CN112630310A - Method for separating and measuring saccharin sodium in levocetirizine hydrochloride drops - Google Patents

Abstract

The invention belongs to the field of analytical chemistry, and discloses a method for separating and measuring saccharin sodium in levocetirizine hydrochloride drops by using a liquid chromatography. The method has the advantages of strong specificity, high accuracy and simple and convenient operation.

Description

Method for separating and measuring saccharin sodium in levocetirizine hydrochloride drops

Technical Field

The invention belongs to the field of analytical chemistry, and particularly relates to a method for separating and determining saccharin sodium in levocetirizine hydrochloride drops by liquid chromatography.

Background

Levocetirizine hydrochloride is a selective histamine H1 receptor antagonist. The traditional Chinese medicine composition is mainly used for relieving allergic symptoms of allergic diseases, and is clinically used for treating skin mucosa allergic diseases such as allergic rhinitis, urticaria, angioneurotic edema and the like; it can also be used for relieving allergy symptoms caused by common cold. The British name is Levocetirizine Hydrochloride, and the chemical name is (R) - (-) -2- [2- (4- (4-chlorphenyl) benzyl) -1-piperazinyl) ethoxy]Acetic acid dihydrochloride of the formula C₂₁H₂₅ClN₂O₃2 HCl. The structural formula of levocetirizine hydrochloride is as follows:

in the process of preparing the levocetirizine hydrochloride drops, in order to keep good mouthfeel and reduce the bitterness of the levocetirizine hydrochloride, a proper amount of sweetener saccharin sodium is required to be added, so that the control of the content of the saccharin sodium is very important. The English name is Saccharin, Sodium Salt Hydrate, the chemical name is Sodium o-benzoylsulfonimide (dihydrate), the molecular formula is C₇H₈NNaO₅And S. The structural formula of saccharin sodium is:

the content of saccharin sodium added in the process of preparing levocetirizine hydrochloride drops needs to be controlled. Therefore, the separation and determination of the saccharin sodium in the levocetirizine hydrochloride drops are realized, and the method has important practical significance in the aspect of quality control of the levocetirizine hydrochloride drops.

Disclosure of Invention

The invention aims to provide a method for analyzing and separating the content of saccharin sodium in levocetirizine hydrochloride drops, so that the separation and the determination of the saccharin sodium in the levocetirizine hydrochloride drops are realized, the content of saccharin sodium in the levocetirizine hydrochloride drops is ensured, and the quality control of final products is realized.

The method for analyzing the content of saccharin sodium in levocetirizine hydrochloride drops by using the liquid chromatography adopts a chromatographic column with octadecylsilane chemically bonded silica as a filler, and takes a buffer salt solution-organic phase in a certain proportion as a mobile phase.

The chromatographic column takes octyl silane bonded silica gel as a filler, and is selected from chromatographic columns with brands of Altima, Kromasil or Phenomenex.

Said organic phase is selected from the group consisting of the following reagents: methanol, acetonitrile, isopropanol and tetrahydrofuran, and the organic phase is preferably acetonitrile.

In the method, the mobile phase buffer salt solution-organic phase adopts gradient elution.

In the above-mentioned method, the buffer salt contained in the buffer salt solution is selected from phosphate, formate, acetate, perchlorate, etc., preferably phosphate; wherein the concentration of the buffer salt solution is 0.05 mol/L; the pH value of the buffer solution is 4.0-6.0, and preferably 4.50.

In the above method, the solvent is a buffered salt solution-organic phase, and the ratio is preferably 85: 15.

The separation and measurement method of the present invention can be realized by the following method:

1) and respectively taking levocetirizine hydrochloride drops and a proper amount of saccharin sodium, dissolving the samples by using a solvent, and preparing a solution containing 0.1-0.3 mg of saccharin sodium per 1 mL.

2) Setting the flow rate of a mobile phase to be 0.5-2.5 mL/min, the detection wavelength to be 205-250 nm, and the temperature of a chromatographic column oven to be 20-40 ℃.

3) And (2) injecting 10-50 mu L of the sample solution obtained in the step 1) into a liquid chromatograph to complete the separation and determination of the saccharin sodium in the levocetirizine hydrochloride drops.

Wherein: the type of the high performance liquid chromatograph has no special requirements, and the chromatograph adopted by the invention is an Shimadzu high performance liquid chromatograph: LC-20AT, CBM-20A, SIL-20AC, SPD-M20A, CTO-10ASvp

A chromatographic column: c18 (Phenomenex, 250X 4.6mm, 5 μm)

Mobile phase: phase A: 0.02mol/L potassium dihydrogen phosphate buffer solution; phase B: acetonitrile;

flow rate: 1.0 mL/min;

detection wavelength: 265 nm;

column temperature: 30 ℃;

sample introduction volume: 20 mu L of the solution;

elution gradient:

T（min）	0.00	10.00	10.01	25.00	25.01	35.00
							B%	15	15	65	65	15	15

the invention adopts the following steps: c18 (Phenomenex, 250X 4.6mm, 5 μm), can effectively separate saccharin sodium in levocetirizine hydrochloride drops. The method solves the problem of separation and determination of saccharin sodium in the levocetirizine hydrochloride drop, thereby ensuring the controllable quality of the levocetirizine hydrochloride drop.

