ANALYSIS METHOD OF 3-METHYL-1-PHENYL-2-PYRAZOLIN-5-ONE ACTIVE PHARMACEUTICAL INGREDIENT, TREATMENT FOR AMYOTROPHIC LATERAL SCLEROSIS, INHIBITION OF PROGRESSION OF AMYOTROPHIC LATERAL SCLEROSIS, AND METHOD OF PRODUCING MEDICAMENT CONTAINING 3-METHYL-1-PHENYL-2-PYRAZOLIN-5-ONE ACTIVE PHARMACEUTICAL INGREDIENT

1 . A method of analyzing a phenylhydrazine content in a 3-methyl-1-phenyl-2-pyrazolin-5-one active pharmaceutical ingredient, comprising:

obtaining a first measured value by measuring a phenylhydrazine content in a phenylhydrazine standard solution including phenylhydrazine or a salt thereof, a first acidic water and a first water-soluble organic solvent and having a phenylhydrazine concentration in a range of 0.01 μg/mL to 10 μg/mL;

obtaining a second measured value by measuring a phenylhydrazine content in a 3-methyl-1-phenyl-2-pyrazolin-5-one sample solution including a 3-methyl-1-phenyl-2-pyrazolin-5-one active pharmaceutical ingredient, a second acidic water and a second water-soluble organic solvent; and

detecting a phenylhydrazine content in a 3-methyl-1-phenyl-2-pyrazolin-5-one active pharmaceutical ingredient based on the first measured value and the second measured value,

wherein the first acidic water is at least one selected from the group consisting of hydrochloric acid, an aqueous acetic acid solution, an aqueous trifluoroacetic acid solution, an aqueous formic acid solution, an aqueous phosphoric acid solution and an aqueous perchloric acid solution, the first water-soluble organic solvent is at least one selected from the group consisting of acetonitrile and methanol, provided that when the first water-soluble organic solvent is methanol, the first acidic water is at least one selected from the group consisting of hydrochloric acid, an aqueous acetic acid solution, an aqueous trifluoroacetic acid solution, an aqueous formic acid solution and an aqueous perchloric acid solution, the second acidic water is at least one selected from the group consisting of hydrochloric acid, an aqueous acetic acid solution, an aqueous trifluoroacetic acid solution, an aqueous formic acid solution, an aqueous phosphoric acid solution and an aqueous perchloric acid solution, and the second water-soluble organic solvent is at least one selected from the group consisting of acetonitrile and methanol, provided that when the second water-soluble organic solvent is methanol, the second acidic water is at least one selected from the group consisting of hydrochloric acid, an aqueous acetic acid solution, an aqueous trifluoroacetic acid solution, an aqueous formic acid solution and an aqueous perchloric acid solution.

2 . The method according to claim 1 , further comprising:

preparing the phenylhydrazine standard solution; and

preparing the 3-methyl-1-phenyl-2-pyrazolin-5-one sample solution.

3 . The method according to claim 1 , wherein the first measured value is obtained by measuring the phenylhydrazine content in the phenylhydrazine standard solution using high performance liquid chromatography, and the second measured value is obtained by measuring the phenylhydrazine content in the 3-methyl-1-phenyl-2-pyrazolin-5-one sample solution using high performance liquid chromatography.

4 . The method according to claim 1 , wherein the first acidic water includes hydrochloric acid in a range of 0.01 mol/L to 5 mol/L, and the second acidic water includes hydrochloric acid in a range of 0.01 mol/L to 5 mol/L.

5 . The method according to claim 1 , wherein a volume ratio of the first acidic water and the first water-soluble organic solvent in the phenylhydrazine standard solution is in a range of 3:1 to 4:5, and a volume ratio of the second acidic water and the second water-soluble organic solvent in the 3-methyl-1-phenyl-2-pyrazoline-5-one sample solution is in a range of 3:1 to 4:5.

