A method for detecting propyl phosphoric acid in 1-propyl phosphoric anhydride

Abstract

The present invention provides a detection method for propyl phosphoric acid in 1-propyl phosphoric anhydride, using high performance liquid chromatography, the stationary phase is octadecylsilane bonded silica gel, and the mobile phase is phosphoric acid aqueous solution or phosphoric acid aqueous solution-acetonitrile system. The present invention separates and detects propyl phosphoric acid in 1-propyl phosphoric anhydride for the first time, thereby effectively controlling the quality of 1-propyl phosphoric anhydride. The detection method of the present invention has strong specificity, good linearity, high sensitivity, accuracy, good repeatability, good stability and good durability, which fills the technical gap of the prior art in the detection of propyl phosphoric acid in 1-propyl phosphoric anhydride, and has practical promotion and application value.

Description

Technical Field

[0001] The invention belongs to the field of drug analysis and detection, and particularly relates to a method for detecting propyl phosphate in 1-propyl phosphoric anhydride. Background Art

[0002] 1-Propyl phosphoric anhydride (T3P), also known as 1-propyl phosphoric cyclic anhydride, is a colorless or light yellow syrupy liquid. It is a new type of condensing agent, dehydrating agent, and coupling agent. As a coupling agent, 1-propyl phosphoric anhydride is mainly used to form amides or esters in peptide synthesis. It can completely replace the traditional HOBt-type explosive and dangerous reagents and is a promising emerging high-efficiency coupling agent. Compared with traditional peptide synthesis coupling agents such as DCC, DIC, EDC, 11BTU, HBTU, etc., the product obtained by 1-propyl phosphoric anhydride has high purity. It is increasingly widely used in the dehydration and coupling reaction of acids and amines in the peptide synthesis process in organic synthesis, and has a trend of gradually replacing DCC, DIC and EDC. As an important condensing agent, 1-propyl phosphoric anhydride has high activity and high yield. It also has excellent reaction selectivity and low diastereoisomerization, as well as the advantages of small dosage, convenient post-treatment, and no sensitization.

[0003] At present, the research on 1-propyl phosphoric acid anhydride mainly focuses on synthesis method and application, such as CN111635434A discloses a synthesis method of 1-propyl phosphoric acid cyclic anhydride, but there is no research report on the quality detection method of 1-propyl phosphoric acid anhydride. The inventor found in the experiment that 1-propyl phosphoric acid anhydride is easy to absorb water during synthesis and storage, and is further hydrolyzed into propyl phosphoric acid, which causes its reaction activity to decrease when used as a condensing agent, affecting the yield of the condensation reaction. However, there is still a lack of corresponding detection methods in the prior art, and the hydrolyzate of the condensing agent cannot be controlled. Therefore, how to efficiently, conveniently, safely and low-costly realize the detection of propyl phosphoric acid in 1-propyl phosphoric acid anhydride becomes a problem to be solved. Summary of the invention

[0004] To solve the above problems, the present invention provides a method for detecting propyl phosphoric acid in 1-propyl phosphoric anhydride, which adopts high performance liquid chromatography, wherein the stationary phase is octadecylsilane bonded silica gel, and the mobile phase is an aqueous phosphoric acid solution or an aqueous phosphoric acid solution-acetonitrile system.

[0005] The above invention is completed based on the following discovery of the inventors: After investigation, it was found that the ultraviolet absorption of the analyte propyl phosphoric acid is very weak, and only has weak absorption at extremely low wavelengths. In this case, if the mobile phase absorption is strong, it will cover the peak of the analyte, or even produce an inverted peak. Therefore, the selection of the mobile phase should not only ensure the separation of the analyte, but also take into account the ultraviolet absorption to avoid interfering with the peak of the analyte. After screening, the inventors found that aqueous phosphoric acid-acetonitrile is the most suitable mobile phase system, which can meet the above two requirements at the same time, and provides the possibility for detecting propyl phosphoric acid in 1-propyl phosphoric anhydride.

[0006] Furthermore, the concentration of the aqueous phosphoric acid solution is 0.05% to 0.15% v / v, preferably 0.08% to 0.12% v / v, and more preferably 0.1% v / v.

