CN111077248A

CN111077248A - Method for detecting (S) -1-amino-3-chloro-2-propanol hydrochloride by UPLC-MS/MS

Info

Publication number: CN111077248A
Application number: CN201911351116.9A
Authority: CN
Inventors: 刘长生; 肖石基
Original assignee: Jiangsu Bioguide Laboratory Co ltd
Current assignee: Jiangsu Bioguide Laboratory Co ltd
Priority date: 2019-12-24
Filing date: 2019-12-24
Publication date: 2020-04-28

Abstract

The invention relates to a method for detecting (S) -1-amino-3-chloro-2-propanol hydrochloride by UPLC-MS/MS, which comprises the following steps: taking 0.01-0.08 g of a sample to be detected, accurately measuring the sample to be detected to 0.01mg, placing the sample into a volumetric flask, adding acetonitrile-water (3-9): 3 to a constant volume of 10-50ml, carrying out vortex mixing for 5-15 min, carrying out ultrasonic extraction for 5-15 min, filtering the mixture through a 0.22 mu m filter membrane, and carrying out sample injection to detect (S) -1-amino-3-chloro-2-propanol hydrochloride in the sample; step (2), detection conditions: the (S) -1-amino-3-chloro-2-propanol hydrochloride is detected by adopting an ultra performance liquid chromatography-tandem mass spectrometry instrument UPLC-MS/MS as a detection instrument. The method can simply, quickly and efficiently carry out qualitative and quantitative detection on the (S) -1-amino-3-chloro-2-propanol hydrochloride in the pharmaceutical raw material medicines, the agrochemicals and the foods by using the UPLC-MS/MS.

Description

Method for detecting (S) -1-amino-3-chloro-2-propanol hydrochloride by UPLC-MS/MS

Technical Field

The invention relates to a detection method of (S) -1-amino-3-chloro-2-propanol hydrochloride, in particular to a method for detecting (S) -1-amino-3-chloro-2-propanol hydrochloride by UPLC-MS/MS.

Background

With the development of society and the advancement of science and technology. Human beings pay more and more attention to health, and medicines play a vital role in ensuring human health. Therefore, the pharmaceutical industry has become one of the most important areas of society. How to accurately and rapidly detect potentially low-limiting amounts of hazardous substances in pharmaceutical agents is a challenge facing the pharmaceutical industry today. The application of advanced modern instrumental analysis technology to meet the challenge is of great significance to the promotion of the safety evaluation of the medicine and the improvement of the commercial value of the medicine.

The instrument analysis is taken as a modern detection main means and is widely applied to the detection in the fields of chemistry, medicines, foods, cosmetics, agricultural chemicals and the like. Instrumental analysis enables the determination of the composition, content of the components and chemical structure of a substance by measuring several physicochemical properties, parameters and changes thereof. With the progress of the scientific technology of instrumental analysis, more and more advanced instrumental analysis techniques are widely applied to various detection fields, wherein the mass spectrometry detection technique is one of the techniques. The mass spectrometry detection is to ionize each component in a sample in an ion source to generate charged ions with different mass-to-charge ratios, after the ions are formed under the action of an accelerating electric field, the ions enter a mass analyzer, and after mass spectrometry signals are detected by the mass analyzer, visible spectrograms are formed on computer software through electronic conversion to be analyzed and researched by scientific research technicians. In order to combine the advantages of chromatographic separation, mass spectrometry is generally used in conjunction with chromatography, resulting in a chromatography-mass spectrometry technique. The chromatography-mass spectrometry combined technology has the advantages of both chromatography and mass spectrometry, namely good selectivity, high sensitivity, less test dosage, easy operation, high analysis speed, good reproducibility, small error, accuracy, reliability and the like. According to different chromatographic techniques, the chromatographic-mass spectrometry technology can be divided into LC-MS and GC-MS. The UPLC-MS/MS is one of the LC-MS used in the invention, the method adopts the tandem mass spectrum as a mass spectrum detector, and is formed by connecting three stages of mass spectrums in series, so that the selectivity of target ions can be effectively improved, the noise interference can be reduced, the signal-to-noise ratio can be further improved, and the detection limit can be further reduced, and the UPLC has the advantages of high separation efficiency, high separation speed, less used reagent and the like. In view of the great advantages of the UPLC-MS/MS technology, the technology is widely applied to the fields of medicines, chemistry, food science, cosmetics and the like.

