A calcium zinc gluconate oral solution composition and its detection method

Abstract

The present invention provides a calcium zinc gluconate oral solution composition and a detection method thereof, belonging to the technical field of pharmaceutical preparations. The calcium zinc gluconate oral solution composition of the present invention comprises calcium gluconate, zinc gluconate, lysine hydrochloride, lactic acid, a preservative, water and other pharmaceutically acceptable excipients. The preservative is potassium sorbate, sodium diacetate and theanine. Through the mutual cooperation of each component, the obtained calcium zinc gluconate oral solution composition has excellent quality and good stability. The detection method of the present invention can significantly distinguish the advantages and disadvantages of this product in various quality attributes, can effectively and comprehensively evaluate the product quality, has high accuracy, good repeatability, short running time, convenient and fast derivatization method, and saves costs.

Description

Technical Field

[0001] The present invention belongs to the technical field of pharmaceutical preparations, and particularly relates to a calcium gluconate and zinc oral solution composition and a detection method thereof. Background Art

[0002] Calcium gluconate and zinc oral solution is a compound preparation composed of calcium gluconate, zinc gluconate, lysine hydrochloride, etc., and is suitable for the treatment of diseases caused by calcium and zinc deficiency. Calcium gluconate and zinc oral solution is the first zinc-calcium supplement product in China to use lysine as an absorption promoter, which can double the promotion of calcium absorption and prevent calcium loss. At present, several calcium gluconate and zinc oral solutions have been approved for marketing in China, but there are the following problems with calcium gluconate and zinc oral solution at present: (1) The product is in a supersaturated state, with poor stability during long-term storage, and is prone to precipitation, which does not meet the legal standards and cannot meet the quality requirements; (2) The quality control of drugs covers all aspects of the production process, and the legal standards and control methods are difficult to meet the quality evaluation of products.

[0003] Chinese Patent CN103191092A discloses a pharmaceutical composition with calcium and zinc supplement effects, including calcium gluconate, zinc gluconate, L-lysine hydrochloride and L-lactic acid. The content of the impurity tyrosine in L-lysine hydrochloride and the content of the impurity DL-lactic acid in L-lactic acid have a direct impact on the quality of the preparation. Chinese Patent CN104800199A provides a stable calcium gluconate and zinc oral solution composition and a preparation method thereof. The composition contains calcium gluconate, zinc gluconate, L-lysine hydrochloride, lactic acid, and flavoring agents, preservatives, and fragrances can also be added. The obtained calcium gluconate and zinc oral solution composition has the advantages of excellent quality, good stability, and simple preparation process.

[0004] Chinese Patent CN114965790A discloses a liquid phase detection method for amino acid impurities in lysine zinc gluconate granules, which can effectively quantify 6 amino acid impurities (including arginine, threonine, proline, phenylalanine, isoleucine, leucine) in lysine zinc gluconate granules. However, due to the differences in excipients, flavors, and processes in the preparation, this method is not applicable to the detection of amino acids in calcium gluconate and zinc oral solution. Chinese Patent CN112666282A discloses a high performance liquid phase detection method for impurities in calcium gluconate and zinc oral solution, but its chromatographic running time is long, the volume of solution preparation is large, which is not convenient for large-scale detection, and the detailed parameters of derivatization are not clear.

[0005] Therefore, it is still necessary to prepare a new calcium gluconate and zinc oral solution composition that can maintain stable quality within the validity period, meet the quality requirements, ensure the safety and effectiveness of the product, and how to sensitively and conveniently detect the key impurities in calcium gluconate and zinc oral solution and make a quality evaluation of the product is also an urgent problem to be solved. Summary of the Invention

[0006] The object of the present invention is to provide a calcium gluconate and zinc gluconate oral solution composition in view of the deficiencies of the prior art, which has good quality and high stability, and no precipitation will occur during long-term storage of the product.

[0007] The present invention also provides a detection method for the calcium gluconate and zinc gluconate oral solution composition, which can significantly distinguish the quality advantages and disadvantages of this product, effectively and comprehensively evaluate the product quality, with high accuracy, good repeatability, short running time, convenient and fast derivatization method, and cost saving.

