Method for detecting phosphate in epinephrine
The phosphate content in adrenaline was detected by the mixing method of ammonium molybdate-sulfuric acid solution and hydrochloric acid-acid stannous chloride solution, and the problem that the prior art cannot be effectively detected is solved, and efficient and accurate drug quality control is achieved.
Patent Information
- Application Number
- CN202411867621.X
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-12-17
- Publication Date
- 2025-05-06
AI Technical Summary
The prior art cannot effectively detect phosphonates in adrenaline, resulting in difficulty in controlling drugs quality and unable to eliminate background interference.
The mixture method of ammonium molybdate-sulfuric acid solution and hydrochloric acid-acid stannous chloride solution was used to determine whether the phosphate content exceeded the standard by comparing the color depth of the test sample and the reference sample.
Accurate detection of phosphate in adrenaline is achieved, which can eliminate background interference, is easy to operate, saves detection time and cost, and is suitable for quality control of adrenaline.
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Abstract
Description
Technical Field
[0001] The invention relates to the technical field of drug analysis, and in particular to a method for detecting phosphonate in adrenaline. Background Art
[0002] Epinephrine (Adrenaline, Epinephrine) is the main neurotransmitter in the adrenal medulla. Its biosynthesis is mainly formed in the medullary chromaffin cells to form norepinephrine, and then further methylated to form epinephrine through the action of phenylethanolamine N-methyl transferase (PNMT). Because the receptors are distributed in the heart, arteries, veins, bronchi, gastrointestinal tract, bladder, uterus, eyes, and glands. It is also involved in the metabolism of glycogen, skeletal muscle glycogen, and fat. It is clinically used in cardiac arrest, anaphylactic shock, bronchial asthma, and is compatible with local anesthetics and local hemostasis. Epinephrine is a drug for emergency and critical illnesses and plays an important role in saving patients' lives. It is contraindicated for hypertension, cerebral arteriosclerosis, organic heart disease, diabetes, and hyperthyroidism.
[0003] As a vasoconstrictor, adrenaline can constrict blood vessels and control blood perfusion in tissues. Adrenaline is added to the local anesthetic solution to counteract the vasodilatory effect of the local anesthetic. The main indicators for adrenaline quality control currently included in the pharmacopoeias of various countries include properties, melting point, specific rotation, clarity and color of acidic solutions, ketone bodies, related substances, loss on drying, ignition residue and content. There is still a blank in the quality control test method for inorganic salts, and the general pharmacopoeia and literature phosphate test methods are not applicable to this variety, and the background interference cannot be eliminated.
[0004] Therefore, a method for accurately detecting phosphate in adrenaline is needed to effectively control the drug quality of adrenaline and ensure drug safety. Summary of the invention
[0005] In order to solve one of the above-mentioned technical problems existing in the prior art, the present invention provides a method for detecting phosphate in epinephrine, which can accurately and efficiently realize quality control of epinephrine products.
[0006] The technical solution of the present invention is as follows:
[0007] A method for detecting phosphate in adrenaline, the method comprising the following steps:
[0008] (1) Prepare test sample solution and reference sample solution;
[0009] (2) mixing the test solution with an ammonium molybdate-sulfuric acid solution, and then adding a hydrochloric acid-acidic stannous chloride solution to obtain a first mixed solution; mixing the reference solution with an ammonium molybdate-sulfuric acid solution, and then adding a hydrochloric acid-acidic stannous chloride solution to obtain a second mixed solution; and judging whether the phosphate content in the test solution exceeds the standard by comparing the color depth of the first mixed solution and the second mixed solution.
[0010] In some embodiments, the preparation of the test solution comprises the following steps:
[0011] The adrenaline sample is dissolved with acid and diluted with water, and then a weak base is added to precipitate the adrenaline, and the solid and liquid are separated to obtain a liquid phase. The liquid phase is decolorized and diluted with water to obtain the test sample.