(S) -1-amino-3-chloro-2-propanolate hydrochloride is a potentially toxic substance. After being inhaled by human body, the medicine can cause the risks of human body allergy or genetic toxicity and the like which are potentially harmful to human life health. Therefore, in order to ensure the safety of the drug, the residual amount of (S) -1-amino-3-chloro-2-propanol hydrochloride in the drug agent must be strictly controlled.

Since (S) -1-amino-3-chloro-2-propanol hydrochloride is easily decomposed at high temperature, has a small molecular weight, weak ultraviolet absorption and a small proportion of carbon and hydrogen, it is difficult to detect the low concentration limit of (S) -1-amino-3-chloro-2-propanol hydrochloride in a sample by using common detection methods such as GC-FID, HPLC-UV and GC-MS.

At present, an ultra-high performance liquid chromatography-tandem mass spectrometry combined method which has high detection sensitivity, is simple, rapid and easy to operate is lacked for effectively detecting the residual (S) -1-amino-3-chloro-2-propanol hydrochloride in the medicine.

Disclosure of Invention

The invention aims to overcome the defects of the prior art and provide a rapid UPLC-MS/MS method for detecting (S) -1-amino-3-chloro-2-propanol hydrochloride, which has good selectivity and high detection sensitivity.

The technical scheme for realizing the purpose of the invention is as follows: a method for detecting (S) -1-amino-3-chloro-2-propanol hydrochloride by UPLC-MS/MS comprises the following steps:

step (1), placing a sample to be detected in a volumetric flask, adding acetonitrile-water (3-9): 3 to a constant volume of 10-50ml, carrying out vortex mixing for 5-15 min, carrying out ultrasonic extraction for 5-15 min, filtering through a 0.22 mu m filter membrane, and carrying out sample injection to detect (S) -1-amino-3-chloro-2-propanol hydrochloride in the sample;

step (2), detecting (S) -1-amino-3-chloro-2-propanol hydrochloride by using an ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) as a detection instrument;

according to the technical scheme, in the step (1), 0.01-0.08 g of a sample to be detected is taken, and the accuracy is 0.01 mg.

In the step (2), the data acquisition mode of the ultra-high performance liquid chromatography-tandem mass spectrometer is to select reaction detection (SRM) and select mass-to-charge ratio

And m/z 110 → 110, m/z 110 → 92, m/z 110 → 56 as qualitative and quantitative ion pairs.

In the step (2), the chromatographic column of the ultra-high performance liquid chromatography-tandem mass spectrometer is a high performance liquid chromatographic column taking C18 bonded silica gel as a stationary phase: the length of the amino column is 50-100 mm, the column diameter is 2.1-4.6 mm, and the particle size of the filler is 1.7-3.5 μm.

In the technical scheme, in the step (2), the flow rate of the ultra-high performance liquid chromatography-tandem mass spectrometry instrument is 0.2-1.0 ml/min.

In the technical scheme, in the step (2), the column temperature of the ultra-high performance liquid chromatography-tandem mass spectrometry combination instrument is 25-45 ℃.

According to the technical scheme, in the step (2), the sample injection volume of the ultra-high performance liquid chromatography-tandem mass spectrometry instrument is 2-20 mu L.

In the step (2), the mass spectrum condition of the ultra-high performance liquid chromatography-tandem mass spectrometer is HESI⁺The ion source has the capillary voltage of 2800-3800 v, the capillary temperature of 250-380 ℃, the ion source temperature of 130-250 ℃, the atomizing gas of nitrogen, the sheath gas pressure of 30-100 psi, the auxiliary gas pressure of 0-10 psi, the ion transmission voltage of-40-90 v, the collision gas of argon, the collision gas pressure of 1.0-2.0 mtorr, the collision energy of-5 v-30 v, and the scanning time of 0.1-1.0 second.

After the technical scheme is adopted, the invention has the following positive effects:

the detection method has the advantages of good selectivity, high sensitivity, simplicity, rapidness and the like. According to the invention, the UPLC-MS/MS can be used for quickly and efficiently carrying out limited test on (S) -1-amino-3-chloro-2-propanol hydrochloride in bulk drugs, pharmaceutical preparations and other samples; the detection sensitivity of the (S) -1-amino-3-chloro-2-propanol hydrochloride can be effectively improved, the matrix interference is effectively avoided, the detection limit of the (S) -1-amino-3-chloro-2-propanol hydrochloride is reduced, and the lowest detection limit is 15 ng/mL; the limit of quantitation is 48.8 ng/mL; the linear correlation coefficient is 0.999 within the concentration range of 48.8ng/mL to 488 ng/mL.

Drawings

In order that the present disclosure may be more readily and clearly understood, reference is now made to the following detailed description of the present disclosure taken in conjunction with the accompanying drawings, in which

FIG. 1 is a qualitative and quantitative ion chromatogram of (S) -1-amino-3-chloro-2-propanoate hydrochloride standard of the present invention;

FIG. 2 is a mass spectrum of a (S) -1-amino-3-chloro-2-propanolate hydrochloride standard of the present invention;

FIG. 3 is a qualitative and quantitative ion chromatogram of a blank solvent of the present invention;

FIG. 4 is a mass spectrum of a blank solvent of the present invention;

FIG. 5 is a qualitative and quantitative ion chromatogram of a blank sample extract according to the present invention;

FIG. 6 is a mass spectrum of a blank sample extract of the present invention;

FIG. 7 is a qualitative and quantitative ion chromatogram of a sample spiked in accordance with the present invention;

FIG. 8 is a mass spectrum of a sample of the present invention with a label;

FIG. 9 is a qualitative and quantitative ion chromatogram of a 15ng/mL (S) -1-amino-3-chloro-2-propanolate hydrochloride standard of the present invention;

FIG. 10 is a mass spectrum of a 15ng/mL (S) -1-amino-3-chloro-2-propanolate hydrochloride standard of the present invention;

FIG. 11 is a qualitative and quantitative ion chromatogram of a 48.8ng/mL (S) -1-amino-3-chloro-2-propanolate hydrochloride standard of the present invention;

FIG. 12 is a mass spectrum of a 48.8ng/mL (S) -1-amino-3-chloro-2-propanolate hydrochloride standard of the present invention;

FIG. 13 is a qualitative and quantitative ion chromatogram of a 98ng/mL (S) -1-amino-3-chloro-2-propanolate hydrochloride standard of the present invention;

FIG. 14 is a mass spectrum of 98ng/mL (S) -1-amino-3-chloro-2-propanolate hydrochloride standard of the present invention;

FIG. 15 is a qualitative and quantitative ion chromatogram of the 146ng/mL (S) -1-amino-3-chloro-2-propanolate hydrochloride standard of the present invention;

FIG. 16 is a mass spectrum of a 146ng/mL (S) -1-amino-3-chloro-2-propanolate hydrochloride standard of the present invention;

FIG. 17 is a qualitative and quantitative ion chromatogram of a 244ng/mL (S) -1-amino-3-chloro-2-propanolate hydrochloride standard of the present invention;

FIG. 18 is a mass spectrum of 244ng/mL (S) -1-amino-3-chloro-2-propanolate hydrochloride standard of the present invention;

FIG. 19 is a qualitative and quantitative ion chromatogram of 488ng/mL (S) -1-amino-3-chloro-2-propanolate hydrochloride standard of the present invention;

FIG. 20 is a mass spectrum of 488ng/mL (S) -1-amino-3-chloro-2-propanolate hydrochloride standard of the present invention;

FIG. 21 is a standard graph of a linear experiment of the present invention.

Detailed Description

In order to make the objects, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are some, but not all, embodiments of the present invention. The components of embodiments of the present invention generally described and illustrated in the figures herein may be arranged and designed in a wide variety of different configurations.

Example 1

The invention provides a method for detecting (S) -1-amino-3-chloro-2-propanol hydrochloride by UPLC-MS/MS, which comprises the following steps:

step (2), detection conditions: the (S) -1-amino-3-chloro-2-propanol hydrochloride is detected by adopting an ultra performance liquid chromatography-tandem mass spectrometry instrument UPLC-MS/MS as a detection instrument.

In the step (1), 0.04g of sample to be detected is taken, the sample is accurate to 0.01mg, the sample is placed in an empty volumetric flask, acetonitrile-water 9:3 is added, the volume is fixed to 20ml, vortex mixing is carried out for 15min, ultrasonic extraction is carried out for 5min, and the sample passes through a 0.22 mu m filter membrane.

In step (2), ultraThe data acquisition mode of the high performance liquid chromatography-tandem mass spectrometer is to select reaction detection SRM, and select mass-to-charge ratio m/z 110 → 110, m/z 110 → 92, m/z 110 → 56 as qualitative and quantitative ion pairs; the chromatographic column of the ultra-high performance liquid chromatography-tandem mass spectrometry instrument is a high performance liquid chromatographic column taking C18 bonded silica gel as a stationary phase: the column length is 50mm, the column diameter is 2.1mm, and the grain diameter of the filler is 1.8 mu m; the flow rate of the ultra-high performance liquid chromatography-tandem mass spectrometry instrument is 0.4 ml/min; the column temperature of the ultra-high performance liquid chromatography-tandem mass spectrometry instrument is 30 ℃; the sample injection volume of the ultra-high performance liquid chromatography-tandem mass spectrometry instrument is 5 mu L; the mass spectrum condition of the ultra-high performance liquid chromatography-tandem mass spectrometer is HESI⁺The ion source has the capillary voltage of 2800-3800 v, the capillary temperature of 250-380 ℃, the ion source temperature of 130-250 ℃, the atomizing gas of nitrogen, the sheath gas pressure of 40psi, the auxiliary gas pressure of 5psi, the ion transmission voltage of-40 to-90 v, the collision gas of argon, the collision gas pressure of 1.0-2.0 mtorr, the collision energy of-5 to-20 v, and the scanning time of 0.5 second.

The following test samples were provided by coffepin pharmaceuticals, inc.

Blank test and specificity test:

as shown in FIGS. 1 to 2, about 25mg of (S) -1-amino-3-chloro-2-propanolate hydrochloride standard was weighed into a 10ml volumetric flask, dissolved with acetonitrile-water 9:3 and brought to volume to the mark. Diluting the (S) -1-amino-3-chloro-2-propanol hydrochloride standard solution to 48.8ng/mL by using acetonitrile-water 9:3, and then sampling and analyzing 1.5mL in a sampling vial to obtain the chromatogram of the (S) -1-amino-3-chloro-2-propanol hydrochloride standard solution. As shown in fig. 3 to 4, are chromatograms of acetonitrile-water 9:3 solvent. 1ml acetonitrile-water 9:3 solvent is taken for sample injection analysis. And the non-interference peak is arranged at the position of the checked target peak.

As shown in fig. 5 to 6, about 40mg of rivaroxaban solid powder is respectively weighed into a 20ml volumetric flask, an appropriate amount of acetonitrile-water 9:3 is added, vortex mixing is carried out for 15min, ultrasonic extraction is carried out for 5min, volume is determined to scale by acetonitrile-water 9:3, and the mixture is uniformly mixed. 2ml of the solution was applied to a 0.22 μm filter and analyzed by injection. And the non-interference peak is arranged at the position of the checked target peak.

Comparing the blank solvent chromatogram and the blank sample chromatogram with the (S) -1-amino-3-chloro-2-propanol hydrochloride standard chromatogram to see that the blank solvent of 9:3 acetonitrile-water and the blank matrix of the sample have no interference peak at the target peak position for 2.5-3.5 min; therefore, it can be considered that the solvent and the sample base do not interfere with the detection of (S) -1-amino-3-chloro-2-propanolate hydrochloride.

Linear experiments:

as shown in fig. 11, fig. 13, fig. 15, fig. 17 and fig. 19, about 25mg of (S) -1-amino-3-chloro-2-propanol hydrochloride standard was weighed into a 10ml volumetric flask, dissolved with 9:3 acetonitrile-water and fixed to the scale. Diluting the (S) -1-amino-3-chloro-2-propanol hydrochloride standard solution with acetonitrile-water 9:3 to prepare a series of standard solutions, and carrying out sample injection analysis. The peak area was plotted against concentration to obtain a linear result. The experimental data obtained are shown in table 1 below, and table 1 is the standard sequence experimental determination data:

TABLE 1

As shown in fig. 21, a standard graph of a linear experiment. The response value of the (S) -1-amino-3-chloro-2-propanol hydrochloride instrument shows a good linear relation in 48.8 ng/mL-488 ng/mL, and the linear correlation coefficient is as follows: 0.999, the requirement of detection is met.

The detection limit and the quantification limit are shown in figure 9 and figure 11, and the chromatogram and the mass spectrogram of the (S) -1-amino-3-chloro-2-propanol hydrochloride standard substance of 15 ng/mL; the signal-to-noise ratio of the detection signal reaches the detection limit requirement of S/N >3 when the signal-to-noise ratio is 11 at 0.6-2.0 min. Chromatogram and mass spectrum of (S) -1-amino-3-chloro-2-propanol hydrochloride standard of 48.8 ng/mL; the signal-to-noise ratio of the method is 44 at 0.6-2.0 min, and the method meets the quantitative limit requirement that S/N is more than 10.

Precision:

selecting a concentration point of (S) -1-amino-3-chloro-2-propanol hydrochloride to perform 6 repeated experiments, wherein 97.6ng/mL is selected as the concentration of (S) -1-amino-3-chloro-2-propanol hydrochloride standard solution for precision experiments. The specific data are shown in the following table 2, and the table 2 shows the precision experimental data of (S) -1-amino-3-chloro-2-propanol hydrochloride:

TABLE 2

As can be seen from the data shown in Table 2, the precision of the experiment can meet the detection requirements.

Standard addition recovery experiment:

three standard adding concentrations are selected in the experiment to carry out three parallel tests respectively, and the standard adding concentrations are as follows: 25.0mg/kg, 50.1mg/kg, 75.1mg/kg i.e. 48.8ng/mL, 7.6ng/mL, 146.4 ng/mL. The experimental data are shown in the following table 3, and the table 3 shows the experimental data of the (S) -1-amino-3-chloro-2-propanol hydrochloride added with the standard and recovered.

TABLE 3

As shown in the table 3, the recovery rate is 70-130% by adding the standard recovery experimental data of (S) -1-amino-3-chloro-2-propanol hydrochloride, and the experimental requirements are met.

Example 2

Example 2 differs from example 1 in that: adding acetonitrile-water 8:3 to a constant volume of 25ml in the step (1), carrying out vortex oscillation for 10min, carrying out ultrasonic extraction for 6min, filtering through a 0.22 mu m filter membrane, and carrying out sample injection to detect (S) -1-amino-3-chloro-2-propanol hydrochloride in a sample; taking a sample to be detected to be 0.05g and accurate to 0.01mg, placing the sample in an empty volumetric flask, adding acetonitrile-water 8:3 to a constant volume of 25ml, carrying out vortex mixing for 10min, and carrying out ultrasonic extraction for 6 min.

