor of bilirubin: The simulated plasma prepared in Example 1 is respectively prepared into simulated plasma with total bilirubin concentrations of 350 μmol / L, 200 μmol / L and 400 μmol / L. According to the second preset cycle parameters in Example 1, the three simulated plasma with different bilirubin concentrations are respectively injected into the second adsorption device for adsorption treatment, and according to the concentration of bilirubin in the simulated plasma before adsorption treatment and the concentration of bilirubin in the simulated plasma after adsorption treatment, the fourth simulated adsorption rate η4 of bilirubin is determined.

[0101] The simulated plasma prepared in Example 1 is respectively prepared into simulated plasma with total bilirubin concentrations of 350 μmol / L, 200 μmol / L and 400 μmol / L. According to the first preset cycle parameters in Example 1, the three simulated plasma with different bilirubin concentrations are respectively injected into the first adsorption device for adsorption treatment, and according to the concentration of bilirubin in the simulated plasma before adsorption treatment and the concentration of bilirubin in the simulated plasma after adsorption treatment, the fifth simulated adsorption rate η5 of bilirubin is determined. The second calibration factor F2 is calculated according to the following formula: F2 = η4 / η5.

[0102] The experiment is repeated for several times to obtain the finally determined F2. The adsorption rate results of the adsorbent and the second calibration factor F2 results obtained under different concentrations of bilirubin are shown in Table 3.

[0103] Table 3 Second calibration factor results table of bilirubin

[0104]

[0105]

[0106] The second calibration factor F2 of bilirubin is finally obtained as F2 = (1.01 + 1.00 + 1.03) / 3 = 1.01.

[0107] (2) The second calibration factor of bile acids was determined using the same method as that used in step (1) to determine the second calibration factor of bilirubin. The experiment was repeated multiple times to obtain the final determined F2. The adsorption rate of the adsorbent and the result of the second calibration factor F2 under different concentrations of bile acids are shown in Table 4.

[0108] Table 4 Results of the Second Calibration Factor for Bile

[0109]

[0110]

[0111] The final second calibration factor for bile acids is F2 = (1.02 + 1.03 + 1.03) / 3 = 1.03.

[0112] Example 4

[0113] This embodiment provides a method for determining a third calibration factor, including the following steps:

[0114] (1) Determine the third calibration factor of bilirubin: According to the first preset cycle parameters in Example 1, the first adsorption device is used to adsorb the plasma of patients with liver disease, and the adsorption rate η7 under real environment is determined according to the concentration of bilirubin in the plasma of patients with liver disease before adsorption treatment and the concentration of bilirubin in the plasma of patients with liver disease after adsorption treatment.

[0115] The simulated plasma prepared in Example 1 was formulated into simulated plasma with the same bilirubin concentration as that in the plasma of patients with liver disease, ensuring that the bilirubin concentration was the same under simulated and real conditions. According to the first preset cycle parameters in Example 1, the simulated plasma was injected into the first adsorption device for adsorption treatment. The adsorption rate η6 under the simulated environment was determined based on the bilirubin concentration in the simulated plasma before adsorption treatment and the bilirubin concentration in the simulated plasma after adsorption treatment. The third calibration factor F3 was calculated according to the following formula: Third calibration factor F3 = η6 / η7.

[0116] The experiment was repeated multiple times to obtain the final determined F3. The adsorption rate results of the adsorbent and the third calibration factor F3 results for different samples are shown in Table 5.

[0117] Table 5 Results of the Third Calibration Factor for Bilirubin

[0118]

[0119]

[0120] The final calibration factor for bilirubin is F3 = (1.63 + 1.69 + 1.56 + 1.64 + 1.73) / 5 = 1.65.

[0121] (2) The third calibration factor of bile acid is determined in the same way as the third calibration factor of bilirubin in step (1), and the experiment is repeated multiple times to obtain the final determined F3. The results of the adsorption rate of the adsorbent obtained under different concentrations of bile acid and the third calibration factor F3 are shown in Table 6.

[0122] Table 6: Third calibration factor results of bile acid

[0123]

[0124] The third calibration factor F3 of bile acid is finally obtained as F3 = (1.25 + 1.27 + 1.28 + 1.25 + 1.26) / 5 = 1.26.

[0125] The accuracy of the method for detecting the adsorption performance of the adsorbent provided in the present embodiment is verified, the adsorption rate of the adsorbent in DPMAS to bilirubin and bile acid in actual application is obtained, and the adsorption rate of the adsorbent to bilirubin and bile acid under simulated conditions under the same conditions is obtained. The adsorption rate results obtained from different samples are shown in Tables 7 and 8.

[0126] Table 7: Accuracy evaluation results of bilirubin adsorption rate

[0127]

[0128] The adsorption rate of bilirubin is calculated by the following formula: Adsorption rate of bilirubin = First simulated adsorption rate of bilirubin / (F1 x F2 x F3) = First simulated adsorption rate of bilirubin / (1.28 x 1.01 x 1.65).

[0129] Table 8: Accuracy evaluation results of bile acid adsorption rate

[0130]

[0131] The adsorption rate of bile acid is calculated by the following formula: Adsorption rate of bile acid = First simulated adsorption rate of bile acid / (F1 x F2 x F3) = First simulated adsorption rate of bile acid / (1.03 x 1.03 x 1.26).

[0132] As can be seen from Table 7, the adsorption rate of bilirubin obtained by the method for detecting the adsorption performance of the adsorbent provided in the present embodiment is close to the actual adsorption rate of bilirubin, and the adsorption rate of bile acid is close to the actual adsorption rate of bile acid. It is shown that the accuracy of the detection method for the adsorption performance of the adsorbent provided in the present embodiment is high, and the effect of the adsorbent in DPMAS on removing bilirubin and bile acid can be truly and accurately evaluated.