[0007] Further, the mobile phase is an aqueous phosphoric acid solution or an aqueous phosphoric acid solution-acetonitrile system for isocratic elution, wherein the volume ratio of the aqueous phosphoric acid solution: acetonitrile is (98-100): (2-0); or, the mobile phase is an aqueous phosphoric acid solution-acetonitrile system for gradient elution, and the gradient elution program is: 0-5min, 0% acetonitrile; 5.01-10min, 30% acetonitrile; 10.01-25min, 0% acetonitrile. Under the above conditions, the analyte can obtain the best separation degree.

[0008] Further, the stationary phase is YMC-Pack ODS-AQ C18 or its equivalent chromatographic column; preferably, the length of YMC-Pack ODS-AQ C18 is 75-300 mm, more preferably 150-250 mm, more preferably 250 mm; preferably, the inner diameter of YMC-Pack ODS-AQ C18 is 1-10 mm, more preferably 2.1-4.6 mm, more preferably 4.6 mm; preferably, the filler particle size of YMC-Pack ODS-AQ C18 is 3-10 μm, more preferably 5 μm.

[0009] Furthermore, the column temperature of the high performance liquid chromatography is set at 20°C to 30°C, preferably 25°C.

[0010] Furthermore, the flow rate of the mobile phase is 0.8-1.2 ml per minute, preferably 1 ml per minute.

[0011] Furthermore, the high performance liquid chromatography method sets the detection wavelength to 190-200 nm, preferably 190 nm.

[0012] Furthermore, the mass volume ratio of 1-propyl phosphoric anhydride: diluent in the test solution is above 50 mg:1 ml, preferably 50-400 mg:1 ml, and more preferably 100 mg:1 ml.

[0013] Furthermore, the mass volume ratio of propyl phosphate: diluent in the reference solution is 0.1-1 mg:1 ml, preferably 0.5 mg:1 ml.

[0014] Furthermore, the injection volume is 5 to 15 μl, more preferably 10 μl.

[0015] Furthermore, the diluent is the phosphoric acid aqueous solution.

[0016] Furthermore, the detection is qualitative detection and / or quantitative detection.

[0017] Furthermore, the quantitative detection is to calculate the content of propyl phosphate by peak area using an external standard method.

[0018] The detection method provided by the present invention realizes the separation and detection of propyl phosphoric acid in 1-propyl phosphoric anhydride for the first time, and quantitatively detects the content of propyl phosphoric acid, thereby effectively controlling the quality of 1-propyl phosphoric anhydride. The methodological verification experiment results show that the system applicability, specificity, linearity, sensitivity, accuracy, stability, etc. of the detection method of the present invention can meet the requirements of accurately detecting propyl phosphoric acid in 1-propyl phosphoric anhydride, filling the technical gap of the prior art for the detection of propyl phosphoric acid in 1-propyl phosphoric anhydride, and having practical promotion and application value.

[0019] Obviously, according to the above contents of the present invention, in accordance with common technical knowledge and customary means in the art, without departing from the above basic technical ideas of the present invention, other various forms of modification, replacement or change may be made.

[0020] The above contents of the present invention are further described in detail below through specific implementation methods in the form of embodiments. However, this should not be understood as the scope of the above subject matter of the present invention being limited to the following examples. All technologies realized based on the above contents of the present invention belong to the scope of the present invention. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] Figure 1 is a typical chromatogram of the blank solvent in Example 1;

[0022] Figure 2 is a typical chromatogram of the reference substance solution in Example 1;

[0023] Figure 3 It is a typical chromatogram of the test solution in Example 1;

[0024] Figure 4 is a typical chromatogram of the mixed solution in Example 1;

[0025] Figure 5 is a typical chromatogram of the propyl phosphoric acid solution in Example 3;

[0026] Figure 6 is a typical chromatogram of the propyl phosphoric acid solution in Example 4;

[0027] Figure 7 It is a typical chromatogram of the propyl phosphoric acid solution in Comparative Example 1;

[0028] Figure 8 It is a typical chromatogram of the propyl phosphoric acid solution in Comparative Example 2. DETAILED DESCRIPTION

[0029] The raw materials and equipment used in the specific embodiments of the present invention are all known products and are obtained through commercial purchase.