[0012] Preferably, in the preparation of the test solution, the acid includes hydrochloric acid, and the hydrochloric acid is preferably dilute hydrochloric acid, for example, the mass concentration of the hydrochloric acid can be 10-15%. In some embodiments, the concentration of the acid is 2.5-3.0 mol / L.
[0013] Preferably, in the preparation of the test solution, the weak base includes one or more of ammonia water, alkali metal carbonates, and alkali metal bicarbonates. Preferably, the weak base includes one or more of ammonia water, sodium carbonate, potassium carbonate, sodium bicarbonate, and potassium bicarbonate. In some preferred embodiments, the weak base includes ammonia water. Preferably, the mass concentration of the ammonia water is 25-28%.
[0014] Preferably, in the preparation of the test solution, activated carbon is used to decolorize the liquid phase.
[0015] Preferably, in the preparation of the test solution, the ratio of the mass of the adrenaline sample to the volume of the acid is 1:(4-6) g / mL.
[0016] Preferably, in the preparation of the test solution, the ratio of the mass of the adrenaline sample to the volume of the weak base is 1:(1-4) g / mL.
[0017] Preferably, in the preparation of the test solution, the adrenaline sample is dissolved with acid and then diluted 70-100 times with water.
[0018] Preferably, in the preparation of the test solution, the liquid phase is decolorized and then diluted 3-7 times with water.
[0019] In some embodiments, the preparation of the reference solution comprises the following steps: dissolving a phosphate standard in water to obtain a reference solution. Preferably, each 1 mL of the reference solution contains 0.70-0.80 μg of phosphate, preferably 0.70-0.75 μg, and more preferably 0.716 μg. Preferably, each 1 mL of the reference solution contains 0.40-0.60 μg, preferably 0.45-0.55 μg, and more preferably 0.5 μg of phosphate. Preferably, the ratio of the mass of phosphate in the reference solution to the mass of the adrenaline sample is not higher than 0.1%, for example, 0.005%, 0.01%, 0.02%, 0.03%, 0.05%, 0.08%, 0.1% or any value therebetween, preferably 0.005%-0.1%.
[0020] Preferably, in the preparation of the reference solution, the phosphate standard includes alkali metal dihydrogen phosphate, preferably one or both of potassium dihydrogen phosphate and sodium dihydrogen phosphate.
[0021] In some embodiments, the concentration of ammonium molybdate in the ammonium molybdate-sulfuric acid solution is 0.2-0.3 g / L, H 2 SO 4 The concentration is 4.0-8.0mol / L.
[0022] In some embodiments, the volume ratio of the test solution to the ammonium molybdate-sulfuric acid solution is (20-30):1, for example, 20:1, 21:1, 22:1, 23:1, 24:1, 25:1, 26:1, 27:1, 28:1, 29:1, 30:1 or any value therebetween. In some embodiments, the volume ratio of the test solution to the ammonium molybdate-sulfuric acid solution is (23-27):1.
[0023] In some embodiments, the volume ratio of the reference solution to the ammonium molybdate-sulfuric acid solution is (20-30):1, such as 20:1, 21:1, 22:1, 23:1, 24:1, 25:1, 26:1, 27:1, 28:1, 29:1, 30:1 or any value therebetween. In some embodiments, the volume ratio of the reference solution to the ammonium molybdate-sulfuric acid solution is (23-27):1.
[0024] In some embodiments, the hydrochloric acid-acidic stannous chloride solution is a mixture of hydrochloric acid and acidic stannous chloride solution. Preferably, the concentration of the hydrochloric acid is 2-5 mol / L. In some embodiments, in the acidic stannous chloride solution, the concentration of stannous chloride is 30-40 μg / mL. Preferably, the hydrochloric acid-acidic stannous chloride solution is a mixture of hydrochloric acid and acidic stannous chloride solution in a volume ratio of (5-15):1, preferably (8-12):1.
[0025] In some embodiments, the volume ratio of the test solution to the hydrochloric acid-acidic stannous chloride solution is 100:(0.05-0.15), preferably 100:(0.08-0.12).
