A method for detecting the boric acid content and / or boron content in compound sulfamethoxazole and zinc oxide ointment

Abstract

The present invention discloses a method for detecting the content of boric acid and / or boron in compound sulfamethoxazole and zinc oxide ointment. The detection method prepares a test solution by hot water extraction of the compound sulfamethoxazole and zinc oxide ointment sample in a water bath at 60°C, and then performs high performance liquid chromatography detection. Compared with the prior art, the detection method of the present invention can more accurately detect the content of boric acid and / or boron in compound sulfamethoxazole and zinc oxide ointment, and avoid the problem that due to the presence of pink rose bengal powder in the matrix of compound sulfamethoxazole and zinc oxide ointment under the current standard, the judgment of the titration end point is interfered, resulting in inaccurate determination. Moreover, this detection method is simple to operate, has high specificity, strong anti-interference ability, good linearity, high precision and recovery rate, and good durability, and can effectively ensure the quality and safety of drugs.

Description

Technical Field

[0001] The present invention relates to the technical field of analytical chemistry, and specifically, to a method for detecting the content of boric acid and / or boron in compound sulfamethoxazole and zinc oxide ointment. Background Art

[0002] Compound sulfamethoxazole and zinc oxide ointment is composed of zinc oxide, boric acid, calamine, sulfamethoxazole, methyl salicylate and a matrix, and has the efficacy of antibacterial and anti-inflammatory, and is used for the treatment of skin diseases such as eczema and wound ulcers. The components such as zinc oxide and boric acid contained therein can better moisten the skin and reduce exudation, and have strong antibacterial activity, and can inhibit the growth of gram-positive and negative bacteria.

[0003] In the "National Drug Standard", the determination standard for boric acid in compound sulfamethoxazole and zinc oxide ointment is to determine the content of boric acid in the drug by titration method. Specifically, it is titrated with sodium hydroxide titrant until it turns pink to judge the end of titration. However, the matrix of compound sulfamethoxazole and zinc oxide ointment contains rose bengal powder. During the titration experiment, rose bengal powder is easily extracted simultaneously during the process of extracting the compound sulfamethoxazole and zinc oxide ointment sample, and the color of rose bengal powder is also pink, which is similar to the color of the titration end point specified in the "National Drug Standard", and it is easy to interfere with the judgment of the titration end point, thereby affecting the judgment of the product quality.

[0004] In the prior art, there are few detection methods for boric acid in compound sulfamethoxazole and zinc oxide ointment. With the widespread application of compound sulfamethoxazole and zinc oxide ointment, better and more perfect quality control methods are also needed to strictly control the production of compound sulfamethoxazole and zinc oxide ointment. Summary of the Invention

[0005] The purpose of the present invention is to overcome the above-mentioned deficiencies of the prior art, and to provide a method for detecting the content of boric acid and / or boron in compound sulfamethoxazole and zinc oxide ointment.

[0006] The first purpose of the present invention is to provide a method for preparing a test solution for detecting the content of boric acid and / or boron in compound sulfamethoxazole and zinc oxide ointment.

[0007] The second purpose of the present invention is to provide a method for detecting the content of boric acid and / or boron in compound sulfamethoxazole and zinc oxide ointment.

[0008] The third purpose of the present invention is to provide the application of the above detection method in detecting the content of boric acid and / or boron in compound sulfamethoxazole and zinc oxide ointment.

[0009] In order to achieve the above purpose, the present invention is realized by the following solutions:

[0010] A method for preparing a test solution for detecting the content of boric acid and / or boron in compound sulfamethoxazole and zinc oxide ointment includes the following steps:

[0011] S1. Mix compound sulfamethoxazole and zinc oxide ointment with water at a ratio of 1 g: 30 - 50 mL, heat and shake under the condition of 60°C - 70°C, cool down to form a solidified matrix, separate the solid and liquid to obtain Solution 1 and the solidified matrix;

[0012] S2. Mix the solidified matrix obtained in Step S1 with water, heat and shake under the condition of 60°C - 70°C, cool down to form a solidified matrix, separate the solid and liquid to obtain Solution 2; the volume of water added is in the ratio of the volume of water: the mass of the compound sulfamethoxazole and zinc oxide ointment in Step S1 = 20 - 40 mL: 1 g;

[0013] S3. Combine Solution 1 obtained in Step S1 and Solution 2 obtained in Step S2, mix well, and dilute with water to a concentration of 0.005 - 0.015 g / mL to obtain the test solution.

[0014] Preferably, the heating and shaking in Step S1 is water bath heating and shaking.

[0015] Preferably, the heating and shaking condition in Step S1 is 60°C.

[0016] Preferably, the heating and shaking condition in Step S2 is 60°C.

[0017] Preferably, the heating and shaking time in Step S1 is 10 - 20 min.

[0018] More preferably, the heating and shaking time in Step S1 is 15 min.

[0019] Preferably, the heating and shaking time in Step S2 is 10 - 20 min.

[0020] More preferably, the heating and shaking time in Step S2 is 10 min.

[0021] A method for detecting the boric acid content and / or boron content in compound sulfamethoxazole and zinc oxide ointment, characterized by comprising the following steps:

[0022] S1. Prepare the test solution according to the above preparation method;

[0023] S2. Prepare boron single-element standard solutions with concentrations ranging from 0.045 - 0.135 mg / mL, including no less than 5 gradient concentrations, conduct high performance liquid chromatography detection, use the lg value of the boron single-element standard solution concentration as the abscissa, and the lg value of the peak area detected by high performance liquid chromatography as the ordinate to draw a standard curve;

[0024] S3. Perform high performance liquid chromatography (HPLC) detection on the test sample solution obtained in step S1, and the content of boric acid and / or boron in Compound Sulfamethoxazole and Zinc Oxide Ointment can be obtained according to the peak areas obtained and the standard curve plotted in step S2.

