Determination method for Labobolifera, Suvorexera, Monebolifera and Amolenet in food

Abstract

The invention discloses a method for determining libosan, suvorexant, nemorexant and amolenet in food, and relates to the technical field of food detection, and particularly relates to a method for determining libosan, suvorexant, nemorexant and amolenet in food. According to the method, a method for determining the contents of the libosan, the suvorexant, the nemorexant and the amolenet in the food is provided, namely candies and beverages are adopted as analysis samples, and the contents of the libosan, the suvorexant, the nemorexant and the amolenet are determined by a high performance liquid chromatography method or a high performance liquid chromatography-tandem mass spectrometry method, so that the content of the libosan, the suvorexant, the nemorexant and the amolenet in the food is determined. And drawing a standard curve by taking the concentration of the standard working solution as an abscissa and the peak area of the chromatographic peak of the compound or quantitative ion as an ordinate, so as to respectively obtain the contents of the lyblogxant, the suvorexant, the nemorexant and the amolenet.

Description

Technical Field

[0001] The present invention relates to the technical field of food detection, and particularly to a method for determining lemborexant, suvorexant, nemorexant and amorexant in food. Background Art

[0002] Lemborexant, suvorexant, nemorexant and amorexant are orexin receptor antagonists, which are used to treat insomnia characterized by difficulty falling asleep and / or difficulty maintaining sleep. Orexin is produced in the hypothalamus and is a small molecule neuropeptide that can regulate the sleep / wake cycle by maintaining wakefulness and contribute to appetite regulation and energy homeostasis. With the increasing prevalence of sleep disorders year by year, the use of sedative-hypnotic drugs is also expanding continuously. As a result, some merchants, in pursuit of profits, add sleep-aiding ingredients to food, giving food some promotional effects that it does not inherently possess, in order to quickly convince consumers. The present invention intends to establish a method for determining the contents of lemborexant, suvorexant, nemorexant and amorexant by high performance liquid chromatography and high performance liquid chromatography-tandem mass spectrometry to ensure clinical medication and food safety. Summary of the Invention

[0003] The purpose of the present invention is to provide a method for determining lemborexant, suvorexant, nemorexant and amorexant in food to solve the problems existing in the prior art.

[0004] To solve the above technical problems, the present invention provides the following technical solution: A method for determining lemborexant, suvorexant, nemorexant and amorexant in food, wherein the method for determining lemborexant, suvorexant, nemorexant and amorexant in food includes a first method and a second method; the first method and the second method are high performance liquid chromatography and high performance liquid chromatography-tandem mass spectrometry respectively.

[0005] Further, the first method or the second method includes the following:

[0006] (1) Preparation of standard stock solution: Weigh accurately lemborexant, suvorexant, nemorexant and amorexant respectively, dissolve them with methanol and make up the volume to 5 mL to prepare standard stock solutions with a concentration of 1000 μg / mL respectively, store them in the dark at -20 °C, and the validity period is 3 months;

[0007] (2) Preparation of standard intermediate working solution;

[0008] (3) Preparation of standard working solution;

[0009] (4) Preparation of sample solutions: Crush candies and solid beverages separately, place them in a 50 mL volumetric flask, warm and dissolve them with water. After the candies are completely dissolved, add methanol, extract ultrasonically for 15 min, cool to room temperature, make up the volume with methanol, transfer to a 50 mL centrifuge tube, centrifuge at 4000 r / min for 5 min, filter the supernatant through a 0.22 μm microporous membrane, take the filtrate, and dilute it with a methanol-water solution to within the linear range according to the actual concentration to obtain candy samples and solid beverage samples respectively; the candies are one or more mixtures of hard candies, crispy candies, gel candies, gum base candies, tablet candies, milk candies, etc.; the solid beverages are one or more mixtures of fruit and vegetable solid beverages, protein solid beverages, coffee solid beverages, tea solid beverages, etc.

[0010] Take the liquid beverage and place it in a 50 mL volumetric flask, add methanol and extract ultrasonically for 15 min, cool to room temperature, make up the volume with methanol, transfer to a 50 mL centrifuge tube, centrifuge at 4000 r / min for 5 min, filter the supernatant through a 0.22 μm microporous membrane, take the filtrate, and appropriately dilute it with a methanol-water solution to within the linear range according to the actual concentration to obtain a liquid beverage sample; the liquid beverage is one or more mixtures of fruit and vegetable juice beverages, protein beverages, carbonated beverages, tea beverages, coffee beverages, plant beverages, etc.