[0026] In some embodiments, the volume ratio of the reference solution to the hydrochloric acid-acidic stannous chloride solution is 100:(0.05-0.15), preferably 100:(0.08-0.12).
[0027] In some embodiments, if the color of the first mixed solution is the same as the color of the second mixed solution, or the color of the first mixed solution is lighter than the color of the second mixed solution, the phosphate content in the test solution does not exceed the standard.
[0028] Compared with the prior art, the present invention has the following beneficial effects:
[0029] The present invention provides a method for detecting phosphate in adrenaline, which is not only specific, can eliminate background interference, has high accuracy, good durability, but also is easy to operate, saves detection time and cost, can be applied to the quality control of adrenaline, is conducive to production and promotion, and has great significance for the clinical safety and effectiveness of adrenaline injection. In addition, in the method of the present invention, the preparation of the test solution is simple and convenient, and the phosphate in adrenaline can be effectively extracted. DETAILED DESCRIPTION
[0030] In order to make the purpose, technical solution and advantages of the present invention more clearly understood, the present invention is further described in detail below in conjunction with embodiments. 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.
[0031] In order to fill the gap in the prior art for the detection method of phosphate in adrenaline, the present invention provides a detection method for phosphate in adrenaline. The method of the present invention comprises the following steps: dissolving an adrenaline sample with an acidic solution, adjusting the pH to alkaline with a weak base such as ammonia water, so that the adrenaline is precipitated, while the phosphate remains in the solution, and then filtering to remove the adrenaline. After the filtrate is decolorized with activated carbon, the phosphate is detected by colorimetry. The method is simple and easy to operate, can effectively remove the interference of the background color of adrenaline, the detection result is accurate and reliable, has strong specificity, good repeatability, and no residual effect, and can be used for the quality control of adrenaline drugs.
[0032] In the present invention, the "phosphate concentration limit" of the control solution refers to the ratio of the mass of phosphate in the control solution to the mass of the adrenaline sample to be tested; for example, "a control solution with a phosphate concentration limit of 0.1%" means that the ratio of the mass of phosphate in the control solution to the mass of the adrenaline sample to be tested is 0.1%.
[0033] The present invention provides a method for detecting phosphate in adrenaline, comprising the following steps:
[0034] (1) Prepare test sample solution and reference sample solution;
[0035] (2) mixing the test solution with an ammonium molybdate-sulfuric acid solution, and then adding a hydrochloric acid-acidic stannous chloride solution to obtain a first mixed solution; mixing the reference solution with an ammonium molybdate-sulfuric acid solution, and then adding a hydrochloric acid-acidic stannous chloride solution to obtain a second mixed solution; and judging whether the phosphate content in the test solution exceeds the standard by comparing the color depth of the first mixed solution and the second mixed solution.
[0036] As a specific implementation, in step (1), the method for preparing the test solution is:
[0037] Take 0.5 g of adrenaline, add 2-3 mL of dilute hydrochloric acid to dissolve, dilute with water to 200 mL, then add 0.5-2 mL of aqueous ammonia, stir for 1-2 minutes, and filter; take the filtrate and add 1-2 g of activated carbon, shake well, let it stand for 1-2 hours, then filter, accurately measure 15-30 mL of the filtrate, place it in a 100 mL volumetric flask, dilute to the scale with water, and shake well.
[0038] Preferably, the amount of dilute hydrochloric acid added is 2 mL.
[0039] Preferably, the concentration of the dilute hydrochloric acid is 2.5-3.0 mol / L, more preferably 2.8 mol / L.
[0040] Preferably, the amount of ammonia water added is 1 mL, and the stirring time is 30 minutes.
[0041] Preferably, the mass concentration of the ammonia water is 25%-28%.
[0042] Preferably, the added amount of the activated carbon is 2 g.
[0043] Preferably, the volume of the subsequent filtrate is 20 mL.