[0030] The test sample 1-propyl phosphoric anhydride in the following experiments is all 1-propyl phosphoric anhydride (50% ethyl acetate solution).

[0031] The experimental materials are prepared as follows:

[0032] Shimadzu LC-2030C PLUS liquid chromatography system;

[0033] Agela Venusil HILIC hydrophilic interaction chromatography column (specifications: 4.6×250mm, 5μm);

[0034] YMC-Pack ODS-AQ C18 chromatographic column (specifications: 4.6×250mm, 5μm);

[0035] One hundred thousandth electronic balance (SECURA225D-1CN, Sartorius);

[0036] Propyl phosphate reference substance: Aladdin, batch number: B2223619;

[0037] 1-Propylphosphonic anhydride 50% ethyl acetate solution sample: Beijing Inokai Technology Co., Ltd., batch number: KYHBO01;

[0038] Acetonitrile was of chromatographic grade and phosphoric acid was of analytical grade.

[0039] Example 1 Detection of Propyl Phosphoric Acid in 1-Propyl Phosphoric Acid Anhydride of the Present Invention

[0040] 1. Chromatographic conditions

[0041] Chromatographic column: A chromatographic column with octadecylsilane bonded silica as filler is used. In this embodiment, a YMC-Pack ODS-AQ C18 chromatographic column is specifically selected, and the specific specifications are 4.6×250mm, 5μm;

[0042] Mobile phase: 0.1% v / v phosphoric acid aqueous solution as mobile phase A, acetonitrile as mobile phase B, gradient elution as shown in the table below;

[0043] The detection wavelength is 190nm;

[0044] The flow rate is 1.0 ml per minute;

[0045] The column temperature was 25°C;

[0046] Injection volume 10 μl;

[0047] Blank solvent / diluent: Mobile phase A (0.1% v / v phosphoric acid in water)

[0048]

[0049] 2. Sample preparation

[0050] 1) Preparation of test solution: Take 1-propylphosphoric anhydride, weigh accurately, dissolve in 0.1% v / v phosphoric acid aqueous solution and quantitatively dilute to make a solution containing 100 mg per 1 ml.

[0051] 2) Preparation of reference solution: Take the propyl phosphate reference, weigh accurately, dissolve in 0.1% v / v phosphoric acid aqueous solution and quantitatively dilute to make a solution containing 0.5 mg per 1 ml.

[0052] 3) Mixed solution: Take 1-propyl phosphoric anhydride and propyl phosphoric acid reference substance, weigh accurately, dissolve and quantitatively dilute with 0.1% v / v phosphoric acid aqueous solution to make a mixed solution containing about 100 mg of 1-propyl phosphoric anhydride and 0.5 mg of propyl phosphoric acid per 1 ml. Pipette blank solvent, reference substance solution, test solution and mixed solution into HPLC, record chromatogram, see Figure 1 to Figure 4 . The experimental results showed that the retention time of the propyl phosphate peak in the chromatogram of the reference solution was 3.214min; the theoretical plate number was calculated based on the impurity propyl phosphate peak to be 9747; the test sample and blank solvent did not interfere with the elution of propyl phosphate; the separation degree between the propyl phosphate peak and the adjacent peak (blank solvent elution) was 1.9. The chromatogram of the test solution showed a chromatographic peak with the same retention time as the propyl phosphate in the chromatogram of the reference solution, confirming that the test sample contained propyl phosphate.

[0053] Example 2 Methodological verification of the detection method of the present invention

[0054] The chromatographic conditions were the same as in Example 1.

[0055] 1 System suitability

[0056] Reference solution: Take an appropriate amount of propyl phosphate reference, weigh accurately, dissolve in 0.1% v / v phosphoric acid aqueous solution and quantitatively dilute to make a solution containing about 0.5 mg per 1 ml.

