Quality control method of cysteine hydrochloride pharmaceutic adjuvant

Abstract

The invention provides a quality control method of cysteine hydrochloride pharmaceutic adjuvants, which adopts high performance liquid chromatography, a mobile phase A adopts an aqueous solution of trifluoroacetic acid with the concentration of 0.1%-0.5%, a mobile phase B adopts an acetonitrile solution of trifluoroacetic acid, and the concentration of trifluoroacetic acid in the mobile phase B is 0.1%-0.5%. According to the quality control method of the cysteine hydrochloride pharmaceutic adjuvant provided by the invention, effective separation of cysteine hydrochloride and related substance tyrosine thereof can be realized, the separation degree is high, further the content of cysteine hydrochloride and related substance tyrosine can be measured at the same time, and the analysis speed is high. The quality control method for the cysteine hydrochloride medicinal auxiliary material has good specificity, accuracy, stability and reliability, the content of cysteine hydrochloride and related substances can be detected at the same time, the related substance tyrosine can be quantitatively detected, and therefore quality control over the cysteine hydrochloride medicinal auxiliary material is achieved.

Description

Technical Field

[0001] The invention relates to the technical field of analysis and detection, and in particular to a quality control method for cysteine ​​hydrochloride pharmaceutical excipient. Background Art

[0002] Cysteine ​​hydrochloride pharmaceutical excipients are an amino acid drug in medicine, which can be used to treat urinary tract diseases and can also be added to freeze-dried powder as an antioxidant. Related substances in cysteine ​​hydrochloride include tyrosine, which can cause side effects such as nausea and vomiting, abnormal nervous system, and allergic reactions. It can be seen that the content of cysteine ​​hydrochloride and related substances have a great impact on the quality, safety and effectiveness of drugs, so it is necessary to control the content of tyrosine and other amino acids in cysteine ​​hydrochloride pharmaceutical excipients.

[0003] At present, the methods for determining the content of cysteine ​​hydrochloride in the auxiliary material include titration and high-performance liquid chromatography. The titration method uses many reagents and the measurement is cumbersome. In the high-performance liquid chromatography method, cysteine ​​hydrochloride needs to be derivatized before testing, and the repeatability is poor. The method for determining other amino acids, tyrosine, is thin-layer chromatography, and the developing solvent is glacial acetic acid-water-n-butanol, which has problems such as long development time. Summary of the invention

[0004] The first purpose of the present invention is to overcome the problem of lack of effective detection of cysteine ​​hydrochloride content and related substances, and provide a quality control method for cysteine ​​hydrochloride pharmaceutical excipients, using high performance liquid chromatography, wherein mobile phase A uses an aqueous solution of trifluoroacetic acid with a concentration of 0.1%vol~0.5%vol, and mobile phase B uses an acetonitrile solution of trifluoroacetic acid, and the concentration of trifluoroacetic acid in the mobile phase B is 0.1%vol~0.5%vol.

[0005] In some embodiments of the present invention, the mobile phase A is an aqueous solution of trifluoroacetic acid with a concentration of 0.2% vol~0.4% vol, and the mobile phase B is an acetonitrile solution of trifluoroacetic acid, and the concentration of trifluoroacetic acid in the mobile phase B is 0.2% vol~0.4% vol.

[0006] The quality control method of cysteine ​​hydrochloride pharmaceutical excipient provided by the invention optimizes the composition of the mobile phase, wherein the mobile phase A adopts an aqueous solution of trifluoroacetic acid with a concentration of 0.1% vol to 0.5% vol, and the mobile phase B adopts an acetonitrile solution of trifluoroacetic acid, wherein the concentration of trifluoroacetic acid in the mobile phase B is 0.1% vol to 0.5% vol, so that effective separation of cysteine ​​hydrochloride and tyrosine can be achieved, the separation degree is large, and the analysis speed is fast.

[0007] In some embodiments of the present invention, the high performance liquid chromatography method uses porous graphitized carbon as a stationary phase. Compared with other conventional fillers, the chromatographic column filled with porous graphitized 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.

[0008] 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.