[0011] (5) Preparation of blank solution: Without adding the sample, process it in the same way as in step (4) to obtain a blank solution.

[0012] Furthermore, the preparation of the standard intermediate working solution in the first method described in step (2) is as follows: Accurately pipette the standard stock solution respectively, dilute it with methanol to 10 mL, shake well, and prepare standard intermediate working solutions of 100 μg / mL respectively, freshly prepared before use; the preparation of the standard intermediate working solution in the second method is as follows: Accurately pipette the standard stock solution, dilute it with methanol to 100 mL, shake well, and prepare a standard working solution of 1 μg / mL, freshly prepared before use.

[0013] Further, the preparation of the standard working solution in the first method described in step (3) is as follows: Accurately pipette the standard intermediate working solution respectively, dilute it with methanol aqueous solution, and shake well to obtain the standard working solution. The concentrations of lemborexant, suvorexant, nemorexant, and amorenone are 0.50 μg / mL, 1.00 μg / mL, 2.00 μg / mL, 5.00 μg / mL, 10.00 μg / mL, and 20.00 μg / mL in sequence. Prepare freshly before use or prepare a standard working solution with an appropriate concentration according to the instrument response. The preparation of the standard working solution in the second method is as follows: Accurately pipette the standard intermediate working solution respectively, dilute it with methanol aqueous solution, and shake well to obtain the standard working solution. The concentrations of lemborexant, suvorexant, nemorexant, and amorenone are 1 μg / L, 5 μg / L, 10 μg / L, 20 μg / L, 50 μg / L, and 100 μg / L in sequence. Prepare freshly before use or prepare a standard working solution with an appropriate concentration according to the instrument response.

[0014] Further, the preparation method of the methanol aqueous solution is as follows: Mix methanol and water in equal volumes; the water is primary water specified in GB / T 6682.

[0015] Further, the determination method of the first method is as follows: Measure the standard working solution according to the instrument reference conditions respectively to obtain the chromatographic peak areas of the corresponding standard solutions; use the concentration of the standard working solution as the abscissa and the peak area of the chromatographic peak of the quantitative ion as the ordinate to plot the standard curve; measure the sample solution according to the instrument reference conditions to obtain the chromatographic peak area of the component to be measured in the sample solution, and obtain the concentration of the component in the test solution according to the standard curve. The number of parallel determinations is not less than two; the blank solution is operated according to the same steps as the sample solution, and then calculate the content of the component to be measured. The determination method of the second method is as follows: Measure the sample and the standard working solution according to the mass spectrometry detection conditions, record the chromatographic retention times of the compounds in the sample and the standard working solution. When a chromatographic peak with a retention time consistent with that of the standard product is detected in the sample, and the change range is within ±2.5%, and the relative ion abundance ratio of the selected monitoring ion pair in the sample chromatogram and the relative ion abundance ratio (k) of the standard solution with a corresponding concentration deviate no more than the allowable relative deviation range, it can be determined that the target substance is detected in the sample; measure the standard working solution according to the instrument reference conditions respectively to obtain the chromatographic peak areas of the corresponding standard solutions. Use the concentration of the standard working solution as the abscissa and the peak area of the chromatographic peak of the quantitative ion as the ordinate to plot the standard curve; measure the sample solution according to the instrument reference conditions to obtain the chromatographic peak area of the component to be measured in the sample solution. Obtain the concentration of the component in the test solution according to the standard curve. The number of parallel determinations is not less than two; the blank solution is operated according to the same steps as the sample solution, and calculate the content of the component to be measured.

[0016] Further, during the determination by the first method, 0.1% formic acid aqueous solution - acetonitrile is used as the mobile phase. The specific preparation method of 0.1% formic acid aqueous solution - acetonitrile is as follows: Pipette 0.5 mL of formic acid, dilute it to 500 mL with water, and mix it with 750 mL of acetonitrile. During the determination by the second method, 0.1% formic acid - 2 mmol / L ammonium acetate water is used as the mobile phase. The specific preparation method of 0.1% formic acid - 2 mmol / L ammonium acetate water is as follows: Accurately pipette 1 mL of formic acid, accurately weigh 0.154 g of ammonium acetate, and dilute it to 1000 mL with water.