[0057] Accurately measure the reference solution and inject it, and record the chromatogram. The chromatogram of the reference solution is shown in Figure 1 The theoretical plate number calculated based on the impurity propyl phosphate peak is 9747, indicating that the column efficiency is high and can meet the detection requirements.

[0058] 2 Exclusiveness

[0059] Propyl phosphate stock solution: Weigh 99.92 mg of impure propyl phosphate reference substance into a 20 ml volumetric flask, add 0.1% v / v phosphoric acid aqueous solution to dissolve and dilute to the scale, and shake well.

[0060] Propyl phosphate reference solution: Accurately pipette 2 ml of propyl phosphate impurity stock solution into a 20 ml volumetric flask, add 0.1% v / v phosphoric acid aqueous solution to dilute to the scale, and shake well.

[0061] Test solution: Weigh 2004.39 mg of 1-propylphosphoric anhydride into a 20 ml volumetric flask, add 0.1% v / v phosphoric acid aqueous solution to dilute to the scale, and shake well.

[0062] Spiked test solution: Weigh 1998.73 mg of 1-propyl phosphoric anhydride and place it in a 20 ml volumetric flask. Then accurately pipette 2 ml of propyl phosphoric acid stock solution and place it in the same 20 ml volumetric flask. Add 0.1% v / v phosphoric acid aqueous solution to dilute to the scale and shake well.

[0063] Accurately measure the blank solvent, reference solution, test solution, and spiked test solution and inject one injection each, and record the chromatogram. Figure 1 to Figure 4 . It can be seen that the solvent does not interfere with the elution of propyl phosphoric acid; other impurities in the test solution do not interfere with the elution of propyl phosphoric acid; the separation degree of propyl phosphoric acid in the spiked test solution and the adjacent chromatographic peak is ≥1.5; the absolute value of the difference between the retention time of the propyl phosphoric acid peak in the spiked test solution and the retention time of the propyl phosphoric acid peak in the reference solution is 0.0min. This shows that the detection method of the present invention has good specificity. (Note: ① In this method, 1-propyl phosphoric anhydride does not present an effective chromatographic peak; ② The two solvent peaks at around 3.5 to 4 minutes in the blank solvent are affected by the pressure disturbance during injection and the slight difference in the gradient regression equilibrium of continuous injection, and the size may fluctuate.)

[0064] 3 Limit of Quantitation and Limit of Detection

[0065] Propyl phosphate stock solution: Weigh 99.66 mg of the impure propyl phosphate reference substance into a 20 ml volumetric flask, add 0.1% v / v phosphoric acid aqueous solution to dissolve and dilute to the scale, and shake well.

[0066] Quantitation limit solution: Dilute the propyl phosphate stock solution to the required concentration so that the signal-to-noise ratio of the impurity peak is 10-20.

[0067] Detection limit solution: Take the quantification limit solution and dilute it until the signal-to-noise ratio of the impurity peak is 3 to 7.

[0068] The quantitative limit solution was accurately measured and injected 6 times continuously, and the detection limit solution was injected 1 time, and the chromatogram was recorded. The experimental results showed that the quantitative limit concentration of propyl phosphate was 47.4620 μg / ml, the detection limit concentration was 23.7310 μg / ml, and the quantitative limit concentration accounted for 0.047% of the test sample concentration, indicating that the detection method of the present invention has good sensitivity.

[0069] 4 Linearity and Range

[0070] Propyl phosphate stock solution: Weigh 99.66 mg of the impure propyl phosphate reference substance into a 20 ml volumetric flask, add 0.1% v / v phosphoric acid aqueous solution to dissolve and dilute to the scale, and shake well.

[0071] Quantitation limit solution: Accurately pipette 0.2 ml of propyl phosphonic acid stock solution into a 20 ml volumetric flask, add 0.1% v / v phosphoric acid aqueous solution to dilute to the scale, and shake well.

[0072] 20% linear solution: Accurately pipette 0.4 ml of propyl phosphonic acid stock solution into a 20 ml volumetric flask, add 0.1% v / v phosphoric acid aqueous solution to dilute to the scale, and shake well.

