A method for separating and preparing 4-aminoindenman using a supercritical fluid chromatography system

Abstract

This invention discloses a method for separating and preparing 4-aminoindane using a supercritical fluid chromatography (SCLC) system, comprising the following steps: dissolving an aminoindane sample; separating the aminoindane sample on a chromatographic column using a SCLC system to prepare 4-aminoindane; the mobile phase of the SCLC system is a mixture of supercritical CO2 and lower alcohols; the stationary phase of the SCLC system includes one or more of the following: silica gel surface-bonded phenyl groups, silica gel surface-bonded urea groups, or silica gel surface-bonded imidazole groups. This invention is the first to discover the use of a SCLC system for the separation and preparation of 4-aminoindane, supplementing existing high-performance liquid chromatography (HPLC); CO2 is more environmentally friendly, reducing harm to human health and the environment, and significantly reducing solvent processing costs; it can reduce the amount of chemical reagents used, reducing costs and increasing efficiency; the separation time is short, the separation effect is good, and the product purity can reach over 99%.

Description

Technical Field

[0001] This invention relates to a method for separating and preparing 4-aminoindenhydrin using a supercritical fluid chromatography system, belonging to the field of chromatographic separation and purification. Background Technology

[0002] 4-Aminoindane is an important pharmaceutical intermediate that can be used to synthesize various active pharmaceutical ingredients. Its molecular structure is shown in formula a. It possesses anti-tumor, antihypertensive, endocrine and metabolic regulation, and heart rate improvement effects. Currently, the main method for producing 4-aminoindane in the market is through direct chemical synthesis, but this easily leads to isomers, such as 5-aminoindane. High-performance liquid chromatography (HPLC) is currently used for separation, but this method suffers from drawbacks such as long separation time and high solvent consumption. The molecular structure of 5-aminoindane is as follows:

[0003] Supercritical fluid chromatography (SFC) uses supercritical carbon dioxide instead of conventional organic reagents. Leveraging the unique physicochemical properties of supercritical carbon dioxide, it has applications in the separation of stereoisomers, the analysis of petroleum products, the separation of lipids and natural products, and pharmaceutical and food analysis. However, there is currently no technology available for separating 4-aminoindenman and 5-aminoindenman using SFC. Summary of the Invention

[0004] Objective of the invention: The technical problem to be solved by the present invention is to provide a method for separating and preparing 4-aminoindenhydride using a supercritical fluid chromatography system.

[0005] Technical Solution: To solve the above-mentioned technical problems, this invention provides a method for separating and preparing 4-aminoindane using a supercritical fluid chromatography system, comprising the following steps: dissolving an aminoindane sample, separating the aminoindane sample on a chromatographic column using a supercritical fluid chromatography system to prepare 4-aminoindane and 5-aminoindane; the mobile phase of the supercritical fluid chromatography system is a mixture of supercritical CO2 and lower alcohols; the stationary phase of the supercritical fluid chromatography system includes one or more of the following: phenyl groups bonded to the surface of silica gel, urea groups bonded to the surface of silica gel, or imidazole groups bonded to the surface of silica gel.

[0006] The back pressure of the supercritical fluid chromatography system is 8–12 MPa.

[0007] The volume ratio of supercritical CO2 to lower alcohol is 90-99:1-10.

[0008] Preferably, the volume ratio of supercritical CO2 to lower alcohol is 90-97:3-10.

[0009] The lower alcohols include methanol or ethanol.

[0010] The aminoindane sample comprises a mixture of 4-aminoindane and 5-aminoindane, and the concentration of the aminoindane sample is 3–273 mg / mL.

[0011] The aminoindane sample comprises a mixture of 4-aminoindane and 5-aminoindane, and the concentration of the aminoindane sample is 3–79.43 mg / mL.

[0012] The column temperature of the supercritical fluid chromatography system is 30–45°C.

[0013] Preferably, the column temperature of the supercritical fluid chromatography system is 30–35°C.

[0014] The detection wavelength of the supercritical fluid chromatography system is 254 nm.

[0015] The flow rate of the mobile phase is 2–8 mL / min.

[0016] The injection volume of the supercritical fluid chromatography system is 6–200 μL.

[0017] The chromatographic column of the supercritical fluid chromatography system is a Hedera Phenyl 5um 4.6*250mm column, a Nucifera MSU 5um 10*250mm column, or a Nucifera Imu 5um 10*250mm column.

[0018] Working principle: Based on the differences in chemical properties between 4-aminoindane and 5-aminoindane, they have different partition coefficients in a system composed of a stationary phase and a mobile phase. When the two phases interact, these substances move together with the mobile phase and undergo repeated partitioning between the two phases, thereby achieving separation of the substances.