[0025] Preferably, the chromatographic conditions for the HPLC detection in step S2 are as follows:

[0026] Chromatographic column: Acclaim TM Trinity TM P2, 100×2.1 mm, 3 μm;

[0027] Drift tube temperature: 30 °C;

[0028] Evaporator temperature: 60 °C;

[0029] Column temperature: 27 °C - 33 °C;

[0030] Flow rate: 0.27 - 0.33 mL / min;

[0031] Injection volume: 2 μL;

[0032] Detector: Evaporative light scattering detector;

[0033] Run time: 10 min;

[0034] Mobile phase: 18 - 22 mmol / L ammonium formate solution.

[0035] More preferably, the chromatographic conditions are as follows:

[0036] Chromatographic column: Acclaim TM Trinity TM P2, 100×2.1 mm, 3 μm;

[0037] Drift tube temperature: 30 °C;

[0038] Evaporator temperature: 60 °C;

[0039] Column temperature: 30 °C;

[0040] Flow rate: 0.3 mL / min;

[0041] Injection volume: 2 μL;

[0042] Detector: Evaporative light scattering detector;

[0043] Run time: 10 min;

[0044] Mobile phase: 20 mmol / L ammonium formate solution.

[0045] The present invention also claims the application of the above detection method in detecting the boric acid content and / or boron content in compound sulfamethoxazole and zinc oxide ointment; the boron content in the compound sulfamethoxazole and zinc oxide ointment is 0.045 mg / mL to 0.135 mg / mL.

[0046] Compared with the prior art, the present invention has the following beneficial effects:

[0047] The present invention provides a detection method for the boric acid content and / or boron content in compound sulfamethoxazole and zinc oxide ointment. Compared with the prior art, this detection method can more accurately detect the boric acid content and / or boron content in compound sulfamethoxazole and zinc oxide ointment. Moreover, this detection method is simple to operate, has high specificity, strong anti-interference ability, good linearity, high precision and recovery rate, and good durability, and can effectively ensure the quality and safety of drugs. BRIEF DESCRIPTION OF THE DRAWINGS

[0048] Figure 1 It is the standard curve graph of boron single element;

[0049] Figure 2 It is the chromatogram of the blank solution;

[0050] Figure 3 It is the chromatogram of the negative blank solution; the time corresponding to the peak is 3.197;

[0051] Figure 4 It is the chromatogram of the reference solution; the time corresponding to the peak is 1.136;

[0052] Figure 5 It is the chromatogram of the test solution 1 of F1017; the times corresponding to the peaks from left to right in the chromatogram are 1.136 and 3.195 in sequence; the peak at 1.136 is the response peak of boron single element in the test solution 1 of F1017; the peak at 3.195 is the response peak of other substances except boron single element in the test solution 1 of F1017;

[0053] Figure 6 It is the chromatogram of the 100% spiked test solution; the times corresponding to the peak data from left to right in the chromatogram are 1.131 and 3.210 in sequence; the peak at 1.131 is the response peak of boron single element in the 100% spiked test solution; the peak at 3.210 is the response peak of other substances except boron single element in the 100% spiked test solution;

[0054] Figure 7 It is the standard curve 2 graph of boron single element;

[0055] Figure 8 It is the determination result graph of detecting boron single element in compound sulfamethoxazole and zinc oxide ointment by CAD method. DETAILED DESCRIPTION OF THE INVENTION

[0056] The present invention will be further elaborated in detail below in conjunction with the accompanying drawings of the specification and specific embodiments. The embodiments are only used to explain the present invention and are not intended to limit the scope of the present invention. The test methods used in the following embodiments are all conventional methods unless otherwise specified; the materials, reagents, etc. used are reagents and materials that can be obtained from commercial channels unless otherwise specified.

[0057] Example 1 Detection method of boric acid in compound sulfamethoxazole and zinc oxide ointment

[0058] 1. Solution preparation

[0059] 1.1. Preparation of test solution

[0060] Preparation of test solution: Accurately weigh about 0.5 g of compound sulfamethoxazole and zinc oxide ointment (F1017), place the compound sulfamethoxazole and zinc oxide ointment in a 100 mL stoppered Erlenmeyer flask, add 20 mL of hot water to the flask, place the stoppered Erlenmeyer flask in a water bath at 60 °C, heat and shake for 15 min, let it cool, and after the matrix solidifies, transfer 1 to 50 mL of the solution to a volumetric flask to obtain the solidified matrix.

[0061] Add 15 mL of hot water to the solidified matrix, place it in a water bath at 60 °C, heat and shake for 10 min, let it cool, and after the matrix solidifies, transfer 2 of the solution to the 50 mL volumetric flask containing solution 1, dilute with water and shake well to obtain the test solution of F1017. The concentration of the test solution of F1017 is 0.01 g / mL (that is, 0.01 g of compound sulfamethoxazole and zinc oxide ointment (F1017) is dissolved in each milliliter of water); prepare two portions in parallel to obtain the test solution 1 and test solution 2 of F1017 respectively.

[0062] Replace compound sulfamethoxazole and zinc oxide ointment (F1017) with compound sulfamethoxazole and zinc oxide ointment (H1019), compound sulfamethoxazole and zinc oxide ointment (H1020), and compound sulfamethoxazole and zinc oxide ointment (H1021) respectively, and prepare the test solution by the same method.

[0063] 1.2. Preparation of standard curve solution

[0064] Taking the detection of F1017 sample as an example, accurately transfer different volumes of boron single-element standard solution (20DC499) to 10 mL volumetric flasks respectively, dilute with water and shake well to obtain boron single-element standard solutions with 5 gradient concentrations ranging from 0.045 to 0.135 mg / mL, as shown in Table 1 specifically.

[0065] Table 1 Boron single-element standard solution

[0066]

[0067]

[0068] 2. Standard Curve Drawing and Sample Detection

[0069] Taking the detection of F1017 sample as an example, the boron single-element standard solution prepared in Step 1.2 was detected by high performance liquid chromatography (HPLC), and the chromatographic conditions are shown in Table 3.

