Detection method of acarbose drug related substances

Abstract

The invention discloses a method for detecting related substances of acarbose drugs. The detection method of the acarbose related substance comprises the following steps: carrying out gradient elution on an acarbose sample to be detected in an amide chromatographic column by adopting a high performance chromatography, a mobile phase is a mixed solution of acetonitrile-ammonium acetate buffer solution; in the mobile phase, the volume ratio of the acetonitrile to the ammonium acetate buffer solution is (76-80): (24-20); in the mobile phase, the concentration of the ammonium acetate buffer solution is 8-12 mM / L; the acarbose to-be-detected product contains acarbose and / or acarbose related substances. According to the detection method, the problem of trailing of main components is solved, the impurity detection capacity is obviously improved, the impurity separation degree is good, and the durability of the chromatographic column is good.

Description

Technical Field

[0001] The present invention relates to a method for detecting acarbose-related substances. Background Art

[0002] The molecular formula of acarbose is C 25 H 43 NO 18 , molecular weight is 645.63, and its structural formula is as follows:

[0003]

[0004] Acarbose is an α-glucosidase inhibitor and a complex oligosaccharide with a structure similar to oligosaccharides. This non-oligosaccharide "pseudo-oligosaccharide" can compete with oligosaccharides at the brush border of cells in the upper small intestine and reversibly bind to α-glucosidase, inhibiting the activity of various α-glucosidases such as maltase, isomaltase, glucoamylase and sucrase, slowing down the decomposition of starch into oligosaccharides such as maltose (disaccharide), maltotriose and dextrin (oligosaccharide) and then into glucose, and slowing down the decomposition of sucrose into glucose and fructose, thus slowing down the absorption of intestinal glucose, thereby alleviating postprandial hyperglycemia and achieving the effect of lowering blood sugar. Long-term use can reduce fasting blood sugar and glycosylated hemoglobin concentrations.

[0005] As a hypoglycemic drug, the quality control of acarbose is particularly important in the production of raw materials. The detection of related substances is a key indicator of drug quality control, such as starting materials, intermediates, isomers, by-products and other substances introduced in the production of raw materials.

[0006] According to literature reports and national drug standards, the analytical methods for acarbose and acarbose tablets include the United States Pharmacopoeia method USP43, the European Pharmacopoeia method EP10.0, and the HPLC method included in the 2020 edition of the Chinese Pharmacopoeia. These three pharmacopoeia methods for HPLC analysis are very similar, as shown in the following table:

[0007]

[0008]

[0009] At present, the detection methods of acarbose all adopt isocratic elution and amino columns. When this method is used to detect related substances, the main peak tailing is relatively serious, which will affect the detection of impurities. In addition, the durability of the chromatographic column is poor. After 200 injections, the column efficiency decreases and the theoretical plate number does not meet the requirements. A new chromatographic column needs to be purchased. Therefore, a new method needs to be developed to detect acarbose and related substances in acarbose tablets. Summary of the invention

[0010] The purpose of the present invention is to solve the problems of serious tailing of the main peak of acarbose, poor impurity detection ability and poor durability of the chromatographic column in the existing detection method of acarbose, and to provide a detection method for acarbose-related substances. The detection method of the present invention solves the problem of tailing of the main component, significantly improves the impurity detection ability, has good impurity separation and good durability of the chromatographic column.

[0011] To achieve the above purpose, the technical solution adopted by the present invention is as follows:

[0012] The present invention provides a method for detecting acarbose-related substances, which comprises the following steps: using high performance liquid chromatography to gradiently elute acarbose to be tested in an amide chromatography column;

[0013] The mobile phase was a mixed solution of acetonitrile-ammonium acetate buffer;

[0014] In the mobile phase, the volume ratio of acetonitrile and ammonium acetate buffer is (76-80):(24-20);

[0015] In the mobile phase, the concentration of ammonium acetate buffer is 8-12 mM / L;

[0016] The acarbose test product contains acarbose and / or acarbose-related substances.

[0017] In the present invention, the pH of the mobile phase may be 8.0-10.0, for example, pH 9.

[0018] In the present invention, the pH of the mobile phase can be adjusted using conventional inorganic bases in the art, such as alkali metal bases (sodium hydroxide).

[0019] In the present invention, the gradient elution can be a two-stage elution.

[0020] Preferably, in the first gradient, the volume ratio of acetonitrile to ammonium acetate buffer is 80:20; in the second gradient, the volume ratio of acetonitrile to ammonium acetate buffer is 76:24.

[0021] Preferably, in the first gradient, the elution time is 35 min, and in the second gradient, the elution time is 25 min (the elution time is the difference between the elution end time and the elution start time of the elution stage).

[0022] In the present invention, the conditions of the gradient elution are preferably:

[0023] Elution stage Elution time Acetonitrile Ammonium acetate buffer First elution stage 35min 80v％ 20v％ Second elution stage 25min 76v％ 24v％

[0024] In the present invention, the amide chromatographic column is selected according to the conventional method in the art. The bonded phase of the amide chromatographic column is a triple-bonded amide-based bonded phase; the length of the amide chromatographic column can be 150-250 mm, such as 150 mm; the inner diameter of the amide chromatographic column can be 1.0-4.6 mm, such as 1.0 mm, 2.1 mm, 3.0 mm or 4.6 mm; the filler particle size in the amide chromatographic column can be 3.5-5 μm, such as 3.5 μm; preferably, the specifications of the amide chromatographic column are: length 150 mm, inner diameter 4.6 mm, filler particle size 3.5 μm.

[0025] In the present invention, the amide chromatography column may be XbridgeAmide, 4.6mm×150mm, 3.5μm, such as the XbridgeAmide chromatography column produced by Waters.

[0026] In the present invention, the column temperature of the amide chromatography column can be 50-60°C, for example 55°C.

[0027] In the present invention, the flow rate of the mobile phase may be 1.0 ml / min-2.0 ml / min according to the conventional practice in the art.

[0028] In the present invention, the injection volume of the HPLC method is conventional in the art and can be 5 μL-10 μL, preferably 5 μL.

[0029] In the present invention, the detection wavelength of the high performance liquid chromatography is conventional in the art, generally 200nm-260nm, preferably 210nm.

[0030] In the present invention, in the detection method, the inner diameter of the amide chromatographic column can be 4.6 mm; the filler particle size of the amide chromatographic column can be 3.5 μm; the length of the amide chromatographic column can be 150-250 mm; the pH of the mobile phase can be 9.0; the mobile phase is a mixed solution of acetonitrile-ammonium acetate buffer; the volume ratio of the acetonitrile and the ammonium acetate buffer can be (76-80): (24-20); the concentration of the ammonium acetate buffer can be 10 mM / L.

[0031] In the present invention, the acarbose test product can be various acarbose products, such as acarbose bulk drug or acarbose preparation.

[0032] In the present invention, the acarbose sample to be tested may be pretreated before testing. Preferably, the pretreatment step comprises the following steps: dissolving the acarbose sample to be tested with a diluent and placing it.

[0033] The diluent may be any diluent solvent that can dissolve the acarbose test product and has no effect on subsequent detection, such as water.

[0034] In the present invention, the acarbose sample to be tested may also contain inorganic impurities.

[0035] In the present invention, the acarbose sample to be tested may further contain residual solvent.

[0036] In the present invention, the acarbose sample to be tested may further contain pharmaceutical excipients.

[0037] In the present invention, the acarbose-related substance is acarbose-related substance in the art, such as one or more of the following structures:

[0038]

[0039] On the basis of being in accordance with the common sense in the art, the above-mentioned preferred conditions can be arbitrarily combined to obtain the preferred embodiments of the present invention.

[0040] The reagents and raw materials used in the present invention are commercially available.

[0041] The positive improvement effect of the present invention is that in the detection method of the present invention, the problem of the tailing factor of the main component is solved, the detection ability of each impurity is significantly improved, the impurity separation degree is good, and after the amide column is used, the column durability is good, and after multiple injections, the column efficiency is still not reduced, and the separation and identification of impurities A, B, C, D, E, F, and G are better achieved. BRIEF DESCRIPTION OF THE DRAWINGS

[0042] Figure 1 This is the liquid chromatogram of the acarbose peak identification solution obtained by analysis in Example 1

[0043] Figure 2 This is the liquid chromatogram of the acarbose peak identification solution obtained by analysis in Comparative Example 1

[0044] Figure 3 This is the liquid chromatogram of the acarbose peak identification solution obtained by analysis in Comparative Example 2

[0045] Figure 4 This is the liquid chromatogram of the acarbose peak identification solution obtained by analysis in Comparative Example 3

[0046] Figure 5 This is the liquid chromatogram of the acarbose peak identification solution obtained by analysis in Comparative Example 4

[0047] Figure 6 This is the liquid chromatogram of the acarbose peak identification solution obtained by analysis in Comparative Example 5

[0048] Figure 7 This is the liquid chromatogram of the acarbose peak identification solution obtained by analysis in Comparative Example 6

[0049] Figure 8This is the liquid chromatogram of the acarbose peak identification solution obtained by analysis in Comparative Example 7

[0050] Fig. 9 This is the liquid chromatogram of the acarbose peak identification solution obtained by analysis in Comparative Example 8

[0051] Fig.10 Solvent blank chromatogram for specificity experiment

[0052] Fig.11 Blank chromatogram of excipients for specificity test DETAILED DESCRIPTION

[0053] The present invention is further described below by way of examples, but the present invention is not limited to the scope of the examples. The experimental methods in the following examples without specifying specific conditions are carried out according to conventional methods and conditions, or selected according to the product specifications.

[0054] The names and structural formulas of the related substances of acarbose in the present invention are shown in the following table

[0055]

[0056] Example 1

[0057] Chromatographic conditions

[0058] Column: Waters Xbridge Amide, 4.6 mm × 150 mm, 3.5 μm

[0059] Mobile phase: Mobile phase A: acetonitrile-buffer (80:20); Mobile phase B: acetonitrile-buffer (76:24); Buffer: 10 mM ammonium acetate buffer; Ammonia test solution was used to adjust the pH to 9.0.

[0060] Column temperature: 55°C

[0061] Flow rate: 1.0ml / min

[0062] Injection volume: 5 μl

[0063] Detection wavelength: 210nm

[0064] Gradient elution program:

[0065]

[0066]

[0067] Preparation of test solution: Take 20 mg of acarbose peak identification reference substance, add 1 ml of purified water to dissolve, and the acarbose peak identification solution is obtained.

[0068] According to the above chromatographic conditions, accurately measure 5 μL of the test solution and inject it into the liquid chromatograph, record the chromatogram, and the test results are as follows: Figure 1 And as shown in Table 1.

[0069] Table 1

[0070]

[0071] Comparative Example 1

[0072] Chromatographic conditions:

[0073] Column: Welch Ultimate XB-NH2 4.6×250mm, 5μm

[0074] Mobile phase: acetonitrile-buffer (75:25), buffer: 0.6 g potassium dihydrogen phosphate and 0.279 g anhydrous disodium hydrogen phosphate, dissolved in water and diluted to 1000 ml, pH 6.8.

[0075] Column temperature: 35°C

[0076] Flow rate: 2ml / min

[0077] Injection volume: 10 μl

[0078] Detection wavelength: 210nm

[0079] Preparation of test solution: Take 20 mg of acarbose peak identification reference substance, add 1 ml of purified water to dissolve, and the acarbose peak identification solution is obtained.

[0080] According to the above chromatographic conditions, accurately measure 10 μL of the test solution and inject it into the liquid chromatograph, perform gradient elution, and record the chromatogram. The test results are as follows: Figure 2 And as shown in Table 2.

[0081] Table 2

[0082]

[0083] Comparative Example 2

[0084] Chromatographic conditions:

[0085] Chromatographic column: Agilent ZorboxNH2 4.6×250mm, 5μm

[0086] Mobile phase: acetonitrile-buffer (75:25), buffer: 0.6 g potassium dihydrogen phosphate and 0.279 g anhydrous disodium hydrogen phosphate, dissolved in water and diluted to 1000 ml, pH 6.8.

[0087] Column temperature: 35°C

[0088] Flow rate: 2ml / min

[0089] Injection volume: 10 μl

[0090] Detection wavelength: 210nm

[0091] Preparation of test solution: Take 20 mg of acarbose peak identification reference substance, add 1 ml of purified water to dissolve, and the acarbose peak identification solution is obtained.

[0092] According to the above chromatographic conditions, accurately measure 10 μL of the test solution and inject it into the liquid chromatograph, isocratically elute, record the chromatogram, and the test results are as follows. Figure 3 And as shown in Table 3.

[0093] Table 3

[0094]

[0095]

[0096] Comparative Example 3

[0097] Chromatographic conditions:

[0098] Chromatographic column: Thermo Fisher Hypersil APS-24.0mm×250mm, 5μm

[0099] Mobile phase: acetonitrile-buffer (75:25), buffer: 0.6 g potassium dihydrogen phosphate and 0.279 g anhydrous disodium hydrogen phosphate, dissolved in water and diluted to 1000 ml, pH 6.8.

[0100] Column temperature: 35°C

[0101] Flow rate: 2ml / min

[0102] Injection volume: 10 μl

[0103] Detection wavelength: 210nm

[0104] Preparation of test solution: Take 20 mg of acarbose peak identification reference substance, add 1 ml of purified water to dissolve, and the acarbose peak identification solution is obtained.

[0105] According to the above chromatographic conditions, accurately measure 10 μL of the test solution and inject it into the liquid chromatograph, isocratically elute, record the chromatogram, and the test results are as follows. Figure 4 And as shown in Table 4.

[0106] Table 4

[0107]

[0108] Comparative Example 4

[0109] Chromatographic conditions:

[0110] Chromatographic column: Waters Spherisorb Amino 4.6mm×150mm, 5μm

[0111] Mobile phase: acetonitrile-buffer (75:25), buffer: 0.6 g potassium dihydrogen phosphate and 0.279 g anhydrous disodium hydrogen phosphate, dissolved in water and diluted to 1000 ml, pH 6.8.

[0112] Column temperature: 35°C

[0113] Flow rate: 1.0ml / min

[0114] Injection volume: 10 μl

[0115] Detection wavelength: 210nm

[0116] Preparation of test solution: Take 20 mg of acarbose peak identification reference substance, add 1 ml of purified water to dissolve, and the acarbose peak identification solution is obtained.

[0117] According to the above chromatographic conditions, accurately measure 10 μL of the test solution and inject it into the liquid chromatograph, isocratically elute, record the chromatogram, and the test results are as follows. Figure 5 And as shown in Table 5.

[0118] Table 5

[0119]

[0120] Comparative Example 5

[0121] Chromatographic conditions:

[0122] Chromatographic column: Fei Nuo Mei LunaNH2 4.6mm×250mm, 5μm

[0123] Mobile phase: acetonitrile-buffer (75:25), buffer: 0.6 g potassium dihydrogen phosphate and 0.279 g anhydrous disodium hydrogen phosphate, dissolved in water and diluted to 1000 ml, pH 6.8.

[0124] Column temperature: 35°C

[0125] Flow rate: 2.0ml / min

[0126] Injection volume: 10 μl

[0127] Detection wavelength: 210nm

[0128] Preparation of test solution: Take 20 mg of acarbose peak identification reference substance, add 1 ml of purified water to dissolve, and the acarbose peak identification solution is obtained.

[0129] According to the above chromatographic conditions, accurately measure 10 μL of the test solution and inject it into the liquid chromatograph, isocratically elute, record the chromatogram, and the test results are as follows. Figure 6 and as shown in Table 6.

[0130] Table 6

[0131]

[0132] Comparative Example 6

[0133] Chromatographic conditions

[0134] Column: Welch Ultimate XB-NH2 4.6×250mm, 5μm

[0135] Mobile phase: acetonitrile-buffer (71:29), buffer: 0.6 g potassium dihydrogen phosphate and 0.279 g anhydrous disodium hydrogen phosphate, dissolve in water and dilute to 1000 ml; adjust pH to 6.0 with 2 mol / L sodium hydroxide solution

[0136] Column temperature: 35°C

[0137] Flow rate: 2.0ml / min

[0138] Injection volume: 10 μl

[0139] Detection wavelength: 210nm

[0140] Preparation of test solution: Take 20 mg of acarbose peak identification reference substance, add 1 ml of purified water to dissolve, and the acarbose peak identification solution is obtained.

[0141] According to the above chromatographic conditions, accurately measure 10 μL of the test solution and inject it into the liquid chromatograph, isocratically elute, record the chromatogram, and the test results are as follows. Figure 7 And as shown in Table 7.

[0142] Table 7

[0143]

[0144] Comparative Example 7

[0145] Chromatographic conditions

[0146] