Quality control method of mannitol raw material

Abstract

The invention provides a mannitol raw material quality control method, which adopts a high performance liquid chromatography, and adopts porous graphite carbon as a stationary phase to fill a chromatographic column. And an aqueous solution of 0-0.5% trifluoroacetic acid is adopted as a mobile phase. According to the mannitol content quality control method provided by the invention, mannitol and sorbitol can be separated, the analysis speed is high, and the separation degree is large. According to the quality control method provided by the invention, the chromatographic column can adopt porous graphite carbon as a stationary phase, so that the analysis accuracy and stability are improved. According to the quality control method provided by the invention, an electrospray detector is adopted as a detector, so that the reproducibility is further improved. The quality control method for the mannitol raw material provided by the invention has better specificity, accuracy, stability and reliability, can be used for detecting the content of mannitol in the mannitol raw material and detecting related substances, and realizes the quality control of the mannitol raw material, especially the quality control of medicinal mannitol.

Description

Technical Field

[0001] The invention relates to the technical field of analysis and detection, and in particular to a quality control method for mannitol raw materials. Background Art

[0002] In the medical field, mannitol is a good diuretic that can reduce intracranial pressure and intraocular pressure. Mannitol can be used as a drug for treating kidney disease, a dehydration drug, a sugar substitute, and also as an excipient for tablets or a diluent for solid and liquid drugs.

[0003] The related substances in mannitol raw materials are mainly sorbitol, an isomer with a similar structure to mannitol, and potential impurities that may be produced. Controlling the sorbitol content in mannitol raw materials can reduce the occurrence of side effects, such as thromboembolism, vasospasm, etc. Therefore, the content of mannitol and related substances in mannitol raw materials has a great impact on the quality, safety and effectiveness of drugs.

[0004] Currently, in Part II of the 2020 edition of the "Chinese Pharmacopoeia", the method for determining the mannitol content in mannitol raw materials is titration, which uses many reagents for measurement, is cumbersome and has poor repeatability; the method for determining related substances is high performance liquid chromatography, and the detector is a differential refractometer, which has problems such as low sensitivity, great influence of environmental temperature and humidity, long equilibrium time and incompatibility with gradient elution.

[0005] The Chinese patent with announcement number CN114660196B discloses a method for determining mannitol in a drug containing a multi-component compound sorbitol solution: using a liquid chromatograph and according to the first chromatographic condition, respectively detecting a mixed standard solution A and a test solution, the first chromatographic condition is as follows: the column temperature is 70°C to 85°C, the chromatographic column is a strong cationic calcium exchange column with a sulfonated cross-linked styrene divinylbenzene copolymer as a filler, and the mobile phase is selected from any one or a combination of two of acetonitrile and water (in a specific embodiment, ultrapure water is used as the mobile phase); and according to the measured peak areas of the mixed standard solution A and the test solution, the content of mannitol in the drug containing the multi-component compound sorbitol solution is calculated.

[0006] Among the above prior arts, the detector used in patent CN114660196B is a differential refractometer, which has the problems of low sensitivity, great influence of environmental temperature and humidity, long equilibrium time and incompatibility with gradient elution; the detection limit and quantification limit of mannitol and other components in CN114660196B are at the mg / ml level, and the detection concentration is high, which is not conducive to the detection of potential low-level impurities. The detection limit and quantification limit of mannitol and sorbitol in the electrospray detector used in the present invention are at the μg / ml level, which can also effectively detect potential low-level impurities. Summary of the invention

[0007] In order to overcome the technical problems in the above prior art that the separation effect of mannitol and sorbitol needs to be further improved and potential low-level impurities cannot be effectively detected, the present invention provides a quality control method for mannitol content, which adopts high performance liquid chromatography for analysis, wherein porous graphite carbon is used as a stationary phase to fill a chromatographic column; an aqueous solution of trifluoroacetic acid with a concentration of 0 to 0.5% vol is used as a mobile phase, and preferably an aqueous solution of trifluoroacetic acid with a concentration of 0.1-0.5% vol or 0.2-0.4% vol is used as the mobile phase.

