A method for detecting the content of panthenol in personal care products

Abstract

The present invention discloses a method for detecting the content of panthenol in personal care products, belonging to the technical field of detection and analysis. The present invention uses benzophenone-4 for demulsification and adopts high performance liquid chromatography for gradient elution separation and detection, which can quickly detect the content of panthenol in personal care products. The operation is simple, and it has good sensitivity, stability and reproducibility, and is easy to operate standardly, having good application prospects.

Description

Technical Field

[0001] The present invention belongs to the technical field of detection and analysis, and particularly relates to a method for detecting and analyzing the content of panthenol in personal care products. Background Art

[0002] Panthenol is an alcohol derivative of pantothenic acid, which is converted into pantothenic acid, namely vitamin B5, in the body. Therefore, panthenol is also known as provitamin B5. Panthenol is included in the pharmacopoeias of various countries. At present, the detection method for its content is the perchloric acid titration method, which is only applicable to the detection of panthenol raw materials.

[0003] The addition amount of panthenol in personal care products is 0.2 - 2.0%, but personal care products have complex components, usually including emulsifiers, oils and fats, conditioners, etc. Obviously, the titration method is difficult to detect the addition amount of panthenol in the products.

[0004] Personal care products are mostly emulsion systems and need to be demulsified first. Commonly used are inorganic salts, methanol, ethanol, etc. Although inorganic salts can demulsify, they have a large dosage and have a thickening effect on some products containing anions, increasing the difficulty of detection. Since panthenol is soluble in methanol, ethanol, etc., methanol and ethanol cannot be used for demulsification. Summary of the Invention

[0005] Object of the Invention: In order to overcome the deficiencies in the prior art, the present invention provides a method for detecting and analyzing the content of panthenol in personal care products, which can quickly detect the content of panthenol in personal care products.

[0006] Technical Solution: To achieve the above object, the present invention adopts the following technical solutions:

[0007] A method for detecting and analyzing the content of panthenol in personal care products includes the following steps:

[0008] Step 1: Pretreatment of personal care products: Prepare an aqueous solution of benzophenone - 4 with a concentration of 0.2 - 0.5% (W / W). Take an appropriate amount of personal care product sample, add it to the aqueous solution of benzophenone - 4 and mix evenly. Use benzophenone - 4 for demulsification, and after centrifugation, filter to obtain the sample solution.

[0009] Step 2: Take the sample solution and the reference solution, inject them into a high - performance liquid chromatograph, and perform gradient elution separation.

[0010] In the above steps, the chromatographic conditions in Step 2 are as follows:

[0011] Mobile phase A is acetonitrile, and mobile phase B is a phosphate buffer solution (accurately weigh 1.03 g of sodium hexanesulfonate monohydrate and 7.35 g of potassium dihydrogen phosphate, add water to 1000 ml, adjust the pH to 2.5 with phosphoric acid, and filter to obtain).

[0012] In the gradient elution program, the change ratio of mobile phase A is 5%-25%, 25%-75%, 75%-8%, 8%; the flow rate is 0.8-1.0 mL / min, the column temperature is 35°C, the detection wavelength is 210 nm, the chromatographic column is Inertsil ODS-3 C18, 5 μm, 4.6*150 mm or a similar chromatographic column; the injection volume is 10 μL; the retention time: 35 min.

[0013] Preparation of the control solution: Weigh accurately 100-150 mg of panthenol reference substance into a 25-ml volumetric flask, dissolve it with purified water and dilute to the mark. Then accurately pipette 1.0 ml of this solution into a 10-ml volumetric flask, dissolve it with purified water and dilute to the mark, and filter to obtain.

[0014] Beneficial effects: The present invention provides a method for detecting and analyzing the content of panthenol in personal care products. In the pretreatment, benzophenone-4 is used for demulsification in advance, and then gradient elution is adopted for separation and detection, which can quickly detect the content of panthenol in personal care products, and has the advantages of simple operation, good sensitivity, stability and reproducibility, easy standardization operation, and good application prospects. Description of the Drawings

[0015] Figure 1 It is the HPLC chromatogram of panthenol reference substance;

[0016] Figure 2 It is the HPLC chromatograms of Example 1, Example 2 and Example 3 of the present invention;

[0017] Figure 3 It is the linear regression graph of panthenol. Detailed Embodiments

[0018] The present invention will be further described in detail below with reference to the drawings and specific embodiments.

