High-concentration osmotic pressure molar concentration standard substance and preparation method thereof
The problem of insufficient concentration of existing standard substances is solved by using sodium chloride and ultrapure water, and the high concentration of standard substances is provided to meet the needs of osmotic pressure control in biological products and pharmaceutical preparations. .
Patent Information
- Application Number
- CN202411717300.1
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-11-27
- Publication Date
- 2025-02-18
AI Technical Summary
The concentrations of existing osmolar molar concentration standard substances are lower than 4000mOsmol/kg, which cannot meet the needs of high concentration standards.
Sodium chloride is used as solute and ultrapure water as solvent, and prepared by weight method to prepare osmotic molar concentration standard substance with a concentration range of (4000-11182) mOsmol/kg, and its standard value is calculated and verified by the freezing point drop method. .
A standard substance with osmolar pressure molar concentration of higher than 4000mOsmol/kg was provided, which filled the gap in high concentration standards. The preparation process was simple, the standard substance was uniform, and the stability period was ≥2 years.
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Abstract
Description
Technical Field
[0001] The invention belongs to the technical field of standard substances, and in particular relates to a high-concentration osmotic pressure molar concentration standard substance and a preparation method thereof. Background Art
[0002] Biological membranes, such as the cell membranes or capillary walls of the human body, generally have the properties of semipermeable membranes. The phenomenon of solvent diffusion from low concentration to high concentration solution through the semipermeable membrane is called osmosis, and the pressure required to prevent osmosis is called osmotic pressure. Osmotic pressure plays an extremely important role in various biological processes involving the diffusion of solutes or the transport of liquids through biological membranes. Therefore, when preparing pharmaceutical preparations such as injections and ophthalmic liquid preparations, their osmotic pressure must be paid attention to. Preparations with osmotic pressure regulators added to the prescription should all have their osmotic pressure molar concentrations controlled.
[0003] The osmotic pressure molarity is usually determined indirectly by measuring the freezing point depression of a solution. In an ideal dilute solution, the freezing point depression conforms to ΔT f =K f ·m, where ΔT f is the freezing point depression, K f is the freezing point depression constant (1.86 when water is the solvent), and m is the molar concentration. The osmotic pressure conforms to the relationship P0=K0·m, where P0 is the osmotic pressure, K0 is the osmotic pressure constant, and m is the molar concentration of the solution. Since the concentrations in the two formulas are the same, the freezing point depression method can be used to determine the osmotic pressure molar concentration of the solution.
[0004] It is often necessary to measure the osmotic pressure molar concentration of solutions or body fluids in the production process of biological products, pharmaceutical preparations, etc. and in clinical diagnosis. Osmotic pressure molar concentration meters have been widely used due to their simple operation, high accuracy, and stable performance. The preparation method of osmotic pressure molar concentration standard solution is specified in the fourth part of the Chinese Pharmacopoeia 0632 Osmotic Pressure Molar Concentration Determination Method published in 2020, but the concentration of osmotic pressure molar concentration standard substances on the market is currently less than 4000mOsmol / kg, and there are no high-concentration osmotic pressure molar concentration standard substances. Summary of the invention
[0005] The object of the present invention is to provide a high-concentration osmotic pressure molar concentration standard substance and a preparation method thereof, so as to fill the blank of high-concentration osmotic pressure molar concentration standard substance.
[0006] The purpose of the present invention can be achieved through the following technical solutions:
[0007] The invention discloses a high-concentration osmotic pressure molar concentration standard substance, which uses sodium chloride, a national first-level purity standard substance, as a solute and ultrapure water as a solvent, and has a concentration range of (4000-11182) mOsmol / kg.
[0008] Furthermore, the stability period of the high-concentration osmotic pressure molar concentration standard substance is ≥ 2 years.
[0009] Furthermore, the relative expanded uncertainty of the high-concentration osmotic pressure molar concentration standard substance is 0.02%.
[0010] According to the colligative properties of the salt concentration of the sodium chloride solution and the freezing point depression temperature, the standard value of the osmotic pressure is calculated by the osmotic pressure P0=537.6×ΔTf (ΔTf is obtained from the empirical measurement table); and its standard value is verified by a high-precision osmotic pressure molar concentration meter.
[0011] Uncertainty assessment factors include: uncertainty of raw material purity, uncertainty of weighing process, uncertainty of freezing point depression empirical determination table (see Appendix 1 of the instruction manual), and uncertainty introduced by sample uniformity and stability.
[0012] A method for preparing a high-concentration osmotic pressure molar concentration standard substance comprises: preparing a solution, wherein the solution is prepared by a gravimetric method:
[0013] (1) Take sodium chloride purity standard material, place it in a crucible, place the crucible in a muffle furnace, dry it at (500±10)°C for 6 h, take it out, place it in a silica gel desiccator, cool it to room temperature and keep it constant in weight;
[0014] (2) This standard substance is prepared by the gravimetric method. Accurately weigh the sodium chloride and ultrapure water, and dissolve the weighed sodium chloride in a volumetric flask with weighed water.
