Method for determining taurine in magnesium acetyltaurate
By combining high-performance liquid chromatography with a UV detector and 2,4-dinitrofluorophenylacetonitrile derivatization, the technical challenge of taurine detection in magnesium acetylatum was solved, achieving detection results with high sensitivity and specificity, reducing detection costs and improving product safety and reliability.
Patent Information
- Application Number
- CN202511341423.4
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-09-19
- Publication Date
- 2025-10-28
AI Technical Summary
The lack of a low-cost, easy-to-operate, highly sensitive method for detecting taurine residues that does not require expensive specialized equipment has resulted in high quality control costs for magnesium acetylacetonate products and difficulty in guaranteeing their safety and effectiveness.
High-performance liquid chromatography (HPLC) combined with a UV detector was used to generate stable derivatized products by reacting magnesium acetylatum with 2,4-dinitrofluorophenylacetonitrile as a derivatizing reagent under alkaline conditions. Gradient elution was performed using phosphate buffer and acetonitrile as the mobile phase, and the derivatized products were finally detected under a UV detector.
This method achieves highly sensitive, specific, and accurate detection of taurine in magnesium acetylatum, reducing detection costs, simplifying the operation process, and ensuring the safety and reliability of the product.
Smart Images
Figure CN120847302A_ABST
Abstract
Description
Technical Field
[0001] This invention belongs to the field of pharmaceutical analysis technology, and specifically relates to a method for determining taurine in magnesium acetylatum. Background Technology
[0002] Magnesium acetylated taurine is a compound that combines magnesium, taurine, and acetyl groups. Through the physiological functions of magnesium and the synergistic effect of taurine, it can supplement magnesium, regulate the nervous system, relieve fatigue, and support cardiovascular health. Clinically, it is often used to improve sleep and relieve muscle tension and anxiety.
[0003] Taurine, a key starting material in the synthesis of magnesium acetylatamate, has a residual level that directly affects the clinical safety and efficacy of the final product. Therefore, strict quality control of raw taurine is necessary. However, existing technologies for taurine detection have significant limitations: 1. The current Chinese Pharmacopoeia primarily uses thin-layer chromatography (TLC) for the detection of taurine-related substances, but this method suffers from low sensitivity; 2. The current Chinese Pharmacopoeia uses post-column derivatization for the determination of taurine content in eye drops, requiring the purchase of an additional derivatization system, resulting in high equipment purchase and maintenance costs; 3. The lack of effective ultraviolet absorption groups in the taurine molecule means that conventional, widely used, and relatively low-cost high-performance liquid chromatography-ultraviolet detection methods cannot be directly applied to the detection of taurine.
[0004] In summary, existing technologies lack a low-cost, easy-to-operate, highly sensitive method for detecting taurine residues that does not rely on expensive specialized equipment. This lack of a suitable method not only increases quality control costs for enterprises but also poses a potential risk to ensuring the final quality, safety, and effectiveness of magnesium acetylatum taurate products. Therefore, there is an urgent need to develop a novel, efficient, and economical method for detecting taurine residues, particularly a method that utilizes conventional high-performance liquid chromatography (HPLC) for direct detection. Summary of the Invention
[0005] The technical problem to be solved by the present invention is to provide a low-cost, easy-to-operate, highly sensitive method for detecting taurine in magnesium acetylatum, which does not rely on expensive special equipment. This method has high sensitivity, good specificity, high accuracy, and can strictly control the quality of magnesium acetylatum, ensuring the safety and reliability of magnesium acetylatum, and has practical significance.
[0006] To solve the above technical problems, the technical solution adopted by the present invention is as follows: This invention provides a method for detecting taurine in magnesium acetylatum, comprising the following steps: (1) Derivatization treatment: Under alkaline conditions, the derivatizing reagent reacts with magnesium acetylatum containing taurine to obtain the derivatized product; (2) Detection: The derivatized products described in step (1) were detected using high performance liquid chromatography-ultraviolet detector; The derivatizing reagent is selected from 2,4-dinitrofluorophenylacetonitrile.
[0007] Preferably, in step (1), the alkaline reagent is selected from one or more of sodium hydroxide, sodium carbonate, and sodium bicarbonate.
[0008] Preferably, the pH value of the alkaline condition is 8.0~10.0, and more preferably 9.0.