[0070] Table 3 Chromatographic Conditions

[0071]

[0072] The HPLC detection results of the boron single-element standard solution are shown in Table 4. Taking this as the standard, with the lg value of the concentration of the boron single-element standard solution as the abscissa (x) and the lg value of the peak area detected by HPLC as the ordinate (y), a standard curve was drawn. The boron single-element standard curve is as Figure 1 shown. The linear equation of the boron single-element standard curve is: y = 1.7317x + 4.3832, and the correlation coefficient r = 0.9990. Boron shows a good linear relationship within the concentration range of 0.045 mg / mL to 0.135 mg / mL, which is equivalent to 50% to 150% of the limit concentration.

[0073] Table 4 HPLC Detection Results of Boron Single-Element Standard Curve Solution

[0074] Solution number Concentration C (mg / mL) Peak area A lgC (mg / mL) lgA STD1 0.045 110.252 -1.3468 2.04239 STD2 0.072 259.002 -1.1427 2.41330 STD3 0.090 384.825 -1.0458 2.58526 STD4 0.108 505.227 -0.9666 2.70349 STD5 0.135 741.310 -0.8697 2.87000

[0075] The test solution 1 of F1017 and the test solution 2 of F1017 prepared in Step 1.1 were detected by HPLC under the chromatographic conditions shown in Table 3. The lg value of the peak area of the chromatographic results was combined into the linear equation of the boron single-element standard curve to calculate the boron concentration, and then the boric acid content was calculated. The specific calculation process is as follows, and the detection results and calculation results are shown in Table 5.

[0076] The formula for calculating the boron content in the test solution is as follows:

[0077] Formula I:

[0078] Ax: Peak area of the target compound in the test solution; K: Slope of the standard curve; b: Intercept of the standard curve; N: Dilution factor of the test solution (50 times); W: Mass (g) of compound sulfamethoxazole and zinc oxide ointment weighed.

[0079] The formula for calculating the boric acid content is as follows:

[0080] Formula II: Boric acid content (mg / g) = (boron content (%)) / 10.81) × 61.8 × 1000;

[0081] Boric acid content (%) = (Boron content (%) / 10.81) × 61.8 × 100%;

[0082] Wherein the boron content (%) is obtained by calculation with Formula I.

[0083] Table 5 Detection results of the test solution of F1017 and calculation of boric acid content

[0084]

[0085] Referring to Step 1.2, prepare the boron single-element standard curve solution 2 in the same manner, and perform the same determination according to the high performance liquid chromatography conditions described in Table 3 and calculate the linear equation of the standard curve 2. The determination results are shown in Table 6.

[0086] Table 6 High performance liquid chromatography detection results of the boron single-element standard curve solution 2

[0087] Solution number Concentration C (mg / mL) Peak area A lgC (mg / mL) lgA STD1 0.045 183.501 -1.3468 2.2636 STD2 0.072 402.053 -1.1249 2.6043 STD3 0.090 577.565 -1.0458 2.7616 STD4 0.108 739.467 -0.9666 2.8689 STD5 0.135 1032.496 -0.8697 3.0139

[0088] The linear equation of the standard curve 2 is: y = 1.5864x + 4.4011, and the correlation coefficient r = 0.9982. x is the lg value of the solution concentration, and y is the lg value of the peak area detected by high performance liquid chromatography.

[0089] Perform high performance liquid chromatography detection on the test solutions of H1019, H1020, and H1021 prepared in Step 1.1 respectively. The chromatography conditions are shown in Table 3. Combine the lg value of the peak area of the chromatography results with the linear equation of the standard curve 2 to calculate the boron concentration, and then calculate the boric acid content. The boric acid content calculation formula is as shown in Formula I and Formula II. The detection results and calculation results are shown in Table 7.

[0090] Table 7 Detection results of the test solutions of H1019, H1020, and H1021 and calculation of boric acid content

[0091]

[0092] The calculation results of the boric acid content of the test solutions of F1017, H1019, H1020, and H1021 are shown in Table 8.

[0093] Table 8 Boric acid content of each test solution

[0094]

[0095] The results show that: the detection results obtained by detecting the boric acid content in compound sulfamethoxazole and zinc oxide ointment using the method described in this example are consistent with the limit range of the boric acid content in compound sulfamethoxazole and zinc oxide ointment. This method can well detect the boric acid content in compound sulfamethoxazole and zinc oxide ointment.

[0096] Methodology Investigation of the Detection Method for Boric Acid in Compound Sulfadiazine Zinc Oxide Ointment in Example 2

[0097] 1. Experimental Method

[0098] 1.1 System Suitability Test

[0099] Reference Solution A1 / B1: Accurately pipette 0.9 mL of boron single-element standard solution into a 50-mL volumetric flask, dissolve with diluent (water) and dilute to volume with shaking to obtain Reference Solution A1. Prepare Reference Solution B1 in parallel. The concentrations of both Reference Solution A1 and Reference Solution B1 are 0.09 mg / mL.

[0100] Set water as the blank solution.

[0101] Inject Reference Solution A1, Reference Solution B1, and the blank solution respectively for high-performance liquid chromatography determination. The chromatographic conditions are shown in Table 3.

[0102] Inject Reference Solution A1 continuously for 6 times. After injecting 6 times, inject Reference Solution A1 once again (denoted as the control standard solution) to investigate the stability of the reference substance.

[0103] Inject Reference Solution B1 continuously for 2 times.

[0104] 1.2 Specificity

[0105] Negative Blank Solution: Weigh approximately 0.5 g of boric acid negative sample (compound sulfadiazine zinc oxide ointment without boric acid), place it in a 100-mL stoppered Erlenmeyer flask, and prepare a boron negative blank solution. The preparation method is the same as that of the test solution in Example 1.

[0106] 100% Spiked Test Solution: Accurately weigh 0.25 g of compound sulfadiazine zinc oxide ointment, place it in a 100-mL stoppered Erlenmeyer flask, then accurately pipette 0.45 mL of boron single-element standard solution into the stoppered Erlenmeyer flask. Add 20 mL of hot water to the flask, place the stoppered Erlenmeyer flask in a 60°C water bath, heat and shake for 15 min. After cooling until the matrix solidifies, transfer a portion of the solution to a 50-mL volumetric flask to obtain the solidified matrix. Add 15 mL of hot water to the solidified matrix, place it in a 60°C water bath, heat and shake for 10 min. After cooling until the matrix solidifies, transfer a portion of the solution (Solution 2) to the 50-mL volumetric flask containing Solution 1, and dilute to volume with water and shake well to obtain the 100% spiked test solution.

[0107] Inject the blank solution, negative blank solution, Reference Solution A1, Test Solution F1017 prepared in Example 1, and 100% spiked test solution respectively for high-performance liquid chromatography determination. The chromatographic conditions are shown in Table 3.

[0108] 1.3 Repeatability

[0109] According to the preparation method of the test solution described in Example 1, using F1017 as the test sample, prepare 6 groups of test solutions in parallel and name them REP-1 to REP-6.

[0110] Inject REP-1 to REP-6 respectively for high performance liquid chromatography determination, and the chromatographic conditions are shown in Table 3.

[0111] Based on the standard curve obtained in Step 2 of Example 1 and combined with the chromatographic detection results of REP-1 to REP-6, the boric acid contents of REP-1 to REP-6 are obtained respectively.

[0112] 1.4 Intermediate precision

[0113] By two different experimenters (Experimenter 1 and Experimenter 2), Experimenter 2, according to the preparation method of the test solution described in Example 1, using F1017 as the test sample, prepare 6 groups of test solutions in parallel and name them REP-IP-1 to REP-IP-6.

[0114] Experimenter 2 refers to the preparation method of the boron single-element standard curve solution in Step 1.2 of Example 1 to prepare and draw the standard curve graph, and calculate the linear equation of the corresponding standard curve. The specific steps refer to Step 2 of Example 1.

[0115] Experimenter 1 uses REP-1 to REP-6.

[0116] The two experimenters inject REP-1 to REP-6 and REP-IP-1 to REP-IP-6 respectively for high performance liquid chromatography determination, and the chromatographic conditions are shown in Table 3.

[0117] Based on the linear equation of the boron single-element standard curve obtained in Step 2 of Example 1 and combined with the chromatographic detection results of REP-1 to REP-6, the boric acid contents are obtained respectively.

[0118] Based on the linear equation of the standard curve graph drawn by Experimenter 2 and combined with the chromatographic detection results of REP-IP-1 to REP-IP-6, the boric acid contents are obtained respectively.

[0119] 1.5 Accuracy (recovery rate)

[0120] 80% Spiked Test Sample Solution: Accurately weigh 0.25 g of Compound Sulfamethoxazole and Zinc Oxide Ointment, place it in a 100 mL stoppered Erlenmeyer flask, then accurately transfer 0.27 mL of boron single-element standard solution into the stoppered Erlenmeyer flask. Add 20 mL of hot water to the flask, place the stoppered Erlenmeyer flask in a 60 °C water bath, heat and shake for 15 min. After cooling and waiting for the matrix to solidify, transfer a portion of the solution to a 50 mL volumetric flask to obtain the solidified matrix. Add 15 mL of hot water to the solidified matrix, place it in a 60 °C water bath, heat and shake for 10 min. After cooling and waiting for the matrix to solidify, transfer a portion of the solution to a 50 mL volumetric flask containing the first solution, dilute with water and shake well to obtain the 80% spiked test sample solution. Prepare three replicates in parallel and name them ACC80-1 to ACC80-3.

[0121] 100% Spiked Test Sample Solution: Accurately weigh 0.25 g of Compound Sulfamethoxazole and Zinc Oxide Ointment, place it in a 100 mL stoppered Erlenmeyer flask, then accurately transfer 0.45 mL of boron single-element standard solution into the stoppered Erlenmeyer flask. Add 20 mL of hot water to the flask, place the stoppered Erlenmeyer flask in a 60 °C water bath, heat and shake for 15 min. After cooling and waiting for the matrix to solidify, transfer a portion of the solution to a 50 mL volumetric flask to obtain the solidified matrix. Add 15 mL of hot water to the solidified matrix, place it in a 60 °C water bath, heat and shake for 10 min. After cooling and waiting for the matrix to solidify, transfer a portion of the solution to a 50 mL volumetric flask containing the first solution, dilute with water and shake well to obtain the 100% spiked test sample solution. Prepare three replicates in parallel and name them ACC100-1 to ACC100-3.

[0122] 120% Spiked Test Sample Solution: Accurately weigh 0.25 g of Compound Sulfamethoxazole and Zinc Oxide Ointment, place it in a 100 mL stoppered Erlenmeyer flask, then accurately transfer 0.63 mL of boron single-element standard solution into the stoppered Erlenmeyer flask. Add 20 mL of hot water to the flask, place the stoppered Erlenmeyer flask in a 60 °C water bath, heat and shake for 15 min. After cooling and waiting for the matrix to solidify, transfer a portion of the solution to a 50 mL volumetric flask to obtain the solidified matrix. Add 15 mL of hot water to the solidified matrix, place it in a 60 °C water bath, heat and shake for 10 min. After cooling and waiting for the matrix to solidify, transfer a portion of the solution to a 50 mL volumetric flask containing the first solution, dilute with water and shake well to obtain the 120% spiked test sample solution. Prepare three replicates in parallel and name them ACC120-1 to ACC120-3.

[0123] Inject the 80% spiked test sample solution, 100% spiked test sample solution, and 120% spiked test sample solution respectively for high performance liquid chromatography determination. The chromatographic conditions are shown in Table 3.

[0124] Perform the same determination for each parallel configuration group of the spiked test sample solution. Calculate the boric acid content respectively based on the linear equation of the boron single-element standard curve obtained in Step 2 of Example 1 and the chromatographic detection results.