[0015] (3) Fill the solution into a clean ampoule and store it at room temperature away from light.
[0016] Furthermore, the weights of sodium chloride and ultrapure water in step (2) are corrected for buoyancy.
[0017] Beneficial effects of the present invention:
[0018] (1) The osmotic pressure molar concentration standard substance provided by the present invention has a concentration higher than 4000mOsmol / kg, filling the gap of high-concentration osmotic pressure molar concentration standard substances in China.
[0019] (2) The preparation process of the osmotic pressure molar concentration standard substance provided by the present invention is simple, and the prepared standard substance has good uniformity and a stable period of ≥2 years.
[0020] (3) The determination method adopted by the present invention is simple and easy to operate, so that the concentration of the standard substance can be determined even in the absence of a comparison standard substance. DETAILED DESCRIPTION
[0021] The following will be combined with the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.
[0022] The implementation method is as follows:
[0023] (1) Take an appropriate amount of sodium chloride purity standard material, place it in a crucible, place the crucible in a muffle furnace, and dry it at (500±10)℃ for 6h. Take it out and place it in a silica gel desiccator to cool to room temperature and constant weight.
[0024] (2) This standard substance is prepared by the gravimetric method. Accurately weigh the weight of sodium chloride and ultrapure water (both corrected for buoyancy), and dissolve the weighed sodium chloride in a 200-mL volumetric flask with weighed water.
[0025] (3) Fill the solution into a clean 2 mL ampoule and store at room temperature away from light.
[0026] (4) Evaluate the value and uncertainty of the product.
[0027] Example 1
[0028] This embodiment provides an osmotic pressure molar concentration standard substance with a concentration of 4000.8mOsmol / kg.
[0029] The preparation method of the above-mentioned osmotic pressure molar concentration standard substance comprises the following steps:
[0030] (1) Take about 30 g of sodium chloride purity standard material and place it in a crucible. Place the crucible in a muffle furnace and dry it at (500±10)℃ for 6 h. Take it out and place it in a silica gel desiccator to cool to room temperature and constant weight.
[0031] (2) This standard substance is prepared by the gravimetric method. The weight of sodium chloride and ultrapure water (both corrected for buoyancy) is accurately weighed, and the weighed sodium chloride is dissolved in a 200 mL volumetric flask with weighed water. The weighed values and buoyancy correction values are shown in Table 1.
[0032] Table 1
[0033]
[0034] (3) Fill the solution into a clean 2 mL ampoule and store at room temperature away from light.
[0035] The sodium chloride salt concentration is calculated based on the sodium chloride correction value and the water correction value, and the freezing point depression temperature (ΔT f ) is 7.442℃, and the osmotic pressure molar concentration calculated according to formula 1 is 4000.8mOsmol / kg.
[0036] P0=537.6×ΔT f Formula 1
[0037] The freezing point depression temperature (ΔT f ) is 7.4426℃, and the osmotic pressure molar concentration is calculated by formula 1 to be 4001.1mOsmol / kg.
[0038] Uniformity test: 10 standard substances were randomly selected and measured three times using a high-precision osmotic pressure molar concentration meter. The measured data were subjected to an F test to evaluate the uniformity of the standard substance. As shown in Table 2, the results showed that the standard substance had good uniformity.
[0039] Table 2
[0040]
[0041]
[0042] Stability test: The stability study of reference materials usually includes short-term stability study and long-term stability study. Short-term stability refers to the stability of the characteristic value of reference materials during transportation under specified transportation conditions, which is related to the additional impact and packaging caused by the transportation of reference materials. Long-term stability refers to the stability of the characteristic value of reference materials under the specific storage conditions required in the reference material certificate, which is related to its production, development, use and storage conditions.
[0043] The standard substance was stored at 60°C, and one sample was taken at each time point on days 0, 1, 2, 3, 5, 9, and 14. The measurements were performed three times in parallel to examine the short-term stability. As shown in Table 3, the results showed that the standard substance had good short-term stability.
[0044] Table 3
[0045]
[0046] The standard substance was stored at room temperature and protected from light. One sample was taken at each time point, namely, 0 month, 1 month, 2 month, 3 month, 6 month, 9 month, 12 month, 18 month and 24 month. The long-term stability was investigated by parallel measurement 3 times (taking the average value). As shown in Table 4, the results showed that the standard substance could be stably stored for 2 years.
