A method for determining related substances of 3-butyl-1(3H)-isobenzofuranone by HPLC

Abstract

The invention provides a detection method for determining 3-butyl-1(3H)-isobenzofuranone-related substances by high performance liquid chromatography (HPLC), the method adopts a phenyl chromatographic column, the detection wavelength is 226nm-230nm, the flow rate is 0.8mL / min-1.2mL / min, the column temperature is 25℃-35℃, the mobile phase is composed of phase A and phase B, phase A is a mixed solution of methanol and acetonitrile with a volume ratio of 50 / 50-40 / 60, 0.01%-0.05% acetic acid is added, and phase B is 0.03%-0.07% acetic acid aqueous solution. The detection method provided by the invention has good specificity, sensitivity, separation degree of each absorption peak, accuracy and stability.

Description

Technical Field

[0001] The invention belongs to the field of drug analysis, and in particular relates to a high performance liquid chromatography (HPLC) detection method for 3-butyl-1(3H)-isobenzofuranone raw material medicine or related substances of its preparation. Background Art

[0002] 3-Butyl-1(3H)-isobenzofuranone is a drug independently researched and developed in my country for the treatment of mild and moderate acute ischemic stroke. In recent years, with the gradual development of science and technology, people have a deeper understanding of the risks of drugs and a stronger awareness of risk control. Domestic and foreign drug review and approval agencies have gradually increased their requirements for drug safety. Therefore, fully recognizing the risks of drugs and taking effective measures to control risks are the key to current drug research and development and production. The first risk that should be controlled during the production and storage of 3-butyl-1(3H)-isobenzofuranone is impurities, including process impurities and degradation impurities. Therefore, developing an effective analytical method suitable for detecting and controlling 3-butyl-1(3H)-isobenzofuranone impurities is crucial to controlling product quality and ensuring product safety.

[0003] This type of compound is unstable and is easily affected by factors such as light, temperature, acid, and alkali, causing changes and mutual transformation. It has the characteristics of many by-products and poor stability. The main process impurities of 3-butyl-1(3H)-isobenzofuranone are similar to its main structure and have similar chemical properties, which makes it difficult to separate and detect. In the patents and literatures reported so far, only some impurities have been studied, such as starting materials, intermediates, alkaline degradation impurities, oxidation impurities, and some impurities introduced by the starting materials. 2-propyl-3-hydroxy-1-indanone, 3-butylisoindolin-1-one and o-pentylbenzoic acid are degradation impurities that are easily produced in the production process. They are difficult to remove in the subsequent refining, resulting in a high risk of residues in the finished product. 3-isopropyl-1(3H)-isobenzofuranone 3-propyl-1(3H)-isobenzofuranone is an impurity in the starting material. During the production process, side reactions will occur and the impurities will be introduced into the finished product. These impurities are difficult to control and remove in the production process of 3-butyl-1(3H)-isobenzofuranone. Effective detection in the finished product should be guaranteed to control product quality. However, the inventors did not find these impurities mentioned in the prior art during their research. This may be because the impurities in the product were not characterized during the research, their sources were not traced, no control was performed, and there was a lack of effective separation methods for separating these impurities.