[0044] As a specific implementation, in step (1), the preparation method of the reference solution is:
[0045] Weigh an appropriate amount of potassium dihydrogen phosphate, dissolve it in water and quantitatively dilute it into a solution containing 0.7-0.8 mg, preferably 0.70-0.75 mg, more preferably 0.716 mg of potassium dihydrogen phosphate per 1 mL. Accurately measure 1 mL of the above solution, place it in a 100 mL volumetric flask, dilute it to the mark with water, shake it well, and use it as a standard solution. Accurately measure 10 mL of the standard solution, place it in a 100 mL volumetric flask, dilute it to the mark with water, and shake it well.
[0046] As a specific embodiment, the method for detecting phosphate in adrenaline comprises the following steps:
[0047] ① Preparation of test solution: Take 0.5g of epinephrine, add 2mL of dilute hydrochloric acid to dissolve, dilute with water to 200mL, add 1mL of ammonia water, stir for 30 minutes, and filter; take the filtrate and add 2g of activated carbon, shake well, let it stand for 1 hour, then filter, accurately measure 20mL of the filtrate, put it in a 100mL volumetric flask, dilute with water to the scale, and shake well;
[0048] Preparation of control solution: Weigh an appropriate amount of potassium dihydrogen phosphate, dissolve it in water and quantitatively dilute it into a solution containing 0.7-0.8 mg, preferably 0.70-0.75 mg, more preferably 0.716 mg of potassium dihydrogen phosphate per 1 mL. Accurately measure 1 mL of the above solution, place it in a 100 mL volumetric flask, dilute it to the mark with water, shake it well, and use it as the standard solution. Accurately measure 10 mL of the standard solution, place it in a 100 mL volumetric flask, dilute it to the mark with water, and shake it well.
[0049] ② Colorimetric detection: Take 100 mL of the above test solution and control solution, add 4 mL of thiomolybdic acid test solution (take 2.5 g of ammonium molybdate, add 20 mL of water, heat to dissolve, add 50 mL of 12 mol / L sulfuric acid solution, dilute to 100 mL with water, shake well; store in polyethylene container), mix well, add 0.1 mL of a mixed solution of 3 mol / L hydrochloric acid solution and 35 μg / mL acidic stannous chloride test solution (volume ratio is hydrochloric acid: acidic stannous chloride test solution = 10:1), mix well. Visually compare colors after 10 minutes.
[0050] In the method of the present invention, the color generated by the test solution after the reaction shall not be darker than the color of the control solution after the reaction, indicating that the phosphate content in the test solution is not higher than the phosphate content in the control solution. In some embodiments, the concentration limit of phosphate in the control solution is not higher than 0.1%, for example, 0.005%, 0.01%, 0.015%, 0.02%, 0.025%, 0.05%, 0.08%, 0.1% or any value therebetween.
[0051] Preferably, the adrenaline is of qualified quality when the phosphate content in the adrenaline does not exceed 0.1%.
[0052] Example 1: Determination of Phosphate in Epinephrine
[0053] The method for determining phosphate in epinephrine is as follows:
[0054] ① Preparation of test solution: Take 0.5g of adrenaline sample, add 2mL of 2.8mol / L dilute hydrochloric acid to dissolve, dilute with water to 200mL, add 1mL of 25% ammonia water, stir for 30 minutes, and filter; take the filtrate and add 2g of activated carbon, shake well, let it stand for 1 hour, then filter, accurately measure 20mL of the filtrate, put it in a 100mL volumetric flask, dilute with water to the scale, and shake well;
[0055] Preparation of a control solution with a phosphate concentration limit of 0.1%: Weigh an appropriate amount of potassium dihydrogen phosphate, dissolve it in water and quantitatively dilute it to a solution containing 0.716 mg per 1 mL. Accurately measure 1 mL of the above solution, place it in a 100 mL volumetric flask, dilute it to the mark with water, shake it well, and use it as a standard solution. Accurately measure 10 mL of the standard solution, place it in a 100 mL volumetric flask, dilute it to the mark with water, shake it well, and obtain a control solution with a phosphate concentration limit of 0.1%.