[0009] In some embodiments of the present invention, gradient elution is used, taking the total volume of the mobile phase as 100%,

[0010] At -5 to 0 min, the volume of mobile phase A 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 is decreased from 100% to 30-50%, and the volume of the mobile phase B is increased from 0% to 50-70%; At 13-16 minutes, the volume of the mobile phase A is 30-50%, and the volume of the mobile phase B is 50-70%; At 16-20 minutes, the volume of the mobile phase A increased from 30-50% to 100%, and the volume of the mobile phase B decreased from 50-70% to 0%.

[0011] The present invention adopts the above-mentioned gradient elution to enhance the retention time of cysteine ​​hydrochloride, effectively separate substances with similar polarity, and improve the separation degree.

[0012] In some embodiments of the present invention, the high performance liquid chromatography method has a flow rate of the mobile phase of 0.4 to 0.8 mL / min. The present invention controls the injection flow rate of the mobile phase to 0.4 to 0.8 mL / min, which is conducive to the retention time of cysteine ​​hydrochloride being within a suitable range; in particular, controlling the injection flow rate of the mobile phase to 0.4-0.6 mL / min is more conducive to the retention time of cysteine ​​hydrochloride being within a suitable range, which can not only achieve effective separation of cysteine ​​hydrochloride and tyrosine, but also control the time of the entire analysis process and improve efficiency.

[0013] 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 particle shape; and / or, the porous graphite carbon has a porosity of 70-80%; and / or, the porous graphite carbon has a pore size of 200-300 Å; and / or, the specific surface area of ​​the porous graphite carbon is 100-140 m 2 / g.

[0014] In some embodiments of the present invention, the column temperature of the chromatographic column in the high performance liquid chromatography is 20°C to 30°C. The column temperature of the chromatographic column used in the present invention is 20-30°C, which can not only achieve effective separation of cysteine ​​hydrochloride and tyrosine, but also ensure that their peaks are not tailed, and improve efficiency. In particular, the column temperature of the chromatographic column in the high performance liquid chromatography is controlled at 25-30°C in the present invention, which can achieve more effective separation of different components.

[0015] In some embodiments of the present invention, the detector of the high performance liquid chromatography is an ultraviolet detector connected in series with an electrospray detector. The present invention uses an ultraviolet detector connected in series with an electrospray detector, which can not only realize the simultaneous detection of analytes of different types and with different response signals, thereby improving the measurement efficiency, but also can mutually verify the substances that respond to both detectors, thereby improving the accuracy.

[0016] In some embodiments of the present invention, the detection wavelength of the ultraviolet detector is 260-300 nm. The present invention sets the detection wavelength of the ultraviolet detector to be 260-300 nm.

[0017] In some embodiments of the present invention, the sampling frequency of the electrospray detector is 3-10 Hz.

[0018] In some embodiments of the present invention, the filtration constant of the electrospray detector is 3-6s.

[0019] In some embodiments of the present invention, the evaporation temperature of the electrospray detector is 35-50°C, preferably the evaporation temperature is 35-45°C.

[0020] The present invention sets the test parameters of the electrospray detector, which can improve the peak shape of cysteine ​​hydrochloride, and is more conducive to improving the reliability of the analysis result and the accuracy of the experiment. In particular, the present invention controls the evaporation temperature to 35-45° C., and the peak shape of cysteine ​​hydrochloride is better.

[0021] In some embodiments of the present invention, the quality control method can detect the content of cysteine ​​hydrochloride and / or its related substance tyrosine in the cysteine ​​hydrochloride pharmaceutical excipient, and can also achieve quantitative detection of the related substance tyrosine, thereby achieving quality control of the cysteine ​​hydrochloride pharmaceutical excipient.

[0022] 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, ultraviolet wavelength and electrospray detector parameters to obtain the quality control method of cysteine ​​hydrochloride pharmaceutical excipient provided by the present invention, which can realize the effective separation of cysteine ​​hydrochloride and tyrosine, thereby realizing the determination of cysteine ​​hydrochloride and tyrosine.

[0023] The quality control method provided by the present invention adopts a high performance liquid chromatograph as the equipment, and adopts the above-mentioned stationary phase and mobile phase to further ensure the effective separation of cysteine ​​hydrochloride and tyrosine, with a large separation degree and a fast analysis speed; and adopts an ultraviolet detector connected in series with an electrospray detector to accurately determine the content of cysteine ​​hydrochloride and related substances.

