Method for simultaneously determining contents of calcium gluconate and calcium glucarate in medicine

Abstract

The invention discloses a method for simultaneously determining the content of calcium gluconate and calcium glucarate in a medicine, the method is a high performance liquid chromatography, and the chromatographic conditions of the high performance liquid chromatography are as follows: a mixed solution of a sodium dihydrogen phosphate solution and a tetrabutylammonium hydrogen sulfate solution is used as a mobile phase, and the detection wavelength is 208-212nm. The method has the beneficial effects that the determination method is high in specificity, high in accuracy, good in repeatability and durability, simple and convenient to operate, rapid and high in sensitivity.

Description

Technical Field

[0001] The invention relates to a method for quantitatively determining calcium gluconate and calcium glucarate, and in particular to a method for simultaneously determining the contents of calcium gluconate and calcium glucarate in medicines by adopting high performance liquid chromatography. Background Art

[0002] Calcium Gluconate is an organic calcium salt that can be used to reduce capillary permeability, increase density, maintain normal excitability of nerves and muscles, strengthen myocardial contractility, and help bone formation. Calcium gluconate is a white crystalline powder or granules, which dissolves in boiling water, slowly dissolves in water, and is insoluble in organic solvents such as anhydrous ethanol, chloroform, and ether. Calcium gluconate is a pharmaceutical excipient, a stabilizer, an acid antacid, and a solubilizing agent. It mainly acts as a stabilizer in calcium gluconate sodium chloride injection.

[0003] The relevant marketed dosage forms of calcium gluconate include tablets, granules, oral solutions, injections, etc. Calcium gluconate sodium chloride injection is mainly suitable for children and adult patients for the treatment of acute symptomatic hypocalcemia. According to the 53rd batch of reference preparations catalog of the State Food and Drug Administration, the reference preparation information of calcium gluconate sodium chloride injection was announced. The license holder is HQ Spencile.Inc of the United States. The specifications are 1g / 50ml (20mg / ml), 2g / 100ml (20mg / ml), 1g / 100ml (10mg / ml), and the reference preparation calcium gluconate dosage is 0.09% (w / v).

[0004] The main method for detecting calcium content is complexometric titration. The above method is often used to detect the single calcium content in drugs. The endpoint is mainly determined by the color change of the indicator by the naked eye. The method is not very sensitive, and it is not applicable to drugs containing two or more organic calcium salts. It is easy to cause interference and cannot accurately measure the calcium content of each component. Calcium gluconate and sodium chloride injection contains two calcium salts, calcium gluconate and calcium glucarate. Accurate quantification cannot be achieved using existing detection methods.

[0005] In summary, it is of great commercial value to develop a method that is easy to operate and can simultaneously and accurately quantify the contents of calcium gluconate and calcium glucarate in calcium gluconate and sodium chloride injection. Summary of the invention

[0006] To solve the above problems, the present invention provides a method and application for simultaneously determining the contents of calcium gluconate and calcium glucarate in medicines, which has strong specificity, high accuracy, good durability, and is simple, fast, and highly sensitive to operate.

[0007] The first aspect of the present invention provides a method for simultaneously determining the content of calcium gluconate and calcium glucarate in a drug, the method being a high performance liquid chromatography method, the chromatographic conditions of the high performance liquid chromatography method being: a mixture of sodium dihydrogen phosphate solution and tetrabutylammonium hydrogen sulfate solution being used as the mobile phase, and a detection wavelength of 208nm-212nm (e.g., 208nm, 210nm, 212nm).

[0008] According to some implementation cases of the method of the present invention,

[0009] The concentration of sodium dihydrogen phosphate in the mobile phase is 0.01-0.04 mol / L (0.01 mol / L, 0.02 mol / L, 0.03 mol / L, 0.04 mol / L), and the concentration of tetrabutylammonium hydrogen sulfate is 0.003-0.007 mol / L (0.003 mol / L, 0.004 mol / L, 0.005 mol / L, 0.006 mol / L, 0.007 mol / L).

[0010] According to some implementation cases of the method of the present invention, the concentration of sodium dihydrogen phosphate in the mobile phase is 0.015-0.035 mol / L, and the concentration of tetrabutylammonium hydrogen sulfate is 0.004-0.006 mol / L.