6 . A method of treating amyotrophic lateral sclerosis, comprising:

obtaining a first measured value by measuring a phenylhydrazine content in a phenylhydrazine standard solution including phenylhydrazine or a salt thereof, a first acidic water and a first water-soluble organic solvent and having a phenylhydrazine concentration in a range of 0.01 μg/mL to 10 μg/mL;

obtaining a second measured value by measuring a phenylhydrazine content in a 3-methyl-1-phenyl-2-pyrazolin-5-one sample solution including a 3-methyl-1-phenyl-2-pyrazolin-5-one active pharmaceutical ingredient, a second acidic water and a second water-soluble organic solvent;

detecting a phenylhydrazine content in a 3-methyl-1-phenyl-2-pyrazolin-5-one active pharmaceutical ingredient based on a based the first measured value and the second measured value to determine whether the phenylhydrazine content in the 3-methyl-1-phenyl-2-pyrazolin-5-one active pharmaceutical ingredient is 20 ppm or less; and

administering a medicament comprising the 3-methyl-1-phenyl-2-pyrazolin-5-one active pharmaceutical ingredient to a patient in need thereof such that the phenylhydrazine content in the 3-methyl-1-phenyl-2-pyrazolin-5-one active pharmaceutical ingredient is 20 ppm or less,

wherein the first acidic water is at least one selected from the group consisting of hydrochloric acid, an aqueous acetic acid solution, an aqueous trifluoroacetic acid solution, an aqueous formic acid solution, an aqueous phosphoric acid solution and an aqueous perchloric acid solution, the first water-soluble organic solvent is at least one selected from the group consisting of acetonitrile and methanol, provided that when the first water-soluble organic solvent is methanol, the first acidic water is at least one selected from the group consisting of hydrochloric acid, an aqueous acetic acid solution, an aqueous trifluoroacetic acid solution, an aqueous formic acid solution and an aqueous perchloric acid solution, the second acidic water is at least one selected from the group consisting of hydrochloric acid, an aqueous acetic acid solution, an aqueous trifluoroacetic acid solution, an aqueous formic acid solution, an aqueous phosphoric acid solution and an aqueous perchloric acid solution, and the second water-soluble organic solvent is at least one selected from the group consisting of acetonitrile and methanol, provided that when the second water-soluble organic solvent is methanol, the second acidic water is at least one selected from the group consisting of hydrochloric acid, an aqueous acetic acid solution, an aqueous trifluoroacetic acid solution, an aqueous formic acid solution and an aqueous perchloric acid solution.

7 . The method according to claim 6 , further comprising:

preparing the phenylhydrazine standard solution; and

preparing the 3-methyl-1-phenyl-2-pyrazolin-5-one sample solution.

8 . The method according to claim 6 , wherein the first measured value is obtained by measuring the phenylhydrazine content in the phenylhydrazine standard solution using high performance liquid chromatography, and the second measured value is obtained by measuring the phenylhydrazine content in the 3-methyl-1-phenyl-2-pyrazolin-5-one sample solution using high performance liquid chromatography.

9 . The method according to claim 6 , wherein the first acidic water includes hydrochloric acid in a range of 0.01 mol/L to 5 mol/L, and the second acidic water includes hydrochloric acid in a range of 0.01 mol/L to 5 mol/L.

10 . The method according to claim 6 , wherein a volume ratio of the first acidic water and the first water-soluble organic solvent in the phenylhydrazine standard solution is in a range of 3:1 to 4:5, and a volume ratio of the second acidic water and the second water-soluble organic solvent in the 3-methyl-1-phenyl-2-pyrazoline-5-one sample solution is in a range of 3:1 to 4:5.

11 . The method according to claim 7 , wherein the first measured value is obtained by measuring the phenylhydrazine content in the phenylhydrazine standard solution using high performance liquid chromatography, and the second measured value is obtained by measuring the phenylhydrazine content in the 3-methyl-1-phenyl-2-pyrazolin-5-one sample solution using high performance liquid chromatography.

12 . The method according to claim 7 , wherein the first acidic water includes hydrochloric acid in a range of 0.01 mol/L to 5 mol/L, and the second acidic water includes hydrochloric acid in a range of 0.01 mol/L to 5 mol/L.

13 . The method according to claim 7 , wherein a volume ratio of the first acidic water and the first water-soluble organic solvent in the phenylhydrazine standard solution is in a range of 3:1 to 4:5, and a volume ratio of the second acidic water and the second water-soluble organic solvent in the 3-methyl-1-phenyl-2-pyrazoline-5-one sample solution is in a range of 3:1 to 4:5.

14 . The method according to claim 8 , wherein the first acidic water includes hydrochloric acid in a range of 0.01 mol/L to 5 mol/L, and the second acidic water includes hydrochloric acid in a range of 0.01 mol/L to 5 mol/L.

15 . The method according to claim 8 , wherein a volume ratio of the first acidic water and the first water-soluble organic solvent in the phenylhydrazine standard solution is in a range of 3:1 to 4:5, and a volume ratio of the second acidic water and the second water-soluble organic solvent in the 3-methyl-1-phenyl-2-pyrazoline-5-one sample solution is in a range of 3:1 to 4:5.

16 . The method according to claim 9 , wherein a volume ratio of the first acidic water and the first water-soluble organic solvent in the phenylhydrazine standard solution is in a range of 3:1 to 4:5, and a volume ratio of the second acidic water and the second water-soluble organic solvent in the 3-methyl-1-phenyl-2-pyrazoline-5-one sample solution is in a range of 3:1 to 4:5.

17 . A method of suppressing progress of amyotrophic lateral sclerosis, comprising:

obtaining a first measured value by measuring a phenylhydrazine content in a phenylhydrazine standard solution including phenylhydrazine or a salt thereof, a first acidic water and a first water-soluble organic solvent and having a phenylhydrazine concentration in a range of 0.01 μg/mL to 10 μg/mL;

obtaining a second measured value by measuring a phenylhydrazine content in a 3-methyl-1-phenyl-2-pyrazolin-5-one sample solution including a 3-methyl-1-phenyl-2-pyrazolin-5-one active pharmaceutical ingredient, a second acidic water and a second water-soluble organic solvent;

detecting a phenylhydrazine content in a 3-methyl-1-phenyl-2-pyrazolin-5-one active pharmaceutical ingredient based on the first measured value and the second measured value to determine whether the phenylhydrazine content in the 3-methyl-1-phenyl-2-pyrazolin-5-one active pharmaceutical ingredient is 20 ppm or less; and

administering a medicament comprising the 3-methyl-1-phenyl-2-pyrazolin-5-one active pharmaceutical ingredient to a patient in need thereof such that the phenylhydrazine content in the 3-methyl-1-phenyl-2-pyrazolin-5-one active pharmaceutical ingredient is 20 ppm or less,

wherein the first acidic water is at least one selected from the group consisting of hydrochloric acid, an aqueous acetic acid solution, an aqueous trifluoroacetic acid solution, an aqueous formic acid solution, an aqueous phosphoric acid solution and an aqueous perchloric acid solution, the first water-soluble organic solvent is at least one selected from the group consisting of acetonitrile and methanol, provided that when the first water-soluble organic solvent is methanol, the first acidic water is at least one selected from the group consisting of hydrochloric acid, an aqueous acetic acid solution, an aqueous trifluoroacetic acid solution, an aqueous formic acid solution and an aqueous perchloric acid solution, the second acidic water is at least one selected from the group consisting of hydrochloric acid, an aqueous acetic acid solution, an aqueous trifluoroacetic acid solution, an aqueous formic acid solution, an aqueous phosphoric acid solution and an aqueous perchloric acid solution, and the second water-soluble organic solvent is at least one selected from the group consisting of acetonitrile and methanol, provided that when the second water-soluble organic solvent is methanol, the second acidic water is at least one selected from the group consisting of hydrochloric acid, an aqueous acetic acid solution, an aqueous trifluoroacetic acid solution, an aqueous formic acid solution and an aqueous perchloric acid solution.

18 . The method according to claim 17 , further comprising:

preparing the phenylhydrazine standard solution; and

preparing the 3-methyl-1-phenyl-2-pyrazolin-5-one sample solution.

19 . The method according to claim 17 , wherein the first measured value is obtained by measuring the phenylhydrazine content in the phenylhydrazine standard solution using high performance liquid chromatography, and the second measured value is obtained by measuring the phenylhydrazine content in the 3-methyl-1-phenyl-2-pyrazolin-5-one sample solution using high performance liquid chromatography.

20 . The method according to claim 17 , wherein the first acidic water includes hydrochloric acid in a range of 0.01 mol/L to 5 mol/L, and the second acidic water includes hydrochloric acid in a range of 0.01 mol/L to 5 mol/L.