[0008] High performance liquid chromatography generally uses acetonitrile or acetonitrile aqueous solution or water as the mobile phase for elution to achieve the separation of mannitol, sorbitol and other potential impurities for analysis; but its separation effect needs to be further improved. In this field, in order to obtain a better separation effect of mannitol, sorbitol and potential impurities, improvements are generally made to the detector and the chromatographic column. Electrospray detectors generally do not recommend the use of pure water as the mobile phase because it may produce higher background noise and cause a decrease in atomization efficiency, affecting the baseline stability and signal accuracy of the detector. The present invention adds trifluoroacetic acid to the mobile phase, which has better effects in terms of baseline stability, separation effect, etc.

[0009] At the same time, the present invention optimizes the concentration of trifluoroacetic acid and adopts an aqueous solution of trifluoroacetic acid with a concentration of 0-0.5%vol as a mobile phase, so that mannitol, sorbitol and other potential impurities can be separated, and the analysis speed is fast and the separation degree is large. Compared with the prior art, the method of the present invention can accurately measure the content of mannitol, sorbitol and potential chemical impurities. In particular, when an aqueous solution of trifluoroacetic acid with a concentration of 0.1-0.5%vol and 0.2-0.4%vol is adopted as a mobile phase, the above effect is more prominent.

[0010] The present invention uses the mobile phase in combination with porous graphite carbon as a stationary phase to further improve the separation degree. Compared with other conventional fillers, the chromatographic column filled with porous graphite carbon used in the present invention has strong separation ability and strong retention ability of polar compounds, improves the separation ability of analytes with similar structures, and has the advantage of being able to maintain stability under extreme pH values ​​and high temperature conditions.

[0011] In some embodiments of the present invention, the stationary phase is 100% porous graphite carbon (i.e., the chromatographic column is filled with porous graphite carbon); and / or, the porous graphite carbon has a particle size of 3-5 μm; and / or, the porous graphite carbon has a spherical shape; and / or, the porosity of the porous graphite carbon is 70-80%; and / or, the pore size of the porous graphite carbon is 200-300 Å; and / or, the specific surface area of ​​the porous graphite carbon is 100-140 m 2 / g.

[0012] In some embodiments of the present invention, the HPLC method uses a chromatographic column of Hypercarb with specifications of 4.6 mm×150 mm and 3 μm.

[0013] In some embodiments of the present invention, the mobile phase injection flow rate is 0.4-0.8 mL / min. The present invention controls the injection flow rate of the mobile phase to be 0.4-0.8 mL / min, which is conducive to the retention time of mannitol within a suitable range; in particular, controlling the injection flow rate of the mobile phase to be 0.4-0.6 mL / min is more conducive to the retention time of mannitol within a suitable range, which can not only achieve effective separation of mannitol, sorbitol and potential chemical impurities, but also control the time of the entire analysis process and improve efficiency.

[0014] In some embodiments of the present invention, the column temperature of the chromatographic column in the high performance liquid chromatography is 20-30°C. The present invention adopts a chromatographic column with a column temperature of 20-30°C, which can not only achieve effective separation of mannitol, sorbitol and potential chemical impurities, but also ensure that the peak shape is not tailed, and improve efficiency. In particular, the present invention controls the column temperature of the chromatographic column in the high performance liquid chromatography to 20-25°C, which can further achieve effective separation of mannitol, sorbitol and potential chemical impurities.

[0015] In some embodiments of the present invention, the high performance liquid chromatography method uses an electrospray detector as a detector. The present invention uses an electrospray detector with high sensitivity, which can achieve low-limit detection of sorbitol and potential low-level impurities, and can detect sorbitol and potential low-level impurities even if their contents are very small; and the electrospray detector has good reproducibility and high reliability.