[0011] According to some implementation examples of the method of the present invention, the concentration of sodium dihydrogen phosphate in the mobile phase is 0.025 mol / L, and the concentration of tetrabutylammonium hydrogen sulfate is 0.005 mol / L. The reagent grade used is an ion pair reagent.

[0012] According to some implementation cases of the method of the present invention, the pH of the mobile phase is 1-4 (for example: pH 1, pH 2, pH 3, pH 4), preferably 2-3, and more preferably 2.5. When the mobile phase is used, the preparation ratio of sodium dihydrogen phosphate and tetrabutylammonium hydrogen sulfate in the mobile phase must be ensured and it should be fully stirred for at least 60 minutes, and the elution method is isocratic elution.

[0013] According to some implementation cases of the method of the present invention, the high performance liquid chromatography uses octadecylsilane bonded silica gel as a filler.

[0014] According to some implementation cases of the method of the present invention, the flow rate of the mobile phase is 0.5-1.5 ml / min (for example: 0.5 ml / min, 0.8 ml / min, 1.0 ml / min, 1.2 ml / min, 1.5 ml / min).

[0015] According to some implementation cases of the method of the present invention, the flow rate of the mobile phase is 0.8-1.2 ml / min.

[0016] According to some implementation cases of the method of the present invention, the flow rate of the mobile phase is 1 ml / min.

[0017] According to some implementation cases of the method of the present invention, the detection wavelength is 210 nm, the column temperature is 20°C-40°C (for example: 20°C, 25°C, 28°C, 30°C, 32°C, 35°C, 40°C), preferably 30°C.

[0018] According to some implementation cases of the method of the present invention, the injection volume during detection is 20 μl.

[0019] The second aspect of the present invention provides the use of the method described in the first aspect of the present invention in determining the content of calcium gluconate and calcium glucarate in calcium gluconate and sodium chloride injection.

[0020] The third aspect of the present invention provides a method for determining the content of calcium gluconate and calcium glucarate in calcium gluconate sodium chloride injection, comprising the following steps:

[0021] (1) Take calcium gluconate and sodium chloride injection, dilute with water to make a solution containing about 2 mg of calcium gluconate and 0.1 mg of calcium glucarate per 1 ml, as the test solution, accurately measure 20 μl, inject into the liquid chromatograph, and record the chromatogram;

[0022] (2) Dissolve calcium gluconate in water and quantitatively dilute to prepare a solution containing about 2 mg of calcium gluconate per 1 ml, as the calcium gluconate reference solution;

[0023] (3) diluting calcium gluconate with water to prepare a solution containing about 0.1 mg of calcium gluconate per 1 ml as the calcium gluconate reference solution;

[0024] (4) Calculate the contents of calcium gluconate and calcium glucarate in calcium gluconate and sodium chloride injection by the external standard method using the peak areas;

[0025] The chromatographic conditions are as follows: octadecylsilane bonded silica gel is used as a filler, a mixture of sodium dihydrogen phosphate solution and tetrabutylammonium hydrogen sulfate solution is used as a mobile phase, the concentration of sodium dihydrogen phosphate in the mobile phase is 0.01-0.04 mol / L, the concentration of tetrabutylammonium hydrogen sulfate is 0.003-0.007 mol / L, and the flow rate is 0.5-1.5 ml / min;

[0026] The detection wavelength is 208nm-212nm, and the column temperature is 20℃-40℃;

[0027] According to some implementation cases of the method of the present invention, the chromatographic conditions are: octadecylsilane bonded silica gel is used as a filler, a mixture of sodium dihydrogen phosphate solution and tetrabutylammonium hydrogen sulfate solution is used as a mobile phase, and the concentration of sodium dihydrogen phosphate in the mobile phase is 0.025 mol / L, the concentration of tetrabutylammonium hydrogen sulfate is 0.005 mol / L, and the flow rate is 1 ml / min;

[0028] The detection wavelength was 210 nm and the column temperature was 30 °C.

[0029] Compared with the prior art, the invention has the following beneficial effects: the invention provides a high performance liquid chromatography method capable of simultaneously and accurately determining the contents of calcium gluconate and calcium glucarate in calcium gluconate sodium chloride injection, which has not only strong specificity, high accuracy, good repeatability, intermediate precision, linearity, range and durability, but also simple and rapid operation, high sensitivity, and can realize the control of each single calcium content and the total calcium content, thus saving detection time and cost, and having very important significance for the quality control and finished product inspection in the production process of the calcium gluconate sodium chloride injection. BRIEF DESCRIPTION OF THE DRAWINGS

[0030] Figure 1 : System suitability calcium gluconate chromatogram in Example 1.