[0133] Although the present disclosure discloses as above, the protection scope of the present disclosure is not limited to this. The person skilled in the art can make various changes and modifications without departing from the spirit and scope of the present disclosure, and these changes and modifications will fall within the protection scope of the present disclosure.

Claims

1. A method of detecting the adsorption performance of an adsorbent, characterized by, The method comprises the following steps: constructing a first adsorption device comprising an anion resin adsorber and a neutral macroporous resin adsorber connected in series; processing the first adsorption device according to a preset scale value to obtain a second adsorption device; obtaining simulated plasma containing a preset concentration of a to-be-tested product, wherein the to-be-tested product comprises bilirubin and / or bile acid; injecting the simulated plasma into the second adsorption device according to preset circulation parameters to perform adsorption processing, and obtaining a first concentration of the to-be-tested product in the simulated plasma; determining a first simulated adsorption rate of the to-be-tested product according to the preset concentration of the to-be-tested product and the first concentration of the to-be-tested product; obtaining a calibration factor, calibrating the first simulated adsorption rate of the to-be-tested product according to the calibration factor, and determining an adsorption rate of the to-be-tested product; the obtaining of the calibration factor comprises: obtaining ex vivo plasma containing the to-be-tested product, and determining a first calibration factor according to the simulated plasma, the ex vivo plasma, and the first adsorption device; determining a second calibration factor according to the simulated plasma, the first adsorption device, and the second adsorption device; obtaining plasma containing the to-be-tested product, and determining a third calibration factor according to the simulated plasma, the plasma, and the first adsorption device; calculating the calibration factor according to the following formula: calibration factor = first calibration factor × second calibration factor × third calibration factor; wherein the determination of the first calibration factor comprises: injecting the simulated plasma into the first adsorption device according to preset circulation parameters to perform adsorption processing, and obtaining a second concentration of the to-be-tested product in the simulated plasma; determining a second simulated adsorption rate of the to-be-tested product according to the preset concentration of the to-be-tested product and the second concentration of the to-be-tested product; before the adsorption processing, obtaining a third concentration of the to-be-tested product in the ex vivo plasma, injecting the ex vivo plasma into the first adsorption device according to preset circulation parameters to perform adsorption processing, and obtaining a fourth concentration of the to-be-tested product in the ex vivo plasma; determining a third simulated adsorption rate of the to-be-tested product according to the third concentration of the to-be-tested product and the fourth concentration of the to-be-tested product; calculating the first calibration factor according to the following formula: first calibration factor = second simulated adsorption rate / third simulated adsorption rate; the determination of the second calibration factor comprises: injecting the simulated plasma into the second adsorption device according to preset circulation parameters to perform adsorption processing, and obtaining a fifth concentration of the to-be-tested product in the simulated plasma; determining a fourth simulated adsorption rate of the to-be-tested product according to the preset concentration of the to-be-tested product and the fifth concentration of the to-be-tested product; injecting the simulated plasma into the first adsorption device according to preset circulation parameters to perform adsorption processing, and obtaining a sixth concentration of the to-be-tested product in the simulated plasma; determining a fifth simulated adsorption rate of the to-be-tested product according to the preset concentration of the to-be-tested product and the sixth concentration of the to-be-tested product; calculating the second calibration factor according to the following formula: second calibration factor = fourth simulated adsorption rate / fifth simulated adsorption rate; the determination of the third calibration factor comprises: According to preset circulation parameters, the simulated blood plasma is injected into the first adsorption device for adsorption treatment, and a seventh concentration of the to-be-tested product in the simulated blood plasma is obtained; According to the preset concentration of the to-be-tested product and the seventh concentration of the to-be-tested product, a sixth simulated adsorption rate of the to-be-tested product is determined; Before the adsorption treatment, an eighth concentration of the to-be-tested product in the blood plasma is obtained, the blood plasma is injected into the first adsorption device according to preset circulation parameters for adsorption treatment, and a ninth concentration of the to-be-tested product in the blood plasma is obtained; According to the eighth concentration of the to-be-tested product and the ninth concentration of the to-be-tested product, a seventh simulated adsorption rate of the to-be-tested product is determined; The third calibration factor is calculated according to the following formula: third calibration factor = sixth simulated adsorption rate / seventh simulated adsorption rate; The preset circulation parameters include first preset circulation parameters and second preset circulation parameters, the first preset circulation parameters and the second preset circulation parameters both include plasma flow rate, plasma volume and circulation time, and the plasma flow rate and the plasma volume in the first preset circulation parameters are n times of the plasma flow rate and the plasma volume in the second preset circulation parameters, and the circulation time in the first preset circulation parameters is equal to the circulation time in the second preset circulation parameters; If the adsorption treatment is performed in the first adsorption device, the concentration of the to-be-tested product after the adsorption treatment is determined according to the first preset circulation parameters; If the adsorption treatment is performed in the second adsorption device, the concentration of the to-be-tested product after the adsorption treatment is determined according to the second preset circulation parameters; The preset concentration of the to-be-tested product in the simulated blood plasma is the same as the third concentration of the to-be-tested product in the ex vivo blood plasma.

2. The method of claim 1, wherein the adsorbent is a zeolite. The preset ratio value is n, and 15≤n≤25.

3. The method of claim 1, wherein the adsorbent is a zeolite. The concentration of the bilirubin in the simulated blood plasma is 100 μmol / L to 1000 μmol / L, and the concentration of the bile acid in the simulated blood plasma is 100 μmol / L to 1000 μmol / L.

4. The method of claim 1, wherein the adsorbent is a zeolite. The anion resin adsorber contains anion exchange resin, and the neutral macroporous resin adsorber contains neutral macroporous adsorption resin.

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