[0019] The main experimental materials used in the present invention and their sources:

[0020] Experimental raw materials Specification Manufacturer Panthenol standard 99％ Aladdin Sodium hexanesulfonate 99.5％ Sinopharm Group Potassium dihydrogen phosphate 99.5％ Sinopharm Group Sodium chloride - Sinopharm Group Perchloric acid - Sinopharm Group Citric acid Citric acid Shandong Lemon Biochemical Co., Ltd. Laboratory-prepared cream Panthenol content: 0.5 - 2.0% - Laboratory-prepared shampoo Panthenol content: 0.5 - 2.0% -

[0021] Experimental instruments: Waters high performance liquid chromatography; Inertsil ODS-3 C18 chromatographic column.

[0022] Example 1

[0023] Preparation of the sample solution: Prepare a 0.2% (W / W) aqueous solution of benzophenone-4. Take 10 g of the cream with 1% panthenol content into a 250-ml volumetric flask, add the aqueous solution of benzophenone-4 to the mark, dissolve it by ultrasonic, centrifuge the solution at 3000 rpm for 15 min, filter, take 5.0 ml of the filtrate into a 10-ml volumetric flask, dilute it to the mark with purified water, and filter to obtain.

[0024] Preparation of reference solution: Weigh accurately 40 - 120 mg of panthenol reference standard into a 25 - ml volumetric flask, dissolve and dilute to the mark with purified water. Then accurately pipette 1.0 ml of this solution into a 10 - ml volumetric flask, dissolve and dilute to the mark with purified water, and filter to obtain the solution.

[0025] Chromatographic conditions: Mobile phase A is acetonitrile, and mobile phase B is phosphate buffer solution (Weigh accurately 1.03 g of sodium hexanesulfonate monohydrate and 7.35 g of potassium dihydrogen phosphate, add water to 1000 ml, adjust the pH to 2.5 with phosphoric acid, and filter to obtain the solution);

[0026] The gradient elution program is as shown in Table 1, the flow rate is 0.8 - 1.0 mL / min, the column temperature is 35 °C; the detection wavelength is 210 nm; the chromatographic column is Inertsil ODS - 3C18, 5 μm, 4.6 * 150 mm or a similar chromatographic column; the injection volume is 10 μL; the retention time is 35 min.

[0027] Table 1 Proportion of mobile phase change in gradient elution for Example 1

[0028] Time, t / min Phase A Phase B 0 5％ 95％ 5 25％ 75％ 12 75％ 25％ 26 75％ 25％ 35 8％ 92％

[0029] Example 2

[0030] Preparation of sample solution: Prepare a 0.35% (W / W) aqueous solution of benzophenone - 4. Take 10 g of the cream with 1% panthenol content into a 250 - ml volumetric flask, add the aqueous solution of benzophenone - 4 to the mark. Dissolve by ultrasonic treatment. Centrifuge the solution at 3000 rpm for 15 min, filter, and then take 5.0 ml of the filtrate into a 10 - ml volumetric flask, dilute to the mark with purified water, and filter to obtain the solution.

[0031] Preparation of reference solution: Weigh accurately 40 - 120 mg of panthenol reference standard into a 25 - ml volumetric flask, dissolve and dilute to the mark with purified water. Then accurately pipette 1.0 ml of this solution into a 10 - ml volumetric flask, dissolve and dilute to the mark with purified water, and filter to obtain the solution.

[0032] Chromatographic conditions: Mobile phase A is acetonitrile, and mobile phase B is phosphate buffer solution (Weigh accurately 1.03 g of sodium hexanesulfonate monohydrate and 7.35 g of potassium dihydrogen phosphate, add water to 1000 ml, adjust the pH to 2.5 with phosphoric acid, and filter to obtain the solution);

[0033] The gradient elution program is as shown in Table 2, the flow rate is 0.8 - 1.0 mL / min, the column temperature is 35 °C; the detection wavelength is 210 nm; the chromatographic column is Inertsil ODS - 3C18, 5 μm, 4.6 * 150 mm or a similar chromatographic column; the injection volume is 10 μL; the retention time is 35 min.

[0034] Table 2 Proportion of mobile phase change in gradient elution for Example 2

[0035] Time, t / min Phase A Phase B 0 15％ 85％ 5 40％ 60％ 15 75％ 25％ 28 50％ 50％ 35 10％ 90％

[0036] Example 3

[0037] Preparation of sample solution: Prepare an aqueous solution of benzophenone-4 at 0.5% (w / w). Take 10 g of the cream containing 1% panthenol and transfer it to a 250-ml volumetric flask. Add the aqueous solution of benzophenone-4 to the scale. Dissolve it by ultrasonic treatment. Centrifuge the solution at 3000 rpm for 15 min, filter, take 5.0 ml and transfer it to a 10-ml volumetric flask, dilute it to the scale with purified water, and filter to obtain the solution.