[0031] Figure 2 : System suitability calcium glucarate chromatogram in Example 1.

[0032] Figure 3 : Specificity chromatogram in Example 2.

[0033] Figure 4 : Calcium gluconate reference substance positioning chromatogram among embodiment 2.

[0034] Figure 5 : Calcium glucarate reference substance positioning chromatogram in Example 2.

[0035] Figure 6 : Calcium gluconate linear diagram in Example 6.

[0036] Figure 7 : Calcium glucarate linear diagram in Example 6.

[0037] Figure 8 : Chromatogram of 220915141 batches of test solution (without ion pair reagent) in Example 7.

[0038] Fig. 9 : Chromatograms of 220915141 batches of test sample solutions in Examples 7 and 8.

[0039] Fig.10: Chromatogram of 220915141 batches of test solution (flow rate 0.4 ml / min) in Example 7.

[0040] Fig.11 : Chromatogram of 10133 batches of test sample solution in Example 8. DETAILED DESCRIPTION

[0041] In order to make the purpose, technical scheme and advantages of the present invention clearer, the present invention is further described in detail below in conjunction with the embodiments and drawings. The specific embodiments described herein are only used to explain the present invention and are not intended to constitute any limitation to the present invention. In addition, in the following description, the description of known structures and technologies is omitted to avoid unnecessary confusion of the concepts of the present disclosure. Such structures and technologies are also described in many publications.

[0042] 1. Chromatographic conditions

[0043] Octadecylsilane bonded silica gel was used as filler, 0.025 mol / L sodium dihydrogen phosphate-0.005 mol / L tetrabutylammonium hydrogen sulfate was used as mobile phase, the detection wavelength was 210 nm, and the column temperature was 30°C.

[0044] 2. Test products

[0045] Calcium Gluconate and Sodium Chloride Injection [Specification] 50ml (calcium gluconate 1g and sodium chloride 0.3375g), batch number: 220915141, 220915142, 220915143, produced by Huaihai Pharmaceutical Factory of Anhui Fengyuan Pharmaceutical Co., Ltd.

[0046] Calcium Gluconate and Sodium Chloride Injection [Specification] 50ml: 1g calcium gluconate and 0.3375g sodium chloride, batch number: 10133, produced by WG Critical Care, LLC.

[0047] The prescription amount of calcium glucarate in both the homemade sample and the reference preparation was 0.09%.

[0048] 3. Reference

[0049] Calcium gluconate: batch number 3.0, content 100%, source EP.

[0050] Calcium glucarate: batch number C2208281, content 100.33%, source Aladdin.

[0051] Example 1: System suitability test

[0052] Calcium gluconate reference solution: Take an appropriate amount of calcium gluconate reference, weigh accurately, dissolve in water and dilute to make a solution containing about 2 mg of calcium gluconate per 1 ml. The chromatogram is as follows: Figure 1 shown.

[0053] Calcium gluconate reference solution: Take an appropriate amount of calcium gluconate reference, weigh it accurately, put it in a beaker, add an appropriate amount of water and heat to dissolve it. After cooling to room temperature, transfer all of it to a volumetric flask, add water to dilute it to make a solution containing about 0.1 mg of calcium gluconate per 1 ml. The chromatogram is as follows Figure 2 shown.

[0054] Accurately measure 20 μl of calcium gluconate reference solution and calcium glucarate reference solution, inject into liquid chromatograph, and record chromatogram. The results show that the peak area RSD (n=6) of calcium gluconate and calcium glucarate is less than 2.0%, and the theoretical plate number of calcium gluconate peak and calcium glucarate peak is greater than 1500, indicating that the system applicability of this method is good. The test results are shown in Table 1.

[0055] Table 1 System suitability results

[0056]

[0057]

[0058] Example 2 Specificity

[0059] Blank solvent: water.

[0060] Blank excipient 1: Accurately measure 1 mL of blank solution without calcium gluconate prepared according to the prescription amount into a 10 mL volumetric flask and dilute to the scale with water.

[0061] Blank excipient 2: Accurately measure 1 mL of blank solution prepared according to the prescription amount without calcium gluconate into a 10 mL volumetric flask and dilute to the scale with water.

[0062] Calcium gluconate positioning solution: Calcium gluconate reference solution: Take an appropriate amount of calcium gluconate reference, weigh accurately, dissolve in water and dilute to make a solution containing about 2 mg of calcium gluconate per 1 ml. The chromatogram is as follows Figure 4 shown.

[0063] Calcium gluconate positioning solution: Take an appropriate amount of calcium gluconate reference substance, weigh it accurately, put it in a beaker, add an appropriate amount of water and heat to dissolve it. After cooling to room temperature, transfer all of it to a volumetric flask, add water to dilute it to make a solution containing about 0.1 mg of calcium gluconate per 1 ml. The chromatogram is as follows Figure 5 shown.

[0064] Specific solution: Take an appropriate amount of calcium gluconate and sodium chloride injection, accurately measure, and dilute with water to make a solution containing about 2 mg of calcium gluconate per 1 ml. The chromatogram is as follows: Figure 3 shown.

[0065] Accurately measure 20 μl of each blank auxiliary material, specific solution, and each positioning solution, inject them into the liquid chromatograph, and record the chromatogram. The results show that the blank solvent and blank auxiliary materials have no interference, the peak purity of calcium gluconate and calcium glucarate is greater than 980, and the separation degree between calcium gluconate and calcium glucarate is 3.60, which is greater than 1.5, indicating that the method has good specificity. The test results are shown in Table 2.

[0066] Table 2 Specificity results

[0067]

[0068] Example 3: Accuracy Test

[0069] Blank excipient (1): Prepare a blank solution without calcium gluconate according to the prescribed amount.

[0070] Blank excipient (2): Prepare a blank solution without calcium glucarate according to the prescribed amount.

[0071] Calcium gluconate reference solution: Take an appropriate amount of calcium gluconate reference, weigh accurately, dissolve in water and dilute to make a solution containing about 2 mg of calcium gluconate per 1 ml;

[0072] Calcium gluconate reference solution: Take an appropriate amount of calcium gluconate reference, weigh it accurately, place it in a beaker, add an appropriate amount of water and heat to dissolve it. After cooling to room temperature, transfer all of it to a volumetric flask and dilute it with water to make a solution containing approximately 0.1 mg of calcium gluconate per 1 ml.

[0073] Recovery solution (calcium gluconate): Take known amounts of calcium gluconate (about 16 mg, 20 mg, and 24 mg), accurately weigh them, and place them in a 10 ml volumetric flask. Add 1 ml of blank auxiliary material (1), dissolve it in water, and dilute to the mark. Shake well and prepare three replicates for each.

[0074] Calcium gluconate stock solution: Take a known amount of calcium gluconate, weigh it accurately, place it in a beaker, add an appropriate amount of water and heat to dissolve it. After cooling to room temperature, transfer it all to a volumetric flask and dilute it with water to make a solution containing about 1 mg of calcium gluconate per 1 ml.

[0075] Recovery solution (calcium gluconate): accurately measure 0.8ml, 1.0ml and 1.2ml of calcium gluconate stock solution into a 10ml volumetric flask, add 1ml of blank auxiliary material (2), dilute to the mark with water, shake well, and prepare three replicates in parallel.

[0076] Take 20 μl of the reference solution and each recovery solution, inject them into the liquid chromatograph, record the chromatogram, and calculate the content by the external standard method. The results show that in 9 recovery solutions of calcium gluconate, the recovery rate is 98% to 101%, the average recovery rate is 99.00%, and the RSD (n=9) is 0.54%, which is less than 2.0%; in 9 recovery solutions of calcium gluconate, the recovery rate of calcium gluconate is 85% to 110%, the average recovery rate is 102.99%, and the RSD (n=9) is 0.20, which is less than 15%; all meet the verification requirements, indicating that the accuracy of this method is good. The test results are shown in Table 3.

[0077] Table 3 Accuracy experimental results

[0078]

[0079]

[0080] Example 4: Repeatability test

[0081] Test solution: Take an appropriate amount of calcium gluconate and sodium chloride injection, accurately measure, and dilute with water to make a solution containing approximately 2 mg of calcium gluconate and 0.1 mg of calcium glucarate per 1 mL.

[0082] Calcium gluconate reference solution: Take an appropriate amount of calcium gluconate reference, weigh accurately, dissolve in water and dilute to make a solution containing approximately 2 mg of calcium gluconate per 1 ml.

[0083] Calcium gluconate reference solution: Take an appropriate amount of calcium gluconate reference, weigh it accurately, place it in a beaker, add an appropriate amount of water and heat to dissolve it. After cooling to room temperature, transfer all of it to a volumetric flask and dilute it with water to make a solution containing approximately 0.1 mg of calcium gluconate per 1 ml.

[0084] Take 20 μl of the above test solution and each reference solution, inject into the liquid chromatograph, and record the chromatogram. The results show that the RSD (n=6) of the calcium gluconate content in the 6 test solutions is 0.10%, less than 1.0%; the RSD (n=6) of the calcium glucarate content is 0.47%, less than 15%, indicating that the method has good repeatability. The test results are shown in Table 4.

[0085] Table 4 Repeatability results

[0086] content 1 2 3 4 5 6 RSD(%) Calcium gluconate (%) 99.22 98.98 99.14 99.02 99.03 98.97 0.10 Calcium glucarate (%) 106.39 106.33 106.28 106.47 105.13 106.25 0.47

[0087] Example 5: Intermediate precision test

[0088] The tests were performed by two analysts on different days using different instruments.

[0089] Test solution: Take an appropriate amount of calcium gluconate and sodium chloride injection, accurately measure, and dilute with water to make a solution containing approximately 2 mg of calcium gluconate and 0.1 mg of calcium glucarate per 1 mL.

[0090] Calcium gluconate reference solution: Take an appropriate amount of calcium gluconate reference, weigh accurately, dissolve and dilute with water to make a solution containing approximately 2 mg of calcium gluconate per 1 ml.

[0091] Calcium gluconate reference solution: Take an appropriate amount of calcium gluconate reference, weigh it accurately, place it in a beaker, add an appropriate amount of water and heat to dissolve, after cooling to room temperature, transfer all to a volumetric bottle, add water to dilute to make a solution containing about 0.1 mg of calcium gluconate per 1 ml.

[0092] Take 20 μl of the above test solution and each reference solution, inject them into the liquid chromatograph, and record the chromatogram. The results show that different personnel, different dates, and different instruments measured 12 test solutions, and the RSD (n=12) of calcium gluconate content was 0.68%, less than 2.0%; the RSD (n=12) of calcium glucarate content was 3.45%, less than 15%, indicating that the intermediate precision of this method is good. The test results are shown in Table 5 below.

[0093] Table 5 Intermediate precision results

[0094]

[0095] Example 6: Linearity and range test

[0096] Calcium gluconate linear solution: Take about 16 mg, 18 mg, 20 mg, 22 mg, and 24 mg of calcium gluconate reference substance respectively, weigh accurately, place in a 10 ml volumetric flask, dissolve with water and dilute to the scale, shake well, and prepare a series of linear solutions with concentration levels of 80%, 90%, 100%, 110%, and 120%. The linear graph is as follows Figure 6 shown.

[0097] Calcium gluconate linear solution: Take about 8 mg, 9 mg, 10 mg, 11 mg, and 12 mg of calcium gluconate reference substance, accurately weigh them, place them in a beaker, add appropriate amount of water and heat to dissolve, cool to room temperature, transfer all to a 100 ml volumetric flask, add water to dilute to the scale, shake well, and prepare a series of linear solutions with concentration levels of 80%, 90%, 100%, 110%, and 120%. The linear graph is shown below. Figure 7 shown.

[0098] Accurately measure 20 μl of each linear solution, inject it into the liquid chromatograph, record the chromatogram, and perform linear regression on the concentration using the peak area. The results show that the linear relationship is good in the concentration range of 1.601-2.409 mg / ml for calcium gluconate and 0.080-0.121 mg / ml for calcium glucarate. The test results are shown in Table 6 below.

[0099] Table 6 Linearity and range test results

[0100]

[0101] Example 7: Durability Test

[0102] The experiments selected column temperatures of 25°C, 30°C, and 35°C; flow rates of 0.8ml / min, 1.0ml / min, and 1.2ml / min; different brands of chromatographic columns, and other corresponding chromatographic conditions remained unchanged.

[0103] Reference solution: Take the reference solution for accuracy test.