[0019] Beneficial Effects: Compared with existing technologies, this invention has the following significant advantages: 1. This invention is the first to discover a method for separating 4-aminoindane and 5-aminoindane using a supercritical fluid chromatography system, supplementing existing high-performance liquid chromatography methods; 2. Using CO2 to replace one of the organic reagents is more environmentally friendly, reducing harm to human health and the environment, and significantly reducing solvent processing costs in subsequent drug production; 3. The critical temperature and pressure of CO2 are compatible with the chromatograph, it is non-destructive to the compounds to be analyzed and purified, has low reactivity and toxicity, is non-flammable, and CO2 itself is abundant and inexpensive; 4. While rapidly processing large quantities of samples, it can reduce the amount of chemical reagents used, achieving less cost input and more benefits; 5. The separation time is short, the separation effect is good, and the purity of 4-aminoindane can reach over 99%; 6. It has broad application prospects in the separation of 4-aminoindane and 5-aminoindane. Attached Figure Description

[0020] Figure 1 The separation spectrum obtained in Example 1;

[0021] Figure 2 The separation spectrum obtained in Example 2;

[0022] Figure 3 The separation spectrum obtained in Example 3;

[0023] Figure 4 The separation spectrum obtained in Example 4;

[0024] Figure 5 The separation spectrum obtained in Example 5;

[0025] Figure 6 The separation spectrum obtained in Example 6;

[0026] Figure 7 The separation spectrum obtained in Example 7;

[0027] Figure 8 The separation spectrum obtained in Example 8;

[0028] Figure 9 The separation spectrum obtained in Example 9;

[0029] Figure 10 The separation spectrum obtained in Example 10;

[0030] Figure 11 The separation spectrum obtained in Example 11;

[0031] Figure 12 The separation spectrum obtained in Comparative Example 1;

[0032] Figure 13 The separation spectrum obtained in Comparative Example 2;

[0033] Figure 14 The separation spectrum obtained in Comparative Example 3;

[0034] Figure 15 The separation spectrum obtained in Comparative Example 4;

[0035] Figure 16 The separation spectrum obtained in Comparative Example 5;

[0036] Figure 17 The separation spectrum obtained in Comparative Example 6;

[0037] Figure 18 The separation spectrum obtained in Comparative Example 7;

[0038] Figure 19 The separation spectrum obtained in Comparative Example 8 is shown. Detailed Implementation

[0039] The technical solution of the present invention will be further described below with reference to the accompanying drawings.

[0040] Example 1

[0041] The chromatograph used in this embodiment is a supercritical fluid chromatograph from Jiangsu Hanbang Technology Co., Ltd., with a column size of 4.6*250mm, a mobile phase flow rate of 2mL / min, a back pressure of 12MPa, a aminoindene sample concentration of 3mg / mL, an injection volume of 8μL, and a detection wavelength of 254nm.

[0042] Table 1-1 lists other relevant conditions used in the method of separating and preparing 4-aminoindenhydride using a supercritical fluid chromatography system in Example 1.

[0043] Table 1-1 SFC separation conditions in Example 1

[0044]

[0045]

[0046] The specific steps corresponding to Example 1 are as follows: The aminoindane sample was dissolved in methanol, sonicated to prepare a 3 mg / mL solution, and filtered through a 0.45 μm nylon membrane. A Hedera Phenyl 5 μm 4.6*250 mm (silica gel surface-bonded phenyl) column was selected as the supercritical fluid chromatograph of Jiangsu Hanbang Technology Co., Ltd. The volume ratio of supercritical carbon dioxide to methanol, total flow rate, back pressure, and column temperature in the mobile phase were controlled according to Table 1-1, while ensuring that the temperature of carbon dioxide entering the pump was 6 °C. The aminoindane sample was separated, and the separation effect is as follows: Figure 1 As shown, baseline separation can be achieved.

[0047] Table 1-2 Figure 1 Retention time and peak area of ​​4-aminoindane and 5-aminoindane

[0048] 4-Aminoindane 5-Aminoindane Retention time (min) 1.80 1.95 Peak area (mAu*s) 2839.1968 4197.0324

[0049] As shown in Table 1-2, Figure 1 The peak with a retention time of 1.80 min was 4-aminoindane. The purity of 4-aminoindane was 99.9%, and the separation time was 1.95 min.

[0050] Example 2

[0051] The chromatograph used in this embodiment is a supercritical fluid chromatograph from Jiangsu Hanbang Technology Co., Ltd., with a column size of 10*250mm, a mobile phase flow rate of 8mL / min, a back pressure of 12MPa, a aminoindene sample concentration of 6.44mg / mL, an injection volume of 20μL, and a detection wavelength of 254nm.

[0052] Table 2-1 lists the other relevant conditions used in the method of separating and preparing 4-aminoindenhydride using a supercritical fluid chromatography system in Example 2.

[0053] Table 2-1 SFC separation conditions in Example 2

[0054]

[0055] The specific steps corresponding to Example 2 are as follows: The aminoindane sample was dissolved in methanol, and sonicated to prepare a solution of 6.44 mg / mL. The solution was then filtered through a 0.45 μm nylon membrane. A Nucifera MSU 5 μm 10*250 mm (silica gel surface-bonded urea group) column was selected as the chromatographic column on a supercritical fluid chromatograph from Jiangsu Hanbang Technology Co., Ltd. The volume ratio of supercritical carbon dioxide to methanol, total flow rate, back pressure, and column temperature in the mobile phase were controlled according to Table 2-1, while ensuring that the temperature of the carbon dioxide entering the pump was 6℃. The aminoindane sample was then separated, and the separation effect was as follows: Figure 2 As shown, the separation effect is good.

[0056] Table 2-2 Figure 2 Retention time and peak area of ​​4-aminoindane and 5-aminoindane

[0057] 4-Aminoindane 5-Aminoindane Retention time (min) 2.94 3.33 Peak area (mAu*s) 2374.4805 4119.2114

[0058] As shown in Table 2-2, Figure 2 The peak with a retention time of 2.94 min was 4-aminoindane. The purity of 4-aminoindane was 99.6%, and the separation time was 3.33 min.