[0125] 1.6 Solution Stability

[0126] The test solution 1 of F1017 prepared in Example 1 and the reference solution A1 prepared in Step 1.2 of this example were respectively injected for high performance liquid chromatography determination, and the chromatographic conditions are shown in Table 3.

[0127] The solution stability was investigated at room temperature. The specific method was as follows: The reference solution A1 was injected for determination at 0 h, 5 h, 9 h and 33 h respectively, and the change rate of the peak area in the chromatogram was calculated with the peak area determined at 0 h.

[0128] The test solution 1 of F1017 was injected for determination at 0 h, 3 h, 7 h and 31 h respectively, and the change rate of the peak area in the chromatogram was calculated with the peak area determined at 0 h.

[0129] 2. Experimental Results

[0130] 2.1 System Suitability Results

[0131] The injection test results of the reference solution A1 are shown in Table 9, and the injection test results of the reference solution B1 are shown in Table 10.

[0132] Table 9 Injection Test Results of the Reference Solution A1

[0133]

[0134] Table 10 Injection Test Results of the Reference Solution B1

[0135]

[0136] According to the results shown in Tables 9 - 10, when the reference solution A1 was continuously injected 6 times, the RSD of the target peak area was 0.8%, and the RSD of the retention time was 0.1%; when the reference solution B1 was continuously injected 2 times, the recovery rate was 101.5%; the RSD of the target peak area of the control standard solution and the reference solution A1 injected 6 times was 4.1%, and the RSD of the retention time was 0.2%.

[0137] The results indicate that the system suitability of the detection method for boric acid in compound sulfamethoxazole and zinc oxide ointment is good.

[0138] 2.2 Specificity Results

[0139] The chromatogram of the blank solution is as Figure 2 shown, the chromatogram of the negative blank solution is as Figure 3 shown, the chromatogram of the reference solution is as Figure 4 shown, the chromatogram of the test solution 1 of F1017 is as Figure 5 shown, the chromatogram of the 100% spiked test solution is asFigure 6 as shown

[0140] The results showed that no target peak was detected in the chromatogram of the blank solution, indicating no interference with the detection; no target peak was detected in the chromatogram of the negative blank solution, indicating no interference with the detection; the retention time of the target peak in the test solution of F1017 was 1.136 min, the retention time of the target peak in the 100% spiked test solution was 1.131 min, and the retention time of the target peak in the reference solution A1 was 1.136 min, showing that the retention times of the target peaks were consistent; the resolution between the target peak in the test solution of F1017 and the adjacent peak was 13.8 ≥ 2.0, and the resolution between the target peak in the 100% spiked test solution and the adjacent peak was 14.1 ≥ 2.0.

[0141] The results indicated that the detection method for boric acid in compound sulfamethoxazole and zinc oxide ointment had good specificity.

[0142] 2.3 Repeatability results

[0143] The determination results of REP-1 to REP-6 are shown in Table 11.

[0144] Table 11 Determination results of REP-1 to REP-6

[0145]

[0146] The average content of boric acid in the test solutions of the 6 groups of parallel preparations in the determination results was 4.8%, and the RSD was 2.5% which was less than 5.0%.

[0147] The results indicated that the detection method for boric acid in compound sulfamethoxazole and zinc oxide ointment had good repeatability.

[0148] 2.4 Intermediate precision results

[0149] The detection results of the boron single-element standard curve solution prepared by Experimenter 2 are shown in Table 12.

[0150] Table 12 Detection results of the boron single-element standard curve solution prepared by Experimenter 2

[0151] Solution number Concentration C (mg / mL) Peak area A lgC (mg / mL) lgA STD1(2) 0.045 110.921 -1.3468 2.04501 STD2(2) 0.072 260.129 -1.1427 2.41519 STD3(2) 0.090 365.684 -1.0458 2.56311 STD4(2) 0.108 500.629 -0.9666 2.69952 STD5(2) 0.135 726.472 -0.8697 2.86122

[0152] The standard curve 2 graph drawn by Experimenter 2 is as Figure 7 shown, and the linear equation corresponding to Standard Curve 2 is y = 1.7039x + 4.3473, with a correlation coefficient r = 0.9992.

[0153] The determination results of REP-1 to REP-6 and REP-IP-1 to REP-IP-6 are shown in Table 13.

[0154] Table 13 Determination Results of REP-1 to REP-6 and REP-IP-1 to REP-IP-6

[0155]

[0156]

[0157] The results showed that the average content of the 6 test solution samples REP-IP-1 to REP-IP-6 of Experimenter 2 was 4.8%, and the RSD was 1.1% < 5%; the RSD of boric acid in the 12 test solution samples of the two experimenters was 1.8%.

[0158] The results indicated that the intermediate precision of the detection method for boric acid in compound sulfamethoxazole and zinc oxide ointment was good.

[0159] 2.5 Accuracy (Recovery) Results

[0160] The determination results of different spiked test solution samples are shown in Table 14.

[0161] Table 14 Determination Results of Spiked Test Solution Samples

[0162]

[0163] The results showed that the recoveries of boric acid in the 80%, 100%, and 120% spiked test solution samples were between 88.13% and 97.37%. The average recovery of boric acid in the 80% spiked test solution sample was 92.2%; the average recovery of boric acid in the 100% spiked test solution sample was 95.5%; the average recovery of boric acid in the 120% spiked test solution sample was 93.1%; the RSD of the 9 spiked test solution samples was 2.8% ≤ 8.0%.

[0164] The results indicated that the accuracy (recovery) of the detection method for boric acid in compound sulfamethoxazole and zinc oxide ointment was good.

[0165] 2.6 Solution Stability Results

[0166] The stability results of the reference solution are shown in Table 15.