[0009] Preferably, the concentration of the 2,4-dinitrofluorophenylacetonitrile is 5~20 mg / mL, and more preferably 10 mg / mL.
[0010] Preferably, in step (1), the reaction time of the derivatization reaction is 40-80 min, more preferably 50-80 min, and even more preferably 60 min.
[0011] Preferably, in step (1), the reaction temperature of the derivatization reaction is 40~80℃; more preferably 50~80℃; and even more preferably 60℃.
[0012] Preferably, in step (1), the derivatization process specifically includes: adding sodium bicarbonate solution (pH adjusted with sodium hydroxide solution) and 2,4-dinitrofluorophenylacetonitrile solution to the taurine-containing magnesium acetyl taurate, shaking well, reacting in a 60°C water bath for 1 hour, taking it out, cooling it, diluting it to the mark with a mobile phase additive, and shaking well.
[0013] Preferably, the pH value of the mobile phase additive is 6.5 to 8.0, and more preferably 7.0.
[0014] Preferably, the mobile phase additive is selected from phosphate buffer.
[0015] Preferably, in step (2), the mobile phase of the high performance liquid chromatography is a mixture of mobile phase A and mobile phase B. Mobile phase A is selected from one or more of phosphate buffer, methanol, tetrahydrofuran, acetonitrile, and water, and mobile phase B is selected from one or more of methanol, tetrahydrofuran, acetonitrile, and water.
[0016] Preferably, the mobile phase A is a mixed solution of phosphate buffer, acetonitrile, and water.
[0017] Preferably, the pH value of the phosphate buffer solution is 6.5 to 8.0, and more preferably 7.0.
[0018] Preferably, the volume ratio of the phosphate buffer-acetonitrile-water is (60~80):(10~20):(10~20), more preferably 70:(10~20):(10~20), and even more preferably 70:15:15.
[0019] Preferably, the mobile phase B is a mixed solution of acetonitrile and water.
[0020] Preferably, the volume ratio of acetonitrile to water is (40~60):(40~60), more preferably 50:50.
[0021] Preferably, in step (2), the detection wavelength of the high performance liquid chromatography is selected from 210~214nm, 263~267nm and 357~361nm, and preferably 360nm.
[0022] Preferably, in step (2), the chromatographic column of the high performance liquid chromatography is a reversed-phase column, preferably an octadecylsilane-bonded silica column, more preferably a GL Sciences Wondasil C18 Superb column or a column with equivalent performance.
[0023] Preferably, the chromatographic column has a length of 150 mm to 250 mm, an inner diameter of 2.7 to 4.6 mm, and a particle size of 3.5 μm to 5 μm; more preferably, it has a length of 250 mm, an inner diameter of 4.6 mm, and a particle size of 5 μm.
[0024] Preferably, the column temperature of the chromatographic column is 30℃~40℃, and more preferably 30℃.
[0025] Preferably, in step (2), the injection flow rate of the high performance liquid chromatography is 0.5 mL / min to 1.5 mL / min, more preferably 0.8 mL / min to 1.2 mL / min, and more preferably 1.0 mL / min.
[0026] Preferably, in step (2), the injection volume of the high performance liquid chromatography is 5 μL to 20 μL, more preferably 10 μL to 20 μL, and even more preferably 10 μL.
[0027] Preferably, in step (2), the gradient elution procedure of the high-performance liquid chromatography is as follows: .
[0028] Due to the adoption of the above technical solutions, the present invention has the following advantages compared with the prior art: This method employs high-performance liquid chromatography (HPLC) combined with a UV detector, eliminating the need for additional derivatization systems and specialized detectors. It offers advantages such as high sensitivity, good specificity, simple operation, high accuracy, good reproducibility, excellent stability, and low cost. By preparing a reference solution and a test solution containing taurine, taurine is quantitatively reacted with 2,4-dinitrofluorobenzene to generate a stable derivatized product, -2,4-dinitrophenyltaurine. The sample is injected into the HPLC system. Gradient elution is performed using phosphate buffer (pH 7.0) and acetonitrile as the mobile phase. After elution, the signal intensity of the test sample is detected using a UV detector, thereby indirectly determining the taurine content in the test sample, acetylated magnesium taurate. This method provides a convenient detection method for monitoring taurine in acetylated magnesium taurate, improves the quality standards for taurine testing in acetylated magnesium taurate, enables strict quality control of taurine in acetylated magnesium taurate, and ensures the safety and reliability of acetylated magnesium taurate, thus possessing practical significance. Attached Figure Description
[0029] Figure 1 This is an HPLC chromatogram of the reference solution that has not undergone derivatization treatment in the specificity test of Example 1 of the present invention.
[0030] Figure 2 This is an HPLC chromatogram of the test solution after derivatization treatment in the specificity test of Example 1 of the present invention.
[0031] Figure 3 This is an HPLC chromatogram of the spiked test solution after derivatization treatment in the specificity test of Example 1 of the present invention. Detailed Implementation
[0032] To make the technical solution and beneficial effects of the present invention more apparent and understandable, a detailed description is provided below in conjunction with the accompanying drawings and specific embodiments. The drawings are not necessarily drawn to scale, and local features may be enlarged or reduced to more clearly show the details of the local features; unless otherwise defined, the technical and scientific terms used herein have the same meanings as those in the technical field to which this application pertains.
[0033] Experimental methods not specified in the following examples are generally performed under standard experimental conditions.
[0034] This invention provides a method for detecting taurine in magnesium acetylatum, comprising the following steps: (1) Derivatization treatment: Under alkaline conditions, the derivatizing reagent reacts with magnesium acetylatum containing taurine to obtain the derivatized product; (2) Detection: The derivatized products described in step (1) were detected using high performance liquid chromatography-ultraviolet detector; The derivatizing reagent is selected from 2,4-dinitrofluorophenylacetonitrile.
[0035] In some embodiments, in step (1), the alkaline reagent is selected from one or more of sodium hydroxide, sodium carbonate, and sodium bicarbonate.
[0036] In some embodiments, in step (1), the alkaline reagent is selected from a mixed solution of sodium hydroxide and sodium bicarbonate.
[0037] In some embodiments, the pH value of the alkaline conditions is 8.0 to 10.0, for example 8.0, 9.0, 10.0.
[0038] In some embodiments, the pH value of the alkaline conditions is 9.0.
[0039] In some embodiments, the concentration of the 2,4-dinitrofluorophenylacetonitrile is 5 to 20 mg / mL, for example, 5 mg / mL, 10 mg / mL, 20 mg / mL, etc.
[0040] In some embodiments, the concentration of the 2,4-dinitrofluorophenylacetonitrile is 10 mg / mL.
[0041] In some embodiments, the reaction time of the derivatization reaction in step (1) is 40 to 80 minutes, for example, 40 minutes, 50 minutes, 60 minutes, or 80 minutes.
[0042] In some embodiments, the reaction time of the derivatization reaction in step (1) is 50-80 min.
[0043] In some embodiments, the reaction time of the derivatization reaction in step (1) is 60 min.
[0044] In some embodiments, in step (1), the reaction temperature of the derivatization reaction is 40~80℃, for example 40℃, 50℃, 60℃, 80℃.
[0045] In some embodiments, the reaction temperature of the derivatization reaction in step (1) is 50~80°C.
[0046] In some embodiments, the reaction temperature of the derivatization reaction in step (1) is 60°C.
[0047] In some embodiments, the derivatization process in step (1) specifically includes: adding sodium bicarbonate solution (pH adjusted with sodium hydroxide solution) and 2,4-dinitrofluorophenylacetonitrile solution to the taurine-containing magnesium acetyl taurate, shaking well, reacting in a 60°C water bath for 1 hour, removing and cooling, diluting to the mark with a mobile phase additive, and shaking well.
[0048] In some embodiments, the pH value of the mobile phase additive is 6.5 to 8.0.
[0049] In some embodiments, the pH of the mobile phase additive is 7.0.
[0050] In some embodiments, the mobile phase additive is selected from phosphate buffer.
[0051] In some embodiments, the mobile phase additive is selected from phosphate buffer (pH 7.0).
[0052] In some embodiments, in step (2), the mobile phase of the high performance liquid chromatography is a mixture of mobile phase A and mobile phase B, wherein mobile phase A is selected from one or more of phosphate buffer, methanol, tetrahydrofuran, acetonitrile, and water, and mobile phase B is selected from one or more of methanol, tetrahydrofuran, acetonitrile, and water.
[0053] In some embodiments, the mobile phase A is a mixed solution of phosphate buffer, acetonitrile, and water.
[0054] In some embodiments, the pH of the phosphate buffer solution is 6.5 to 8.0.
[0055] In some embodiments, the pH of the phosphate buffer solution is 7.0.
[0056] In some embodiments, the mobile phase A is a mixture of phosphate buffer (pH 7.0)-acetonitrile-water.
[0057] In some embodiments, the volume ratio of the phosphate buffer-acetonitrile-water is (60~80):(10~20):(10~20).
[0058] In some embodiments, the volume ratio of the phosphate buffer (pH 7.0)-acetonitrile-water is (60~80):(10~20):(10~20).
[0059] In some embodiments, the volume ratio of the phosphate buffer-acetonitrile-water is 70:(10~20):(10~20).
[0060] In some embodiments, the volume ratio of the phosphate buffer (pH 7.0)-acetonitrile-water is 70:(10~20):(10~20).
[0061] In some embodiments, the volume ratio of the phosphate buffer-acetonitrile-water is 70:15:15.
[0062] In some embodiments, the volume ratio of the phosphate buffer (pH 7.0)-acetonitrile-water is 70:15:15.
[0063] In some embodiments, the phosphate buffer solution comprises sodium hydroxide.
[0064] In some embodiments, the phosphate buffer solution is a mixed solution of sodium hydroxide and phosphate.
[0065] In some embodiments, the phosphate is selected from one or more of ammonium dihydrogen phosphate, potassium dihydrogen phosphate, sodium dihydrogen phosphate, diammonium hydrogen phosphate, dipotassium hydrogen phosphate, and disodium hydrogen phosphate.
[0066] In some embodiments, the phosphate is selected from potassium dihydrogen phosphate.
[0067] In some embodiments, the phosphate buffer solution is a mixed solution of sodium hydroxide and potassium dihydrogen phosphate.
[0068] In some embodiments, the mobile phase B is a mixed solution of acetonitrile and water.
[0069] In some embodiments, the volume ratio of acetonitrile to water is (40~60):(40~60).
[0070] In some embodiments, the volume ratio of acetonitrile to water is 50:50.
[0071] In some embodiments, in step (2), the detection wavelength of the high-performance liquid chromatography is selected from 210~214nm, 263~267nm, and 357~361nm. The derivatized product is analyzed by full-wavelength ultraviolet detection using a DAD full-wavelength UV detector, and three maximum absorption peaks are obtained at 212nm, 265nm, and 359nm. The deviation may be ±2nm depending on the instrument.
[0072] In some embodiments, in step (2), the detection wavelength of the high performance liquid chromatography is selected from 357~361nm, for example, 357nm, 358nm, 359nm, 360nm, 361nm.
[0073] In some embodiments, in step (2), the detection wavelength of the high performance liquid chromatography is 360 nm.
[0074] In some embodiments, in step (2), the chromatographic column of the high performance liquid chromatography is a reversed-phase column.
[0075] In some embodiments, in step (2), the chromatographic column of the high performance liquid chromatography is an octadecylsilane-bonded silica column.
[0076] In some embodiments, in step (2), the chromatographic column of the high performance liquid chromatography is a GLSciences Wondasil C18 Superb column or a column with equivalent performance.
[0077] In some embodiments, the chromatographic column has a length of 150 mm to 250 mm, an inner diameter of 2.7 to 4.6 mm, and a particle size of 3.5 μm to 5 μm.
[0078] In some embodiments, the chromatographic column has a length of 250 mm, an inner diameter of 4.6 mm, and a particle size of 5 μm.
[0079] In some implementations, the column temperature is 30°C to 40°C.
[0080] In some implementations, the column temperature is 30°C.
[0081] In some embodiments, in step (2), the injection flow rate of the high performance liquid chromatography is 0.5 mL / min to 1.5 mL / min, for example 0.5 mL / min, 0.8 mL / min, 1.0 mL / min, 1.2 mL / min, or 1.5 mL / min.
[0082] In some embodiments, in step (2), the injection flow rate of the high performance liquid chromatography is 0.8 mL / min to 1.2 mL / min.
[0083] In some embodiments, in step (2), the injection flow rate of the high performance liquid chromatography is 1.0 mL / min.
[0084] In some embodiments, in step (2), the injection volume of the high performance liquid chromatography is 5 μL to 20 μL.
[0085] In some embodiments, in step (2), the injection volume of the high performance liquid chromatography is 10 μL to 20 μL.
[0086] In some embodiments, in step (2), the injection volume of the high-performance liquid chromatography is 10 μL.
[0087] In some embodiments, the gradient elution procedure of the high-performance liquid chromatography is as follows: .
[0088] In the specific embodiments of this invention, the instruments and taurine reference standards used are all known products. The magnesium acetylated taurine is produced by our company, and the instruments and taurine reference standards are obtained by purchasing commercially available products.
[0089] Table 1. Instruments and reagents used in this detection method.
[0090] Table 2 Samples used in this testing method
[0091] Example 1 1. The following conditions were used for all experiments in this embodiment: Instrument: High-performance liquid chromatograph; Chromatographic column: GL Sciences Wondasil C18 superb, 4.6 mm × 250 mm, particle size 5 μm; Detector: Ultraviolet detector; Mobile phase A: phosphate buffer (pH 7.0) - acetonitrile - water (70:15:15); Mobile phase B: Acetonitrile-water (50:50); Flow rate: 1.0 mL / min; Column temperature: 30℃; Detection wavelength: 360nm; Injection volume: 10 μL; Perform gradient elution according to the table below; .
[0092] 2. Solution preparation 1% 2,4-dinitrofluorophenylacetonitrile solution: Weigh approximately 200 mg of 2,4-dinitrofluorophenyl accurately, place it in a 20 mL volumetric flask, dissolve and dilute to the mark with acetonitrile to obtain a solution with a concentration of 10 mg / mL.
[0093] Phosphate buffer (pH 7.0): Take 0.68 g of potassium dihydrogen phosphate, add 29.1 mL of 0.1 mol / L sodium hydroxide solution, and dilute with water to 100 mL.
[0094] Test solution: Weigh approximately 300 mg of this product accurately, place it in a 10 mL volumetric flask, add water to dissolve and dilute to the mark, and shake well to obtain the test solution.
[0095] Reference solution: Weigh approximately 30 mg of taurine reference standard dried to constant weight at 105℃, place it in a 100 mL volumetric flask, dissolve and dilute with water to the mark, and shake well to obtain the solution.
[0096] Spiked test solution: Weigh approximately 300 mg of this product accurately, place it in a 10 mL volumetric flask, add reference solution to dissolve and dilute to the mark, shake well, and the solution is ready.
[0097] Taurine derivatization: Accurately measure 1 mL of the sample solution and place it in a 10 mL volumetric flask. Add 1 mL of 0.5 mol / L sodium bicarbonate solution (adjust the pH to 9.0 with 1 mol / L sodium hydroxide solution) and 0.5 mL of 1% 2,4-dinitrofluorophenylacetonitrile solution. Shake well and react in a 60℃ water bath for 1 hour. Remove from the water, cool, and dilute to the mark with phosphate buffer (pH 7.0). Shake well to obtain the final product.
[0098] Prepare the test solution, reference solution, and spiked test solution by following the "Taurine Derivatization Treatment" under "2. Solution Preparation" to obtain the derivatized test solution, derivatized reference solution, and derivatized spiked test solution, respectively.
[0099] 3. Limit of Quantitation and Preparation of Linear Solutions Limit of Quantification Solution: The reference solution was serially diluted with phosphate buffer (pH 7.0) with a signal-to-noise ratio of not less than 10. The solution was prepared according to the "Taurine Derivatization Treatment" under "2. Solution Preparation" to prepare the limit of quantification solution.
[0100] Linear stock solution: Accurately weigh 30.15 mg of taurine reference standard dried to constant weight at 105℃, place it in a 100 mL volumetric flask, dissolve and dilute with water to the mark, shake well, and prepare a linear stock solution containing 29.85 μg per mL.
[0101] Linear solution 1: Accurately transfer 1 mL of the linear stock solution into a 20 mL volumetric flask, dilute with water to the mark, shake well, and prepare linear solution 1 containing 1.492 μg per mL according to the "2. Preparation of solutions" section under "Taurine derivatization treatment".