Drawings

FIG. 1 is a sample HPLC plot at the time of example 1;

FIG. 2 is a HPLC chart of a control at the time of example 1;

FIG. 3 is a diagram of an empty silica (except saccharin sodium) HPLC at the time of example 1;

FIG. 4 is a solvent HPLC plot for example 2;

FIG. 5 is a sample HPLC plot at the time of example 2;

FIG. 6 is a HPLC chart of a control in example 2;

FIG. 7 is a sample HPLC plot at the time of example 3;

FIG. 8 is a HPLC chart of the control of example 3.

The specific implementation mode is as follows:

the method for detecting the content of saccharin sodium in levocetirizine hydrochloride drops according to the present invention is further described in detail by way of examples below, but it should not be construed that the scope of the above-described subject matter of the present invention is limited to the following examples, and all the technologies realized based on the above-described contents of the present invention fall within the scope of the present invention.

Example 1

Apparatus and conditions

Shimadzu high performance liquid chromatograph: LC-20AT, CBM-20A, SIL-20AC, SPD-M20A, CTO-10 ASvp;

a chromatographic column: c18 (Phenomenex, 250X 4.6mm, 5 μm)

Mobile phase: phase A: 0.02mol/L potassium dihydrogen phosphate buffer solution (pH 4.5); phase B: acetonitrile;

elution gradient:

T（min）	0.00	10.00	10.01	25.00	25.01	35.00
							B%	15	15	65	65	15	15

flow rate: 1.0mL/min

Detection wavelength: 265nm

Column temperature: 30 deg.C

Sample introduction volume: 20 μ L

Experimental procedure

And respectively taking levocetirizine hydrochloride drops and a proper amount of saccharin sodium, dissolving the samples by using a solvent, and preparing a solution containing 0.1-0.3 mg of saccharin sodium per 1 mL. And preparing a proper amount of empty adjuvant (except saccharin sodium) into an empty adjuvant solution. Performing high performance liquid chromatography analysis according to the conditions, and recording a chromatogram. The result is shown in attached figures 1-4, wherein figure 1 is a sample solution chromatogram, and the chromatographic peak with retention time of 7.691min in the chromatogram is saccharin sodium; FIG. 2 is a chromatogram of a control sample, wherein the chromatographic peak with retention time of 7.698min is saccharin sodium; FIG. 3 is a chromatogram of empty sodium salt (excluding saccharin sodium); FIG. 4 is a solvent chromatogram.

Example 2

Apparatus and conditions

Shimadzu high performance liquid chromatograph: LC-20AT, CBM-20A, SIL-20AC, SPD-M20A, CTO-10 ASvp;

a chromatographic column: c18 (Phenomenex, 250X 4.6mm, 5 μm)

Mobile phase: phase A: 0.05mol/L potassium dihydrogen phosphate buffer solution (pH 4.0); phase B: acetonitrile

Elution gradient:

T（min）	0.00	10.00	10.01	25.00	25.01	35.00
							B%	10	15	60	60	15	15

flow rate: 1.0mL/min

Detection wavelength: 265nm

Column temperature: 20 deg.C

Sample introduction volume: 20 μ L

Experimental procedure

And respectively taking levocetirizine hydrochloride drops and a proper amount of saccharin sodium, dissolving the samples by using a solvent, and preparing a solution containing 0.1-0.3 mg of saccharin sodium per 1 mL. And preparing a proper amount of empty adjuvant (except saccharin sodium) into an empty adjuvant solution. Performing high performance liquid chromatography analysis according to the conditions, and recording a chromatogram. The result is shown in figures 5-6, figure 5 is a sample solution chromatogram, and the chromatographic peak with retention time of 9.937min in the chromatogram is saccharin sodium; FIG. 6 is a chromatogram of a control sample, wherein the chromatographic peak with retention time of 10.077min is saccharin sodium.