[0017] Further, the calculation method for the content of the component to be measured in the sample is as follows:

[0018]

[0019] In the formula:

[0020] X—the content of the analyte in the sample, in milligrams per kilogram (mg / kg);

[0021] c—the concentration of the analyte in the test solution read from the standard curve, in micrograms per liter (μg / L);

[0022] V—the final volume of the sample solution after volume fixation, in milliliters (mL);

[0023] m—the mass represented by the sample solution, in grams (g);

[0024] f—the dilution factor;

[0025] The calculation result is expressed as the arithmetic mean of two independent determination results obtained under repeatability conditions, and the result is retained to three significant figures.

[0026] Further, the allowable relative deviation range is as follows: When k > 50%, the allowable relative deviation is ±20%; when 50% ≥ k > 20%, the allowable relative deviation is ±25%; when 20% ≥ k > 10%, the allowable relative deviation is ±30%; when k ≤ 10%, the allowable relative deviation is ±30%.

[0027] Further, during the determination by the first method, when the sample weight of candies and solid beverages is 1 g, the quantitative limits of lemborexant, suvorexant, nemorexant, and amorenate are all 50 mg / kg; when the sample weight of liquid beverage samples is 5 g, the quantitative limits of lemborexant, suvorexant, nemorexant, and amorenate are all 10 mg / kg; during the determination by the second method, when the sample weight of candies and solid beverage samples is 1 g, the quantitative limits of lemborexant, suvorexant, nemorexant, and amorenate are all 0.05 mg / kg; when the sample weight of liquid beverage samples is 5 g, the quantitative limits of lemborexant, suvorexant, nemorexant, and amorenate are all 0.01 mg / kg.

[0028] The beneficial effects achieved by the present invention are as follows: The present invention adopts the method of dissolving with warm water and ultrasonic extraction with methanol. The operation steps are simple and the cost is low. Different from the traditional determination methods of lemborexant or suvorexant, especially using 0.1% formic acid aqueous solution - acetonitrile or 0.1% formic acid - 2 mmol / L ammonium acetate water, which is beneficial to the separation of the components to be measured in food. And through the calculation formula of the content of the components to be measured in the sample, the content of the components to be measured can be accurately calculated; moreover, this method can simultaneously determine the contents of four components, namely lemborexant, suvorexant, nemorexant, and amorenate in food, and the detection is rapid, and the results are accurate and stable. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] The drawings are used to provide a further understanding of the present invention, and constitute a part of the specification. Together with the embodiments of the present invention, they are used to explain the present invention, and do not constitute a limitation to the present invention. In the drawings:

[0030] Figure 1 is the liquid chromatography diagram of 1.00 μg / mL lemborexant, suvorexant, nemorexant, and amorenate standard solutions in Example 1 of the present application;

[0031] Figure 2 is the multiple reaction monitoring (MRM) mass chromatogram of 1.00 μg / L lemborexant, suvorexant, nemorexant, and amorenate in Example 2 of the present application. DETAILED DESCRIPTION OF THE EMBODIMENTS

[0032] Next, the technical solutions in the embodiments of the present invention will be clearly and completely described in conjunction with the embodiments of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative work shall fall within the protection scope of the present invention.

[0033] Example 1

[0034] (1) Preparation of standard stock solutions: Accurately weigh 5.00 mg of lemborexant, suvorexant, nemorexant, and amolent respectively, dissolve them with methanol and make up the volume to 5 mL to prepare standard stock solutions with a concentration of 1000 μg / mL respectively. Store them in the dark at -20°C, and the validity period is 3 months.

[0035] (2) Preparation of standard intermediate working solutions: Accurately pipette 1.0 mL of the standard stock solution respectively, dilute it to 10 mL with methanol, shake well, and prepare standard intermediate working solutions with a concentration of 100 μg / mL respectively. Prepare fresh before use.

[0036] (3) Preparation of standard working solutions: Accurately pipette the standard intermediate working solutions respectively, dilute them with methanol aqueous solution, shake well, and use them as standard working solutions. The concentrations of lemborexant, suvorexant, nemorexant, and amolent are all 1.00 μg / mL. Prepare fresh before use or prepare standard working solutions with appropriate concentrations according to the instrument response. The volume ratio of methanol to water in the methanol aqueous solution is 1:1; the water is the first-class water specified in GB / T 6682.

[0037] (4) Preparation of sample solutions: Crush candies and solid beverages respectively, accurately weigh 1 g and place it in a 50 mL volumetric flask, add 5 mL of water, warm it to dissolve, wait until completely dissolved, add methanol and extract by ultrasound for 15 min, cool to room temperature, make up the volume to 50 mL with methanol, transfer it to a 50 mL centrifuge tube, centrifuge at 4000 r / min for 5 min, filter the supernatant through a 0.22 μm microporous filter membrane, take the filtrate, dilute it to the linear range with methanol aqueous solution according to the actual concentration, and set aside;

[0038] Take 5 g of liquid beverage and place it in a 50 mL volumetric flask, add methanol and extract by ultrasound for 15 min, cool to room temperature, make up the volume to 50 mL with methanol, transfer it to a 50 mL centrifuge tube, centrifuge at 4000 r / min for 5 min, filter the supernatant through a 0.22 μm microporous filter membrane, take the filtrate, dilute it to the linear range with methanol aqueous solution according to the actual concentration, and set aside;

[0039] (5) Preparation of blank solutions: Without adding samples, process them in the same way as in step (4) to obtain blank solutions.

[0040] (6) Measure the standard working solutions respectively according to the instrument reference conditions to obtain the chromatographic peak areas of the corresponding standard solutions; take the concentration of the standard working solutions as the abscissa and the peak area of the chromatographic peak of the quantitative ion as the ordinate to draw a standard curve.

[0041] (7) Measure the sample solutions according to the instrument reference conditions to obtain the chromatographic peak areas of the components to be measured in the sample solutions. Obtain the concentrations of the components in the test solutions according to the standard curve, and the number of parallel determinations is not less than two.

[0042] (8) The blank solution was measured following the same procedure as in step (7), and the content of the analyte to be determined was calculated.

[0043] Furthermore, when the sample weight for the determination of candy and solid beverage samples is 1 g, the quantification limits of lemborexant, suvorexant, nemorexant, and amorexant are all 50 mg / kg; when the sample weight for liquid beverage samples is 5 g, the quantification limits of lemborexant, suvorexant, nemorexant, and amorexant are all 10 mg / kg.

[0044] Furthermore, the calculation method for the content of the analyte to be determined in the test sample is as follows:

[0045]

[0046] In the formula:

[0047] X—the content of the analyte in the test sample, unit: milligram per kilogram (mg / kg);

[0048] c—the concentration of the analyte in the test solution read from the standard curve, unit: microgram per liter (μg / L);

[0049] V—the final volume of the sample solution after volume fixation, unit: milliliter (mL);

[0050] m—the mass represented by the test sample solution, unit: gram (g);

[0051] f—the dilution factor;

[0052] The calculation result is expressed as the arithmetic mean of two independent determination results obtained under repeatability conditions, and the result is retained to three significant figures.

[0053] Furthermore, during the determination process, the absolute difference between two independent determination results obtained under repeatability conditions shall not exceed 10% of the arithmetic mean.

[0054] Example 2

[0055] (1) Preparation of the standard stock solution: Accurately weigh 5 mg of lemborexant, suvorexant, nemorexant, and amorexant, dissolve with methanol and fix the volume to 5 mL to prepare a standard stock solution with a concentration of 1000 μg / mL, store in the dark at -20 °C, and the validity period is 3 months;

[0056] (2) Preparation of the standard intermediate working solution: Accurately pipette 0.1 mL of the standard stock solution, dilute with methanol to 100 mL, shake well to prepare a standard working solution of 1 μg / mL, and prepare fresh for immediate use;

[0057] (3) Preparation of standard working solutions: Accurately pipette the standard intermediate working solutions respectively, dilute with methanol aqueous solution, shake well, and use as standard working solutions. The concentrations of lemborexant, suvorexant, nemorexant and amolant are all 1 μg / L. Prepare freshly before use or prepare standard working solutions with appropriate concentrations according to the instrument response;