[0024] The quality control method provided by the present invention can simultaneously determine the content of cysteine ​​hydrochloride and related substances. According to the "9101 Validation Guidelines for Analytical Methods" of the Chinese Pharmacopoeia 2020, methodological validation 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 cysteine ​​hydrochloride content and related substances. The establishment of the quality control method provided by the present invention realizes the accurate detection of cysteine ​​hydrochloride content and related substances. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] 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.

[0026] Figure 1 This is one of the blank solution chromatograms involved in the quality control method for cysteine ​​hydrochloride pharmaceutical excipients in Example 1 provided by the present invention.

[0027] Figure 2 This is one of the chromatograms of the cysteine ​​hydrochloride reference solution involved in the quality control method for the cysteine ​​hydrochloride pharmaceutical excipient in Example 1 provided by the present invention.

[0028] Figure 3This is one of the electrospray ionization detector (CAD) chromatograms of the tyrosine reference solution involved in the quality control method for the cysteine ​​hydrochloride pharmaceutical excipient in Example 1 provided by the present invention.

[0029] Figure 4 The present invention provides one of the ultraviolet detector (UV) chromatograms of the tyrosine reference solution involved in the quality control method of the cysteine ​​hydrochloride pharmaceutical excipient in Example 1.

[0030] Figure 5 This is one of the chromatograms of the cystine reference solution involved in the quality control method for cysteine ​​hydrochloride pharmaceutical excipients in Example 1 provided by the present invention.

[0031] Figure 6 It is one of the chromatograms of the cysteine ​​hydrochloride test solution involved in the quality control method of the cysteine ​​hydrochloride pharmaceutical excipient in Example 1 provided by the present invention.

[0032] Figure 7 This is one of the system suitability solution chromatograms involved in the quality control method for the cysteine ​​hydrochloride pharmaceutical excipient in Example 1 provided by the present invention.

[0033] Figure 8 This is one of the linear and range chromatograms of cysteine ​​hydrochloride involved in the quality control method of cysteine ​​hydrochloride pharmaceutical excipient in Example 1 provided by the present invention.

[0034] Fig. 9 This is one of the chromatograms of the cysteine ​​hydrochloride reference solution involved in the quality control method for the cysteine ​​hydrochloride pharmaceutical excipient in Comparative Example 1 provided by the present invention.

[0035] Fig.10 This is one of the chromatograms of the cysteine ​​hydrochloride reference solution involved in the quality control method for cysteine ​​hydrochloride pharmaceutical excipients in Comparative Example 2 provided by the present invention.

[0036] Fig.11 This is one of the chromatograms of the cysteine ​​hydrochloride reference solution involved in the quality control method for cysteine ​​hydrochloride pharmaceutical excipients in Comparative Example 3 provided by the present invention. DETAILED DESCRIPTION

[0037] 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.

[0038] In the following examples, the instruments and reagents involved are as follows: Ultimate 3000 high performance liquid chromatograph, Thermo Fisher Scientific; XPR205 / A electronic balance, Mettler; HPLC column, Hypercarb, 4.6 mm × 150 mm, 3 μm; Cysteine ​​hydrochloride reference substance, China Food and Drug Inspection Institute; Trifluoroacetic acid (chromatographic grade), Aladdin; Acetonitrile (chromatographic grade), Wokai; Cysteine ​​hydrochloride Pharmaceutical excipients: Merck KGaA; Ultrapure water was prepared by a laboratory ultrapure water machine.

[0039] In the following examples, unless otherwise specified, the solutions involved were prepared according to the following methods.