[0167] Table 15 Stability Results of the Reference Solution

[0168]

[0169] Note: "NA" indicates not applicable

[0170] Table 16 Stability Results of Test Solution 1

[0171]

[0172] Note: "NA" indicates not applicable

[0173] Results show that the change rate of the boric acid peak area of the reference solution A1 from 0 h to 33 h was calculated with that at 0 h, and the change rate was between 10.5% and 12.8%, and the results met the requirements; the change rate of the boric acid peak area of the test solution 1 of F1017 from 0 h to 31 h was calculated with that at 0 h, and the change rate was between 12.2% and 12.7%, and the results met the requirements.

[0174] The results indicate that the solution in the detection method of boric acid in compound sulfamethoxazole and zinc oxide ointment has good stability.

[0175] Example 3 Durability Investigation of the Detection Method of Boric Acid in Compound Sulfamethoxazole and Zinc Oxide Ointment

[0176] 1. Experimental Method

[0177] The control group was set as follows: The peak areas and retention times of the reference solution A1 and the reference solution B1 were determined under the chromatographic conditions described in Table 3. Among them, the reference solution A1 was continuously injected 6 times, and after 6 injections, the reference solution A1 (recorded as the control standard solution) was injected once again to investigate the stability of the reference substance.

[0178] The boron single-element standard curve solution prepared in Step 1.2 of Example 1 was selected, and the linear equation of the corresponding standard curve and the corresponding detection range were calculated according to the method described in Step 2 of Example 1 under the chromatographic conditions described in Table 3; the test solution of F1017 obtained in Example 1 was injected to determine the boric acid content.

[0179] The difference between Treatment Group 1 and the control group was that the column temperature in Table 3 was adjusted to 33 °C;

[0180] The difference between Treatment Group 2 and the control group was that the column temperature in Table 3 was adjusted to 27 °C;

[0181] The difference between Treatment Group 3 and the control group was that the flow rate in Table 3 was adjusted to 0.33 mL / min;

[0182] The difference between Treatment Group 4 and the control group was that the flow rate in Table 3 was adjusted to 0.27 mL / min;

[0183] The difference between Treatment Group 5 and the control group was that the ammonium formate concentration in Table 3 was adjusted to 18 mmol / L;

[0184] The difference between Treatment Group 6 and the control group was that the ammonium formate concentration in Table 3 was adjusted to 22 mmol / L;

[0185] The difference between Treatment Group 7 and the control group was that the chromatographic column in Table 3 was replaced with GB-L-21-07-251.

[0186] According to the measurement method of the control group, the treatment groups 1 to 7 were treated equally, and the corresponding results were measured.

[0187] 2. Experimental Results

[0188] During the high performance liquid chromatography (HPLC) determination of the control group and each treatment group, the peak areas of the reference solution A1 are shown in Table 17; the retention times of the reference solution A1 during the HPLC determination of the control group and each treatment group are shown in Table 18; the recovery rates of the reference solution B1 during the HPLC determination of the control group and each treatment group are shown in Table 19; the linear equations, detection ranges, and the determined contents of the test solution F017 for the HPLC of the control group and each treatment group are shown in Table 20.

[0189] Table 17 Results of the Peak Areas of the Reference Solution A1 under Different Conditions

[0190]

[0191] Table 18 Retention Times of the Reference Solution A1 under Different Conditions

[0192]

[0193]

[0194] Table 19 Results of the Recovery Rates of the Reference Solution B1 under Different Conditions

[0195]

[0196] Table 20 Linear Equations, Detection Ranges, and Determined Contents of the HPLC under Different Conditions

[0197]

[0198] The results show that when the column temperature varies within the range of 27°C to 33°C, the flow rate varies within the range of 0.27 mL / min to 0.33 mL / min, the mobile phase ammonium formate varies within the range of 18 mmol / L to 22 mmol / L, and different numbered chromatographic columns are replaced, the relative standard deviation (RSD) range of the peak areas of the target compounds in the reference solution A1 is 0.6% to 2.9%, and the RSD range of the retention times is 0.1% to 0.3%; the recovery rate range of the reference solution B1 is 98.1% to 102.3%; the range of the correlation coefficient r of the standard curve of the target compounds is 0.9990 to 0.9999, showing a good linear relationship; the RSD range of the determined content of the target compounds in the test solution F1017 and the content measured under the standard conditions is 1.8% to 3.6%.

[0199] The results indicate that the durability of the detection method for boric acid in compound sulfamethoxazole and zinc oxide ointment is good.

[0200] Control Example 1 Determination of Boric Acid Content in Compound Sulfamethoxazole and Zinc Oxide Ointment by Atomic Absorption Spectrometry

[0201] 1. Experimental Method

[0202] (1) Preparation of Standard Solution

[0203] Reference Stock Solution: Accurately pipette 0.7 mL of boron single element standard solution (5000 μg / mL) into a 10 mL volumetric flask, dilute to the scale line with 1% nitric acid solution (v / v) to obtain the reference stock solution (350 μg / mL) for standby use.

[0204] Preparation of Calibration Curve Solution: Accurately pipette 2 mL of the reference stock solution into a 10 mL volumetric flask, dilute to the scale line with 1% nitric acid solution (v / v) to obtain the standard stock solution for standby use (70 μg / mL); accurately pipette 0.00 mL, 0.25 mL, 0.40 mL, 0.50 mL, 0.60 mL, and 0.75 mL of the standard stock solution into 10 mL volumetric flasks respectively, and add 1% nitric acid solution (v / v) to prepare calibration curve solutions with concentrations of 0 μg / mL, 1.75 μg / mL, 2.80 μg / mL, 3.5 μg / mL, 4.2 μg / mL, and 5.25 μg / mL in sequence, and name them sample blank, sample 1, sample 2, sample 3, sample 4, and sample 5.

[0205] (2) Preparation of Test Solution for Atomic Absorption Spectrometry

[0206] Accurately weigh 0.2 g of compound sulfamethoxazole and zinc oxide ointment (F1017), place it in a pressure-resistant and high-temperature-resistant microwave digestion tank, add 5 mL of nitric acid, heat it on a hot plate for pre-digestion until no yellow smoke emerges, take it out and let it cool, add 5 mL of nitric acid again, cover the inner lid of the digestion tank, seal it, put the digestion tank into the microwave digestion system, and the microwave digestion program is shown in Table 21.