[0104] Test solution: Take an appropriate amount of calcium gluconate and sodium chloride injection, accurately measure, and dilute with water to make a solution containing approximately 2 mg of calcium gluconate and 0.1 mg of calcium glucarate per 1 mL.

[0105] Take 20 μl of the reference solution and the test solution, inject them into the liquid chromatograph, record the chromatogram, and calculate the content by the external standard method. The test shows that this method has good durability. The results are shown in Table 7 and Table 8.

[0106] Table 7 Calcium gluconate content determination durability test results

[0107]

[0108]

[0109] Table 8 Calcium gluconate content determination durability test results

[0110]

[0111]

[0112] Example 7: Comparative test

[0113] 1. In the experiment, a salt solution containing an ion pair reagent (tetrabutylammonium hydrogen sulfate) and a salt solution not containing an ion pair reagent (tetrabutylammonium hydrogen sulfate) were selected as mobile phases, and other corresponding chromatographic conditions remained unchanged.

[0114] Test solution: Take an appropriate amount of calcium gluconate and sodium chloride injection (batch number: 220915141), accurately measure, and dilute with water to make a solution containing approximately 2 mg of calcium gluconate and 0.1 mg of calcium glucarate per 1 mL.

[0115] Take 20 μl of the above test solution, inject it into the liquid chromatograph, record the chromatogram, and observe the peaks of calcium gluconate and calcium glucarate.

[0116] The experiment showed that the chromatogram of the mobile phase with a salt solution that does not contain an ion pair reagent (tetrabutylammonium hydrogen sulfate) showed that the retention time of the calcium gluconate peak was advanced, and due to the interference of the blank solvent, the calcium gluconate peak did not appear (the chromatogram is shown in Figure 8 ); The chromatogram of the mobile phase containing the salt solution of the ion pair reagent (tetrabutylammonium hydrogen sulfate) shows that both the calcium gluconate peak and the calcium glucarate peak can be effectively detected, with good specificity (the chromatogram is shown in Fig. 9 ).

[0117] 2. In the experiment, flow rates of 0.4 ml / min, 1.0 ml / min and 2.0 ml / min were selected respectively, and other corresponding chromatographic conditions remained unchanged.

[0118] Test solution: Take an appropriate amount of calcium gluconate and sodium chloride injection (batch number: 220915141), accurately measure, and dilute with water to make a solution containing approximately 2 mg of calcium gluconate and 0.1 mg of calcium glucarate per 1 mL.

[0119] Take 20 μl of the above test solution, inject it into the liquid chromatograph, record the chromatogram, and observe the peaks of calcium gluconate and calcium glucarate.

[0120] The experiment showed that changing the flow rate had a significant effect on the peak shape of the calcium gluconate peak and the calcium glucarate peak. The peak shape was good in the chromatogram at a flow rate of 1.0 ml / min (see the chromatogram for details). Fig.10 ), flow rate 0.4ml / min (the color spectrum is shown in Fig.10 ) and 2.0 ml / min, the two main peaks are wrapped together and cannot be effectively separated.

[0121] 3. In the experiment, column temperatures of 20℃, 30℃ and 40℃ were selected respectively, and other corresponding chromatographic conditions remained unchanged.

[0122] Test solution: Take an appropriate amount of calcium gluconate and sodium chloride injection (batch number: 220915141), accurately measure, and dilute with water to make a solution containing approximately 2 mg of calcium gluconate and 0.1 mg of calcium glucarate per 1 mL.

[0123] Take 20 μl of the above test solution, inject it into the liquid chromatograph, record the chromatogram, and observe the peaks of calcium gluconate and calcium glucarate.

[0124] The experiment showed that changing the column temperature had a significant effect on the determination of calcium gluconate and calcium glucarate. The contents of calcium gluconate and calcium glucarate were normal at a column temperature of 30°C; the content of calcium glucarate was about 10% lower at a column temperature of 20°C; and the content of calcium gluconate was about 10% higher at a column temperature of 40°C.

[0125] Example 8: Sample determination

[0126] According to the above-mentioned high performance liquid chromatography detection method, the batch numbers of different manufacturers were respectively 220915141 (the chromatograms are shown in FIG. Fig. 9 ), 220915142, 220915143, 10133 (the chromatogram is as shown in Fig.11 The contents of calcium gluconate and calcium glucarate in the (shown) batches of calcium gluconate and sodium chloride injections were measured. The results are shown in Table 9.