[0056] ② Colorimetric detection: Take 100 mL of the above test solution and control solution, add 4 mL of thiomolybdic acid test solution (take 2.5 g of ammonium molybdate, add 20 mL of water, heat to dissolve, add 50 mL of 12 mol / L sulfuric acid solution, dilute to 100 mL with water, shake well; store in polyethylene container), mix well, add 0.1 mL of a mixed solution of 3 mol / L hydrochloric acid solution and 35 μg / mL acidic stannous chloride test solution (volume ratio is hydrochloric acid: acidic stannous chloride test solution = 10:1), mix well. Visually compare colors after 10 minutes.
[0057] According to the above method, the phosphate in 9 batches of epinephrine samples were tested using control solutions with phosphate concentration limits of 0.01% and 0.02%, respectively. The results are shown in Table 1. The preparation method of the control solutions with phosphate concentration limits of 0.01% and 0.02% is as follows:
[0058] Preparation of a control solution with a phosphate concentration limit of 0.01%: Accurately measure 1 mL of the above-prepared control solution with a phosphate concentration limit of 0.1%, place it in a 10 mL volumetric flask, dilute it to the scale with water, shake well, and obtain a control solution with a phosphate concentration limit of 0.01%.
[0059] Preparation of a control solution with a phosphate concentration limit of 0.02%: Accurately measure 2 mL of the above-prepared control solution with a phosphate concentration limit of 0.1%, place it in a 10 mL volumetric flask, dilute to the scale with water, shake well, and obtain a control solution with a phosphate concentration limit of 0.2%.
[0060] Table 1 Phosphate detection results in adrenaline samples
[0061]
[0062] Example 2: Specificity
[0063] Blank interference test:
[0064] Blank solution: Take 100mL of water, add 4mL of thiomolybdic acid test solution (take 2.5g of ammonium molybdate, add 20mL of water, heat and dissolve, add 50mL of 12mol / L sulfuric acid solution, dilute to 100mL with water, shake well; store in a polyethylene container), mix well, add 0.1mL of a mixed solution of 3mol / L hydrochloric acid solution and 35μg / mL acidic stannous chloride test solution (volume ratio is hydrochloric acid: acidic stannous chloride test solution = 10:1), mix well. Observe the color after 10 minutes.
[0065] Positive solution: Take 1mL of potassium dihydrogen phosphate stock solution, add 2mL of 2.8mol / L dilute hydrochloric acid to dissolve, dilute with water to 200mL, add 1mL of 25% ammonia water, stir for 30 minutes, and filter; take the filtrate and add 2g of activated carbon, shake well, let it stand for 1 hour and then filter, accurately measure 20mL of the filtrate, put it in a 100mL volumetric flask, dilute with water to the scale, and shake well.
[0066] Potassium dihydrogen phosphate stock solution: Take 0.716 g of potassium dihydrogen phosphate, place it in a 1000 mL volumetric flask, dilute to the scale with water, and shake well.
[0067] Control solution: Accurately measure 1 mL of potassium dihydrogen phosphate stock solution, place it in a 100 mL volumetric flask, dilute it to the mark with water, shake well, and use it as the standard solution. Accurately measure 10 mL of the standard solution, place it in a 100 mL volumetric flask, dilute it to the mark with water, shake well, and obtain a control solution with a phosphate concentration limit of 0.1%.
[0068] Determination method: Take 100mL of the above positive solution and control solution, add 4mL of thiomolybdic acid test solution (take 2.5g of ammonium molybdate, add 20mL of water, heat and dissolve, add 50mL of 12mol / L sulfuric acid solution, dilute to 100mL with water, shake well; store in polyethylene container), mix well, add 0.1mL of a mixed solution of 3mol / L hydrochloric acid solution and 35μg / mL acidic stannous chloride test solution (volume ratio is hydrochloric acid: acidic stannous chloride test solution = 10:1) prepared just before use, mix well. Visually compare the colors after 10 minutes.