[0058] (4) Preparation of sample solutions: Crush candies and solid beverages respectively, accurately weigh 1 g and place it in a 50 mL volumetric flask, add 5 mL of water, warm and dissolve. After complete dissolution, add methanol and ultrasonically extract for 15 min. Cool to room temperature, make up the volume to 50 mL with methanol, transfer to a 50 mL centrifuge tube, centrifuge at 4000 r / min for 5 min. Filter the supernatant through a 0.22 μm microporous membrane, take the filtrate, and dilute it to the linear range with methanol aqueous solution according to the actual concentration for standby;

[0059] Take 5 g of liquid beverage and place it in a 50 mL volumetric flask, add methanol and ultrasonically extract for 15 min. Cool to room temperature, make up the volume to 50 mL with methanol, transfer to a 50 mL centrifuge tube, centrifuge at 4000 r / min for 5 min. Filter the supernatant through a 0.22 μm microporous membrane, take the filtrate, and dilute it to the linear range with methanol aqueous solution according to the actual concentration for standby;

[0060] (5) Preparation of blank solutions: Without adding samples, treat in the same way as in step (4) to obtain blank solutions;

[0061] (6) Determine the samples and standard working solutions according to the mass spectrometry detection conditions, record the chromatographic retention times of the compounds in the samples and standard solutions. When a chromatographic peak with the same retention time as the standard product chromatographic peak (within the range of ±2.5%) is detected in the sample, and the deviation of the relative ion abundance ratio of the selected monitoring ion pairs in the sample chromatogram from the relative ion abundance ratio (k) of the standard solution with the corresponding concentration does not exceed the allowable relative deviation range, the target substance can be determined to be detected in the sample; the allowable relative deviation range is: when k > 50%, the allowable relative deviation is ±20%; when 50% ≥ k > 20%, the allowable relative deviation is ±25%; when 20% ≥ k > 10%, the allowable relative deviation is ±30%, when k ≤ 10%, the allowable relative deviation is ±30%;

[0062] (7) Determine the standard working solutions according to the instrument reference conditions respectively to obtain the chromatographic peak areas of the corresponding standard solutions; use the concentration of the standard working solutions as the abscissa and the peak area of the chromatographic peak of the quantitative ion as the ordinate to draw a standard curve;

[0063] (8) Determine the sample solution according to the instrument reference conditions to obtain the chromatographic peak area of the component to be measured in the sample solution. Obtain the concentration of the component in the test solution according to the standard curve, and the number of parallel determinations is not less than two;

[0064] (9) The blank solution was assayed following the same procedure as in step (8), and the content of the analyte to be determined was calculated.

[0065] Furthermore, during the determination, when the sample weight of candies and solid beverages is 1 g, the quantification limits of lemborexant, suvorexant, nemorexant, and amorexant are all 0.05 mg / kg; when the sample weight of liquid beverages is 5 g, the quantification limits of lemborexant, suvorexant, nemorexant, and amorexant are all 0.01 mg / kg.

[0066] Furthermore, the calculation method for the content of the analyte to be determined in the test sample is as follows:

[0067]

[0068] In the formula:

[0069] X—the content of the analyte in the test sample, unit: milligram per kilogram (mg / kg);

[0070] c—the concentration of the analyte in the test solution read from the standard curve, unit: microgram per liter (μg / L);

[0071] V—the final volume of the sample solution after constant volume, unit: milliliter (mL);

[0072] m—the mass represented by the test sample solution, unit: gram (g);

[0073] f—the dilution factor;

[0074] The calculation result is expressed as the arithmetic mean of two independent determinations obtained under repeatability conditions, and the result is retained to three significant figures.

[0075] Furthermore, during the determination, the absolute difference between two independent determinations obtained under repeatability conditions shall not exceed 20% of the arithmetic mean.

[0076] For those skilled in the art, it is obvious that the present invention is not limited to the details of the above exemplary embodiments, and the present invention can be implemented in other specific forms without departing from the spirit or basic characteristics of the present invention. Therefore, from any point of view, the embodiments should be regarded as exemplary and non-limiting. The scope of the present invention is defined by the appended claims rather than the above description. Therefore, all changes falling within the meaning and scope of the equivalent elements of the claims are intended to be encompassed within the present invention. Any reference signs in the claims should not be construed as limiting the claimed rights.

Claims

1. A method for the determination of lemborexant, suvorexant, nemorexant and amolent in food, characterized in that, The determination methods of lemborexant, suvorexant, nemorexant and amorexant in the food include the first method and the second method; the first method and the second method are high performance liquid chromatography and high performance liquid chromatography-tandem mass spectrometry respectively.