[0016] In some embodiments of the present invention, the sampling frequency of the electrospray detector is 3-10 Hz; and / or, the filter constant of the electrospray detector is 3-6 s; and / or, the evaporation temperature of the electrospray detector is 35-50° C. The test parameters of the electrospray detector are set in the present invention to improve the peak shape of mannitol, which is more conducive to improving the reliability of the analysis results and the accuracy of the experiment. In particular, the present invention controls the evaporation temperature to 35-45° C., and the peak shape of mannitol is better.

[0017] In some embodiments of the present invention, the mannitol raw material is selected from at least one of medicinal mannitol, food mannitol, and feed mannitol; the medicinal mannitol can be a mannitol raw material, a mannitol pharmaceutical excipient, etc., and the food mannitol can be a food additive, such as a sweetener, an anti-sticking agent, etc.

[0018] In some embodiments of the present invention, the quality control method can detect the content of mannitol and / or its related substances (sorbitol, other potential impurities) in the mannitol raw material, and can achieve quantitative detection of its related substances, thereby achieving quality control of the mannitol raw material (such as pharmaceutical excipients) and reducing side effects caused by related substances sorbitol and potential impurities.

[0019] The beneficial effects of the present invention are as follows: the present invention optimizes chromatographic parameters (mobile phase, flow rate, column temperature, etc.), chromatographic column parameters and electrospray ionization detector parameters to obtain the quality control method of mannitol raw materials provided by the present invention, which can achieve effective separation of mannitol, sorbitol and other potential impurities, thereby achieving the determination of mannitol and sorbitol.

[0020] The quality control method provided by the present invention uses a high performance liquid chromatograph as the equipment. The above-mentioned stationary phase and mobile phase can further ensure the effective separation of mannitol and sorbitol, with a large separation degree and a fast analysis speed. Furthermore, on the basis of the stationary phase and mobile phase, the present invention also uses an electrospray detector to improve the reproducibility of the quality control method. The electrospray detector is a new type of universal detector, which has high sensitivity to substances with no or weak ultraviolet absorption, such as mannitol and sorbitol. Moreover, the difference between the detector and the evaporative light scattering detector is that the evaporative light scattering detector is affected by the spatial structure of the compound due to the scattering, and even the same concentration response value of isomers (mannitol / sorbitol) will have a large difference, while the response factor of the electrospray detector is consistent, independent of the compound structure, and has good reproducibility, and can accurately measure the content of mannitol and related substances.

[0021] The quality control method provided by the present invention can simultaneously determine the content of mannitol and related substances. According to the 2020 edition of the Chinese Pharmacopoeia "9101 Analytical Method Verification Guidelines", methodological verification of specificity, linearity and range, precision, and accuracy was carried out, proving that this method has good specificity, accuracy, stability, and reliability, and can be used for the determination and control of mannitol content and related substances. The establishment of the quality control method of the present invention realizes the accurate detection of mannitol content and related substances. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] In order to more clearly illustrate the technical solutions in the present invention or the prior art, the following briefly introduces the drawings required for use in the embodiments or the description of the prior art. Obviously, the drawings described below are some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0023] Figure 1It is one of the blank solution chromatograms involved in the quality control method for mannitol raw materials in Example 1 provided by the present invention.

[0024] Figure 2 It is one of the chromatograms of the mannitol reference solution involved in the quality control method of the mannitol raw material in Example 1 provided by the present invention.

[0025] Figure 3 It is one of the chromatograms of the sorbitol reference solution involved in the quality control method of mannitol raw material in Example 1 provided by the present invention.

[0026] Figure 4 It is one of the system applicability solution chromatograms involved in the quality control method of mannitol raw material in Example 1 provided by the present invention.

[0027] Figure 5 It is one of the chromatograms of the mannitol test solution involved in the quality control method of the mannitol raw material in Example 1 provided by the present invention.

[0028] Figure 6 It is one of the linear and range chromatograms of mannitol involved in the quality control method of mannitol raw material in Example 1 provided by the present invention.

[0029] Figure 7 It is one of the chromatograms of the test sample solution for determining related substances involved in the quality control method for mannitol raw materials in Example 1 provided by the present invention.