[0038] Preparation of reference solution: Weigh accurately 40 - 120 mg of panthenol reference substance and transfer it to a 25-ml volumetric flask. Dissolve and dilute it to the scale with purified water. Then accurately pipette 1.0 ml of this solution into a 10-ml volumetric flask, dissolve and dilute it to the scale with purified water, and filter to obtain the solution.

[0039] Chromatographic conditions: Mobile phase A is acetonitrile, and mobile phase B is phosphate buffer solution (accurately weigh 1.03 g of sodium hexanesulfonate monohydrate and 7.35 g of potassium dihydrogen phosphate, add water to 1000 ml, adjust the pH to 2.5 with phosphoric acid, and filter to obtain).

[0040] Gradient elution program is shown in Table 3, the flow rate is 0.8 - 1.0 mL / min, the column temperature is 35 °C; the detection wavelength is 210 nm; the chromatographic column is Inertsil ODS-3 C18, 5 μm, 4.6 * 150 mm or a similar chromatographic column; the injection volume is 10 μL; the retention time is 35 min.

[0041] Table 3 Proportion of mobile phase change in gradient elution of Example 3

[0042] Time, t / min Phase A Phase B 0 10％ 90％ 5 30％ 70％ 15 65％ 35％ 30 65％ 35％ 35 10％ 90％

[0043] Example 4

[0044] Preparation of sample solution: Prepare an aqueous solution of benzophenone-4 at 0.35% (w / w). Take 20 g of the cream containing 1% panthenol and transfer it to a 250-ml volumetric flask. Add the aqueous solution of benzophenone-4 to the scale. Dissolve it by ultrasonic treatment. Centrifuge the solution at 3000 rpm for 15 min, filter, take 5.0 ml and transfer it to a 10-ml volumetric flask, dilute it to the scale with purified water, and filter to obtain the solution.

[0045] Others are the same as in Example 1.

[0046] Example 5

[0047] Preparation of sample solution: Prepare an aqueous solution of benzophenone-4 at 0.2% (W / W). Take 15 g to 250 ml volumetric flask of the cream with 1% panthenol content, add the aqueous solution of benzophenone-4 to the scale. Dissolve by ultrasonic. Centrifuge the solution at 3000 rpm for 15 min, take 5.0 ml to a 10 ml volumetric flask after filtration, dilute to the scale with purified water, and filter to obtain.

[0048] Others are the same as in Example 1.

[0049] Example 6

[0050] Preparation of sample solution: Prepare an aqueous solution of benzophenone-4 at 0.5% (W / W). Take 20 g to 250 ml volumetric flask of the cream with 1% panthenol content, add the aqueous solution of benzophenone-4 to the scale. Dissolve by ultrasonic. Centrifuge the solution at 3000 rpm for 15 min, take 5.0 ml to a 10 ml volumetric flask after filtration, dilute to the scale with purified water, and filter to obtain.

[0051] Others are the same as in Example 1.

[0052] Comparative example

[0053] Comparative example 1

[0054] Preparation of sample solution: Take 10 g to 250 ml volumetric flask of the paste with 1% panthenol content, add purified water to the scale. Dissolve by ultrasonic. Centrifuge the solution at 3000 rpm for 15 min, take 5.0 ml to a 10 ml volumetric flask after filtration, dilute to the scale with purified water, and filter to obtain.

[0055] Others are the same as in Example 1.

[0056] Comparative example 2

[0057] Preparation of sample solution: Take 10 g to 250 ml volumetric flask of the paste with 1% panthenol content, add ethanol to the scale. Dissolve by ultrasonic. Centrifuge the solution at 3000 rpm for 15 min, take 5.0 ml to a 10 ml volumetric flask after filtration, dilute to the scale with purified water, and filter to obtain.

[0058] Others are the same as in Example 1.

[0059] Comparative example 3

[0060] Preparation of sample solution: Take 10 g to 250 ml volumetric flask of the cream with 1% panthenol content, add 20% sodium chloride solution to the scale. Dissolve by ultrasonic. Centrifuge the solution at 3000 rpm for 15 min, take 5.0 ml to a 10 ml volumetric flask after filtration, dilute to the scale with purified water, and filter to obtain.

[0061] Others are the same as in Example 1.