[0047] Table 4
[0048]
[0049] Uncertainty assessment: The osmotic pressure molar concentration standard material is prepared by the weight method, and the standard value is obtained by the osmotic pressure molar concentration calculation formula. The main sources of uncertainty are as follows: (1) uncertainty introduced by raw material purity; (2) uncertainty introduced by the weighing process; (3) uncertainty introduced by the empirical table for freezing point depression; (4) uncertainty introduced by uniformity; (5) uncertainty introduced by stability.
[0050] (1) Uncertainty introduced by raw material purity: The uncertainty of the raw material certificate is 0.0080%, k = 2, then the uncertainty introduced by the raw material is 0.0040%.
[0051] (2) The uncertainty introduced by the weighing process is
[0052] (3) The uncertainty of the freezing point depression empirical measurement table is 0.02%.
[0053] (4) The uncertainty introduced by uniformity is 0.10%.
[0054] (5) The uncertainty introduced by stability is 0.0017%.
[0055] The relative uncertainty components are synthesized to obtain the relative composite standard uncertainty u of the standard substance. rel =0.1%, taking inclusion factor k=2, U rel =0.1%×2=0.2%.
[0056] The above results show that the relative expanded uncertainty of the osmotic pressure molar concentration standard substance of 4000.8mOsmol / kg is 0.2% (k=2).
[0057] Example 2
[0058] This embodiment provides an osmotic pressure molar concentration standard substance with a concentration of 7473.7 mOsmol / kg.
[0059] The preparation method of the above-mentioned osmotic pressure molar concentration standard substance comprises the following steps:
[0060] (1) Take about 40 g of sodium chloride purity standard material and place it in a crucible. Place the crucible in a muffle furnace and dry it at (500±10)℃ for 6 h. Take it out and place it in a silica gel desiccator to cool to room temperature and constant weight.
[0061] (2) This standard substance is prepared by the gravimetric method. The weight of sodium chloride and ultrapure water (both corrected for buoyancy) is accurately weighed, and the weighed sodium chloride is dissolved in a 200 mL volumetric flask with weighed water. The weighed values and buoyancy correction values are shown in Table 5.
[0062] Table 5
[0063]
[0064] (3) Fill the solution into a clean 2 mL ampoule and store at room temperature away from light.
[0065] The sodium chloride salt concentration is calculated based on the sodium chloride correction value and the water correction value, and the freezing point depression temperature (ΔT f ) is 13.902℃, and the osmotic pressure molar concentration calculated according to formula 1 is 7473.7mOsmol / kg.
[0066] The freezing point depression temperature (ΔT f ) is 13.9045℃, and the osmotic pressure molar concentration is calculated by formula 1 to be 7475.1mOsmol / kg.
[0067] Homogeneity test: According to the homogeneity test method in Example 1, the osmotic pressure molar concentration standard substance with a concentration of 7473.7mOsmol / kg was tested, and F was 0.77, which was less than 2.39 (limit value). The homogeneity of the standard substance was good.
[0068] Stability test: According to the stability test method in Example 1, the osmotic pressure molar concentration standard substance with a concentration of 7473.7mOsmol / kg was tested for short-term stability and long-term stability. As shown in Table 6, the results show that the short-term stability of the standard substance meets the requirements and can be stored at room temperature in the dark for 2 years.
[0069] Table 6
[0070]
[0071] Uncertainty assessment: The uncertainty of the osmotic pressure molar concentration standard substance with a concentration of 7473.7 mOsmol / kg was assessed according to the uncertainty assessment method in Example 1.
[0072] (1) Uncertainty introduced by raw material purity: The uncertainty of the raw material certificate is 0.0080%, k = 2, then the uncertainty introduced by the raw material is 0.0040%.
[0073] (2) The uncertainty introduced by the weighing process is 0.00088%.
[0074] (3) The uncertainty of the freezing point depression empirical measurement table is 0.02%.
[0075] (4) The uncertainty introduced by uniformity is 0.085%.
[0076] (5) The uncertainty introduced by stability is 0.0036%.
[0077] The relative uncertainty components are synthesized to obtain the relative composite standard uncertainty u of the standard substance. rel =0.09%, taking inclusion factor k=2, U rel =0.09%×2=0.2%.
[0078] The above results show that the relative expanded uncertainty of the osmotic pressure molar concentration standard material of 7473.7mOsmol / kg is 0.2%.
[0079] Example 3
[0080] This embodiment provides an osmotic pressure molar concentration standard substance with a concentration of 11182.1 mOsmol / kg.
[0081] The preparation method of the above-mentioned osmotic pressure molar concentration standard substance comprises the following steps:
[0082] (1) Take about 50 g of sodium chloride purity standard material and place it in a crucible. Place the crucible in a muffle furnace and dry it at (500±10)℃ for 6 h. Take it out and place it in a silica gel desiccator to cool to room temperature and constant weight.