[0004] For example, Chinese patent application CN201610395891.4 discloses a method for simultaneously determining related substances. This method solves the problem that phthalic acid, intermediate I [2-(α-carbonylpentyl)benzoic acid], intermediate II [sodium 2-(α-hydroxypentyl)benzoate], 3-propyl-1(3H)-isobenzofuranone, 3-butylene-1(3H)-isobenzofuranone, and dibutylphthalide impurities are difficult to separate from the main component, but the detection method described in the patent application does not include the separation of 2-benzofuranone and 3-n-butyl-3-hydroxy-1(3H)-isobenzofuranone, and cannot separate and determine 2-benzofuranone, 3-n-butyl-3-hydroxy-1(3H)-isobenzofuranone and other derivative impurities. Chinese patent application CN201910466352.9 discloses a method for related substances. The method solves the problem that 2-benzofuranone and 3-n-butyl-3-hydroxy-1(3H)-isobenzofuranone cannot be separated in the prior art, and at the same time increases verification of good separation effect on impurities such as o-carboxybenzyl alcohol, o-carboxybenzaldehyde, 3-methyl-1(3H)-isobenzofuranone, 3-ethyl-1(3H)-isobenzofuranone, 3-pentyl-1(3H)-isobenzofuranone, 3-isobutyl-1(3H)-isobenzofuranone, 3-sec-butyl-1(3H)-isobenzofuranone, 3-tert-butyl-1(3H)-isobenzofuranone, 3-n-butyl-3-hydroxy-1(3H)-isobenzofuranone and 3-butyl-1(3H)-isobenzofuranone. However, the patent application does not cover the key process impurities in the production process, such as 2-propyl-3-hydroxy-1-indanone, o-pentylbenzoic acid, 3-butylisoindolin-1-one and 3-isopropyl-1(3H)-isobenzofuranone 3-propyl-1(3H)-isobenzofuranone. In addition, the inventors used the method of the patent application to investigate the above impurities and found that the method was poor in separating 3-isopropyl-1(3H)-isobenzofuranone 3-propyl-1(3H)-isobenzofuranone from 3-butylisoindolin-1-one, 3-isobutyl-1(3H)-isobenzofuranone from 3-butyl-1(3H)-isobenzofuranone, with separation degrees of 0.65 and 1.38, respectively, which could not meet the requirements of accurate detection of impurities and quality control (see Comparative Example 1 below for specific results).

[0005] It can be seen that the analytical methods currently reported for detecting related substances in 3-butyl-1(3H)-isobenzofuranone APIs and their preparations are mostly HPLC methods, but the impurities studied by these methods cannot cover the main process impurities and degradation impurities of 3-butyl-1(3H)-isobenzofuranone, and different chromatographic systems have certain limitations in the separation and detection of impurities, which cannot meet the current pharmaceutical industry's requirements for product quality control. Summary of the invention

[0006] In order to solve the shortcomings of the existing analytical methods for detecting impurities in 3-butyl-1(3H)-isobenzofuranone raw materials and preparations thereof, the present invention develops an HPLC analytical method that can more comprehensively separate and detect 3-butyl-1(3H)-isobenzofuranone impurities, thereby solving the technical problem that the existing technology cannot effectively separate impurities.

[0007] Specifically, the present invention is realized through the following technical solutions:

[0008] The invention provides a detection method for determining 3-butyl-1(3H)-isobenzofuranone-related substances by using HPLC. The method adopts a phenyl chromatographic column, a detector is an ultraviolet detector, a detection wavelength is 226nm-230nm, a flow rate is 0.8mL / min-1.2mL / min, a column temperature is 25°C-35°C, a mobile phase is composed of a phase A and a phase B, the phase A is a mixed solution of methanol and acetonitrile in a volume ratio of 50 / 50-40 / 60, 0.01%-0.05% acetic acid is added, and the phase B is a 0.03%-0.07% acetic acid aqueous solution;

[0009] The related substances are selected from one or more of the following: 3-methyl-1(3H)-isobenzofuranone, 2-propyl-3-hydroxy-1-indanone, potassium 2-(α-hydroxypentyl)benzoate, 3-ethyl-1(3H)-isobenzofuranone, 2-(α-carbonylpentyl)benzoic acid, 3-isopropyl-1(3H)-isobenzofuranone, 3-propyl-1(3H)-isobenzofuranone, 3-butylisoindolin-1-one, 3-propyl-1(3H)-isobenzofuranone, 3-n-butyl-3-hydroxy-1(3H)-isobenzofuranone, 3-isobutyl-1(3H)-isobenzofuranone, 3-pentyl-1(3H)-isobenzofuranone and o-pentylbenzoic acid;

[0010] The linear gradient elution conditions were as follows:

[0011]

[0012] Preferably, the 3-butyl-1(3H)-isobenzofuranone is a bulk drug or a preparation thereof, and more preferably, the 3-butyl-1(3H)-isobenzofuranone preparation is a soft capsule or an injection.

[0013] As an optional manner, in the detection method, the amount of acetic acid added to mobile phase A is 0.02%-0.04%, and the amount of acetic acid added to mobile phase B is 0.04%-0.06%.