[0059] Example 3

[0060] The chromatograph used in this embodiment is a supercritical fluid chromatograph from Jiangsu Hanbang Technology Co., Ltd., with a column size of 10*250mm, a mobile phase flow rate of 8mL / min, a back pressure of 12MPa, a concentration of aminoindene in the sample of 79.43mg / mL, an injection volume of 200μL, and a detection wavelength of 254nm.

[0061] Table 3-1 lists the other relevant conditions used in the method of separating and preparing 4-aminoindenhydride using a supercritical fluid chromatography system in Example 3.

[0062] Table 3-1 SFC separation conditions in Example 3

[0063]

[0064] The specific steps corresponding to Example 3 are as follows: The aminoindane sample was dissolved in methanol, and sonicated to prepare a solution of 79.43 mg / mL. The solution was then filtered through a 0.45 μm nylon membrane. A Nucifera Imu 5 μm 10*250 mm (silica gel surface-bonded imidazole group) column was selected as the chromatographic column on a supercritical fluid chromatograph from Jiangsu Hanbang Technology Co., Ltd. The volume ratio of supercritical carbon dioxide to methanol, total flow rate, back pressure, and column temperature in the mobile phase were controlled according to Table 3-1, while ensuring that the temperature of the carbon dioxide entering the pump was 6℃. The aminoindane sample was then separated, and the separation effect was as follows: Figure 3 As shown, the separation effect is good.

[0065] Table 3-2 Figure 3 Retention time and peak area of ​​4-aminoindane and 5-aminoindane

[0066] 4-Aminoindane 5-Aminoindane Retention time (min) 5.16 7.60 Peak area (mAu*s) 34809.1385 59425.5867

[0067] As shown in Table 3-2 Figure 3 The peak with a retention time of 5.16 min was 4-aminoindane. The purity of 4-aminoindane was 99.5%, and the separation time was 7.60 min.

[0068] Example 4

[0069] The chromatograph used in this embodiment is a supercritical fluid chromatograph from Jiangsu Hanbang Technology Co., Ltd., with a column size of 10*250mm, a mobile phase flow rate of 8mL / min, a back pressure of 12MPa, a aminoindene sample concentration of 273mg / mL, an injection volume of 10μL, and a detection wavelength of 254nm.

[0070] Table 4-1 lists the other relevant conditions used in the method of separating and preparing 4-aminoindenhydride using a supercritical fluid chromatography system in Example 4.

[0071] Table 4-1 SFC separation conditions in Example 4

[0072]

[0073] The specific steps corresponding to Example 4 are as follows: The aminoindane sample was dissolved in methanol, sonicated to prepare a solution of 273 mg / mL, and filtered through a 0.45 μm nylon membrane. A Nucifera Imu 5 μm 10*250 mm (silica gel surface-bonded imidazole group) column was selected as the chromatographic column on a supercritical fluid chromatograph from Jiangsu Hanbang Technology Co., Ltd. The volume ratio of supercritical carbon dioxide to ethanol, total flow rate, back pressure, and column temperature in the mobile phase were controlled according to Table 4-1, while ensuring that the temperature of carbon dioxide entering the pump was 6℃. The aminoindane sample was separated, and the separation effect was as follows: Figure 4 As shown, the separation effect is average, with obvious trailing phenomenon.

[0074] Table 4-2 Figure 4 Retention time and peak area of ​​4-aminoindane and 5-aminoindane

[0075] 4-Aminoindane 5-Aminoindane Retention time (min) 5.25 7.83 Peak area (mAu*s) 14129.9417 24101.2841

[0076] As shown in Table 4-2 Figure 4 The 4-aminoindane was retained for 5.25 min, with a purity of 99.5% and a separation time of 7.83 min.

[0077] Example 5

[0078] The chromatograph used in this embodiment is a supercritical fluid chromatograph from Jiangsu Hanbang Technology Co., Ltd., with a column size of 10*250mm, a mobile phase flow rate of 8mL / min, a back pressure of 8MPa, a concentration of 273mg / mL for the aminoindene sample, an injection volume of 6μL, and a detection wavelength of 254nm.

[0079] Table 5-1 lists the other relevant conditions used in the method of separating and preparing 4-aminoindenhydride using a supercritical fluid chromatography system in Example 5.

[0080] Table 5-1 SFC separation conditions in Example 5

[0081]

[0082] The specific steps corresponding to Example 5 are as follows: The aminoindane sample was dissolved in methanol, sonicated to prepare a solution of 273 mg / mL, and filtered through a 0.45 μm nylon membrane. A Nucifera Imu 5 μm 10*250 mm (silica gel surface-bonded imidazole group) column was selected as the chromatographic column on a supercritical fluid chromatograph from Jiangsu Hanbang Technology Co., Ltd. The volume ratio of supercritical carbon dioxide to methanol, total flow rate, back pressure, and column temperature in the mobile phase were controlled according to Table 5-1, while ensuring that the temperature of carbon dioxide entering the pump was 6℃. The aminoindane sample was separated, and the separation effect was as follows: Figure 5 As shown, the separation effect is average, with obvious trailing phenomenon.

[0083] Table 5-2 Figure 5 Retention time and peak area of ​​4-aminoindane and 5-aminoindane

[0084] 4-Aminoindane 5-Aminoindane Retention time (min) 5.11 7.26 Peak area (mAu*s) 9511.3267 16799.5693

[0085] As shown in Table 5-2 Figure 5The retention time was 5.11 min for 4-aminoindane, and the purity of 4-aminoindane was 99.6%, with a separation time of 7.26 min.

[0086] Example 6

[0087] The chromatograph used in this embodiment is a supercritical fluid chromatograph from Jiangsu Hanbang Technology Co., Ltd., with a column size of 10*250mm, a mobile phase flow rate of 8mL / min, a back pressure of 12MPa, a aminoindene sample concentration of 273mg / mL, an injection volume of 6μL, and a detection wavelength of 254nm.

[0088] Table 6-1 lists the other relevant conditions used in the method of separating and preparing 4-aminoindenhydrin using a supercritical fluid chromatography system in Example 6.

[0089] Table 6-1 SFC separation conditions in Example 6

[0090]

[0091]

[0092] The specific steps corresponding to Example 6 are as follows: The aminoindane sample was dissolved in methanol, sonicated to prepare a solution of 273 mg / mL, and filtered through a 0.45 μm nylon membrane. A Nucifera Imu 5 μm 10*250 mm (silica gel surface-bonded imidazole group) column was selected as the chromatographic column on a supercritical fluid chromatograph from Jiangsu Hanbang Technology Co., Ltd. The volume ratio of supercritical carbon dioxide to methanol, total flow rate, back pressure, and column temperature in the mobile phase were controlled according to Table 6-1, while ensuring that the temperature of carbon dioxide entering the pump was 6℃. The aminoindane sample was separated, and the separation effect was as follows: Figure 6 As shown, the separation effect is good.

[0093] Table 6-2 Figure 6 Retention time and peak area of ​​4-aminoindane and 5-aminoindane

[0094] 4-Aminoindane 5-Aminoindane Retention time (min) 3.28 4.11 Peak area (mAu*s) 9593.5934 12368.0620

[0095] As shown in Table 6-2 Figure 6 The retention time was 3.28 min for 4-aminoindane, the purity of the collected 4-aminoindane was 99.4%, and the separation time was 4.11 min.

[0096] Example 7

[0097] The chromatograph used in this embodiment is a supercritical fluid chromatograph from Jiangsu Hanbang Technology Co., Ltd., with a column size of 10*250mm, a mobile phase flow rate of 8mL / min, a back pressure of 12MPa, a aminoindene sample concentration of 273mg / mL, an injection volume of 6μL, and a detection wavelength of 254nm.

[0098] Table 7-1 lists the other relevant conditions used in the method of separating and preparing 4-aminoindenhydride using a supercritical fluid chromatography system in Example 7.

[0099] Table 7-1 SFC separation conditions in Example 7

[0100]

[0101] The specific steps corresponding to Example 7 are as follows: The aminoindane sample was dissolved in methanol, sonicated to prepare a solution of 273 mg / mL, and filtered through a 0.45 μm nylon membrane. A Nucifera Imu 5 μm 10*250 mm (silica gel surface-bonded imidazole group) column was selected as the chromatographic column on a supercritical fluid chromatograph from Jiangsu Hanbang Technology Co., Ltd. The volume ratio of supercritical carbon dioxide to methanol, total flow rate, back pressure, and column temperature in the mobile phase were controlled according to Table 7-1, while ensuring that the temperature of carbon dioxide entering the pump was 6℃. The aminoindane sample was separated, and the separation effect was as follows: Figure 7 As shown, the separation effect is poor and the tailing phenomenon is severe.

[0102] Table 7-2 Figure 7 Retention time and peak area of ​​4-aminoindane and 5-aminoindane

[0103] 4-Aminoindane 5-Aminoindane Retention time (min) 4.85 6.27 Peak area (mAu*s) 5837.0356 7351.7949

[0104] As shown in Table 7-2 Figure 7 The retention time was 4.85 min for 4-aminoindane, and the purity of 4-aminoindane was 99.1%, with a separation time of 6.27 min.

[0105] Example 8

[0106] The chromatograph used in this embodiment is a supercritical fluid chromatograph from Jiangsu Hanbang Technology Co., Ltd., with a column size of 10*250mm, a mobile phase flow rate of 8mL / min, a back pressure of 12MPa, a aminoindene sample concentration of 273mg / mL, an injection volume of 6μL, and a detection wavelength of 254nm.

[0107] Table 8-1 lists the other relevant conditions used in the method of separating and preparing 4-aminoindenhydride using a supercritical fluid chromatography system in Example 8.

[0108] Table 8-1 SFC separation conditions in Example 8

[0109]

[0110] The specific steps corresponding to Example 8 are as follows: The aminoindane sample was dissolved in methanol, sonicated to prepare a solution of 273 mg / mL, and filtered through a 0.45 μm nylon membrane. A Nucifera MSU 5 μm 10*250 mm (silica gel surface-bonded urea group) column was selected as the chromatographic column on a supercritical fluid chromatograph from Jiangsu Hanbang Technology Co., Ltd. The volume ratio of supercritical carbon dioxide to ethanol, total flow rate, back pressure, and column temperature in the mobile phase were controlled according to Table 8-1, while ensuring that the temperature of carbon dioxide entering the pump was 6℃. The aminoindane sample was separated, and the separation effect was as follows: Figure 8 As shown, the separation effect is good, but there is a trailing phenomenon.

[0111] Table 8-2 Figure 8 Retention time and peak area of ​​4-aminoindane and 5-aminoindane

[0112] 4-Aminoindane 5-Aminoindane Retention time (min) 4.16 5.68 Peak area (mAu*s) 13348.1080 30553.8583

[0113] As shown in Table 8-2 Figure 8 The retention time was 4.16 min for 4-aminoindane, and the purity of 4-aminoindane was 99.6%, with a separation time of 5.68 min.

[0114] Example 9

[0115] The chromatograph used in this embodiment is a supercritical fluid chromatograph from Jiangsu Hanbang Technology Co., Ltd., with a column size of 10*250mm, a mobile phase flow rate of 8mL / min, a back pressure of 8MPa, a concentration of 273mg / mL for the aminoindene sample, an injection volume of 6μL, and a detection wavelength of 254nm.

[0116] Table 9-1 lists the other relevant conditions used in the method of separating and preparing 4-aminoindenhydride using a supercritical fluid chromatography system in Example 9.

[0117] Table 9-1 SFC separation conditions in Example 9

[0118]

[0119] The specific steps corresponding to Example 9 are as follows: The aminoindane sample was dissolved in methanol, sonicated to prepare a solution of 273 mg / mL, and filtered through a 0.45 μm nylon membrane. A Nucifera MSU 5 μm 10*250 mm (silica gel surface-bonded urea group) column was selected as the chromatographic column on a supercritical fluid chromatograph from Jiangsu Hanbang Technology Co., Ltd. The volume ratio of supercritical carbon dioxide to methanol, total flow rate, back pressure, and column temperature in the mobile phase were controlled according to Table 9-1, while ensuring that the temperature of carbon dioxide entering the pump was 6°C. The aminoindane sample was separated, and the separation effect was as follows: Figure 9 As shown, the separation effect is good.

[0120] Table 9-2 Figure 9 Retention time and peak area of ​​4-aminoindane and 5-aminoindane

[0121] 4-Aminoindane 5-Aminoindane Retention time (min) 3.83 5.07 Peak area (mAu*s) 15005.2967 32589.0822

[0122] As shown in Table 9-2 Figure 9 The retention time was 3.83 min for 4-aminoindane, and the purity of 4-aminoindane was 99.1%, with a separation time of 5.07 min.

[0123] Example 10

[0124] The chromatograph used in this embodiment is a supercritical fluid chromatograph from Jiangsu Hanbang Technology Co., Ltd., with a column size of 10*250mm, a mobile phase flow rate of 8mL / min, a back pressure of 12MPa, a aminoindene sample concentration of 273mg / mL, an injection volume of 6μL, and a detection wavelength of 254nm.

[0125] Table 10-1 lists other relevant conditions used in the method for separating and preparing 4-aminoindenhydride using a supercritical fluid chromatography system in Example 10.

[0126] Table 10-1 SFC separation conditions in Example 10

[0127]

[0128] The specific steps corresponding to Example 10 are as follows: The aminoindane sample was dissolved in methanol, sonicated to prepare a solution of 273 mg / mL, and filtered through a 0.45 μm nylon membrane. A Nucifera MSU 5 μm 10*250 mm (silica gel surface-bonded urea group) column was selected as the chromatographic column on a supercritical fluid chromatograph from Jiangsu Hanbang Technology Co., Ltd. The volume ratio of supercritical carbon dioxide to methanol, total flow rate, back pressure, and column temperature in the mobile phase were controlled according to Table 10-1, while ensuring that the temperature of carbon dioxide entering the pump was 6℃. The aminoindane sample was separated, and the separation effect was as follows: Figure 10 As shown, the separation effect is good, but there is a trailing phenomenon.

[0129] Table 10-2 Figure 10 Retention time and peak area of ​​4-aminoindane and 5-aminoindane

[0130] 4-Aminoindane 5-Aminoindane Retention time (min) 2.73 3.22 Peak area (mAu*s) 12120.7262 22630.0171

[0131] As shown in Table 10-2, Figure 10 The retention time was 2.73 min for 4-aminoindane, and the purity of 4-aminoindane was 99.2%, with a separation time of 3.22 min.

[0132] Example 11

[0133] The chromatograph used in this embodiment is a supercritical fluid chromatograph from Jiangsu Hanbang Technology Co., Ltd., with a column size of 10*250mm, a mobile phase flow rate of 8mL / min, a back pressure of 12MPa, a aminoindene sample concentration of 273mg / mL, an injection volume of 6μL, and a detection wavelength of 254nm.

[0134] Table 11-1 lists other relevant conditions used in the method of separating and preparing 4-aminoindenhydride using a supercritical fluid chromatography system in Example 11.

[0135] Table 11-1 SFC separation conditions in Example 11

[0136]

[0137]

[0138] The specific steps corresponding to Example 11 are as follows: The aminoindane sample was dissolved in methanol, sonicated to prepare a solution of 273 mg / mL, and filtered through a 0.45 μm nylon membrane. A Nucifera MSU 5 μm 10*250 mm (silica gel surface-bonded urea group) column was selected as the chromatographic column on a supercritical fluid chromatograph from Jiangsu Hanbang Technology Co., Ltd. The volume ratio of supercritical carbon dioxide to methanol, total flow rate, back pressure, and column temperature in the mobile phase were controlled according to Table 11-1, while ensuring that the temperature of carbon dioxide entering the pump was 6℃. The aminoindane sample was separated, and the separation effect was as follows: Figure 11 As shown, the separation effect is good, but there is obvious tailing phenomenon.

[0139] Table 11-2 Figure 11 Retention time and peak area of ​​4-aminoindane and 5-aminoindane

[0140] 4-Aminoindane 5-Aminoindane Retention time (min) 3.98 4.81 Peak area (mAu*s) 11567.7326 25988.3172

[0141] As shown in Table 11-2, Figure 11 The retention time was 3.98 min for 4-aminoindane, and the purity of 4-aminoindane was 99.5%, with a separation time of 4.81 min.