Example 3

Example 3 differs from example 1 in that: adding acetonitrile-water 6:3 to 50ml of constant volume in the step (1), carrying out vortex oscillation for 10min, carrying out ultrasonic extraction for 8min, filtering through a 0.22 mu m filter membrane, and carrying out sample injection to detect (S) -1-amino-3-chloro-2-propanol hydrochloride in a sample; taking a sample to be detected as 0.08g, accurately measuring to 0.01mg, placing in an empty volumetric flask, adding acetonitrile-water 6:3 to a constant volume of 50ml, carrying out vortex mixing for 10min, and carrying out ultrasonic extraction for 8 min.

The above-mentioned embodiments are intended to illustrate the objects, technical solutions and advantages of the present invention in further detail, and it should be understood that the above-mentioned embodiments are only exemplary embodiments of the present invention, and are not intended to limit the present invention, and any modifications, equivalents, improvements and the like made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims

1. A method for detecting (S) -1-amino-3-chloro-2-propanol hydrochloride by UPLC-MS/MS is characterized by comprising the following steps:

and (2) detecting the (S) -1-amino-3-chloro-2-propanol hydrochloride by using an ultra-performance liquid chromatography-tandem mass spectrometry instrument UPLC-MS/MS as a detection instrument.

2. The method for detecting (S) -1-amino-3-chloro-2-propanol hydrochloride by UPLC-MS/MS according to claim 1, wherein the method comprises the following steps: in the step (1), 0.01-0.08 g of a sample to be detected is taken, and the accuracy is 0.01 mg.

3. The method for detecting (S) -1-amino-3-chloro-2-propanol hydrochloride by UPLC-MS/MS according to claim 1, wherein the method comprises the following steps: in the step (2), the data acquisition mode of the ultra-high performance liquid chromatography-tandem mass spectrometer is to select a reaction detection SRM and select a mass-to-charge ratio

4. The method for detecting (S) -1-amino-3-chloro-2-propanol hydrochloride by UPLC-MS/MS according to claim 1, wherein the method comprises the following steps: in the step (2), the chromatographic column of the ultra-high performance liquid chromatography-tandem mass spectrometer is a high performance liquid chromatographic column taking C18 bonded silica gel as a stationary phase: the length of the amino column is 50-100 mm, the column diameter is 2.1-4.6 mm, and the particle size of the filler is 1.7-3.5 μm.

5. The method for detecting (S) -1-amino-3-chloro-2-propanol hydrochloride by UPLC-MS/MS according to claim 1, wherein the method comprises the following steps: in the step (2), the flow rate of the ultra-high performance liquid chromatography-tandem mass spectrometry instrument is 0.2-1.0 ml/min.

6. The method for detecting (S) -1-amino-3-chloro-2-propanol hydrochloride by UPLC-MS/MS according to claim 1, wherein the method comprises the following steps: in the step (2), the column temperature of the ultra-high performance liquid chromatography-tandem mass spectrometry instrument is 25-45 ℃.

7. The method for detecting (S) -1-amino-3-chloro-2-propanol hydrochloride by UPLC-MS/MS according to claim 1, wherein the method comprises the following steps: in the step (2), the sample injection volume of the ultra-high performance liquid chromatography-tandem mass spectrometry instrument is 2-20 mu L.

8. The method for detecting (S) -1-amino-3-chloro-2-propanol hydrochloride by UPLC-MS/MS according to claim 1, wherein the method comprises the following steps: in the step (2), the mass spectrum condition of the ultra-high performance liquid chromatography-tandem mass spectrometer is HESI⁺The ion source has the capillary voltage of 2800-3800 v, the capillary temperature of 250-380 ℃, the ion source temperature of 130-250 ℃, the atomizing gas of nitrogen, the sheath gas pressure of 30-100 psi, the auxiliary gas pressure of 0-10 psi, the ion transmission voltage of-40-90 v, the collision gas of argon, the collision gas pressure of 1.0-2.0 mtorr, the collision energy of-5 v-30 v, and the scanning time of 0.1-1.0 second.