[0040] A) Prepare blank solution

[0041] Prepare an aqueous solution containing 0.3% trifluoroacetic acid; or use the aqueous solution as a blank solution; B) Preparation of cysteine ​​hydrochloride reference solution

[0042] Weigh 12.5 mg of cysteine ​​hydrochloride reference substance, dissolve it in the blank solution, and make the volume to 250 ml to prepare a 50.0 μg / mL reference substance solution; C) Preparation of tyrosine reference solution

[0043] Weigh 12.5 mg of tyrosine reference substance, dissolve it in the blank solution, and make the volume to 250 ml to prepare a 50.0 μg / mL reference substance solution; D) Preparation of cystine reference solution

[0044] Weigh 12.5 mg of cystine reference substance, dissolve it in the blank solution, and make the volume to 250 ml to prepare a 50.0 μg / mL reference substance solution; E) Prepare system suitability solution

[0045] Mix equal volumes of cysteine ​​hydrochloride reference solution, tyrosine reference solution, and cystine reference solution to prepare a system suitability solution; F) Preparation of cysteine ​​hydrochloride test solution

[0046] Weigh 12.5 mg of cysteine ​​hydrochloride pharmaceutical excipient, dissolve it in the blank solution, and make the volume to 250 ml to prepare a 50.0 μg / mL cysteine ​​hydrochloride test solution; G) Preparation of test solution for determination of related substances

[0047] Weigh 100 mg of cysteine ​​hydrochloride pharmaceutical excipient, dissolve it in the blank solution, and dilute to 10 ml to prepare a test solution for related substance determination; H) Preparation of reference solution for determination of related substances

[0048] Take 1 mL of the test solution for related substance determination and dilute it with 200 mL of blank solution to prepare the reference solution for related substance determination; Example 1

[0049] In this embodiment, a quality control method for a cysteine ​​hydrochloride pharmaceutical excipient is provided, specifically, the content of cysteine ​​hydrochloride in the cysteine ​​hydrochloride pharmaceutical excipient is detected by high performance liquid chromatography.

[0050] This embodiment uses a high performance liquid chromatograph to perform the above quality control method, wherein: Chromatographic column: porous graphite carbon is used as filler, and the chromatographic column specification is Hypercarb (4.6mm×150mm, 3μm), in which the porous graphite carbon particles are spherical and completely porous; 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.

[0051] Column temperature: 30°C; Mobile phase A: 0.3% trifluoroacetic acid in water; Mobile phase B: 0.3% trifluoroacetic acid in acetonitrile; Elution gradient: At -5 to 0 min, the volume of mobile phase A 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 13-16 min, the volume of the mobile phase A was 45%, and the volume of the mobile phase B was 55%; From 16 to 20 minutes, the volume of the mobile phase A increased from 45% to 100%, and the volume of the mobile phase B decreased from 55% to 0%.

[0052] Mobile phase flow rate: 0.6 mL / min; Detector: UV detector connected in series with electrospray detector, the detection wavelength of UV detector is 280 nm, the sampling frequency of electrospray detector is 5 Hz, the filtration constant is 3.6 s, and the evaporation temperature is 35 °C.

[0053] Effect verification

[0054] According to the 2020 edition of the Chinese Pharmacopoeia "9101 Analytical Method Validation Guidelines", the method described in Example 1 was subjected to methodological validation of specificity, linearity and range, precision, and accuracy, as follows: Specificity test

[0055] Take 15 μl of each of the blank solution, cysteine ​​hydrochloride reference solution, tyrosine reference solution, cystine reference solution, system suitability solution, and cysteine ​​hydrochloride test solution and inject them into the HPLC instrument 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 cysteine ​​hydrochloride reference solution is shown in Figure 2 , CAD chromatogram of tyrosine reference solution is shown in Figure 3 , the UV chromatogram of tyrosine reference solution is shown in Figure 4 , cystine reference solution chromatogram is shown in Figure 5 , the chromatogram of cysteine ​​hydrochloride test solution is shown in Figure 6 , the system suitability solution chromatogram is shown in 7.

[0056] Table 1 Specificity

[0057] Specificity conclusion: the blank solvent has no interference at the position of the cysteine ​​hydrochloride chromatographic peak; the separation of cysteine ​​hydrochloride, tyrosine and cystine in the system suitability solution meets the requirement of ≥1.5; the retention time of cysteine ​​hydrochloride in the cysteine ​​hydrochloride test solution is consistent with the retention time of the cysteine ​​hydrochloride reference solution; this shows that the quality control method for determining cysteine ​​hydrochloride pharmaceutical excipients by high performance liquid chromatography provided in Example 1 has good specificity.