[0142] Comparative Example 1

[0143] The chromatograph used in this comparative example was a supercritical fluid chromatograph from Jiangsu Hanbang Technology Co., Ltd., with a column size of 4.6*250mm, a mobile phase flow rate of 2mL / min, a back pressure of 12MPa, a aminoindene sample concentration of 3mg / mL, an injection volume of 4μL, and a detection wavelength of 254nm.

[0144] Table 12 shows the other relevant conditions used in the method of separating and preparing 4-aminoindenhydrin using a supercritical fluid chromatography system in Comparative Example 1.

[0145] Table 12 SFC separation conditions in Comparative Example 1

[0146]

[0147] The specific steps for Comparative Example 1 were as follows: The aminoindane sample was dissolved in methanol, sonicated to prepare a 3 mg / mL solution, and filtered through a 0.45 μm nylon membrane. A Nucifera C18P 5 μm 4.6*250 mm (silica gel surface bonded with octadecyl groups) column was selected as the supercritical fluid chromatograph used by Jiangsu Hanbang Technology Co., Ltd. The volume ratio of supercritical carbon dioxide to methanol, total flow rate, back pressure, and column temperature in the mobile phase were controlled according to Table 12, while ensuring that the temperature of carbon dioxide entering the pump was 6 °C. The aminoindane sample was separated, and the separation effect was as follows: Figure 12 As shown, the sample has no resolution and is not retained on the chromatographic column.

[0148] Comparative Example 2

[0149] The chromatograph used in this comparative example was a supercritical fluid chromatograph from Jiangsu Hanbang Technology Co., Ltd., with a Hedera Phenyl 5um 10*250mm column, a mobile phase flow rate of 8mL / min, a back pressure of 12MPa, an aminoindene sample concentration of 273mg / mL, an injection volume of 2μL, and a detection wavelength of 254nm.

[0150] Table 13 shows the other relevant conditions used in the method of separating and preparing 4-aminoindenhydrin using a supercritical fluid chromatography system in Comparative Example 2.

[0151] Table 13 SFC separation conditions in Comparative Example 2

[0152]

[0153]

[0154] The specific steps for Comparative Example 2 were as follows: The aminoindane sample was dissolved in methanol, sonicated to prepare a solution of 273 mg / mL, and filtered through a 0.45 μm nylon membrane. A Hedera Phenyl 5 μm 10*250 mm (silica gel surface-bonded phenyl) column was used as the column in a supercritical fluid chromatograph manufactured by Jiangsu Hanbang Technology Co., Ltd. The volume ratio of supercritical carbon dioxide to methanol, total flow rate, back pressure, and column temperature in the mobile phase were controlled according to Table 13, while ensuring that the temperature of carbon dioxide entering the pump was 6℃. The aminoindane sample was then separated, and the separation effect was as follows: Figure 13 As shown, there is no separation effect.

[0155] Comparative Example 3

[0156] The chromatograph used in this comparative example was a supercritical fluid chromatograph from Jiangsu Hanbang Technology Co., Ltd., with a column size of 10*250mm, a mobile phase flow rate of 8mL / min, a back pressure of 12MPa, a aminoindene sample concentration of 273mg / mL, an injection volume of 4μL, and a detection wavelength of 254nm.

[0157] Table 14-1 shows the other relevant conditions used in the method of separating and preparing 4-aminoindenhydrin using a supercritical fluid chromatography system in Comparative Example 3.

[0158] Table 14-1 SFC separation conditions in Comparative Example 3

[0159]

[0160] The specific steps for Comparative Example 3 were as follows: The aminoindane sample was dissolved in methanol, sonicated to prepare a solution of 273 mg / mL, and filtered through a 0.45 μm nylon membrane. A Hedera Phenyl 5 μm 10*250 mm (silica gel surface-bonded phenyl) column was used as the column in a supercritical fluid chromatograph manufactured by Jiangsu Hanbang Technology Co., Ltd. The volume ratio of supercritical carbon dioxide to ethanol, total flow rate, back pressure, and column temperature in the mobile phase were controlled according to Table 14-1, while ensuring that the temperature of carbon dioxide entering the pump was 6℃. The aminoindane sample was then separated, and the separation effect was as follows: Figure 14 As shown, the separation effect is average, with obvious trailing phenomenon.

[0161] Table 14-2 Figure 14 Retention time and peak area of ​​4-aminoindane and 5-aminoindane

[0162] 4-Aminoindane 5-Aminoindane Retention time (min) 2.35 2.57 Peak area (mAu*s) 4893.4285 9996.4721

[0163] As shown in Table 14-2 Figure 14 The retention time of 4-aminoindane was 2.35 min, and the purity of 4-aminoindane was 98.8%, with a separation time of 2.57 min.

[0164] Comparative Example 4

[0165] The chromatograph used in this comparative example was a supercritical fluid chromatograph from Jiangsu Hanbang Technology Co., Ltd., with a column size of 10*250mm, a mobile phase flow rate of 8mL / min, a back pressure of 8MPa, a concentration of 273mg / mL for the aminoindene sample, an injection volume of 2μL, and a detection wavelength of 254nm.

[0166] Table 15-1 shows the other relevant conditions used in the method of separating and preparing 4-aminoindenman using a supercritical fluid chromatography system in Comparative Example 4.