[0008] To achieve the above object, the technical solutions adopted by the present invention are as follows:

[0009] A calcium gluconate and zinc gluconate oral solution composition, comprising calcium gluconate, zinc gluconate, lysine hydrochloride, lactic acid, preservatives, water and other pharmaceutically acceptable excipients; wherein,

[0010] The content of the lactic acid is 6.50 - 16.00 mg / mL;

[0011] The preservatives are potassium sorbate, sodium diacetate and theanine.

[0012] Furthermore, the content of lactic acid in the calcium gluconate and zinc gluconate oral solution composition is 8.00 - 12.50 mg / mL.

[0013] Furthermore, among the preservatives, the weight ratio of potassium sorbate, sodium diacetate and theanine is 1:0.4 - 0.6:0.1 - 0.3.

[0014] Preferably, among the preservatives, the weight ratio of potassium sorbate, sodium diacetate and theanine is 1:0.5:0.2.

[0015] Furthermore, the water refers to purified water, distilled water or water for injection, preferably purified water.

[0016] Furthermore, the other pharmaceutically acceptable excipients are one or a mixture of several of flavoring agents, fragrances, pH regulators, and thickening agents.

[0017] Even further, the flavoring agent is one or a mixture of several of sorbitol, xylitol, mannitol, sucrose, sucralose, stevioside, sodium saccharin, aspartame, and acesulfame potassium. Preferably, the flavoring agent is acesulfame potassium and / or acesulfame potassium.

[0018] Furthermore, the essence includes but is not limited to one or a mixture of more than one of apple essence, strawberry essence, grape essence, mango essence, pineapple essence, blueberry essence, milk essence, peach essence, cantaloupe essence, orange essence, tangerine essence, and lemon essence. Preferably, the essence is grape essence, strawberry essence, lemon essence, orange essence, peach essence, and / or apple essence.

[0019] Furthermore, the pH regulator includes but is not limited to one or a mixture of more than one of hydrochloric acid, phosphoric acid and its phosphates, acetic acid and its acetates, citric acid and its citrates, carbonic acid and its carbonates, boric acid and its borates, malic acid, sodium hydroxide, and potassium hydroxide. Preferably, the pH regulator is sodium carbonate and / or disodium hydrogen phosphate.

[0020] Furthermore, the thickener includes but is not limited to one or a mixture of more than one of glycerol, hypromellose, polyethylene glycol, polyvinylpyrrolidone, polyvinyl alcohol, β-cyclodextrin, carbomer, and propylene glycol. Preferably, the thickener is glycerol and / or polyvinyl alcohol.

[0021] Further, the calcium zinc gluconate oral solution composition includes 55.0 - 70.0 mg / mL of calcium gluconate, 2.5 - 3.5 mg / mL of zinc gluconate, 8.5 - 11.0 mg / mL of lysine hydrochloride, 6.5 - 16.0 mg / mL of lactic acid, 1.0 - 2.0 mg / mL of preservative, 0 - 1.6 mg / mL of flavoring agent, and 0 - 0.5 mg / mL of essence, with the balance being water.

[0022] Furthermore, the calcium zinc gluconate oral solution composition includes 60.0 - 65.0 mg / mL of calcium gluconate, 3.0 - 3.5 mg / mL of zinc gluconate, 9.5 - 10.0 mg / mL of lysine hydrochloride, 6.5 - 8 mg / mL of lactic acid, 1.0 - 1.5 mg / mL of preservative, 0.8 - 1.6 mg / mL of flavoring agent, and 0.3 - 0.5 mg / mL of essence, with the balance being water.

[0023] Preferably, the calcium zinc gluconate oral solution composition includes 60.0 mg / mL of calcium gluconate, 3.0 mg / mL of zinc gluconate, 1.0 mg / mL of lysine hydrochloride, 6.5 mg / mL of lactic acid, 1.4 mg / mL of preservative, 1.6 mg / mL of flavoring agent, and 0.3 mg / mL of essence, with the balance being water.