[0062] Comparative example 4

[0063] Preparation of sample solution: Take 10 g of the cream with 1% panthenol content into a 250 ml volumetric flask, add purified water to the scale. Dissolve by ultrasonic. Centrifuge the solution at 3000 rpm for 15 min, take 5.0 ml after filtration into a 10 ml volumetric flask, dilute to the scale with purified water, and filter to obtain. Detect the content of panthenol according to the titration detection method of USP.

[0064] The HPLC results of Example 1, Example 2, and Example 3 are as Figure 2 shown. It can be seen from the results that compared with Example 2 and Example 3, gradient 1 can better separate panthenol and other impurities in the personal care product, and the retention time is appropriate. Therefore, the final gradient elution is determined as: mobile phase A 5% - 25%, 25% - 75%, 75% - 8%, 8%.

[0065] Conduct a methodological investigation on gradient 1

[0066] 1. System suitability

[0067] As shown by the chromatograms of the sample solution and the reference solution in Example 1, the sample chromatogram and the reference chromatogram have corresponding absorption peaks at the same time, and the peaks are well classified, indicating good system suitability. The results are as Figure 1 and Figure 2 shown. The sample chromatogram and the reference chromatogram have corresponding absorption peaks at the same time, and the peaks are well classified, indicating good system suitability.

[0068] 2. Linear relationship

[0069] Prepare a reference solution of panthenol with a known concentration, and dilute it with purified water to different concentrations according to Table 4.

[0070] Table 4 Concentrations for investigating the linear relationship

[0071] Sample Panthenol concentration, μg / mL 1 480.2 2 240.1 3 192.08 4 96.04 5 48.02 6 24.01

[0072] Measure the peak area according to the chromatographic conditions of gradient 1, and take the peak area (Y) as the ordinate and the concentration (X) as the abscissa to plot the standard curve and perform linear regression calculation. The regression equation of panthenol is y = 321.83x - 493.18, R 2 = 0.9994. Figure 3 It is the linear relationship spectrum, indicating that panthenol shows good linearity in the concentration range of 24.01 μg / mL to 480.20 μg / mL.

[0073] 3. Repeatability

[0074] Take 6 portions of 100 mg panthenol reference standards of the same batch (content 99.6%), prepare solutions S1 - S6 according to the preparation method of the reference solution, and conduct repeatability verification according to the elution procedure in Example 1. The results of the repeatability verification are shown in Table 5.

[0075] Table 5 Results of Repeatability Verification

[0076] Sample Panthenol sample weight, mg Panthenol test result, % S1 100.02 99.81 S2 100.11 99.65 S3 100.15 99.6 S4 99.98 99.6 S5 100.08 99.57 S6 99.97 99.75 RSD, % 0.10

[0077] The RSD of the panthenol content measured in the repeatability test was 0.10%, indicating that the method has good repeatability.

[0078] 4. Precision

[0079] Take the reference solution S1, inject samples continuously 6 times repeatedly according to the chromatographic conditions described in Example 1, record the peak areas of each peak and calculate the RSD value. The results are shown in the table. The RSD is less than 2%, indicating that the instrument has good precision.

[0080] Table 6 Results of Precision Test

[0081] Injection times Panthenol peak area 1 128516 2 129110 3 130320 4 131278 5 130416 6 131709 RSD, % 0.94

[0082] 5. Stability

[0083] Take the solution S1, inject samples at different times within 24 h according to the chromatographic conditions described in Example 1, record the peak areas of each peak and calculate the RSD value. The results are shown in Table 7 below. The RSD is less than 2%, indicating that the test solution is stable within 24 h.

[0084] Table 7 Results of Stability Test

[0085]

[0086]

[0087] 6. Spiked Recovery

[0088] Take 6 portions of 10 g each of the shampoo sample with known panthenol content (panthenol content 11.2 mg / g), prepare the sample solution according to Example 2, add 1 mL of 224 μg / mL panthenol reference solution respectively, determine according to the chromatographic conditions under Example 1, and calculate the recovery rate. The results are shown in Table 8 below. It shows that the spiked recovery rate meets the requirements, indicating that this detection method is reliable.

[0089] Table 8 Results of Spiked Recovery

[0090]

[0091] The spiked recovery test proves that the average recovery rate of panthenol is 99.79%, which is within the range of 95% - 105% of the recovery rate, indicating that this method is stable and reliable.

[0092] Detection and Comparison of Panthenol Content in Examples and Comparative Examples

[0093] Table 9 Detection Results of Panthenol Content in Examples and Comparative Examples

[0094] Panthenol content, % Example 1 0.82 Example 2 0.98 Example 3 0.97 Example 4 0.92 Example 5 0.90 Example 6 0.88 Comparative Example 1 0.58 Comparative Example 2 0.72 Comparative Example 3 0.80 Comparative Example 4 No response

[0095] As can be seen from Table 9, by adding benzophenone-4 solution, demulsification can be quickly achieved, enabling panthenol to enter the aqueous phase, so as to accurately detect the content of panthenol. Its detection accuracy is significantly better than that of salt demulsification and organic solvent demulsification.