2. The method for determining lemborexant, suvorexant, nemorexant and amolent in a food according to claim 1, characterized in that, The first method or the second method includes the following: (1) Preparation of standard stock solutions: Accurately weigh lemborexant, suvorexant, nemorexant and amorexant respectively, dissolve them with methanol and make up the volume to 5 mL to prepare standard stock solutions with a concentration of 1000 μg / mL respectively, store them in the dark at -20°C, and the validity period is 3 months; (2) Preparation of standard intermediate working solutions; (3) Preparation of standard working solutions; (4) Preparation of sample solutions: Crush candies and solid beverages respectively, place them in a 50 mL volumetric flask, warm and dissolve them with water. After the candies are completely dissolved, add methanol, ultrasonically extract for 15 min, cool to room temperature, make up the volume with methanol, transfer to a 50 mL centrifuge tube, centrifuge at 4000 r / min for 5 min, filter the supernatant through a 0.22 μm microporous membrane, take the filtrate, and dilute it with a methanol aqueous solution to within the linear range according to the actual concentration to obtain candy samples and solid beverage samples respectively; the candies are one or more mixtures of hard candies, crispy candies, gel candies, gum base candies, tablet candies, milk candies, etc.; the solid beverages are one or more mixtures of fruit and vegetable solid beverages, protein solid beverages, coffee solid beverages, tea solid beverages, etc.; Take liquid beverages and place them in a 50 mL volumetric flask, add methanol and ultrasonically extract for 15 min, cool to room temperature, make up the volume with methanol, transfer to a 50 mL centrifuge tube, centrifuge at 4000 r / min for 5 min, filter the supernatant through a 0.22 μm microporous membrane, take the filtrate, and appropriately dilute it with a methanol aqueous solution to within the linear range according to the actual concentration to obtain liquid beverage samples; the liquid beverages are one or more mixtures of fruit and vegetable juice beverages, protein beverages, carbonated beverages, tea beverages, coffee beverages, plant beverages, etc.; (5) Preparation of blank solutions: Without adding samples, treat them in the same way as in step (4) to obtain blank solutions.

3. The method for determining lemborexant, suvorexant, nemorexant and amolent in a food according to claim 2, characterized in that, The preparation of the standard intermediate working solutions in the first method in step (2) is as follows: Accurately pipette the standard stock solutions respectively, dilute them with methanol to 10 mL, shake well, and prepare standard intermediate working solutions with a concentration of 100 μg / mL respectively, freshly prepared before use; the preparation of the standard intermediate working solutions in the second method is as follows: Accurately pipette the standard stock solutions, dilute them with methanol to 100 mL, shake well, and prepare a standard working solution with a concentration of 1 μg / mL, freshly prepared before use.

4. The method for determining lemborexant, suvorexant, nemorexant and amorexant in food according to claim 2, characterized in that, The preparation of the standard working solution in the first method described in step (3) is as follows: Accurately pipette the standard intermediate working solution respectively, dilute it with methanol aqueous solution, and shake well to obtain the standard working solution. The concentrations of lemborexant, suvorexant, nemorexant, and amorenone are 0.50 μg / mL, 1.00 μg / mL, 2.00 μg / mL, 5.00 μg / mL, 10.00 μg / mL, and 20.00 μg / mL in sequence. Prepare fresh before use or prepare a standard working solution with an appropriate concentration according to the instrument response. The preparation of the standard working solution in the second method is as follows: Accurately pipette the standard intermediate working solution respectively, dilute it with methanol aqueous solution, and shake well to obtain the standard working solution. The concentrations of lemborexant, suvorexant, nemorexant, and amorenone are 1 μg / L, 5 μg / L, 10 μg / L, 20 μg / L, 50 μg / L, and 100 μg / L in sequence. Prepare fresh before use or prepare a standard working solution with an appropriate concentration according to the instrument response.

5. The method for determining lemborexant, suvorexant, nemorexant and amolent in a food according to claims 2 and 4, characterized in that, The preparation method of the methanol aqueous solution is: Mix methanol and water in equal volumes; the water is the first-class water specified in GB / T 6682.