[0014] As an optional manner, in the detection method, the amount of acetic acid added to mobile phase A is 0.03%, and the amount of acetic acid added to mobile phase B is 0.05%.

[0015] As an option, in the detection method, the ratio of methanol to acetonitrile in mobile phase A is 55 / 45.

[0016] As an alternative, in the detection method, the linear gradient elution conditions are as follows:

[0017]

[0018] As an alternative, in the detection method, the linear gradient elution conditions are as follows:

[0019]

[0020] As an optional manner, in the detection method, the column temperature is 30°C, the flow rate is 0.9mL / min-1.1mL / min, preferably 1.0mL / min, the injection volume is 10-50μL, preferably 10μL, and the detection wavelength is 228nm.

[0021] As an optional manner, in the detection method, the column length of the phenyl chromatographic column is 4.6×250 mm, and the filler particle size is 5 μm. Preferably, the phenyl chromatographic column is an Ultimate XB-Phenyl column.

[0022] As an optional embodiment, in the detection method, the related substances are composed of: 3-methyl-1(3H)-isobenzofuranone, 2-propyl-3-hydroxy-1-indanone, potassium 2-(α-hydroxypentyl)benzoate, 3-ethyl-1(3H)-isobenzofuranone, 2-(α-carbonylpentyl)benzoic acid, 3-isopropyl-1(3H)-isobenzofuranone, 3-propyl-1(3H)-isobenzofuranone, 3-butylisoindolin-1-one, 3-propyl-1(3H)-isobenzofuranone, 3-n-butyl-3-hydroxy-1(3H)-isobenzofuranone, 3-isobutyl-1(3H)-isobenzofuranone, 3-pentyl-1(3H)-isobenzofuranone and o-pentylbenzoic acid.

[0023] As an optional embodiment, in the detection method, the separation degree of 3-isopropyl-1(3H)-isobenzofuranone, 3-propyl-1(3H)-isobenzofuranone and 3-butylisoindolin-1-one, and the separation degree of 3-isobutyl-1(3H)-isobenzofuranone and 3-butyl-1(3H)-isobenzofuranone are both greater than 1.5.

[0024] Compared with the prior art, the present invention has the following beneficial effects:

[0025] (1) The present invention develops an HPLC analysis method that can more comprehensively separate and detect 3-butyl-1(3H)-isobenzofuranone impurities, solving the technical problem that the current existing technology cannot effectively separate the key process impurities of 3-butyl-1(3H)-isobenzofuranone, 2-propyl-3-hydroxy-1-indanone, o-pentylbenzoic acid, 3-butylisoindolin-1-one and 3-isopropyl-1(3H)-isobenzofuranone 3-propyl-1(3H)-isobenzofuranone.

[0026] (2) The detection method of the present invention has the advantages of good specificity, high sensitivity, good repeatability, etc., and can be used for simultaneous monitoring of multiple related substances during the industrial production, preparation and storage of 3-butyl-1(3H)-isobenzofuranone. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] Figure 1 : HPLC spectrum of impurities of the present invention detected by the method of Example 1 in patent CN201910466352.9.

[0028] Figure 2 : HPLC spectrum of impurity separation in the method of embodiment 1 of the present invention.

[0029] Figure 3 : HPLC spectrum of impurity separation in the method of embodiment 2 of the present invention.

[0030] Figure 4 : HPLC spectrum of impurity separation in the method of embodiment 3 of the present invention.

[0031] Figure 5 : HPLC spectrum of impurity separation in the method of embodiment 4 of the present invention.

[0032] Figure 6 : HPLC spectrum of impurity separation in the method of embodiment 5 of the present invention.

[0033] Figure 7 : HPLC spectrum of impurity separation in the method of embodiment 6 of the present invention.

[0034] Figure 8 : HPLC spectrum of impurity separation in the method of embodiment 7 of the present invention.

[0035] Fig. 9 : HPLC spectrum of impurity separation in the method of Example 8 of the present invention.

[0036] Fig.10 : HPLC spectrum of impurity separation in the method of embodiment 9 of the present invention. DETAILED DESCRIPTION

[0037] The present invention will be further described with reference to specific embodiments. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not intended to limit the scope of the present invention.