[0073] 50% linear solution: Accurately pipette 1 ml of propyl phosphonic acid stock solution into a 20 ml volumetric flask, add 0.1% v / v phosphoric acid aqueous solution to dilute to the scale, and shake well.

[0074] 100% linear solution: Accurately pipette 2 ml of propyl phosphate stock solution into a 20 ml volumetric flask, add 0.1% v / v phosphoric acid aqueous solution to dilute to the scale, and shake well.

[0075] 150% linear solution: Accurately pipette 3 ml of propyl phosphonic acid stock solution into a 20 ml volumetric flask, add 0.1% v / v phosphoric acid aqueous solution to dilute to the scale, and shake well.

[0076] 200% linear solution: Accurately pipette 4 ml of propyl phosphonic acid stock solution into a 20 ml volumetric flask, add 0.1% v / v phosphoric acid aqueous solution to dilute to the scale, and shake well.

[0077] Precisely measure LOQ, 20%, 50%, 100%, 150%, 200% linear solutions and inject one needle respectively, and record the chromatogram. The experimental results show that the correlation coefficient r is 1.000; the ratio of the absolute value of the Y-axis intercept to the 100% linear concentration level response value is 2.0%; the response value RSD is 5.2%.

[0078] 5. Accuracy

[0079] Propyl phosphate stock solution: Weigh 99.92 mg of impure propyl phosphate reference substance into a 20 ml volumetric flask, add 0.1% v / v phosphoric acid aqueous solution to dissolve and dilute to the scale, and shake well.

[0080] Propyl phosphate reference solution: Accurately pipette 2 ml of propyl phosphate impurity stock solution into a 20 ml volumetric flask, add 0.1% v / v phosphoric acid aqueous solution to dilute to the scale, and shake well.

[0081] 50% recovery solution: Accurately weigh about 2000 mg of 1-propyl phosphoric anhydride and place it in a 20 ml volumetric flask. Transfer 1 ml of propyl phosphoric acid stock solution to the same 20 ml volumetric flask, add solvent to dissolve and dilute to scale, shake well, and you have it. (Prepare 3 copies in parallel)

[0082] 100% recovery solution: Weigh about 2000 mg of 1-propyl phosphoric anhydride and place it in a 20 ml volumetric flask. Transfer 2 ml of propyl phosphoric acid stock solution to the same 20 ml volumetric flask, add solvent to dissolve and dilute to scale, shake well, and you have it. (Prepare 3 copies in parallel)

[0083] 150% recovery solution: Weigh about 2000 mg of 1-propyl phosphoric anhydride and place it in a 20 ml volumetric flask. Transfer 3 ml of propyl phosphoric acid stock solution to the same 20 ml volumetric flask, add solvent to dissolve and dilute to scale, shake well, and you have it. (Prepare 3 copies in parallel)

[0084] Accurately measure each recovery solution and inject one injection, and record the chromatogram. The experimental results show that the recovery of propyl phosphate in 9 spiked test solutions is between 91.0% and 95.8%, and the recovery RSD is 1.7%.

[0085] 6 Solution stability

[0086] Propyl phosphate stock solution: Weigh 99.92 mg of impure propyl phosphate reference substance into a 20 ml volumetric flask, add 0.1% v / v phosphoric acid aqueous solution to dissolve and dilute to the scale, and shake well.

[0087] Propyl phosphate reference solution: Accurately pipette 2 ml of propyl phosphate impurity stock solution into a 20 ml volumetric flask, add 0.1% v / v phosphoric acid aqueous solution to dilute to the scale, and shake well.

[0088] Spiked test solution: Weigh 1998.73 mg of 1-propyl phosphoric anhydride and place it in a 20 ml volumetric flask. Then accurately pipette 2 ml of propyl phosphoric acid stock solution and place it in the same 20 ml volumetric flask. Add solvent to dilute to the scale and shake well.

[0089] Accurately measure the reference solution and the spiked test solution and inject one injection at different time points (0h, 5h, 9h, 16h, 24h) to record the chromatogram. The experimental results show that the ratio of the peak area of ​​the reference solution injected at each time point within 24h to the peak area at 0h is between 99.2% and 99.8%; the ratio of the peak area of ​​the spiked test solution injected at each time point within 24h to the peak area at 0h is between 100.2% and 100.7%.