6. A method for determining lemborexant, suvorexant, nemorexant, and amorexant in food according to claims 2 to 5, characterized in that, The determination method of the first method is as follows: Measure the standard working solution respectively according to the instrument reference conditions to obtain the chromatographic peak areas of the corresponding standard solutions; Use the concentration of the standard working solution as the abscissa and the peak area of the chromatographic peak of the quantitative ion as the ordinate to draw a standard curve; Measure the sample solution according to the instrument reference conditions to obtain the chromatographic peak area of the component to be measured in the sample solution. Obtain the concentration of the component in the test solution according to the standard curve, and the number of parallel determinations is not less than two; The blank solution is operated in the same manner as the sample solution determination steps, and then calculate the content of the component to be measured. The determination method of the second method is as follows: Measure the sample and the standard working solution according to the mass spectrometry detection conditions, record the chromatographic retention times of the compounds in the sample and the standard working solution. When a chromatographic peak with a retention time consistent with that of the standard product is detected in the sample, and the variation range is within ±2.5%, and the deviation of the relative ion abundance ratio of the selected monitoring ion pairs in the sample chromatogram from the relative ion abundance ratio (k) of the standard solution with a corresponding concentration does not exceed the allowable relative deviation range, it can be determined that the target substance is detected in the sample; Measure the standard working solution respectively according to the instrument reference conditions to obtain the chromatographic peak areas of the corresponding standard solutions. Use the concentration of the standard working solution as the abscissa and the peak area of the chromatographic peak of the quantitative ion as the ordinate to draw a standard curve; Measure the sample solution according to the instrument reference conditions to obtain the chromatographic peak area of the component to be measured in the sample solution. Obtain the concentration of the component in the test solution according to the standard curve, and the number of parallel determinations is not less than two; The blank solution is operated in the same manner as the sample solution determination steps, and calculate the content of the component to be measured.

7. The method for determining lemborexant, suvorexant, nemorexant and amolent in a food according to claim 6, characterized in that, In the determination process of the first method, 0.1% formic acid aqueous solution - acetonitrile is used as the mobile phase. The specific preparation method of 0.1% formic acid aqueous solution - acetonitrile is as follows: Pipette 0.5 mL of formic acid, dilute it with water to 500 mL, and mix it with 750 mL of acetonitrile. In the determination process of the second method, 0.1% formic acid - 2 mmol / L ammonium acetate water is used as the mobile phase. The specific preparation method of 0.1% formic acid - 2 mmol / L ammonium acetate water is as follows: Accurately pipette 1 mL of formic acid, accurately weigh 0.154 g of ammonium acetate, and dilute it with water to 1000 mL.

8. The method for determining lemborexant, suvorexant, nemorexant and amolent in a food according to claim 6, wherein The calculation method for the content of the component to be measured in the sample is as follows: Where: X—the content of the analyte in the sample, in milligrams per kilogram (mg / kg); c—the concentration of the analyte in the test solution read from the standard curve, in micrograms per liter (μg / L); V—the final volume of the sample solution after volume fixation, in milliliters (mL); m—the mass represented by the sample solution, in grams (g); f—the dilution factor; The calculation result is expressed as the arithmetic mean of two independent determination results obtained under repeatability conditions, and the result is retained to three significant figures.

9. The method for determining lemborexant, suvorexant, nemorexant and amolent in a food according to claim 6, characterized in that, The allowable relative deviation range is as follows: When k > 50%, the allowable relative deviation is ±20%; when 50% ≥ k > 20%, the allowable relative deviation is ±25%; when 20% ≥ k > 10%, the allowable relative deviation is ±30%; when k ≤ 10%, the allowable relative deviation is ±30%.

10. A method for determining lemborexant, suvorexant, nemorexant and amorexant in food according to claims 1 to 9, characterized in that, In the determination process of the first method, when the sample weight is 1 g for candy and solid beverage samples, the quantitative limits of lemborexant, suvorexant, nemorexant, and amorenone are all 50 mg / kg; when the sample weight is 5 g for liquid beverage samples, the quantitative limits of lemborexant, suvorexant, nemorexant, and amorenone are all 10 mg / kg. In the determination process of the second method, when the sample weight is 1 g for candy and solid beverage samples, the quantitative limits of lemborexant, suvorexant, nemorexant, and amorenone are all 0.05 mg / kg; when the sample weight is 5 g for liquid beverage samples, the quantitative limits of lemborexant, suvorexant, nemorexant, and amorenone are all 0.01 mg / kg.

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