[0069] Conclusion: The blank solution does not show blue color and there is no color interference; the positive solution is consistent with the control solution (0.1%) in color.
[0070] Example 3: Detection Limit
[0071] Referring to the detection method of Example 2, blank solutions and control solutions of a series of limits were prepared to the lowest limit that can be reliably detected. The results are shown in Table 2.
[0072] Table 2 Detection limit results
[0073]
[0074] Conclusion: The 0.005% limit of the control solution is visually distinguishable from the blank solution, which is the minimum limit that can be reliably detected, indicating that the detection limit of the method of the present invention can be as low as 0.005%.
[0075] Example 4: Accuracy Test
[0076] Potassium dihydrogen phosphate stock solution: Take 0.716 g of potassium dihydrogen phosphate, place it in a 1000 mL volumetric flask, dilute to the scale with water, and shake well.
[0077] Standard solution: Accurately measure 1 mL of potassium dihydrogen phosphate stock solution, place it in a 100 mL volumetric flask, dilute to the scale with water, and shake well.
[0078] Accuracy solution: Take 0.5g of adrenaline sample, add 1mL of potassium dihydrogen phosphate stock solution, add 2mL of 2.8mol / L dilute hydrochloric acid to dissolve, dilute with water to 200mL, add 1mL of 25% ammonia water, stir for 30 minutes, and filter; take the filtrate and add 2g of activated carbon, shake well, let it stand for 1 hour and then filter, accurately measure 20mL of the filtrate, put it in a 100mL volumetric flask, dilute with water to the scale, and shake well.
[0079] Test sample stock solution: Take 0.5 g of adrenaline sample, add 2 mL of 2.8 mol / L dilute hydrochloric acid to dissolve, dilute with water to 200 mL, then add 1 mL of 25% ammonia water, stir for 30 minutes, and filter; take the filtrate and add 2 g of activated carbon, shake well, let it stand for 1 hour, and then filter.
[0080] Control solution: Accurately measure 20 mL of the test sample stock solution and place it in a 100 mL volumetric flask, add 10 mL of the standard solution, dilute to the scale with water, and shake well.
[0081] Determination: Take 100mL of the accuracy solution and control solution, add 4mL of thiomolybdic acid test solution (take 2.5g of ammonium molybdate, add 20mL of water, heat and dissolve, add 50mL of 12mol / L sulfuric acid solution, dilute to 100mL with water, shake well; store in polyethylene container), mix well, add 0.1mL of a mixed solution of 3mol / L hydrochloric acid solution and 35μg / mL acidic stannous chloride test solution (volume ratio is hydrochloric acid: acidic stannous chloride test solution = 10:1) prepared just before use, mix well. Visually compare colors after 10 minutes.
[0082] Conclusion: The color of the accuracy solution is consistent with that of the control solution, and the accuracy is good.
[0083] Example 5: Durability
[0084] According to the detection method in Example 1, the influence of changing the amount of the thiomolybdic acid test solution and the mixed solution of the freshly prepared 3 mol / L hydrochloric acid solution and 35 μg / mL acidic stannous chloride test solution was investigated respectively. The results are shown in Table 3.
[0085] Table 3 Durability results
[0086]
[0087] Conclusion: When the dosage of thiomolybdic acid test solution and mixed solution was changed, the color of the test solution did not change significantly compared with normal conditions, and the durability was good.
[0088] The technical solution of the present invention is not limited to the above-mentioned specific embodiments. All technical variations made according to the technical solution of the present invention fall within the protection scope of the present invention.
Claims
1. A method for detecting phosphate in adrenaline, comprising the following steps: (1) Prepare test sample solution and reference sample solution; (2) mixing the test solution with an ammonium molybdate-sulfuric acid solution, and then adding a hydrochloric acid-acidic stannous chloride solution to obtain a first mixed solution; mixing the reference solution with an ammonium molybdate-sulfuric acid solution, and then adding a hydrochloric acid-acidic stannous chloride solution to obtain a second mixed solution; and judging whether the phosphate content in the test solution exceeds the standard by comparing the color depth of the first mixed solution and the second mixed solution.