[0167] Table 15-1 SFC separation conditions in Comparative Example 4

[0168]

[0169] The specific steps for Comparative Example 4 were as follows: The aminoindane sample was dissolved in methanol, sonicated to prepare a solution of 273 mg / mL, and filtered through a 0.45 μm nylon membrane. A Hedera Phenyl 5 μm 10*250 mm (silica gel surface-bonded phenyl) column was used as the column in a supercritical fluid chromatograph manufactured by Jiangsu Hanbang Technology Co., Ltd. The volume ratio of supercritical carbon dioxide to methanol, total flow rate, back pressure, and column temperature in the mobile phase were controlled according to Table 15-1, while ensuring that the temperature of carbon dioxide entering the pump was 6℃. The aminoindane sample was then separated, and the separation effect was as follows: Figure 15 As shown, the separation effect is average, with obvious trailing phenomenon.

[0170] Table 15-2 Figure 15 Retention time and peak area of ​​4-aminoindane and 5-aminoindane

[0171] 4-Aminoindane 5-Aminoindane Retention time (min) 2.38 2.58 Peak area (mAu*s) 4830.0629 9094.4257

[0172] As shown in Table 15-2, Figure 15 The peak with a retention time of 2.38 min was 4-aminoindane. The purity of 4-aminoindane collected and detected was 98.6%, and the separation time was 2.58 min.

[0173] Comparative Example 5

[0174] The chromatograph used in this comparative example was a supercritical fluid chromatograph from Jiangsu Hanbang Technology Co., Ltd., with a column size of 10*250mm, a mobile phase flow rate of 8mL / min, a back pressure of 12MPa, a aminoindene sample concentration of 273mg / mL, an injection volume of 2μL, and a detection wavelength of 254nm.

[0175] Table 16-1 shows the other relevant conditions used in the method of separating and preparing 4-aminoindenman using a supercritical fluid chromatography system in Comparative Example 5.

[0176] Table 16-1 SFC separation conditions in Comparative Example 5

[0177]

[0178] The specific steps for Comparative Example 5 were as follows: The aminoindane sample was dissolved in methanol, sonicated to prepare a solution of 273 mg / mL, and filtered through a 0.45 μm nylon membrane. A Hedera Phenyl 5 μm 10*250 mm (silica gel surface-bonded phenyl) column was used as the column in a supercritical fluid chromatograph manufactured by Jiangsu Hanbang Technology Co., Ltd. The volume ratio of supercritical carbon dioxide to methanol, total flow rate, back pressure, and column temperature in the mobile phase were controlled according to Table 16-1, while ensuring that the temperature of carbon dioxide entering the pump was 6℃. The aminoindane sample was then separated, and the separation effect was as follows: Figure 16 As shown, the separation effect is average, with obvious trailing phenomenon.

[0179] Table 16-2 Figure 16 Retention time and peak area of ​​4-aminoindane and 5-aminoindane

[0180] 4-Aminoindane 5-Aminoindane Retention time (min) 2.39 2.55 Peak area (mAu*s) 3571.4881 7574.3402

[0181] As shown in Table 16-2 Figure 16 The retention time was 2.39 min for 4-aminoindane, the purity of which was 98.5% and the separation time was 2.55 min.

[0182] Comparative Example 6

[0183] The chromatograph used in this comparative example was a supercritical fluid chromatograph from Jiangsu Hanbang Technology Co., Ltd., with a column size of 10*250mm, a mobile phase flow rate of 8mL / min, a back pressure of 12MPa, a aminoindene sample concentration of 273mg / mL, an injection volume of 6μL, and a detection wavelength of 254nm.

[0184] Table 17-1 lists other relevant conditions used in the method for separating and preparing 4-aminoindenhydrin using a supercritical fluid chromatography system in Comparative Example 6.

[0185] Table 17-1 SFC separation conditions in Comparative Example 6

[0186]

[0187] The specific steps for Comparative Example 6 were as follows: The aminoindane sample was dissolved in methanol, sonicated to prepare a solution of 273 mg / mL, and filtered through a 0.45 μm nylon membrane. A Nucifera Imu 5 μm 10*250 mm (silica gel surface-bonded imidazole group) column was selected as the supercritical fluid chromatograph at Jiangsu Hanbang Technology Co., Ltd. The volume ratio of supercritical carbon dioxide to methanol, total flow rate, back pressure, and column temperature in the mobile phase were controlled according to Table 17-1, while ensuring that the temperature of carbon dioxide entering the pump was 6℃. The aminoindane sample was then separated, and the separation effect was as follows: Figure 17 As shown, the separation effect is mediocre and impurities cannot be separated.

[0188] Table 17-2 Figure 17 Retention time and peak area of ​​4-aminoindane and 5-aminoindane

[0189] 4-Aminoindane 5-Aminoindane Retention time (min) 2.57 3.01 Peak area (mAu*s) 9371.4346 11092.9277

[0190] As shown in Table 17-2 Figure 17 The retention time was 2.57 min for 4-aminoindane, and the purity of 4-aminoindane was 98.2%, with a separation time of 3.01 min.

[0191] Comparative Example 7

[0192] The chromatograph used in this comparative example was a supercritical fluid chromatograph from Jiangsu Hanbang Technology Co., Ltd., with a column size of 10*250mm, a mobile phase flow rate of 8mL / min, a back pressure of 12MPa, a aminoindene sample concentration of 273mg / mL, an injection volume of 6μL, and a detection wavelength of 254nm.