[0102] Linear solution 2: Accurately transfer 1 mL of the linear stock solution into a 10 mL volumetric flask, dilute with water to the mark, shake well, and prepare linear solution 2 containing 2.895 μg per mL according to the "2. Preparation of solutions" section under "Taurine derivatization treatment".
[0103] Linear solution 3: Accurately transfer 2 mL of the linear stock solution into a 10 mL volumetric flask, dilute with water to the mark, shake well, and prepare linear solution 3 containing 5.970 μg per mL according to the "2. Preparation of solutions" section under "Taurine derivatization treatment".
[0104] Linear solution 4: Accurately transfer 5 mL of the linear stock solution into a 10 mL volumetric flask, dilute with water to the mark, shake well, and prepare linear solution 4 containing 14.92 μg per mL according to the "2. Preparation of solutions" section under "Taurine derivatization treatment".
[0105] Linear solution 5: Take the linear stock solution and prepare linear solution 5 containing 29.85 μg per 1 mL by "taurine derivatization treatment" under "2. Solution preparation".
[0106] 4. Preparation of accuracy solution Accuracy solution: Take 9 portions of the test sample, each approximately 300 mg, accurately weigh them, and place them in 10 mL volumetric flasks. Accurately add 0.5 mL, 1.0 mL, and 5.0 mL of taurine reference solution with a concentration of 29.85 μg / mL (3 portions of each volume), respectively. Dissolve and dilute with water to the mark, shake well, and prepare the accuracy solution according to the "Taurine Derivatization Treatment" under "2. Solution Preparation".
[0107] Methodological study of the detection method of the present invention 1. Specificity test Accurately measure 10 μL of the underivatized reference solution, the derivatized test solution, and the derivatized spiked test solution prepared above, and inject them separately into the liquid chromatograph, recording the chromatograms. Figures 1-3 The results are shown in Table 3.
[0108] Table 3 Specificity test results
[0109] According to the experimental results (Table 3), the chromatographic conditions of this detection method can separate 2,4-dinitrofluorobenzene, taurine, and magnesium acetylatum, which meets the requirements. The detection method of this invention has good specificity.
[0110] 2. Sensitivity Test Accurately measure 10 μL of the solution prepared above and inject it into the liquid chromatograph. Record the chromatogram. The results are shown in Table 4.
[0111] Table 4 Sensitivity Test Results
[0112] According to the test results (Table 4), the detection method of the present invention has good sensitivity.
[0113] 3. Linearity and Range Accurately measure 10 μL of the above-prepared limit of quantitation solution, linear solution 1, linear solution 2, linear solution 3, linear solution 4 and linear solution 5, inject them into the liquid chromatograph, record the chromatograms, and the results are shown in Table 5.
[0114] Table 5. Results of Linearity and Range Tests
[0115] According to the experimental results (Table 5), the linear equation of this detection method is as follows within the concentration range of 0.5970 μg / mL to 29.85 μg / mL: y = 82335787.6402 x - 862654.9190, r = 1.0000 (n = 6), indicating good linearity.
[0116] 4. Stability test Accurately measure 10 μL of the derivatized reference solution and the derivatized test solution prepared above, and inject them into the liquid chromatograph at different time points (0h, 25h, 33h, 41h). Record the chromatograms. The results are shown in Table 6.
[0117] Table 6 Stability Test Results
[0118] According to the test results (Table 6), the control solution and the test solution remained stable within 41 hours.
[0119] 5. Accuracy test (recovery rate test) Accurately measure 10 μL of the accuracy solution prepared above, inject it into the liquid chromatograph, record the chromatogram, and the results are shown in Table 7.
[0120] Table 7 Accuracy Test Results
[0121] According to the experimental results (Table 7), the recovery rate is between 98.7% and 100.3%, and the RSD of the 9 recovery rate data is 4.5%, indicating that the detection method of the present invention has good accuracy.
[0122] It should be understood that the above embodiments are exemplary and are not intended to encompass all possible implementations included in the claims. Various modifications and changes can be made to the above embodiments without departing from the scope of this disclosure. Similarly, the various technical features of the above embodiments can be arbitrarily combined to form other embodiments of the present invention that may not be explicitly described. Therefore, the above embodiments only illustrate several implementations of the present invention and do not limit the scope of protection of this patent.
Claims
1. A method for detecting taurine in magnesium acetylatum, characterized in that, Includes the following steps: (1) Derivatization treatment: Under alkaline conditions, the derivatizing reagent reacts with magnesium acetylatum containing taurine to obtain the derivatized product; (2) Detection: The derivatized products described in step (1) were detected using high performance liquid chromatography-ultraviolet detector; The derivatizing reagent is selected from 2,4-dinitrofluorophenylacetonitrile.
2. The detection method according to claim 1, characterized in that: In step (1), the alkaline reagent is selected from one or more of sodium hydroxide, sodium carbonate, and sodium bicarbonate; Preferably, the pH value of the alkaline condition is 8.0~10.0, and more preferably 9.0; Preferably, the concentration of the 2,4-dinitrofluorophenylacetonitrile is 5~20 mg / mL, and more preferably 10 mg / mL.
3. The detection method according to claim 1, characterized in that: In step (1), the reaction time of the derivatization reaction is 40-80 min, preferably 50-80 min, and more preferably 60 min; Preferably, in step (1), the reaction temperature of the derivatization reaction is 40~80℃; more preferably 50~80℃; and even more preferably 60℃.
4. The detection method according to any one of claims 1 to 3, characterized in that, In step (1), the derivatization process specifically includes: adding sodium bicarbonate solution (pH adjusted with sodium hydroxide solution) and 2,4-dinitrofluorophenylacetonitrile solution to the taurine-containing magnesium acetyl taurate, shaking well, reacting in a 60°C water bath for 1 hour, taking it out, cooling it, diluting it to the mark with a mobile phase additive, and shaking well. Preferably, the pH value of the mobile phase additive is 6.5~8.0, more preferably 7.0; Preferably, the mobile phase additive is selected from phosphate buffer.
5. The detection method according to claim 1, characterized in that, In step (2), the mobile phase of the high performance liquid chromatography is a mixture of mobile phase A and mobile phase B. Mobile phase A is selected from one or more of phosphate buffer, methanol, tetrahydrofuran, acetonitrile, and water, and mobile phase B is selected from one or more of methanol, tetrahydrofuran, acetonitrile, and water. Preferably, the mobile phase A is a mixed solution of phosphate buffer, acetonitrile, and water; Preferably, the pH value of the phosphate buffer solution is 6.5~8.0, more preferably 7.0; Preferably, the volume ratio of the phosphate buffer-acetonitrile-water is (60~80):(10~20):(10~20), more preferably 70:(10~20):(10~20), and even more preferably 70:15:15; Preferably, the mobile phase B is a mixed solution of acetonitrile and water; Preferably, the volume ratio of acetonitrile to water is (40~60):(40~60), more preferably 50:
50.
6. The detection method according to claim 1, characterized in that, In step (2), the detection wavelength of the high performance liquid chromatography is selected from 210~214nm, 263~267nm and 357~361nm, preferably 360nm.
7. The detection method according to claim 1, characterized in that, In step (2), the chromatographic column of the high performance liquid chromatography is a reversed phase column, preferably an octadecylsilane bonded silica column, more preferably a GL Sciences Wondasil C18 Superb column or a column with equivalent performance. Preferably, the chromatographic column has a length of 150 mm to 250 mm, an inner diameter of 2.7 to 4.6 mm, and a particle size of 3.5 μm to 5 μm; more preferably, it has a length of 250 mm, an inner diameter of 4.6 mm, and a particle size of 5 μm. Preferably, the column temperature of the chromatographic column is 30℃~40℃, and more preferably 30℃.
8. The detection method according to claim 1, characterized in that, In step (2), the injection flow rate of the high performance liquid chromatography is 0.5 mL / min to 1.5 mL / min, preferably 0.8 mL / min to 1.2 mL / min, and more preferably 1.0 mL / min.
9. The detection method according to claim 1, characterized in that, In step (2), the injection volume of the high performance liquid chromatography is 5 μL to 20 μL, preferably 10 μL to 20 μL, and more preferably 10 μL.
10. The detection method according to claim 1, characterized in that, In step (2), the gradient elution procedure of the high-performance liquid chromatography is as follows: 。