[0024] The present invention also provides a preparation method of the above calcium zinc gluconate oral solution composition, including the following steps:

[0025] (1) Dissolve calcium gluconate and zinc gluconate in water at 80 - 95 °C to obtain Solution I;

[0026] (2) Cool the Solution I in step (2) to 45°C or below, add other components, and dissolve to obtain Solution II;

[0027] (3) Make the volume of Solution II in step (2) up to the full volume, mix well, filter, and seal to obtain the product.

[0028] Furthermore, the filtration refers to filtration with a microporous membrane having a pore size ≤ 1 μm, preferably filtration with a microporous membrane having a pore size of 0.8 μm.

[0029] Preferably, the preparation method of the calcium zinc gluconate oral solution composition comprises the following steps:

[0030] (1) Take purified water accounting for 80% of the total mass of calcium gluconate and zinc gluconate, heat it to 90°C, add calcium gluconate and zinc gluconate, and dissolve to obtain Solution I;

[0031] (2) Cool the Solution I in step (2) to 45°C or below, add other components, and dissolve to obtain Solution II;

[0032] (3) Make the volume of Solution II in step (2) up to 1 L, mix well, filter, and seal to obtain the product.

[0033] The present invention also provides an HPLC detection method for related substances of a calcium zinc gluconate oral solution, comprising the following steps:

[0034] (1) Perform derivatization treatment on the above-mentioned calcium zinc gluconate oral solution composition to obtain a test solution;

[0035] (2) Analyze the test solution in step (1) by high performance liquid chromatography, and the analysis conditions are as follows:

[0036] Chromatographic column: C18 chromatographic column (4.6 mm × 250 cm, 5 μm)

[0037] Mobile phase: Mobile phase A is a 40 - 55 mmol / L sodium acetate buffer solution containing 1% N,N-dimethylformamide, pH = 4.6 - 5.2; Mobile phase B is an acetonitrile-methanol-aqueous solution; Mobile phase C is an acetonitrile-aqueous solution;

[0038] Detection wavelength: 360 nm;

[0039] Column temperature: 25 - 30°C;

[0040] Flow rate: 1 - 1.5 mL / min;

[0041] The mobile phase running gradient is as follows:

[0042]

[0043] Further, in step (1), the derivatization treatment step includes:

[0044] Mix the calcium zinc gluconate oral solution composition with sodium carbonate solution and 2,4-dinitrofluorobenzeneacetonitrile solution, heat in the dark, cool, dilute to the mark with phosphate buffer solution, and shake well to obtain.

[0045] Furthermore, the volume ratio of the calcium zinc gluconate oral solution composition, sodium carbonate solution, and 2,4-dinitrofluorobenzeneacetonitrile solution is 1:1 - 2:1 - 1.2. Preferably, the volume ratio of the calcium zinc gluconate oral solution composition, sodium carbonate solution, and 2,4-dinitrofluorobenzeneacetonitrile solution is 1:1:1.

[0046] Furthermore, the mass concentration of the sodium carbonate solution is 4 - 6%; preferably, the concentration of the sodium carbonate solution is 5%.

[0047] Furthermore, the volume concentration of the 2,4-dinitrofluorobenzeneacetonitrile solution is 1 - 2%; preferably, the volume concentration of the 2,4-dinitrofluorobenzeneacetonitrile solution is 1%.

[0048] Furthermore, the temperature of the heat treatment is 50 - 60°C, and the time is 40 - 60 min; preferably, the temperature of the heat treatment is 60°C, and the time is 60 min.

[0049] Preferably, the pH of the phosphate buffer solution is 7.0.

[0050] Preferably, the derivatization treatment step includes:

[0051] Precisely measure 1.0 mL of the calcium zinc gluconate oral solution composition, place it in a 10 mL volumetric flask, precisely measure 1.0 mL of 5% sodium carbonate solution and 1.0 mL of 2,4-dinitrofluorobenzeneacetonitrile solution, place them in the same volumetric flask, shake well, place in the dark, heat in a water bath at 60°C for 60 min, take out, cool, dilute to the mark with phosphate buffer solution with a pH of 7.0, and shake well to obtain.

[0052] Further, in step (2), the chromatographic column is a Kromasil 100 - 5 - C18 chromatographic column.