[0207] Table 21 Microwave Digestion Program

[0208] Step Temperature (°C) Pressure (bar) Insulation time (min) 1 120 10 2 2 150 20 3 3 180 30 3 4 200 40 15

[0209] After complete digestion, cool the digestion solution to below 60 °C, take out the digestion tank, remove the inner digestion tank and heat it on a hot plate at 100 °C for 20 min, let it cool, open the inner lid until the red-brown smoke dissipates, transfer the digestion solution to a 25 mL volumetric flask, wash the inner lid and the digestion tank 3 times with a small amount of water, combine the washing solutions into the volumetric flask, dilute to the scale line with water, shake well to obtain the dilution solution, accurately pipette 0.5 mL of the dilution solution into a 10 mL volumetric flask, dilute to the scale with water, and shake well to obtain the test solution 1 for atomic absorption spectrometry.

[0210] Prepare 5 portions of the test solution for atomic absorption spectrometry in parallel, and label them as the test solutions for atomic absorption spectrometry 2 to 6 respectively.

[0211] The preparation method of the blank solution is different from that of the test solution for atomic absorption spectrometry in that: the compound sulfamethoxazole and zinc oxide ointment (F1017) is not added to the blank solution.

[0212] (3) Test method

[0213] The test conditions are as follows:

[0214] Method: Graphite furnace method; Type: Zeeman; Element: B; Sampling mode: Automatic preparation; Measurement mode: Peak height; Calibration mode: Concentration; Wavelength: 249.8 nm.

[0215] Drying temperature: 85°C - 120°C, drying time: 55 s, ashing temperature: 1000°C, ashing time: 8 s, atomization temperature: 3000°C, atomization time: 5.0 s.

[0216] The test procedure of the graphite furnace method is shown in Table 22.

[0217] Table 22 Graphite furnace method procedure

[0218]

[0219]

[0220] Remark: R: Reading step; S: Signal storage

[0221] Measure the blank standard and standards 1 to 5 respectively and calculate the linear equation.

[0222] Repeatability investigation: Select the test solutions for atomic absorption spectrometry 1 to 6 prepared in step (2), and detect the boric acid concentration under the above conditions respectively. Each sample is detected in parallel twice.

[0223] 2. Experimental results

[0224] The test results of the blank standard and standards 1 to 5 are shown in Table 23, and the results of the repeatability investigation are shown in Table 24.

[0225] Table 23 Test results of the blank standard and standards 1 to 5

[0226]

[0227] Table 24 Results of repeatability investigation

[0228]

[0229] The results showed that the theoretical detection concentration of the sample should be approximately 3.5 μg / mL. After repeated testing, the graphite tube was damaged, the atomization hole increased significantly, and the stability was poor. This should be caused by a relatively high atomization temperature. If the atomization temperature was reduced, the response would immediately drop linearly and fail to meet the sensitivity requirements.

[0230] Determination of boric acid content in compound sulfamethoxazole and zinc oxide ointment by comparative example 2 CAD method

[0231] 1. Experimental method

[0232] The preparation method of the test solution for the CAD method was the same as that of the test solution of F1017 in Example 1; the preparation methods of the blank solution, negative blank solution, 100% spiked test solution, and reference solution were the same as those described in Example 2 for solution preparation.

[0233] The experimental conditions are shown in Table 25.

[0234] Table 25 Experimental conditions for the CAD method

[0235] Instrument information Agilent Chromatographic column <![CDATA[Acclaim TM Trinity TM P2, 100×2.1mm, 3μm]]> Nebulizer temperature 50℃ Column temperature 30℃ Flow rate 0.3 mL / min Injection volume 2 μl Running time 10 min Mobile phase 20 mmol / l ammonium formate solution

[0236] The blank solution, negative blank solution, reference solution, and 100% spiked test solution were used for determination in sequence.

[0237] 2. Experimental results

[0238] The measurement results are as Figure 8 shown. The results showed that the target peak was at 3.8 minutes in the figure. The black color was the measurement result of the blank solution, and the other two colors were the measurement results of the reference solution. It can be seen that there was interference between the measurement results of different solutions, affecting the detection accuracy.

[0239] Determination of boric acid content in compound sulfamethoxazole and zinc oxide ointment by comparative example 3 derivatization method

[0240] 1. Experimental method

[0241] Acidic methanol: Take methanol solution and adjust the pH value to pH = 4 with acetic acid.

[0242] Curcumin glacial acetic acid solution: Weigh 125 mg of curcumin and dissolve it in glacial acetic acid, and dilute it to 100 mL with glacial acetic acid to obtain curcumin glacial acetic acid solution.

[0243] Sulfuric acid glacial acetic acid mixture: Mix sulfuric acid and glacial acetic acid according to the volume ratio of sulfuric acid: glacial acetic acid = 1:1 to obtain sulfuric acid glacial acetic acid mixture.

[0244] Reference solution: Weigh accurately 30 mg of boric acid reference standard, place it in a 200 mL volumetric flask, dissolve it with acidic methanol solution and dilute to the mark, shake well to obtain boric acid reference standard solution.

[0245] Precisely measure 1 mL of boric acid standard solution and place it in a 25-mL volumetric flask. Add 4 mL of curcumin glacial acetic acid solution, shake well, then add 2 mL of sulfuric acid glacial acetic acid mixture, shake well and let stand for 30 min. Dilute to the mark with methanol, shake well to obtain the stock solution of the reference substance. Precisely measure 2.5 mL of the stock solution of the reference substance and place it in a 25-mL volumetric flask. Dilute to the mark with methanol, shake well to obtain the reference solution.