[0030] Figure 8 It is one of the chromatograms of the test sample solution for determining related substances involved in the quality control method for mannitol raw materials in Example 1 provided by the present invention (enlarged view).

[0031] Fig. 9 It is one of the blank solution chromatograms involved in the quality control method for mannitol raw materials in Example 3 provided by the present invention.

[0032] Fig.10 It is one of the system applicability solution chromatograms involved in the quality control method of mannitol raw material in Example 3 provided by the present invention.

[0033] Fig.11 It is one of the system applicability solution chromatograms involved in the quality control method of mannitol raw materials in Comparative Example 2 provided by the present invention.

[0034] Fig.12 It is one of the chromatograms of the mannitol reference solution involved in the quality control method of the mannitol raw material in Comparative Example 3 provided by the present invention.

[0035] Fig.13 It is one of the chromatograms of the mannitol reference solution involved in the quality control method of the mannitol raw material in Comparative Example 4 provided by the present invention. DETAILED DESCRIPTION

[0036] In order to make the purpose, technical solution and advantages of the present invention clearer, the technical solution of the present invention will be clearly and completely described below in conjunction with the drawings of the present invention. Obviously, the described embodiments are part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0037] In the following examples, the information of instruments and reagents is as follows: Ultimate 3000 high performance liquid chromatograph, Thermo Fisher Scientific; XPR205 / A electronic balance, Mettler; Mannitol reference substance: Aladdin; Sorbitol reference substance: Sinopharm Group; Mannitol pharmaceutical excipients: sigma-aldrich; Trifluoroacetic acid (chromatographic grade): Aladdin; Ultrapure water was prepared by a laboratory ultrapure water machine.

[0038] Unless otherwise specified, the solutions involved in the following examples were prepared according to the following methods.

[0039] (A) Preparation of blank solution

[0040] Prepare an aqueous solution containing 0.3% trifluoroacetic acid; or use the aqueous solution as a blank solution; (B) Preparation of mannitol reference solution 1

[0041] Weigh 12.5 mg of mannitol reference substance, dissolve it in the blank solution, and dilute to 250 ml to prepare a 50.0 ug / mL reference substance solution; (C) Preparation of sorbitol reference solution

[0042] Weigh 12.5 mg of sorbitol reference substance, dissolve it in the blank solution, and dilute to 250 ml to prepare a 50.0 ug / mL reference substance solution; (D) Preparation of system suitability solution

[0043] Mix equal volumes of mannitol reference solution 1 and sorbitol reference solution to prepare a system suitability solution; (E) Preparation of mannitol test solution

[0044] Weigh 12.5 mg of mannitol pharmaceutical excipient, dissolve it in the blank solution, and dilute to 250 ml to prepare a mannitol test solution; (F) Preparation of test solution for determination of related substances

[0045] Weigh 25 mg of mannitol reference substance, dissolve it in the blank solution, and make the volume to 5 ml to prepare a 5 mg / mL test solution for related substance determination; (G) Preparation of reference solution for determination of related substances

[0046] Take 1mL of the test solution for related substance determination and dilute it with 100mL of blank solution to prepare the reference solution for related substance determination; or directly take the mannitol test solution.

[0047] Example 1

[0048] In this embodiment, a method for quality control of mannitol pharmaceutical excipients is provided, and the content of mannitol in the mannitol pharmaceutical excipients is detected by high performance liquid chromatography.

[0049] The mobile phase was a trifluoroacetic acid aqueous solution with a volume concentration of 0.3%; the mobile phase injection flow rate was 0.4 mL / min; and isocratic elution was used.

[0050] The chromatographic column uses porous graphitized carbon as a filler, the column temperature is 20°C, the chromatographic column specification is Hypercarb (4.6mm×150mm, 3μm), the particle shape of the porous graphite carbon filler of the chromatographic column stationary phase is spherical and completely porous, the porosity of the porous graphite carbon is 75%, the pore size of the porous graphite carbon is 250 Å, and the specific surface area of ​​the porous graphite carbon is 120 m 2 / g. .