[0053] Further, in step (2), the mobile phase is: Mobile phase A is 50 mmol / L sodium acetate buffer solution containing 1% N,N-dimethylformamide, pH = 5.0; Mobile phase B is acetonitrile - methanol - water solution; Mobile phase C is acetonitrile - water solution.

[0054] Further, the volume percentage of the organic phase (acetonitrile, methanol) in the mobile phase B is 48-52%; preferably, the volume ratio of acetonitrile, methanol and water in the mobile phase B is 25:25:50.

[0055] Further, the volume percentage of acetonitrile in the mobile phase C is 48-52%; preferably, the volume ratio of acetonitrile and water in the mobile phase C is 50:50.

[0056] Further, in step (2), the column temperature is 27 °C.

[0057] Further, in step (2), the flow rate is 1.2 mL / min.

[0058] The beneficial effects of the present invention are as follows:

[0059] (1) The calcium zinc gluconate oral solution composition provided by the present invention improves the product stability and quality by controlling the lactic acid content in the prescription. When stored at 60 °C and 2-8 °C for 30 days, and at 40 °C and 25 °C for 6 months, the product remains a colorless or slightly yellowish clear liquid, the solution is clear without precipitation, the color is not darker than the yellow No. 3 standard colorimetric solution, and in the related substances, threonine ≤ 0.2%, leucine ≤ 0.2%, phenylalanine ≤ 0.7%, meeting the requirements of the internal control quality standard and the standard WS1-XG-008-2001;

[0060] (2) The present invention uses potassium sorbate, sodium diacetate and theanine as preservatives, which not only have an antiseptic and antibacterial effect, but also can improve the stability of the product to a certain extent and extend the storage period;

[0061] (3) In the process of preparing the calcium zinc gluconate oral solution composition of the present invention, calcium gluconate and zinc gluconate are first heated under high temperature conditions, and then other components are added after cooling, which helps to improve the stability of the product;

[0062] (4) The high performance liquid chromatography detection method for the related substances of the calcium zinc gluconate oral solution provided by the present invention has a short running time, high accuracy, good repeatability, and a convenient, fast and cost-saving derivatization method; a quality evaluation method and standard for this product have been established, which can comprehensively and effectively evaluate the product quality and ensure the safety and effectiveness of clinical medication. Specific Embodiments

[0063] The following non-limiting examples can enable those of ordinary skill in the art to understand the present invention more comprehensively, but do not limit the present invention in any way. The following content is only an exemplary illustration of the scope claimed by the present invention. Those skilled in the art can make various changes and modifications to the invention according to the disclosed content, and it should also fall within the scope claimed by the present invention.

[0064] The present invention will be further described below by way of specific embodiments. All kinds of chemical reagents used in the embodiments of the present invention are obtained through conventional commercial channels unless otherwise specified.

[0065] Example 1

[0066] Calcium Gluconate and Zinc Gluconate Oral Solution Composition

[0067] Prescription, see Table 1 below:

[0068] Table 1

[0069] Calcium gluconate 60g Zinc gluconate 3g Lysine hydrochloride 10g Lactic acid 6.5g Preservative 1.4g Acesulfame potassium 0.8g Aspartame 0.8g Peach essence 0.3g Purified water Weigh to 1 L

[0070] Among them, the preservative is potassium sorbate, sodium diacetate and theanine with a weight ratio of 1:0.5:0.2.

[0071] Preparation method:

[0072] (1) Take 80% of the purified water in the prescription amount, heat it to 90 °C, add calcium gluconate and zinc gluconate to dissolve it to obtain Solution I;

[0073] (2) Cool the Solution I in step (1) to 45 °C or below, add other components, dissolve it to obtain Solution II;

[0074] (3) Make up the purified water to 1 L in the Solution II in step (2), mix well, filter and seal to obtain the product.

[0075] Example 2

[0076] Calcium Gluconate and Zinc Gluconate Oral Solution Composition

[0077] Prescription, see Table 2 below:

[0078] Table 2

[0079] Calcium gluconate 60g Zinc gluconate 3g Lysine hydrochloride 10g Lactic acid 8g Preservative 1.4g Aspartame 0.8g Acesulfame potassium 0.8g Acesulfame potassium 0.8g Purified water Weigh to 1 L

[0080] Among them, the preservative is potassium sorbate, sodium diacetate and theanine with a weight ratio of 1:0.5:0.2.

[0081] The preparation method is the same as that of Example 1.

[0082] Example 3

[0083] Calcium Gluconate and Zinc Gluconate Oral Solution Composition

[0084] Prescription, see Table 3 below:

[0085] Table 3

[0086]

[0087]

[0088] Among them, the preservative is potassium sorbate, sodium diacetate and theanine with a weight ratio of 1:0.5:0.2. The preparation method is the same as that of Example 1.

[0089] Example 4

[0090] Calcium Gluconate and Zinc Oral Solution Composition

[0091] Prescription, see Table 4 below:

[0092] Table 4

[0093] Calcium gluconate 60g Zinc gluconate 3g Lysine hydrochloride 10g Lactic acid 16g Preservative 1.4g Aspartame 0.8g Acesulfame potassium 0.8g Peach essence 0.3g Add purified water to Weigh to 1 L

[0094] Among them, the preservative is potassium sorbate, sodium diacetate and theanine with a weight ratio of 1:0.5:0.2. The preparation method is the same as that of Example 1.

[0095] Comparative Example 1

[0096] Calcium Gluconate and Zinc Oral Solution Composition

[0097] Prescription, see Table 5 below:

[0098] Table 5

[0099]

[0100]

[0101] Among them, the preservative is potassium sorbate, sodium diacetate and theanine with a weight ratio of 1:0.5:0.2. The preparation method is the same as that of Example 1.

[0102] Comparative Example 2

[0103] Calcium Gluconate and Zinc Oral Solution Composition

[0104] Prescription, see Table 6 below:

[0105] Table 6

[0106] Calcium gluconate 60g Zinc gluconate 3g Lysine hydrochloride 10g Lactic acid 20g Preservative 1.4g Aspartame 0.8g Acesulfame potassium 0.8g Peach essence 0.3g Purified water Weigh to 1 L

[0107] Among them, the preservative is potassium sorbate, sodium diacetate and theanine with a weight ratio of 1:0.5:0.2.

[0108] The preparation method is the same as that of Example 1.

[0109] Comparative Example 3

[0110] Calcium Gluconate and Zinc Oral Solution Composition

[0111] Prescription, see Table 7 below:

[0112] Table 7

[0113]

[0114]

[0115] Among them, the preservative is sodium diacetate.

[0116] The preparation method is the same as that of Example 1.

[0117] Comparative Example 4

[0118] Calcium zinc gluconate oral solution composition

[0119] Prescription, see Table 8 below:

[0120] Table 8

[0121] Calcium gluconate 60g Zinc gluconate 3g Lysine hydrochloride 10g Lactic acid 8g Preservative 1.4g Aspartame 0.8g Acesulfame potassium 0.8g Peach essence 0.3g Purified water Weigh to 1 L

[0122] Among them, the preservative is potassium sorbate.

[0123] The preparation method is the same as that of Example 1.

[0124] Comparative Example 5

[0125] Calcium zinc gluconate oral solution composition

[0126] Prescription, see Table 9 below:

[0127] Table 9

[0128]

[0129]

[0130] Among them, the preservative is L-theanine.

[0131] The preparation method is the same as that of Example 1.

[0132] Comparative Example 6

[0133] Calcium zinc gluconate oral solution composition

[0134] Prescription, see Table 10 below:

[0135] Table 10

[0136] Calcium gluconate 60g Zinc gluconate 3g Lysine hydrochloride 10g Lactic acid 8g Preservative 1.4g Aspartame 0.8g Acesulfame potassium 0.8g Peach essence 0.3g Purified water Weigh to 1 L

[0137] Among them, the preservative is sodium benzoate.

[0138] The preparation method is the same as that of Example 1.