[0096] Application Examples

[0097] Detection Results of Panthenol Content in O / W Cream

[0098] Prepare sample test solutions and reference substance test solutions according to the methods of Example 2 and Comparative Examples 1-4, and detect the panthenol content in the cream. The results are shown in Table 10.

[0099] Table 10 Detection of Panthenol Content in O / W Cream

[0100] Panthenol addition amount Example 2 (HPLC) Comparative Example 1 Comparative Example 2 Comparative Example 3 Comparative Example 4 0.51％ 0.47％ 0.12％ 0.35％ 0.40％ No response 1.0％ 0.98％ 0.44％ 0.78％ 0.85％ No response 2.1％ 1.96％ 0.92％ 1.66％ 1.81％ 0.14％

[0101] Detection Results of Panthenol Content in Shampoo

[0102] Prepare sample test solutions and reference substance test solutions according to the methods of Example 2 and Comparative Examples 1-4, and detect the panthenol content in the shampoo. The results are shown in Table 11.

[0103] Table 11 Detection of Panthenol Content in Shampoo

[0104] Panthenol addition amount Example 2 (HPLC) Comparative Example 1 Comparative Example 2 Comparative Example 3 Comparative Example 4 0.52％ 0.52％ 0.22％ 0.38％ 0.42％ No response 1.0％ 0.98％ 0.51％ 0.76％ 0.72％ 0.25％ 2.1％ 2.04％ 1.02％ 1.49％ 1.79％ 0.47％

[0105] As can be seen from Table 10 and Table 11, with benzophenone-4 as the demulsifier and combined with the HPLC detection method, the panthenol content in shampoo and cream can be accurately detected. The present invention discloses a method for detecting the panthenol content in personal care products. The method established by the present invention is simple to operate, has good sensitivity, stability and reproducibility, is easy to standardize the operation, and has good application prospects.

[0106] The above are only the preferred embodiments of the present invention. It should be pointed out that for those of ordinary skill in the art, without departing from the principle of the present invention, several improvements and refinements can still be made, and these improvements and refinements should also be regarded as the protection scope of the present invention.

Claims

1. A method for detecting and analyzing the content of panthenol in personal care products, characterized in that, It includes the following steps: Step 1: Pretreatment of personal care products: Prepare an aqueous solution of benzophenone-4 at 0.2-0.5% W / W. Take an appropriate amount of the oil-in-water system personal care product sample, add the aqueous solution of benzophenone-4 and mix evenly. After centrifugation, filter to obtain the sample solution; Step 2: Take the sample solution and the reference solution, and inject them into a high-performance liquid chromatograph respectively for gradient elution separation and detection.

2. According to the detection and analysis method of the panthenol content in the personal care product in claim 1, it is characterized in that, In Step 2, the mobile phase A of the high-performance liquid chromatograph is acetonitrile, and the mobile phase B is a phosphate buffer solution.

3. The detection and analysis method for the content of panthenol in the personal care product according to claim 1 or 2, characterized in that, The high-performance liquid chromatography conditions in Step 2 are as follows: the flow rate is 0.8-1.0 mL / min, the column temperature is 35 °C, the detection wavelength is 210 nm, and the injection volume is 10 μL; the retention time is 35 min.

4. According to the detection and analysis method of the content of panthenol in the personal care product in claim 2, it is characterized in that, The preparation process of the mobile phase B is as follows: Weigh 1.03 g of sodium hexanesulfonate monohydrate and 7.35 g of potassium dihydrogen phosphate, add water to 1000 ml, adjust the pH to 2.5 with phosphoric acid, and filter to obtain it.

5. The detection and analysis method for the content of panthenol in the personal care product according to claim 2, characterized in that, In the gradient elution program, the change ratio of the mobile phase A is 5%-25%, 25%-75%, 75%-8%, 8%.

6. According to the detection and analysis method of the panthenol content in the personal care product of claim 1, it is characterized in that, The preparation process of the reference solution is as follows: Accurately weigh 100-150 mg of the panthenol reference standard into a 25-ml volumetric flask, dissolve it with purified water and dilute to the mark. Then accurately transfer 1.0 ml of this solution into a 10-ml volumetric flask, dissolve it with purified water and dilute to the mark, and filter to obtain it.

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