[0051] The detector used is an electrospray detector, the sampling frequency of the electrospray detector is 5 Hz, the filtration constant is 3.6 s, and the evaporation temperature is 35°C.

[0052] According to the "9101 Validation Guidelines for Analytical Methods" of the Chinese Pharmacopoeia 2020 edition, the method described in Example 1 was subjected to methodological validation tests for specificity, linearity and range, precision, and accuracy.

[0053] Specificity test

[0054] Take 5 μl of each of the blank solution, mannitol reference solution 1, sorbitol reference solution, system suitability solution, and mannitol test solution, and inject them into the HPLC to record the chromatogram. The test results are shown in Table 1, and the blank solution chromatogram is shown in Figure 1 , the chromatogram of mannitol reference solution 1 is shown in Figure 2 , chromatogram of sorbitol reference solution is shown in Figure 3 , the system suitability solution chromatogram is shown in Figure 4 The chromatogram of mannitol test solution is shown in Figure 5 .

[0055] Table 1 Specificity test results

[0056] Specificity test conclusion: the blank solvent has no interference at the mannitol chromatographic peak position; the separation degree of mannitol and sorbitol in the system suitability solution meets the requirement of ≥1.5; the retention time of mannitol in the mannitol test solution is consistent with the retention time of mannitol reference solution 1; it shows that the quality control method for determining mannitol raw materials by high performance liquid chromatography provided in Example 1 has good specificity.

[0057] Linearity and range tests

[0058] Prepare mannitol reference solution 2: Take mannitol reference and dilute it with blank solution to different concentrations, as shown in Table 2.

[0059] Table 2 Mannitol reference solution 2 in linearity and range test

[0060] Take 5 μl of each mannitol reference solution 2 of different concentrations, inject them into the high performance liquid chromatograph, and record the chromatogram. Use the concentration of mannitol as the horizontal axis and the peak area of ​​mannitol as the vertical axis to make a quadratic function regression equation; the results are shown in Table 3, and the linear and range chromatograms of mannitol are shown in Figure 6 .

[0061] Table 3 Linearity and range

[0062] Linearity and range test conclusion: Mannitol is in the range of 7.81~500.0 μg / mL, and the regression equation is y=-0.00005x 2 +0.055x+0.8826, the correlation coefficient is 0.998, which meets the requirement of r≥0.990, and the peak area has a good linear relationship with the concentration.

[0063] Precision

[0064] Take 1,5 μl of 20ug / mL mannitol reference solution, inject 6 injections into the high performance liquid chromatograph, and record the chromatogram. The results are shown in Table 4.

[0065] Table 4 Precision results

[0066] Conclusion of precision test: In the precision test, the retention time and peak area RSD (%) of mannitol peak were both ≤2.0, indicating that the precision results were good.

[0067] Accuracy

[0068] Mannitol reference solution 3: Weigh 10.65 mg of mannitol reference, dissolve it in the blank solution, dilute it 200 times, and prepare 53.2 μg / mL mannitol reference solution 3.

[0069] Take 5 μl of the above solution, inject 6 injections into the high performance liquid chromatograph, record the chromatogram, substitute the peak area into the linear equation in Test Example 2, calculate the actual concentration, and calculate the ratio of the actual concentration to the theoretical preparation concentration to calculate the recovery rate of mannitol reference solution 3.

[0070] Accuracy conclusion: The recovery rate determined by this method is between 85% and 110%, indicating good accuracy results.

[0071] Mannitol content

[0072] Mannitol test solution: weigh 12.52 mg of mannitol pharmaceutical excipient, dissolve it in the blank solution, and make the volume to 250 ml to prepare a mannitol test solution. The concentration of the mannitol test solution is 50.08 μg / mL.

[0073] Take 5 μl of the above solution, inject it into the HPLC with one injection, and record the chromatogram. Figure 5 The peak area y of mannitol and the linear regression equation y=-0.00005x 2 +0.055x+0.8826, we can calculate the test concentration of mannitol in the mannitol test sample as x=49.18μg / mL, and further calculate the content of mannitol in the mannitol pharmaceutical excipient as 98.2%.