[0246] Test solution: Take 0.6 g of the test sample (Compound Sulfamethoxazole and Zinc Oxide Ointment (F1017)), place it in a 200-mL beaker, add 25 - 50 mL of acidic methanol, heat in a water bath at 80 °C until the test sample is completely dissolved. Transfer the solution in the beaker to a 200-mL volumetric flask, rinse the beaker with acidic methanol, and transfer the rinsed solution to the 200-mL volumetric flask as well. Make up to the mark with acidic methanol and shake well. Filter, precisely measure 1 mL of the continued filtrate and place it in a 25-mL volumetric flask. Add 4 mL of curcumin glacial acetic acid solution, shake well, then add 2 mL of sulfuric acid glacial acetic acid mixture, shake well and let stand for 30 min. Dilute to the mark with methanol, shake well to obtain the stock solution of the test sample. Precisely measure 2.5 mL of the stock solution of the test sample and place it in a 25-mL volumetric flask. Dilute to the mark with methanol, shake well to obtain the test solution.

[0247] The experimental conditions are shown in Table 27.

[0248] Table 27 Experimental Conditions for the Derivatization Method

[0249] Instrument information High performance liquid chromatograph Chromatographic column Octadecylsilyl bonded silica column, 4.6 mm × 250 mm, 5 μm Column temperature 30℃ Flow rate 0.8 mL / min Injection volume 10 μl Detection wavelength 550 nm Mobile phase Methanol: 0.012 mol / L tetrabutylammonium bromide solution = 80:20 (v / v)

[0250] Inject the test solution for detection and record the corresponding detection results.

[0251] Inject for detection again and record them as Test Sample - 1 and Test Sample - 2 respectively.

[0252] 2. Experimental Results

[0253]

[0254] Boric acid itself has no absorption under ultraviolet light. In this example, curcumin is added to the acetic acid solution of boric acid and curcumin to carry out derivatization treatment at room temperature to form a complex, and this complex is analyzed by high-performance liquid chromatography to determine its content. There are a large amount of pigments, paraffin, and petrolatum in Compound Sulfamethoxazole and Zinc Oxide Ointment, which interfere with the derivatization reaction, affect the complexation effect, and thus affect the determination results.

[0255] The results show that when using the derivatization method to determine boric acid in Compound Sulfamethoxazole and Zinc Oxide Ointment, the measured boric acid content is only 4.4%, compared with the actual content of 5%, only 88% of the actual content can be detected.

[0256] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention rather than limit the protection scope of the present invention. For those of ordinary skill in the art, other different forms of changes or modifications can be made based on the above description and ideas. It is not necessary and impossible to enumerate all implementation manners here. Any modifications, equivalent replacements, improvements, etc. made within the spirit and principle of the present invention shall be included in the protection scope of the claims of the present invention.

Claims

1. A method for detecting the boric acid content and / or boron content in compound sulfamethoxazole and zinc oxide ointment, characterized in that, It includes the following steps: S1. Prepare the test solution according to the following preparation method: S11. Mix compound sulfamethoxazole and zinc oxide ointment with water at a ratio of 1 g: 30 - 50 mL, heat and shake under the condition of 60°C - 70°C, cool down to form a solidified matrix, separate the solid and liquid to obtain solution 1 and the solidified matrix; S12. Mix the solidified matrix obtained in step S11 with water, heat and shake under the condition of 60°C - 70°C, cool down to form a solidified matrix, separate the solid and liquid to obtain solution 2; the volume of water added is based on the volume of water: the mass of compound sulfamethoxazole and zinc oxide ointment in step S1 = 20 - 40 mL: 1 g; S13. Combine solution 1 obtained in step S11 and solution 2 obtained in step S12, mix well, and dilute with water to a concentration of 0.005 - 0.015 g / mL of the solution to obtain the test solution; S2. Prepare boron single-element standard solutions with concentrations ranging from 0.045 - 0.135 mg / mL, including no less than 5 gradient concentrations, conduct high-performance liquid chromatography detection, use the lg value of the boron single-element standard solution concentration as the abscissa, and the lg value of the peak area detected by high-performance liquid chromatography as the ordinate to draw a standard curve; Among them, the chromatographic conditions for high-performance liquid chromatography detection are: Chromatographic column: Acclaim TM Trinity TM P2, 100×2.1 mm, 3 μm; Drift tube temperature: 30°C; Evaporator temperature: 60°C; Column temperature: 27°C - 33°C; Flow rate: 0.27 - 0.33 mL / min; Injection volume: 2 μL; Detector: Evaporative light scattering detector; Running time: 10 min; Mobile phase: 18 - 22 mmol / L ammonium formate solution; S3. Conduct high-performance liquid chromatography detection on the test solution obtained in step S1, and obtain the content of boric acid and / or boron in the compound sulfamethoxazole and zinc oxide ointment according to the peak area obtained from the detection and the standard curve drawn in step S2.

2. The detection method according to claim 1, characterized in that, The heating and shaking in step S11 is water bath heating and shaking.

3. The detection method according to claim 1, wherein The heating and shaking condition in step S11 is 60°C.

4. The detection method according to claim 1, wherein The heating and shaking condition in step S12 is 60°C.

5. The detection method according to claim 1, wherein The heating and shaking time in step S11 is 10 - 20 min.

6. The detection method according to claim 1, wherein The heating and shaking time in step S12 is 10 - 20 min.

7. The detection method according to claim 1, wherein The chromatographic conditions are: Chromatographic column: Acclaim TM Trinity TM P2, 100×2.1 mm, 3 μm; Drift tube temperature: 30°C; Evaporator temperature: 60°C; Column temperature: 30°C; Flow rate: 0.3 mL / min; Injection volume: 2 μL; Detector: Evaporative light scattering detector; Running time: 10 min; Mobile phase: 20 mmol / L ammonium formate solution.

8. Application of the detection method according to any one of claims 1 - 7 in detecting the content of boric acid and / or boron in compound sulfamethoxazole and zinc oxide ointment; the boron content in the compound sulfamethoxazole and zinc oxide ointment is 0.045 mg / mL - 0.135 mg / mL.

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