[0090] Based on the above experimental results, the detection method provided by the present invention has good system applicability, specificity, sensitivity, linearity, accuracy, and solution stability, and can accurately determine propyl phosphate in 1-propyl phosphoric anhydride.

[0091] Example 3 Detection by isocratic elution using a phosphoric acid aqueous solution-acetonitrile system

[0092] 1. Chromatographic conditions

[0093] A chromatographic column using octadecylsilane bonded silica as a filler (YMC-Pack ODS-AQ C18, 4.6×250mm, 5μm or a chromatographic column of equivalent performance);

[0094] The mobile phase was 0.1% v / v aqueous phosphoric acid-acetonitrile (98:2, v / v);

[0095] The detection wavelength is 190nm;

[0096] The flow rate is 1 ml per minute;

[0097] The column temperature was 30°C;

[0098] Injection volume: 10 μl.

[0099] 2. Solution preparation

[0100] Propyl phosphate solution: Accurately weigh 19.21 mg of propyl phosphate into a 20 ml volumetric flask, add 0.1% v / v phosphoric acid aqueous solution to dissolve and dilute to the scale, and shake well.

[0101] Chromatogram see Figure 5 As can be seen from the figure, this method adopts isocratic elution with a mobile phase system containing a lower proportion of organic phase. The experimental results show that the separation degree between the solvent peak and the propyl phosphate peak is 1.6, which can basically meet the detection requirements, but the separation effect is not as good as Example 1.

[0102] Example 4 Detection by isocratic elution with phosphoric acid aqueous solution

[0103] 1. Chromatographic conditions

[0104] A chromatographic column using octadecylsilane bonded silica as a filler (YMC-Pack ODS-AQ C18, 4.6×250mm, 5μm or a chromatographic column of equivalent performance);

[0105] Isocratic elution was performed using 0.1% v / v phosphoric acid aqueous solution as the mobile phase;

[0106] The detection wavelength is 190nm;

[0107] The flow rate is 1.0 ml per minute;

[0108] The column temperature was 30°C;

[0109] Injection volume: 10 μl.

[0110] 2. Solution preparation

[0111] Propyl phosphate solution: Accurately weigh 20.13 mg of propyl phosphate into a 20 ml volumetric flask, add 0.1% v / v phosphoric acid aqueous solution to dissolve and dilute to the scale, and shake well.

[0112] Chromatogram see Figure 6 As can be seen from the figure, this method can achieve effective separation of propyl phosphoric acid and adjacent peaks by isocratic elution with pure water phase, with a separation degree of 1.8. However, it was found in the experiment that under the above conditions, subsequent components such as 1-propyl phosphoric anhydride were not completely eluted, which would interfere with the detection of subsequent continuous injection samples, and the elution effect was not as good as the gradient elution method used in Example 1.

[0113] Comparative Example 1 Effect of fixed phase on detection results of propyl phosphoric acid in 1-propyl phosphoric anhydride

[0114] 1. Chromatographic conditions

[0115] A hydrophilic interaction chromatography column (CAgela Venusil HILIC, 4.6 × 250 mm, 5 μm) was used;

[0116] The mobile phase was 0.1% v / v aqueous phosphoric acid-acetonitrile (20:80, v / v) for isocratic elution;

[0117] The detection wavelength is 190nm;

[0118] The flow rate is 1 ml per minute;

[0119] The column temperature was 30°C;

[0120] Injection volume: 10 μl.

[0121] 2. Solution preparation

[0122] Propyl phosphate solution: Accurately weigh 19.21 mg of propyl phosphate into a 20 ml volumetric flask, add acetonitrile to dissolve and dilute to the scale, and shake well.

[0123] Chromatogram see Figure 7 As can be seen from the figure, the solvent interferes with the elution of propyl phosphate under this method, and the separation degree between the two is only 0.8, which cannot be effectively detected and is not applicable.