[0083] (2) This standard substance is prepared by the gravimetric method. The weight of sodium chloride and ultrapure water (both corrected for buoyancy) is accurately weighed, and the weighed sodium chloride is dissolved in a 200 mL volumetric flask with weighed water. The weighed values and buoyancy correction values are shown in Table 7.
[0084] Table 7
[0085]
[0086] (3) Fill the solution into a clean 2 mL ampoule and store at room temperature away from light.
[0087] The sodium chloride salt concentration is calculated based on the sodium chloride correction value and the water correction value, and the freezing point depression temperature (ΔTf ) is 20.800°C, and the osmotic pressure molar concentration calculated according to Formula 1 is 11182.1 mOsmol / kg.
[0088] The freezing point depression temperature (ΔT f ) is 20.7987℃, and the osmotic pressure molar concentration calculated by formula 1 is 11181.4mOsmol / kg.
[0089] Homogeneity test: According to the homogeneity test method in Example 1, the osmotic pressure molar concentration standard substance with a concentration of 11182.1mOsmol / kg was tested, and F was 1.10, which was less than 2.39 (limit value), and the standard substance had good homogeneity.
[0090] Stability test: According to the stability test method in Example 1, the osmotic pressure molar concentration standard substance with a concentration of 11182.1mOsmol / kg was tested for short-term stability and long-term stability. As shown in Table 8, the results show that the short-term stability of the standard substance meets the requirements and can be stored at room temperature and protected from light for 2 years.
[0091] Table 8
[0092]
[0093] Uncertainty assessment: The uncertainty of the osmotic pressure molar concentration standard substance with a concentration of 11182.1 mOsmol / kg was assessed according to the uncertainty assessment method in Example 1.
[0094] (1) Uncertainty introduced by raw material purity: The uncertainty of the raw material certificate is 0.0080%, k = 2, then the uncertainty introduced by the raw material is 0.0040%.
[0095] (2) The uncertainty introduced by the weighing process is 0.00085%.
[0096] (3) The uncertainty of the freezing point depression empirical measurement table is 0.02%.
[0097] (4) The uncertainty introduced by uniformity is 0.11%.
[0098] (5) The uncertainty introduced by stability is 0.0041%.
[0099] The relative uncertainty components are synthesized to obtain the relative composite standard uncertainty u of the standard substance. rel =0.1%, taking inclusion factor k=2, U rel =0.1%×2=0.2%.
[0100] The above results show that the relative expanded uncertainty of the osmotic pressure molar concentration standard material of 11182.1mOsmol / kg is 0.2%.
[0101] The empirical determination table of freezing point depression is shown in Table 10 below.
[0102] Table 10
[0103]
[0104]
[0105] Table 10 - continued
[0106]
[0107]
[0108] It should be noted that, in this article, relational terms such as first and second, etc. are only used to distinguish one entity or operation from another entity or operation, and do not necessarily require or imply any such actual relationship or order between these entities or operations. Moreover, the terms "include", "comprise" or any other variants thereof are intended to cover non-exclusive inclusion, so that a process, method, article or device including a series of elements includes not only those elements, but also other elements not explicitly listed, or also includes elements inherent to such process, method, article or device.
[0109] Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that various changes, modifications, substitutions and variations may be made to the embodiments without departing from the principles and spirit of the present invention, and that the scope of the present invention is defined by the appended claims and their equivalents.
Claims
1. A high concentration osmotic pressure molar concentration standard substance, characterized in that, Sodium chloride, a national first-level purity standard substance, is used as the solute and ultrapure water is used as the solvent. The concentration range is (4000~11182)mOsmol / kg.
2. A high concentration osmotic pressure molar concentration standard substance according to claim 1, characterized in that: The stability period of the high-concentration osmotic pressure molar concentration standard substance is ≥2 years.
3. A high concentration osmotic pressure molar concentration standard substance according to claim 1, characterized in that: The relative expanded uncertainty of the high concentration osmotic pressure molar concentration standard material is 0.02%.
4. A method for preparing a high-concentration osmotic pressure molar concentration standard substance as claimed in any one of claims 1 to 3, characterized in that: include: The solution is prepared by a weight method and comprises the following steps: (1) Take sodium chloride purity standard material, place it in a crucible, place the crucible in a muffle furnace, dry it at (500±10)°C for 6 h, take it out, place it in a silica gel desiccator, cool it to room temperature and keep it constant weight; (2) This standard substance is prepared by the gravimetric method. Accurately weigh the sodium chloride and ultrapure water, and dissolve the weighed sodium chloride in a volumetric flask with weighed water. (3) Fill the solution into a clean ampoule and store it at room temperature away from light.
5. The method for preparing a high-concentration osmotic pressure molar concentration standard substance according to claim 4, characterized in that: The weights of sodium chloride and ultrapure water in step (2) are corrected for buoyancy.