[0058] Linearity and range tests

[0059] Cysteine ​​hydrochloride reference solution II: Take cysteine ​​hydrochloride reference and dilute it with blank solution to concentrations of 7.8125, 15.625, 31.25, 62.5, 125, and 250 μg / ml. Take 15 μl of cysteine ​​hydrochloride reference solution II of different concentrations and inject it into the high performance liquid chromatograph. Record the chromatogram. Use the concentration of cysteine ​​hydrochloride as the horizontal axis and the peak area of ​​cysteine ​​hydrochloride as the vertical axis to make a quadratic function regression equation. The results are shown in Table 2. The linear and range chromatograms of cysteine ​​hydrochloride are shown in Figure 8 .

[0060] Table 2 Linearity and range

[0061] Linearity and range conclusion: Cysteine ​​hydrochloride is in the range of 7.8125~250μg / mL, and the regression equation is y=-0.00006x 2 +0.032x+0.2987, the correlation coefficient is 0.998, which meets the requirement of r≥0.998, and the peak area has a good linear relationship with the concentration.

[0062] Precision

[0063] Take 15 μl of 150 μg / ml cysteine ​​hydrochloride reference solution, inject it into the high performance liquid chromatograph by 6 injections continuously, and record the chromatogram. The results are shown in Table 3.

[0064] Table 3 Precision results

[0065] Precision conclusion: In the precision test, the peak retention time and peak area RSD (%) of cysteine ​​hydrochloride were both ≤2.0%, indicating that the quality control method provided in Example 1 had good precision results.

[0066] Accuracy

[0067] Weigh 31.25 mg of cysteine ​​hydrochloride reference substance, dissolve it in the blank solution, and make the volume to 250 ml to prepare a 125 μg / mL reference substance solution;

[0068] Take 15 μl of each of the above solutions, inject 6 injections into the high performance liquid chromatograph, and record the chromatogram. The results are shown in Table 4.

[0069] Table 4 Accuracy

[0070] Accuracy conclusion: The quality control method provided in Example 1 measured a total of 6 solutions, and the recovery rates were between 90% and 108%, indicating that the accuracy results were good.

[0071] Content of Cysteine ​​Hydrochloride

[0072] Weigh 12.48 mg of cysteine ​​hydrochloride pharmaceutical excipient reference substance, dissolve it in the blank solution, and dilute to 250 ml to prepare a 49.92 μg / mL test solution. Take 15 μl of the above solution, inject it into the high performance liquid chromatograph with one injection, and record the chromatogram. Figure 6 The peak area y of cysteine ​​hydrochloride and the linear regression equation y=-0.00006x 2 +0.032x+0.2987, we can calculate that the cysteine ​​hydrochloride concentration in the cysteine ​​hydrochloride test sample is x=49.20μg / mL, and further we can calculate that the content of cysteine ​​hydrochloride pharmaceutical excipient is 98.6%.

[0073] Example 2

[0074] In this embodiment, a quality control method for cysteine ​​hydrochloride pharmaceutical excipient is provided, and tyrosine, a related substance in cysteine ​​hydrochloride pharmaceutical excipient, is detected by high performance liquid chromatography.

[0075] In this embodiment, a high performance liquid chromatograph is used to perform the above quality control method, wherein: Chromatographic column: porous graphite carbon is used as filler, and the chromatographic column specification is Hypercarb (4.6mm×150mm, 3μm), in which the porous graphite carbon particles are spherical and completely porous; 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. ; Chromatographic column temperature: 30°C; Mobile phase A: 0.3% trifluoroacetic acid in water; Mobile phase B: 0.3% trifluoroacetic acid in acetonitrile; Elution gradient: At -5 to 0 min, the volume of mobile phase A 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 13-16 min, the volume of the mobile phase A was 45%, and the volume of the mobile phase B was 55%; From 16 to 20 minutes, the volume of the mobile phase A increased from 45% to 100%, and the volume of the mobile phase B decreased from 55% to 0%; Mobile phase flow rate: 0.6 mL / min; Detector: UV detector connected in series with electrospray ionization (CAD) detector, the detection wavelength of the UV detector is 280 nm, the sampling frequency of the electrospray ionization detector is 5 Hz, the filtration constant is 3.6 s, and the evaporation temperature is 35 °C.