[0139] Comparative Example 7

[0140] Calcium Gluconate and Zinc Gluconate Oral Solution Composition

[0141] The prescription is the same as that of Example 2;

[0142] Preparation method:

[0143] (1) Take 80% of the purified water in the prescription amount, heat it to 90 °C, add calcium gluconate, zinc gluconate, lysine hydrochloride, lactic acid, preservative, aspartame, acesulfame potassium and peach essence and dissolve to obtain Solution I;

[0144] (2) Cool the Solution I in step (1) to 45 °C or below, make up the volume to 1 L with purified water, mix well, filter and seal to obtain the product.

[0145] Comparative Example 8

[0146] Calcium Gluconate and Zinc Gluconate Oral Solution Composition

[0147] The prescription is the same as that of Example 2;

[0148] Preparation method:

[0149] (1) Take 80% of the purified water in the prescription amount, add lactic acid and dissolve to obtain Solution I;

[0150] (2) Heat the material I in step (1) to 90 °C, add calcium gluconate and dissolve, stop heating to obtain Material II;

[0151] (3) Add other components to the Material II in step (2), dissolve, make up the volume to 1 L with purified water, stir evenly to obtain the medicinal liquid;

[0152] (4) Filter the medicinal liquid and then seal to obtain the product.

[0153] Experimental Example 1

[0154] Stability Study of Calcium Gluconate and Zinc Gluconate Oral Solution Composition

[0155] 1. Place the calcium gluconate and zinc gluconate oral solution compositions obtained in Examples 1-4 and Comparative Examples 1-8 at 60 °C for 30 days. Use the indicators of appearance, pH value, clarity and color, and related substances to comprehensively evaluate the quality of this product during the stability period in combination with the standard limit requirements. The appearance, pH value, clarity and color are detected with reference to the general rules of the Chinese Pharmacopoeia, and the related substances are detected according to the method described in Example 5 below. The measured results are shown in Table 11 below.

[0156] Table 11. Results of 60 °C Stability Experiment

[0157]

[0158]

[0159]

[0160] The calcium zinc gluconate oral solution compositions obtained in Examples 1-4 and Comparative Examples 1-8 were placed under the conditions of 40°C and a humidity of 25% RH for 6 months, and the quality during the stability period of this product was comprehensively evaluated by using the indicators of appearance, clarity and related substances in combination with the standard limit requirements. The results are shown in Table 12 below.

[0161] Table 12. Results of the stability experiment at 40°C

[0162]

[0163]

[0164] The calcium zinc gluconate oral solution compositions obtained in Examples 1-4 and Comparative Examples 1-8 were placed under the conditions of 25°C and a humidity of 65% RH for 6 months and under the conditions of 2-8°C for 30 days, and the quality during the stability period of this product was comprehensively evaluated by using the indicators of appearance and clarity in combination with the standard limit requirements. The results are shown in Table 13 below.

[0165] Table 13. Results of the stability experiments at 25°C and 2-8°C

[0166]

[0167]

[0168] The results show that the appearance, pH value, clarity and color, and related substances of the calcium zinc gluconate oral solution of the examples meet the limit requirements after being placed at 60°C for 30 days, 40°C for 6 months, 25°C for 6 months, and 2-8°C for 30 days, with good quality and high stability; while the calcium zinc gluconate oral solution of the comparative examples shows phenomena such as deepening of color, precipitation, and exceeding the limit of related substances under the above conditions, with poor stability and quality not meeting the standard. It can be seen that the calcium zinc gluconate oral solution provided by the present invention has high stability.

[0169] Example 5

[0170] The HPLC method for detecting related substances of calcium zinc gluconate oral solution comprises the following steps:

[0171] (1) Derivative treatment: Measure 1.0 mL of the calcium zinc gluconate oral solution composition, place it in a 10 mL volumetric flask, accurately measure 1.0 mL of 5% sodium carbonate solution and 1.0 mL of 2,4-dinitrofluorobenzene acetonitrile solution, place them in the same volumetric flask, shake well, take it out after heating in a water bath at 60 °C for 60 min, let it cool, dilute to the scale with phosphate buffer solution with a pH of 7.0, shake well to obtain the test solution;

[0172] (2) Analyze the test solution in step (1) by high performance liquid chromatography, and the analysis conditions are as follows:

[0173] Chromatographic column: Kromasil 100-5-C18 (4.6 mm × 250 cm, 5 μm);