2. The detection method according to claim 1, characterized in that: The preparation of the test solution comprises the following steps: The adrenaline sample is dissolved with an acid and then diluted with water, and then a weak base is added to precipitate the adrenaline, and the solid and liquid are separated to obtain a liquid phase, and the liquid phase is decolorized and then diluted with water to obtain the test sample; Preferably, the acid comprises hydrochloric acid, and the concentration of the acid is preferably 2.5 to 3.0 mol / L; Preferably, the weak base includes one or more of ammonia water, alkali metal carbonates, and alkali metal bicarbonates, more preferably includes one or more of ammonia water, sodium carbonate, potassium carbonate, sodium bicarbonate, and potassium bicarbonate; further preferably includes ammonia water; Preferably, the mass concentration of the weak base is 25-28%; Preferably, the liquid phase is decolorized using activated carbon.
3. The detection method according to claim 2, characterized in that: The ratio of the mass of the adrenaline sample to the volume of the acid is 1:(4-6) g / mL; and / or, The ratio of the mass of the adrenaline sample to the volume of the weak base is 1:(1-4) g / mL.
4. The detection method according to claim 2 or 3, characterized in that: Dissolve the adrenaline sample in acid and then dilute it 70-100 times with water; and / or, The liquid phase is decolorized and then diluted 3-7 times with water.
5. The detection method according to any one of claims 1 to 4, characterized in that: The preparation of the reference solution comprises the following steps: dissolving a phosphate standard in water to obtain a reference solution; Preferably, each 1 mL of the reference solution contains 0.40-0.60 μg of phosphate, preferably 0.45-0.55 μg, more preferably 0.5 μg; Preferably, the ratio of the mass of phosphate in the reference solution to the mass of the adrenaline sample is not higher than 0.1%, preferably 0.005%-0.1%; Preferably, the phosphate standard comprises alkali metal dihydrogen phosphate, more preferably comprises one or both of potassium dihydrogen phosphate and sodium dihydrogen phosphate.
6. The detection method according to any one of claims 1 to 5, characterized in that: In the ammonium molybdate-sulfuric acid solution, the concentration of ammonium molybdate is 0.2-0.3 g / L, and the concentration of H2SO4 is 4.0-8.0 mol / L.
7. The detection method according to any one of claims 1 to 6, characterized in that: The volume ratio of the test solution to the ammonium molybdate-sulfuric acid solution is (20-30):1, preferably (23-27):1; and / or, The volume ratio of the reference solution to the ammonium molybdate-sulfuric acid solution is (20-30):1, preferably (23-27):
1.
8. The detection method according to any one of claims 1 to 7, characterized in that: The hydrochloric acid-acidic stannous chloride solution is a mixture of hydrochloric acid and acidic stannous chloride solution, wherein the concentration of the hydrochloric acid is 2-5 mol / L, and the concentration of stannous chloride in the acidic stannous chloride solution is 30-40 μg / mL; Preferably, the hydrochloric acid-acidic stannous chloride solution is a mixed solution of hydrochloric acid and acidic stannous chloride solution in a volume ratio of (5-15):1, preferably (8-12):
1.
9. The detection method according to any one of claims 1 to 8, characterized in that: The volume ratio of the test solution to the hydrochloric acid-acidic stannous chloride solution is 100:(0.05-0.15), preferably 100:(0.08-0.12); and / or, The volume ratio of the reference solution to the hydrochloric acid-acidic stannous chloride solution is 100:(0.05-0.15), preferably 100:(0.08-0.12).
10. The detection method according to any one of claims 1 to 9, characterized in that: If the color of the first mixed solution is the same as the color of the second mixed solution, or the color of the first mixed solution is lighter than the color of the second mixed solution, the phosphate content in the test solution does not exceed the standard.