21 . The method according to claim 17 , wherein a volume ratio of the first acidic water and the first water-soluble organic solvent in the phenylhydrazine standard solution is in a range of 3:1 to 4:5, and a volume ratio of the second acidic water and the second water-soluble organic solvent in a 3-methyl-1-phenyl-2-pyrazoline-5-one sample solution is in a range of 3:1 to 4:5.

22 . A medicament for treating amyotrophic lateral sclerosis or suppressing progress of amyotrophic lateral sclerosis, wherein the medicament is produced by a process comprising:

obtaining a first measured value by measuring a phenylhydrazine content in a phenylhydrazine standard solution including phenylhydrazine or a salt thereof, a first acidic water and a first water-soluble organic solvent and having a phenylhydrazine concentration in a range of 0.01 μg/mL to 10 μg/mL;

obtaining a second measured value by measuring a phenylhydrazine content in a 3-methyl-1-phenyl-2-pyrazolin-5-one sample solution including a 3-methyl-1-phenyl-2-pyrazolin-5-one active pharmaceutical ingredient, a second acidic water and a second water-soluble organic solvent;

detecting a phenylhydrazine content in a 3-methyl-1-phenyl-2-pyrazolin-5-one active pharmaceutical ingredient based on the first measured value and the second measured value to determine whether the phenylhydrazine content in the 3-methyl-1-phenyl-2-pyrazolin-5-one active pharmaceutical ingredient is 20 ppm or less; and

mixing the 3-methyl-1-phenyl-2-pyrazolin-5-one active pharmaceutical ingredient with at least one pharmacologically and pharmaceutically acceptable additive such that the phenylhydrazine content in the 3-methyl-1-phenyl-2-pyrazolin-5-one active pharmaceutical ingredient is 20 ppm or less,

wherein the first acidic water is at least one selected from the group consisting of hydrochloric acid, an aqueous acetic acid solution, an aqueous trifluoroacetic acid solution, an aqueous formic acid solution, an aqueous phosphoric acid solution and an aqueous perchloric acid solution, and the first water-soluble organic solvent is at least one selected from the group consisting of acetonitrile and methanol, provided that when the first water-soluble organic solvent is methanol, the first acidic water is at least one selected from the group consisting of hydrochloric acid, an aqueous acetic acid solution, an aqueous trifluoroacetic acid solution, an aqueous formic acid solution and an aqueous perchloric acid solution, the second acidic water is at least one selected from the group consisting of hydrochloric acid, an aqueous acetic acid solution, an aqueous trifluoroacetic acid solution, an aqueous formic acid solution, an aqueous phosphoric acid solution and an aqueous perchloric acid solution, and the second water-soluble organic solvent is at least one selected from the group consisting of acetonitrile and methanol, provided that when the second water-soluble organic solvent is methanol, the second acidic water is at least one selected from the group consisting of hydrochloric acid, an aqueous acetic acid solution, an aqueous trifluoroacetic acid solution, an aqueous formic acid solution and an aqueous perchloric acid solution.

23 . The medicament according to claim 22 , further comprising:

preparing the phenylhydrazine standard solution; and

preparing the 3-methyl-1-phenyl-2-pyrazolin-5-one sample solution.

24 . The medicament according to claim 22 , wherein the first measured value is obtained by measuring the phenylhydrazine content in the phenylhydrazine standard solution using high performance liquid chromatography, and the second measured value is obtained by measuring the phenylhydrazine content in the 3-methyl-1-phenyl-2-pyrazolin-5-one sample solution using high performance liquid chromatography.

25 . The medicament according to claim 22 , wherein the first acidic water includes hydrochloric acid in a range of 0.01 mol/L to 5 mol/L, and the second acidic water includes hydrochloric acid in a range of 0.01 mol/L to 5 mol/L.

26 . The medicament according to claim 22 , wherein a volume ratio of the first acidic water and the first water-soluble organic solvent in the phenylhydrazine standard solution is in a range of 3:1 to 4:5, and a volume ratio of the second acidic water and the second water-soluble organic solvent in the 3-methyl-1-phenyl-2-pyrazoline-5-one sample solution is in a range of 3:1 to 4:5.

27 . A 3-methyl-1-phenyl-2-pyrazolin-5-one active pharmaceutical ingredient obtained by a process comprising the method of claim 1 .