[0174] Mobile phase: Mobile phase A is 50 mmol / L sodium acetate buffer solution containing 1% N,N-dimethylformamide, pH = 5.0; Mobile phase B is acetonitrile-methanol-water solution (the volume ratio of acetonitrile, methanol and water is 25:25:50); Mobile phase C is acetonitrile-water solution (the volume ratio of acetonitrile and water is 50:50);

[0175] Detection wavelength: 360 nm;

[0176] Column temperature: 27 °C;

[0177] Flow rate: 1.2 mL / min;

[0178] The running gradient is as shown in Table 14:

[0179] Table 14. Running gradient

[0180]

[0181] Experimental Example 3

[0182] Study on the HPLC detection method for related substances of calcium zinc gluconate oral solution

[0183] 1. Repeatability

[0184] Preparation of reference solution: Accurately weigh 10 mg of threonine reference substance, 35 mg of phenylalanine reference substance and 10 mg of leucine reference substance and place them in a 100 mL volumetric flask, dissolve with water and dilute to the scale, shake well to obtain the reference stock solution; Take 5 mL of the reference stock solution respectively and place them in 100 mL volumetric flasks, dilute with water and make up to the scale, shake well and then carry out derivative treatment (the method is the same as that in Example 5) to obtain the reference solution.

[0185] Preparation of test sample solution: Accurately measure appropriate amounts of the control solutions of threonine, phenylalanine, and leucine, place them in a 100 mL volumetric flask, add the composition of Example 1, shake well, take 5 mL from it and transfer it to a 100 mL volumetric flask, then dilute to the scale with the composition of Example 1, shake well and perform derivatization treatment (the same method as in Example 5) to obtain the test sample solution.

[0186] Prepare 6 parallel test sample solutions, inject the control solution and the test sample solution into the liquid chromatograph respectively, and perform analysis according to the analysis conditions of Example 5. Calculate the impurity content and RSD value by the external standard method based on the peak area. The results are shown in Table 15 below.

[0187] Table 15. Results of repeatability experiment

[0188] Name 1 2 3 4 5 6 Average value RSD Threonine 0.19％ 0.19％ 0.19％ 0.19％ 0.19％ 0.19％ 0.19％ 0.00％ Phenylalanine 0.70％ 0.71％ 0.71％ 0.71％ 0.71％ 0.71％ 0.71％ 0.00％ Leucine 0.19％ 0.20％ 0.20％ 0.20％ 0.20％ 0.19％ 0.20％ 2.63％

[0189] The results show that the content differences of threonine, phenylalanine, and leucine in the 6 test sample solutions to be measured are small, and the RSD value is small. It can be seen that the HPLC detection method for related substances of the calcium zinc gluconate oral solution provided by the present invention has good repeatability.

[0190] 2. Linearity

[0191] Preparation of linear solution: Accurately measure 5 mg of threonine control, 18 mg of phenylalanine control, and 5 mg of leucine control, place them in a 100 mL volumetric flask, dissolve with water and dilute to the scale, shake well to obtain the control stock solution; then take 0.5 mL, 1 mL, 2 mL, 3 mL, 4 mL, and 6 mL of the control stock solution respectively and place them in 100 mL volumetric flasks, dilute to the scale with water, shake well and perform derivatization treatment (the same method as in Example 5) to obtain the linear solution.

[0192] Inject the linear solution into the liquid chromatograph respectively, the analysis conditions are the same as in Example 5, and record the chromatogram. Perform linear regression with the concentration as the abscissa and the peak area as the ordinate to obtain the linear equations of the three amino acids as follows:

[0193] Threonine: The linear regression equation is y = 0.8818x + 0.0008; it has good linearity in the concentration range of 0.1005 - 1.5123 μg / mL, and the quantitative limit concentration is 0.1005 μg / mL.

[0194] Leucine: The linear regression equation is y = 1.1050x - 0.0020; it has good linearity in the concentration range of 0.0255 - 1.5393 μg / mL, and the quantitative limit concentration is 0.0255 μg / mL.

[0195] Phenylalanine: The linear regression equation is y = 0.9057x - 0.0055; it has a good linearity in the concentration range of 0.0347 - 5.2818 μg / mL, and the quantitative limit concentration is 0.0347 μg / mL.

[0196] 3. Accuracy

[0197] Accurately weigh 10 mg of phenylalanine into a 100-mL volumetric flask, dissolve it with the composition of Example 1 and dilute to the mark, shake well. Take 5 mL from it and place it in a 100-mL volumetric flask, dilute it to the mark with the composition of Example 1, and shake well to obtain the recovery reserve solution.

[0198] Take 1 mL, 2 mL, and 3 mL of the recovery reserve solution, each in 3 portions, into 10-mL volumetric flasks, make up the volume to the mark with the composition of Example 1, mix well and then perform derivatization treatment (the same method as in Example 5) to obtain low, medium, and high concentration solutions. Inject the reference solution (prepared in the same way as in Experimental Example 2) and the low, medium, and high concentration solutions into the liquid chromatograph respectively, and the analysis conditions are the same as in Example 5. The results are shown in Table 16 below.

[0199] Table 16. Results of the accuracy experiment

[0200]

[0201] In this invention, phenylalanine was taken as an example to investigate the accuracy of the HPLC detection method for the related substances of calcium zinc gluconate oral solution. The results showed that the recoveries of phenylalanine at various concentrations were between 98 - 103% at three concentrations, and the RSD% was 1.40%, indicating that the detection method of this invention has good recoveries and high accuracy.

[0202] The above description of the embodiments is for the convenience of those of ordinary skill in the art to understand and use the invention. It is obvious that those skilled in the art can easily make various modifications to these embodiments and apply the general principles described herein to other embodiments without creative labor. Therefore, the present invention is not limited to the above embodiments, and the improvements and modifications made by those skilled in the art without departing from the scope of the present invention should be within the protection scope of the present invention.

Claims

1. A calcium zinc gluconate oral solution composition, characterized in that, Containing 55.0 - 70.0 mg / mL calcium gluconate, 2.5 - 3.5 mg / mL zinc gluconate, 8.5 - 11.0 mg / mL lysine hydrochloride, 6.5 - 16.0 mg / mL lactic acid, 1.0 - 2.0 mg / mL preservative, 0 - 1.6 mg / mL flavoring agent and 0 - 0.5 mg / mL essence, with the balance being water; In the said preservative, the weight ratio of potassium sorbate, sodium diacetate and theanine is 1:0.4 - 0.6:0.1 - 0.3; The preparation method of the calcium gluconate and zinc gluconate oral solution composition comprises the following steps: (1) Dissolve calcium gluconate and zinc gluconate in water at 80 - 95 °C to obtain Solution I; (2) Cool the Solution I in step (1) to 45 °C or below, add other components, dissolve to obtain Solution II; (3) Make up the volume of Solution II in step (2) to the full volume, mix well, filter and seal to obtain the product.

2. The calcium zinc gluconate oral solution composition according to claim 1, characterized in that, The content of the said lactic acid is 8.00 - 12.50 mg / mL.

3. The calcium zinc gluconate oral solution composition according to claim 2, characterized in that, In the said preservative, the weight ratio of potassium sorbate, sodium diacetate and theanine is 1:0.5:0.

2.

4. The calcium zinc gluconate oral solution composition according to claim 1, wherein Containing 60.0 - 65.0 mg / mL calcium gluconate, 3.0 - 3.5 mg / mL zinc gluconate, 9.5 - 10.0 mg / mL lysine hydrochloride, 6.5 - 8 mg / mL lactic acid, 1.0 - 1.5 mg / mL preservative, 0.8 - 1.6 mg / mL flavoring agent and 0.3 - 0.5 mg / mL essence, with the balance being water.

5. The preparation method of the calcium zinc gluconate oral solution composition according to any one of claims 1-4, characterized in that, Including the following steps: (1) Dissolve calcium gluconate and zinc gluconate in water at 80 - 95 °C to obtain Solution I; (2) Cool the Solution I in step (1) to 45 °C or below, add other components, dissolve to obtain Solution II; (3) Make up the volume of Solution II in step (2) to the full volume, mix well, filter and seal to obtain the product.

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