[0124] Comparative Example 2 Effect of Mobile Phase Ratio on the Detection Results of Propyl Phosphoric Acid in 1-Propyl Phosphoric Anhydride

[0125] 1. Chromatographic conditions

[0126] A chromatographic column using octadecylsilane bonded silica as a filler (YMC-Pack ODS-AQ C18, 4.6×250mm, 5μm or a chromatographic column of equivalent performance);

[0127] The mobile phase was 0.1% v / v aqueous phosphoric acid-acetonitrile (95:5, v / v) for isocratic elution;

[0128] The detection wavelength is 190nm;

[0129] The flow rate is 1 ml per minute;

[0130] The column temperature was 30°C;

[0131] Injection volume: 10 μl.

[0132] 2. Solution preparation

[0133] Propyl phosphate solution: Accurately weigh 19.21 mg of propyl phosphate into a 20 ml volumetric flask, add 0.1% v / v phosphoric acid aqueous solution to dissolve and dilute to the scale, and shake well.

[0134] Chromatogram see Figure 8 , it can be seen from the figure that the resolution between the solvent peak and the propyl phosphate peak is 1.3 under this method. The resolution has improved, but if accurate quantitative detection is to be performed, the resolution needs to be further improved.

Claims

1. A method for detecting propyl phosphoric acid in 1-propyl phosphoric anhydride, characterized in that: The high performance liquid chromatography method is adopted, the stationary phase of the chromatographic column is octadecylsilane bonded silica gel, and the mobile phase is a phosphoric acid aqueous solution-acetonitrile system with a volume ratio of (98-100): (2-0) for isocratic elution; or, the mobile phase is a phosphoric acid aqueous solution-acetonitrile system for gradient elution, and the gradient elution program is: 0-5min, 0% acetonitrile; 5.01-10min, 30% acetonitrile; 10.01-25min, 0% acetonitrile.

2. The detection method according to claim 1, characterized in that: The concentration of the phosphoric acid aqueous solution is 0.05% to 0.15% v / v.

3. The detection method according to claim 2, characterized in that: The concentration of the phosphoric acid aqueous solution is 0.08% to 0.12% v / v.

4. The detection method according to claim 3, characterized in that: The concentration of the phosphoric acid aqueous solution is 0.1% v / v.

5. The detection method according to claim 1, characterized in that: The chromatographic column is YMC-Pack ODS-AQ C18.

6. The detection method according to claim 5, characterized in that: The YMC-Pack ODS-AQ C18 has a length of 250 mm, an inner diameter of 4.6 mm, and a filler particle size of 5 μm.

7. The detection method according to claim 1, characterized in that: The column temperature of the high performance liquid chromatography method is set at 20°C to 30°C.

8. The detection method according to claim 7, characterized in that: The column temperature of the HPLC was set at 25°C.

9. The detection method according to claim 1, characterized in that: The flow rate of the mobile phase is 0.8-1.2 ml per minute.

10. The detection method according to claim 9, characterized in that: The flow rate of the mobile phase was 1 ml per minute.

11. The detection method according to claim 1, characterized in that: The high performance liquid chromatography method is set to have a detection wavelength of 190-200 nm.

12. The detection method according to claim 11, characterized in that: The high performance liquid chromatography method is set with a detection wavelength of 190 nm.

13. The detection method according to claim 1, characterized in that: The mass volume ratio of 1-propyl phosphoric anhydride: diluent in the test solution is above 50 mg:1 ml; and / or, the mass volume ratio of propyl phosphoric acid: diluent in the reference solution is 0.1~1 mg:1 ml; and / or, the injection volume is 5~15 μl.

14. The detection method according to claim 13, characterized in that: The mass volume ratio of 1-propyl phosphoric anhydride: diluent in the test solution is 50~400mg:1ml, and / or, the mass volume ratio of propyl phosphoric acid: diluent in the reference solution is 0.5mg:1ml; and / or, the injection volume is 10μl.

15. The detection method according to claim 13 or 14, characterized in that: The diluent is a phosphoric acid aqueous solution.

16. The detection method according to claim 1, characterized in that: The detection is qualitative detection and / or quantitative detection.

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