[0076] Take 15 μl of the test solution for related substance determination and the reference solution for related substance determination, respectively, inject them into the liquid chromatograph, and record the chromatogram. No tyrosine peak is detected in the chromatogram of the UV or CAD detector of the test solution for related substance determination.

[0077] Conclusion: The determination results of relevant substances meet the requirements.

[0078] When the qualitative analysis result shows that the related substance tyrosine exists in the cysteine ​​hydrochloride pharmaceutical excipient, the technicians in this field can conduct further experiments and calculations on this basis, such as linearity and range tests, and then obtain a linear equation for the corresponding relationship between the concentration of the related substance and the peak area, and conduct quantitative analysis on the related substances in the cysteine ​​hydrochloride pharmaceutical excipient.

[0079] Comparative Example 1

[0080] This comparative example provides a quality control method for cysteine ​​hydrochloride pharmaceutical excipient, which is basically the same as Example 1, except that the mobile phase is water, the elution is isocratic, and the chromatogram of cysteine ​​hydrochloride is shown in Fig. 9 As shown, the retention time of cysteine ​​hydrochloride is 1.814 min, which is too short.

[0081] Comparative Example 2

[0082] This comparative example provides a quality control method for cysteine ​​hydrochloride pharmaceutical excipients. The difference from Example 1 is that the chromatographic column specification is Hypercarb (2.1mm×100mm, 5μm), and the mobile phase adopts ① or ② or ③: ① 0.3% trifluoroacetic acid aqueous solution and 0.3% trifluoroacetic acid acetonitrile solution, gradient elution; ② a certain proportion of water and acetonitrile solution, isocratic or gradient elution; ③ different proportions of acetonitrile / water solution with trifluoroacetic acid, perfluoropentanoic acid or formic acid added, isocratic or gradient elution. The chromatogram of cysteine ​​hydrochloride is shown in Fig.10 or Fig.11 As shown, there are problems such as too short retention time, peak spurs or peak bifurcation, and too small theoretical plate number.

[0083] 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 quality control method for cysteine ​​hydrochloride pharmaceutical excipient, using high performance liquid chromatography, characterized in that: Mobile phase A and mobile phase B are used as mobile phases, wherein the mobile phase A is an aqueous solution of trifluoroacetic acid with a concentration of 0.1% vol to 0.5% vol, and the mobile phase B is an acetonitrile solution of trifluoroacetic acid, and the concentration of trifluoroacetic acid in the mobile phase B is 0.1% vol to 0.5% vol.

2. The quality control method according to claim 1, characterized in that: The mobile phase A is an aqueous solution of trifluoroacetic acid with a concentration of 0.2% vol to 0.4% vol, and the mobile phase B is an acetonitrile solution of trifluoroacetic acid, wherein the concentration of trifluoroacetic acid in the mobile phase B is 0.2% vol to 0.4% vol.

3. The quality control method according to claim 1 or 2, characterized in that: In the high performance liquid chromatography method, the chromatographic column uses porous graphitized carbon as the stationary phase, the length of the chromatographic column is 150 mm, and the diameter is 3.0-4.6 mm.

4. The quality control method according to claim 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: Adopt gradient elution, taking the total volume of the mobile phase as 100%, At -5 to 0 min, the volume of mobile phase A 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 is decreased from 100% to 30-50%, and the volume of the mobile phase B is increased from 0% to 50-70%; At 13-16 minutes, the volume of the mobile phase A is 30-50%, and the volume of the mobile phase B is 50-70%; At 16-20 minutes, the volume of the mobile phase A increased from 30-50% to 100%, and the volume of the mobile phase B decreased from 50-70% to 0%.

6. The quality control method according to any one of claims 1 to 5, characterized in that: In the high performance liquid chromatography method, the flow rate of the mobile phase is 0.4 to 0.8 mL / min; preferably, the flow rate of the mobile phase is 0.4 to 0.6 mL / min.

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

8. The quality control method according to any one of claims 1 to 7, characterized in that: In the high performance liquid chromatography method, the detector is an ultraviolet detector connected in series with an electrospray detector.

9. The quality control method according to claim 8, characterized in that: The detection wavelength of the ultraviolet detector is 260-300 nm; And / or, 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 salt quality control method comprises detecting the content of cysteine ​​hydrochloride and / or tyrosine therein.

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