[0038] If no specific techniques or conditions are specified in the examples, the techniques or conditions described in the literature in the field or the product instructions are used. If no manufacturer is specified for the reagents or instruments used, they are all conventional products that can be purchased through regular channels.

[0039] The experimental methods in the following examples are conventional methods unless otherwise specified. The experimental materials used in the following examples are commercially available products unless otherwise specified.

[0040] Unless otherwise specified, when the present invention relates to the percentage between liquids, the percentage is volume / volume percentage; when the present invention relates to the percentage between liquids and solids, the percentage is volume / weight percentage; when the present invention relates to the percentage between solids and liquids, the percentage is weight / volume percentage; the rest are weight / weight percentages.

[0041] Comparative Example 1: Using the method of Example 1 in patent CN201910466352.9

[0042] Chromatographic column: Agilent ZORBAX plus Pheny-Hexyl, 4.6mm×250mm, 5μm

[0043] Mobile phase: Phase A is 0.1% acetic acid in water (volume ratio); Phase B is acetonitrile:methanol (1:1, volume ratio)

[0044] Detection wavelengths were 227 nm and 280 nm, flow rate was 1.0 mL / min, and column temperature was 30 °C.

[0045] The gradient elution conditions are shown in Table 1 below:

[0046] Time (minutes) 0 5 40 50 50.1 60 A％ 62 62 45 30 62 62 B％ 38 38 55 70 38 38

[0047] Solution preparation and determination: 3-methyl-1(3H)-isobenzofuranone Z01, 3-ethyl-1(3H)-isobenzofuranone Z02, 3-propyl-1(3H)-isobenzofuranone Z03, Z04, 3-isopropyl-1(3H)-isobenzofuranone 3-propyl-1(3H)-isobenzofuranone Z05, 3-isobutyl-1(3H)-isobenzofuranone Z06, 3-butylisoindolin-1-one (Z07), 2-propyl-3-hydroxy-1-indanone (Z08), 3-isopropyl-1(3H)-isobenzofuranone Z09, 3-isobutyl-1(3H)-isobenzofuranone Z100, 3-isobutyl-1(3H)-isobenzofuranone Z111, 3-isobutyl-1(3H)-isobenzofuranone Z121, 3-isobutyl-1(3H)-isobenzofuranone Z132, 3-isobutyl-1(3H)-isobenzofuranone Z143, 3-isobutyl-1(3H)-isobenzofuranone Z154, 3-isobutyl-1(3H)-isobenzofuranone Z165, 3-isobutyl-1(3H)-isobenzofuranone Z170, 3-isobutyl-1(3H)-isobenzofuranone Z181, 3-isobutyl-1(3H)-isobenzofuranone Z192, 3-isobutyl-1(3H)-isobenzofuranone Z113, 3-isobutyl-1(3H)-isobenzofuranone Z114 09), o-pentylbenzoic acid (Z10), 3-n-butyl-3-hydroxy-1(3H)-isobenzofuranone (Z11), 2-(α-carbonylpentyl)benzoic acid (Z12), potassium 2-(α-hydroxypentyl)benzoate (Z14), and 3-butyl-1(3H)-isobenzofuranone were accurately weighed to prepare a mixed sample solution containing 50 μg / mL of each impurity and 1 mg / mL of 3-butyl-1(3H)-isobenzofuranone. The results were as shown in Tables 2 and 3. Figure 1 As shown:

[0048]

[0049] The results showed that the method was poorly effective in separating 3-isopropyl-1(3H)-isobenzofuranone, 3-propyl-1(3H)-isobenzofuranone and 3-butylisoindolin-1-one, and 3-isobutyl-1(3H)-isobenzofuranone and 3-butyl-1(3H)-isobenzofuranone, with separation degrees of 0.65 and 1.38, respectively, which could not meet the requirements of accurate impurity detection and quality control.

[0050] Embodiment 1:

[0051] The impurities in Comparative Example 1 were determined by HPLC.

[0052] Column: Ultimate XB-Phenyl, 4.6×250mm, 5μm

[0053] Mobile phase: A: methanol / acetonitrile (55 / 45) + 0.03% acetic acid; B: 0.05% acetic acid in water Flow rate: 1.0 mL / min; Wavelength 228 nm; Column temperature 30°C

[0054] The gradient elution conditions are shown in Table 3 below:

[0055]

[0056] The results are shown in Table 4 and Figure 2 As shown:

[0057]

[0058] It can be seen from the analysis results that when the ratio of methanol to acetonitrile in phase A is 1.2:1 and the initial ratio is 40%, each component can achieve a good separation effect.

[0059] Embodiment 2:

[0060] The impurities in Comparative Example 1 were determined by HPLC.

[0061] Column: Ultimate XB-Phenyl, 4.6×250mm, 5μm

[0062] Mobile phase: A: methanol / acetonitrile (55 / 45) + 0.03% acetic acid; B: 0.03% acetic acid aqueous solution

[0063] Flow rate: 1.0 mL / min; wavelength 228 nm; column temperature 30 °C

[0064] The gradient elution conditions are shown in Table 5 below:

[0065]

[0066] The results are shown in Table 6 and Figure 3 As shown:

[0067]

[0068] It can be seen from the analysis results that when the proportion of acetic acid in phase B is 0.03%, a better separation effect can be achieved.

[0069] Embodiment 3:

[0070] The impurities in Comparative Example 1 were determined by HPLC.

[0071] Column: Ultimate XB-Phenyl, 4.6×250mm, 5μm

[0072] Mobile phase: A: methanol / acetonitrile (55 / 45) + 0.02% acetic acid; B: 0.03% acetic acid in water

[0073] Flow rate: 1.0 mL / min; wavelength 228 nm; column temperature 30 °C

[0074] The gradient elution conditions are shown in Table 7 below:

[0075]

[0076] The results are shown in Table 8 and Figure 4 As shown:

[0077]

[0078] It can be seen from the analysis results that when the proportion of acetic acid in phase A is 0.02%, a better separation effect can be achieved.

[0079] Embodiment 4:

[0080] The impurities in Comparative Example 1 were determined by HPLC.

[0081] Column: Ultimate XB-Phenyl, 4.6×250mm, 5μm

[0082] Mobile phase: A: methanol / acetonitrile (55 / 45) + 0.01% acetic acid; B: 0.04% acetic acid in water

[0083] Flow rate: 1.0 mL / min; wavelength 228 nm; column temperature 30 °C

[0084] The gradient elution conditions are shown in Table 9 below:

[0085]

[0086] The results are shown in Table 10 and Figure 5 As shown:

[0087]

[0088] It can be seen from the analysis results that when the proportion of acetic acid in phase A is 0.01%, a better separation effect can be achieved.

[0089] Embodiment 5:

[0090] The impurities in Comparative Example 1 were determined by HPLC.

[0091] Column: Ultimate XB-Phenyl, 4.6×250mm, 5μm

[0092] Mobile phase: A: methanol / acetonitrile (55 / 45) + 0.05% acetic acid; B: 0.07% acetic acid in water

[0093] Flow rate: 1.0 mL / min; wavelength 228 nm; column temperature 30 °C

[0094] The gradient elution conditions are shown in Table 11 below:

[0095]

[0096]

[0097] The results are shown in Table 12 and Figure 6 As shown:

[0098]

[0099] The analysis results show that a better separation effect can be achieved when the acetic acid ratio of phase A is 0.05% and that of phase B is 0.07%.

[0100] Embodiment 6:

[0101] The impurities in Comparative Example 1 were determined by HPLC.

[0102] Column: Ultimate XB-Phenyl, 4.6×250mm, 5μm

[0103] Mobile phase: A: methanol / acetonitrile (50 / 50) + 0.03% acetic acid; B: 0.05% acetic acid in water

[0104] Flow rate: 1.0 mL / min; wavelength 228 nm; column temperature 30 °C

[0105] The gradient elution conditions are shown in Table 13 below:

[0106]

[0107]

[0108] The results are shown in Table 14 and Figure 7 As shown:

[0109]

[0110] It can be seen from the analysis results that when the ratio of methanol to acetonitrile in phase A is 1:1, a better separation effect can be achieved.