[0127] Table 9 Sample test results

[0128]

[0129] The preferred embodiments of the present invention are described in detail above, but the present invention is not limited thereto. Within the technical concept of the present invention, the technical solution of the present invention can be subjected to a variety of simple modifications, including the combination of various technical features in any other suitable manner, and these simple modifications and combinations should also be regarded as the contents disclosed by the present invention and belong to the protection scope of the present invention.

Claims

1. A method for simultaneously determining the content of calcium gluconate and calcium glucarate in medicines, It is characterized in that The method is a high performance liquid chromatography method, and the chromatographic conditions of the high performance liquid chromatography method are: a mixed solution of sodium dihydrogen phosphate solution and tetrabutylammonium hydrogen sulfate solution is used as a mobile phase, and a detection wavelength is 208nm-212nm.

2. The method according to claim 1, It is characterized in that The concentration of sodium dihydrogen phosphate in the mobile phase is 0.01-0.04 mol / L, and the concentration of tetrabutylammonium hydrogen sulfate is 0.003-0.007 mol / L; Preferably, the concentration of sodium dihydrogen phosphate in the mobile phase is 0.015-0.035 mol / L, and the concentration of tetrabutylammonium hydrogen sulfate is 0.004-0.006 mol / L; More preferably, the concentration of sodium dihydrogen phosphate in the mobile phase is 0.025 mol / L, and the concentration of tetrabutylammonium hydrogen sulfate is 0.005 mol / L.

3. The method according to claim 1, It is characterized in that The pH of the mobile phase is 1-4, preferably 2-3, and more preferably 2.

5.

4. The method according to claim 1 or 2, It is characterized in that The high performance liquid chromatography uses octadecylsilane bonded silica gel as a filler; and / or The flow rate of the mobile phase is 0.8-1.2 ml / min, preferably, the flow rate of the mobile phase is 0.9-1.0 ml / min.

5. The method according to claim 4, It is characterized in that The flow rate of the mobile phase is 1 ml / min.

6. The method according to claim 4, It is characterized in that The detection wavelength is 210 nm; the column temperature is 20°C-40°C, preferably 30°C.

7. The method according to claim 4, It is characterized in that The injection volume during detection was 20 μl.

8. Application of the method according to any one of claims 1 to 7 in measuring the content of calcium gluconate and calcium glucarate in calcium gluconate and sodium chloride injection.

9. A method for determining the content of calcium gluconate and calcium glucarate in calcium gluconate and sodium chloride injection, It is characterized in that The following steps are involved: (1) Take calcium gluconate and sodium chloride injection, dilute with water to make a solution containing about 2 mg of calcium gluconate and 0.1 mg of calcium glucarate per 1 ml, as the test solution, accurately measure 20 μl, inject into the liquid chromatograph, and record the chromatogram; (2) Dissolve calcium gluconate in water and quantitatively dilute to prepare a solution containing about 2 mg of calcium gluconate per 1 ml, as the calcium gluconate reference solution; (3) diluting calcium gluconate with water to prepare a solution containing about 0.1 mg of calcium gluconate per 1 ml as the calcium gluconate reference solution; (4) Calculate the contents of calcium gluconate and calcium glucarate in calcium gluconate and sodium chloride injection by the external standard method using the peak areas; The chromatographic conditions are as follows: octadecylsilane bonded silica gel is used as filler, a mixture of sodium dihydrogen phosphate solution and tetrabutylammonium hydrogen sulfate solution is used as mobile phase, and the concentration of sodium dihydrogen phosphate in the mobile phase is 0.01-0.04 mol / L, the concentration of tetrabutylammonium hydrogen sulfate is 0.003-0.007 mol / L, the flow rate is 0.8-1.2 ml / min; the detection wavelength is 208 nm-212 nm, and the column temperature is 20°C-40°C; Preferably, the chromatographic conditions are: using octadecylsilane bonded silica gel as a filler, using a mixture of sodium dihydrogen phosphate solution and tetrabutylammonium hydrogen sulfate solution as a mobile phase, wherein the concentration of sodium dihydrogen phosphate in the mobile phase is 0.025 mol / L, the concentration of tetrabutylammonium hydrogen sulfate is 0.005 mol / L, and the flow rate is 1 ml / min; The detection wavelength was 210 nm and the column temperature was 30 °C.

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