Example 3

Apparatus and conditions

Shimadzu high performance liquid chromatograph: LC-20AT, CBM-20A, SIL-20AC, SPD-M20A, CTO-10 ASvp;

a chromatographic column: c18 (Kromasil, 250X 4.6mm, 5 μm)

Mobile phase: phase A: 0.05mol/L sodium perchlorate buffer solution (pH 4.0); phase B: acetonitrile

Elution gradient:

T（min）	0.00	10.00	10.01	25.00	25.01	35.00
							B%	20	20	65	65	15	15

flow rate: 1.2mL/min

Detection wavelength: 265nm

Column temperature: 30 deg.C

Sample introduction volume: 20 μ L

Experimental procedure

And respectively taking levocetirizine hydrochloride drops and a proper amount of saccharin sodium, dissolving the samples by using a solvent, and preparing a solution containing 0.1-0.3 mg of saccharin sodium per 1 mL. And preparing a proper amount of empty adjuvant (except saccharin sodium) into an empty adjuvant solution. Performing high performance liquid chromatography analysis according to the conditions, and recording a chromatogram. The result is shown in figures 7-8, figure 7 is a sample solution chromatogram, and the chromatographic peak with retention time of 6.215min in the chromatogram is saccharin sodium; FIG. 8 is a chromatogram of a control sample, wherein the chromatographic peak with retention time of 6.137min is saccharin sodium.

The analysis method for the content of saccharin sodium in the levocetirizine hydrochloride drops is verified by the following items:

system suitability test

According to the chromatographic conditions determined in the above example 1, the levocetirizine hydrochloride drop sample solution, the saccharin sodium reference solution and the empty sodium salt (saccharin sodium removed) solution are respectively used for analyzing whether the chromatographic conditions meet the requirements. As can be seen from figure 1, the separation degree between the saccharin sodium and each peak meets the requirement, and both the peak purity and the single-point threshold value meet the requirement.

Stability of solution

Taking the system applicability solution, injecting samples at 0, 2, 4, 6 and 8 hours respectively according to the chromatographic conditions described in example 1, and examining the solution stability of the product at 4 ℃ when the quantitative determination is carried out, and the result shows that the sample solution is stable within 12 hours at 4 DEG C

Saccharin sodium salt	0H	2H	4H	6H	8H	10H	12H	Average	RSD(%)
										Retention time (min)	7.691	7.683	7.688	7.695	7.687	7.695	7.690	7.69	0.06
Peak area (A)	1226963	1227153	1223998	1225960	1225621	1228491	1225961	1226307	0.12

Durability

Since the chromatographic conditions of the product are gradient elution and the corresponding chromatographic column model, column temperature, flow rate, pH value and the like are specified, the conditions are correspondingly finely adjusted to investigate the durability of the method. As a result, the method is found to have good durability under the conditions of chromatographic columns of different brands, column temperature variation of +/-5 ℃, flow rate variation of +/-0.2 mL/min, pH value variation of +/-0.2 and the like. Under the conditions of chromatographic columns of different brands, different column temperatures, different flow rates, different pH values and the like, the retention time of saccharin sodium peaks in levocetirizine hydrochloride drops is not obviously changed, and the levocetirizine hydrochloride drops can be effectively separated.

Claims (10)

1. The method for separating and measuring the saccharin sodium in the levocetirizine hydrochloride drops is characterized by comprising the following steps: a chromatographic column using octadecylsilane chemically bonded silica as a filler, and a buffer salt solution-organic phase with a certain proportion as a mobile phase.

2. The separation assay of claim 1, wherein the chromatographic column is selected from the group consisting of chromatographic columns sold under the brand names alttima, Kromasil, or Phenomenex.

3. The separation assay of claim 1, wherein the organic phase is selected from one or more of the following reagents: methanol, acetonitrile, isopropanol, tetrahydrofuran.

4. The separation assay method of claim 3, the organic phase is preferably acetonitrile.

5. The isolation assay of claim 1, wherein said buffered salt solution is selected from the group consisting of: phosphates, formates, acetates, perchlorates and the like, with phosphates being preferred.

6. The separation assay method of claim 5, wherein the concentration of said buffered salt solution is preferably 0.05 mol/L.

7. The separation assay method according to claim 5, wherein the pH of the buffer solution is 4.0 to 6.0, preferably 4.50.

8. The separation assay of claim 1, wherein the solvent is a buffered saline solution-organic phase, preferably in a ratio of 85: 15.

9. The separation assay method of claim 1, comprising the steps of:

1) taking a proper amount of saccharin sodium sample, dissolving the sample by using a mobile phase, and preparing a sample solution containing 0.1-0.3 mg of saccharin sodium in every 1 mL;

2) setting the flow rate of a mobile phase to be 0.5-2.5 mL/min, the detection wavelength to be 205-250 nm, and the temperature of a chromatographic column incubator to be 20-40 ℃;

3) and (2) injecting 10-50 mu L of the sample solution obtained in the step 1) into a liquid chromatograph to complete the separation and determination of the saccharin sodium in the levocetirizine hydrochloride drops.

10. The separation and analysis method according to claim 9, wherein the flow rate of said mobile phase in step 2) is preferably 1.0mL/min, and the measurement wavelength is preferably 265 nm.

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