[0111] Embodiment 7:

[0112] The impurities in Comparative Example 1 were determined by HPLC.

[0113] Column: Ultimate XB-Phenyl, 4.6×250mm, 5μm

[0114] Mobile phase: A: methanol / acetonitrile (55 / 45); B: 0.02% acetic acid in water

[0115] Flow rate: 1.0 mL / min; wavelength 228 nm; column temperature 30 °C

[0116] The gradient elution conditions are shown in Table 15 below:

[0117]

[0118]

[0119] The results are shown in Table 16 and Figure 8 As shown:

[0120]

[0121] It can be seen from the results that when the acetic acid ratio in phase A is lower than 0.01%, the separation effect of potassium 2-(α-hydroxypentyl)benzoate and 3-ethyl-1(3H)-isobenzofuranone becomes poor and baseline separation cannot be achieved. Therefore, the acetic acid ratio in the mobile phase cannot be too small.

[0122] Embodiment 8:

[0123] The impurities in Comparative Example 1 were determined by HPLC.

[0124] Column: Ultimate XB-Phenyl, 4.6×250mm, 5μm

[0125] Mobile phase: A: methanol / acetonitrile (60 / 40) + 0.03% acetic acid; B: 0.05% acetic acid aqueous solution

[0126] Flow rate: 1.0 mL / min; wavelength 228 nm; column temperature 30 °C

[0127] The gradient elution conditions are shown in Table 17 below:

[0128]

[0129] The results are shown in Table 18 and Fig. 9 As shown:

[0130]

[0131] It can be seen from the results that when the initial ratio is adjusted to 45%, potassium 2-(α-hydroxypentyl)benzoate and 3-ethyl-1(3H)-isobenzofuranone overlap and cannot be separated, which shows that the initial ratio cannot be higher than 40%.

[0132] Embodiment 9:

[0133] The impurities in Comparative Example 1 were determined by HPLC.

[0134] Column: Ultimate XB-Phenyl, 4.6×250mm, 5μm

[0135] Mobile phase: A: methanol / acetonitrile (60 / 40) + 0.05% acetic acid; B: 0.12% acetic acid in water

[0136] Flow rate: 1.0 mL / min; wavelength 228 nm; column temperature 30 °C

[0137] The gradient elution conditions are shown in Table 19 below:

[0138]

[0139] The results are shown in Table 20 and Fig.10 As shown:

[0140]

[0141]

[0142] It can be seen from the results that as the proportion of acetic acid in phase B is increased, the separation of 2-(α-carbonylpentyl)benzoic acid and 3-isopropyl-1(3H)-isobenzofuranone and 3-propyl-1(3H)-isobenzofuranone becomes worse, and 3-butylisoindolin-1-one and 3-propyl-1(3H)-isobenzofuranone overlap, and the impurity control requirements cannot be met.

[0143] Based on the research results of the above examples, the detection method for determining the related substances of 3-butyl-1(3H)-isobenzofuranone developed by our company can separate and determine the process impurities and degradation impurities of 3-butyl-1(3H)-isobenzofuranone. At the same time, we have carried out methodological verification of this method, and the verification method is as follows:

[0144] Chromatographic conditions: phenylsilane bonded silica gel was used as filler; 0.03% acetic acid-methanol / acetonitrile (55 / 45) was used as mobile phase A, 0.05% acetic acid aqueous solution was used as mobile phase B, gradient elution was performed according to the table below, column temperature was 30°C, flow rate was 1.0 ml / min, detection wavelength was 228 nm, and injection volume was 20 μl.

[0145]

[0146] The validation results of the analytical method showed that the method had strong specificity, high accuracy, good solution stability, and its linear range and sensitivity could meet the requirements for the determination of butylphthalide impurities.

[0147] Obviously, those skilled in the art can make various changes and modifications to the present invention without departing from the spirit and scope of the present invention. Thus, if these modifications and variations of the present invention fall within the scope of the claims of the present invention and their equivalents, the present invention is also intended to include these modifications and variations.

Claims

1. A method for determining related substances of 3-butyl-1(3H)-isobenzofuranone by HPLC, characterized in that: The method adopts a phenyl chromatographic column, a detector is an ultraviolet detector, a detection wavelength of 226 nm-230 nm, a flow rate of 0.8 mL / min-1.2 mL / min, a column temperature of 25°C-35°C, and a mobile phase composed of a phase A and a phase B, wherein the phase A is a mixed solution of methanol and acetonitrile in a volume ratio of 50 / 50-40 / 60 with 0.01%-0.05% acetic acid added, and the phase B is a 0.03%-0.07% acetic acid aqueous solution; The related substances are selected from one or more of the following: 3-methyl-1(3H)-isobenzofuranone, 2-propyl-3-hydroxy-1-indanone, potassium 2-(α-hydroxypentyl)benzoate, 3-ethyl-1(3H)-isobenzofuranone, 2-(α-carbonylpentyl)benzoic acid, 3-isopropyl-1(3H)-isobenzofuranone, 3-propyl-1(3H)-isobenzofuranone, 3-butylisoindolin-1-one, 3-propyl-1(3H)-isobenzofuranone, 3-n-butyl-3-hydroxy-1(3H)-isobenzofuranone, 3-isobutyl-1(3H)-isobenzofuranone, 3-pentyl-1(3H)-isobenzofuranone and o-pentylbenzoic acid; The linear gradient elution conditions were as follows: 。 2. The detection method according to claim 1, characterized in that: The 3-butyl-1(3H)-isobenzofuranone is a 3-butyl-1(3H)-isobenzofuranone raw material drug or a preparation thereof.

3. The detection method according to claim 2, characterized in that: The preparation is a soft capsule or an injection.

4. The detection method according to claim 1, characterized in that: The amount of acetic acid added to mobile phase A is 0.02%-0.04%, and the amount of acetic acid added to mobile phase B is 0.04%-0.06%.

5. The detection method according to claim 1, characterized in that: The amount of acetic acid added to mobile phase A was 0.03%, and the amount of acetic acid added to mobile phase B was 0.05%.

6. The detection method according to claim 1, characterized in that: The ratio of methanol to acetonitrile in mobile phase A was 55 / 45.

7. The detection method according to claim 1, characterized in that: The linear gradient elution conditions were as follows: 。 8. The detection method according to claim 1, characterized in that: The linear gradient elution conditions were as follows: 。 9. The detection method according to claim 1, characterized in that: The column temperature is 30° C., the flow rate is 0.9 mL / min-1.1 mL / min, the injection volume is 10-50 μL, and the detection wavelength is 228 nm.

10. The detection method according to claim 9, characterized in that: The flow rate was 1.0 mL / min, and the injection volume was 10 μL.

11. The detection method according to claim 1, characterized in that: The column length of the phenyl chromatographic column is 4.6×250 mm, and the filler particle size is 5 μm.

12. The detection method according to claim 11, characterized in that: The phenyl chromatographic column is an Ultimate XB-Phenyl column.

13. The detection method according to claim 1, characterized in that: The related substances are composed of: 3-methyl-1(3H)-isobenzofuranone, 2-propyl-3-hydroxy-1-indanone, potassium 2-(α-hydroxypentyl)benzoate, 3-ethyl-1(3H)-isobenzofuranone, 2-(α-carbonylpentyl)benzoic acid, 3-isopropyl-1(3H)-isobenzofuranone, 3-propyl-1(3H)-isobenzofuranone, 3-butylisoindolin-1-one, 3-propyl-1(3H)-isobenzofuranone, 3-n-butyl-3-hydroxy-1(3H)-isobenzofuranone, 3-isobutyl-1(3H)-isobenzofuranone, 3-pentyl-1(3H)-isobenzofuranone and o-pentylbenzoic acid.

14. The detection method according to claim 1, characterized in that: The resolution of 3-isopropyl-1(3H)-isobenzofuranone, 3-propyl-1(3H)-isobenzofuranone and 3-butylisoindolin-1-one, and that of 3-isobutyl-1(3H)-isobenzofuranone and 3-butyl-1(3H)-isobenzofuranone were all greater than 1.5.

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