[0193] Table 18-1 shows the other relevant conditions used in the method for separating and preparing 4-aminoindenhydrin using a supercritical fluid chromatography system in Comparative Example 7.

[0194] Table 18-1 SFC separation conditions in Comparative Example 7

[0195]

[0196] The specific steps for Comparative Example 7 were as follows: The aminoindane sample was dissolved in methanol, sonicated to prepare a solution of 273 mg / mL, and filtered through a 0.45 μm nylon membrane. A Nucifera MSU 5 μm 10*250 mm (silica gel surface-bonded urea group) column was selected as the supercritical fluid chromatograph used by Jiangsu Hanbang Technology Co., Ltd. The volume ratio of supercritical carbon dioxide to methanol, total flow rate, back pressure, and column temperature in the mobile phase were controlled according to Table 18-1, while ensuring that the temperature of carbon dioxide entering the pump was 6℃. The aminoindane sample was then separated, and the separation effect was as follows: Figure 18 As shown, the separation effect is poor and complete separation is not possible.

[0197] Table 18-2 Figure 18 Retention time and peak area of ​​4-aminoindane and 5-aminoindane

[0198] 4-Aminoindane 5-Aminoindane Retention time (min) 2.25 2.51 Peak area (mAu*s) 12326.0839 20981.2110

[0199] As shown in Table 18-2 Figure 18 The retention time of 4-aminoindane was 2.25 min, the purity of 4-aminoindane was 97.8%, and the separation time was 2.51 min.

[0200] Comparative Example 8

[0201] The chromatograph used in this embodiment is a high-performance liquid chromatograph from Huapu Scientific Instruments Technology Co., Ltd., with a column size of 4.6*250mm, a mobile phase flow rate of 1mL / min, an aminoindene sample concentration of 79.3mg / mL, an injection volume of 5μL, and a detection wavelength of 254nm.

[0202] Table 19-1 lists other relevant conditions used in the method for separating and preparing 4-aminoindenymium using a high-performance liquid chromatography system in Comparative Example 8.

[0203] Table 19-1 HPLC separation conditions in Comparative Example 8

[0204]

[0205] The specific steps for Comparative Example 8 were as follows: The aminoindane sample was dissolved in methanol, sonicated to prepare a solution of 79.3 mg / mL, and filtered through a 0.45 μm nylon membrane. A Nucifera C18A5um 4.6*250 mm (silica gel surface bonded with octadecyl groups) column was selected as the chromatographic column on a high-performance liquid chromatograph (HPLC) system manufactured by Huapu Scientific Technology Co., Ltd. The volume ratio of acetonitrile to 0.05 M sodium dihydrogen phosphate aqueous solution, the total flow rate, and the column temperature were controlled according to Table 19-1. The aminoindane was then separated, and the separation effect was as follows: Figure 19As shown, the sample has a certain degree of separation but poor peak shape.

[0206] Table 19-2 Figure 19 Retention time and peak area of ​​4-aminoindane and 5-aminoindane

[0207] 4-Aminoindane 5-Aminoindane Retention time (min) 7.807 14.120 Peak area (mAu*s) 24545.92 26592.81

[0208] As shown in Table 19-2 Figure 19 The peak with a retention time of 14.120 min is 4-aminoindane. The purity of 4-aminoindane collected and detected is 99.9%. In the high-performance liquid chromatograph of Huapu Scientific Instruments Technology Co., Ltd., it takes 15 minutes for all substances to be completely separated, while in the supercritical fluid chromatography system, all impurities are separated in at least 8 minutes, which shows the short separation time of supercritical fluid chromatography.

Claims

1. A method for separating and preparing 4-aminoindan using a supercritical fluid chromatography system, characterized by, The method comprises the following steps: Dissolving an aminoindane sample, separating the aminoindane sample on a chromatographic column by using a supercritical fluid chromatography system to prepare 4-aminoindane; a mobile phase of the supercritical fluid chromatography system is a mixture of supercritical CO2 and a lower alcohol; a stationary phase of the supercritical fluid chromatography system comprises one or more of silica gel surface-bonded phenyl or silica gel surface-bonded urea or silica gel surface-bonded imidazole.

2. The method of claim 1, wherein, The back pressure of the supercritical fluid chromatography system is 8-12 MPa.

3. The method of claim 1, wherein, The volume ratio of the supercritical CO2 to the lower alcohol is 90-99:1-10.

4. The method of claim 1, wherein, The lower alcohol comprises methanol or ethanol.

5. The method of claim 1, wherein, The aminoindane sample comprises a mixture of 4-aminoindane and 5-aminoindane, and the concentration of the aminoindane sample is 3-273 mg / mL.

6. The method of claim 1, wherein, The aminoindane sample comprises a mixture of 4-aminoindane and 5-aminoindane, and the concentration of the aminoindane sample is 3-79.43 mg / mL.

7. The method of claim 1, wherein, The temperature of the chromatographic column of the supercritical fluid chromatography system is 30-45 DEG C.

8. The method of claim 1, wherein, The chromatographic column of the supercritical fluid chromatography system is Hedera Phenyl 5um 4.6*250mm or Nucifera MSU 5um 10*250mm or Nucifera Imu 5um 10*250mm.

9. The method of claim 1, wherein, The flow rate of the mobile phase is 2-8 mL / min.

10. The method of claim 1, wherein, The injection volume of the supercritical fluid chromatography system is 6-200 mu L.

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