[0074] Example 2

[0075] In this embodiment, a quality control method for mannitol pharmaceutical excipient is provided, and in this embodiment, it is achieved by detecting the related substance sorbitol. In this embodiment, the related substance sorbitol in the mannitol raw material is detected by using the quality control method, and is determined by high performance liquid chromatography.

[0076] In this embodiment, a high performance liquid chromatograph is used to perform the above quality control method, wherein: The chromatographic column uses porous graphitized carbon as a filler, and the chromatographic column specifications are Hypercarb (4.6mm×150mm, 3μm); the filler of the chromatographic column stationary phase is 100% porous graphite carbon; the particle shape of the porous graphite carbon is spherical; the porosity of the porous graphite carbon is 75%; the pore size of the porous graphite carbon is 250 Å; the specific surface area of ​​the porous graphite carbon is 120 m 2 / g.

[0077] Chromatographic column temperature: 20℃; Mobile phase: 0.3% trifluoroacetic acid in water; Mobile phase flow rate: 0.4 mL / min; isocratic elution; Detector: Electrospray detector, the sampling frequency of the electrospray detector is 5 Hz, the filtration constant is 3.6 s, and the evaporation temperature is 35 °C.

[0078] Determination of related substances: Take 5 μl of system suitability solution, test solution for related substance determination and reference solution for related substance determination, respectively, and inject them into the liquid chromatograph, record the chromatogram, and the results of the test solution for related substance determination are shown in Figure 7 and Figure 8 For the chromatogram of the reference substance solution, see Figure 3 , the chromatogram of the system suitability solution is shown in Figure 4 .

[0079] Qualitative conclusion of related substance determination: The sum of the peak areas of impurities in the test solution for related substance determination is smaller than the peak area of ​​the reference solution for related substance determination, and the separation degrees of sorbitol and each potential impurity from mannitol all meet the requirement of being greater than 1.5. The determination results of related substances meet the requirements.

[0080] On this basis, those skilled in the art can conduct further experiments and calculations, such as linearity and range tests, to obtain a linear equation for the corresponding relationship between the concentration of the relevant substance and the peak area, and conduct a quantitative analysis of the relevant substances in the mannitol pharmaceutical excipient.

[0081] Example 3

[0082] In this embodiment, a method for quality control of mannitol raw materials is provided, which is substantially the same as that in Example 1, except that the mobile phase is a pure aqueous solution, wherein the chromatogram of the solvent water is as shown in FIG. Fig. 9 As shown in the figure, it can be seen that when water is used as the mobile phase, the baseline stability is poor. The chromatogram of the system suitability solution test is shown in Figure 10. The separation degree of mannitol and sorbitol is 2.32, which is greater than 1.5.

[0083] In the present embodiment, the content of mannitol obtained by calculation is 98.3%.

[0084] Example 4

[0085] In this embodiment, a quality control method for mannitol raw materials is provided, which is basically the same as that in Example 1, except that a 0.5% trifluoroacetic acid aqueous solution is used as the mobile phase, and the chromatogram is similar to that in Example 1.

[0086] In the present embodiment, the content of mannitol obtained by calculation is 98.5%.

[0087] Comparative Example 1

[0088] This comparative example provides a quality control method for mannitol raw materials, which is basically the same as Example 1, except that the column temperature is 35°C, and the retention time and separation degree of mannitol and sorbitol in the system suitability solution are 1.32, which is less than 1.5.

[0089] Comparative Example 2

[0090] This comparative example provides a method for controlling the quality of mannitol raw materials, which is basically the same as Example 1, except that the mobile phase is that mobile phase A uses an aqueous solution and mobile phase B uses an acetonitrile solution, and the total volume of the mobile phase is 100%: At 0-2 min, the volume of mobile phase A was 100%; From the 2nd to the 13th minute, the volume of the mobile phase A decreased from 100% to 45%, and the volume of the mobile phase B increased from 0% to 55%; At 13-22 minutes, the volume of the mobile phase A was increased from 45% to 100%, and the volume of the mobile phase B was decreased from 55% to 0%; the peak shape and tailing factor of mannitol and sorbitol met the requirements, but the separation degree of mannitol and sorbitol in the system suitability solution was 1.18, which was less than 1.5. The chromatogram was shown in FIG. Fig.11 .

[0091] Comparative Example 3

[0092] This comparative example provides a method for controlling the quality of mannitol raw materials, which is basically the same as Example 3, except that the chromatographic column specification is Hypercarb (2.1 mm × 100 mm, 5 μm). Fig.12 , the retention time difference, peak bifurcation and theoretical plate number of the mannitol peak are too low.

[0093] Comparative Example 4

[0094] This comparative example provides a method for controlling the quality of mannitol raw materials, which is basically the same as Example 1, except that the chromatographic column specification is Hypercarb (2.1 mm × 100 mm, 5 μm), and the mobile phase adopts ① or ②: ① acetonitrile / water solution in different proportions or ② acetonitrile / water solution in different proportions with trifluoroacetic acid, perfluoropentanoic acid or formic acid added. See the chromatogram Fig.13 , the retention time of mannitol is poor, the peak has multiple spurs and the theoretical plate number is too low.

[0095] The effects of the embodiments and comparative examples are compared in Table 5.

[0096] Table 5 Effect comparison

[0097] In summary, it can be seen that the separation degree of mannitol and sorbitol, the peak shape and tailing factor of mannitol and the theoretical plate number in Examples 1, 3 and 4 all meet the requirements, while the comparative example has the problem that the separation degree is lower than 1.5 or the peak shape is poor or the theoretical plate number is too low.

[0098] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the aforementioned embodiments, or make equivalent replacements for some of the technical features therein. However, these modifications or replacements do not deviate the essence of the corresponding technical solutions from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. A method for quality control of mannitol raw materials, using high performance liquid chromatography, characterized in that: The high performance liquid chromatography method adopts porous graphite carbon as the stationary phase to fill the chromatographic column; and adopts 0-0.5%vol trifluoroacetic acid aqueous solution as the mobile phase.

2. The quality control method according to claim 1, characterized in that: The mobile phase is an aqueous solution of 0.2-0.4% vol trifluoroacetic acid.

3. The quality control method according to claim 1 or 2, characterized in that: The chromatographic column has a length of 150 mm and a diameter of 3.0 to 4.6 mm.

4. The quality control method according to any one of claims 1 to 3, characterized in that: The stationary phase is 100% porous graphite carbon; And / or, the porous graphite carbon has a particle size of 3-5 μm; And / or, the porous graphite carbon particles are in the shape of spheres; and / or, the porosity of the porous graphite carbon is 70-80%; And / or, the pore size of the porous graphite carbon is 200-300 Å; And / or, the specific surface area of ​​the porous graphite carbon is 100-140 m 2 / g.

5. The quality control method according to any one of claims 1 to 4, characterized in that: The mobile phase injection flow rate is 0.4-0.8 mL / min.

6. The quality control method according to claim 5, characterized in that: The mobile phase injection flow rate is 0.4-0.6 mL / min.

7. The quality control method according to any one of claims 1 to 6, characterized in that: The column temperature of the chromatographic column is 20-30°C; preferably, the column temperature of the chromatographic column is 20-25°C.

8. The quality control method according to any one of claims 1 to 7, characterized in that: The high performance liquid chromatography method uses an electrospray ionization detector as a detector.

9. The quality control method according to any one of claims 1 to 8, characterized in that: The sampling frequency of the electrospray detector is 3-10 Hz; And / or, the filtration constant of the electrospray detector is 3-6s; And / or, the evaporation temperature of the electrospray detector is 35-50°C, preferably the evaporation temperature is 35-45°C.

10. The quality control method according to any one of claims 1 to 9, characterized in that: The mannitol raw material is selected from at least one of medicinal mannitol, food mannitol, and feed mannitol; Preferably, the quality control method comprises detecting the mannitol content and / or sorbitol in the mannitol raw material.

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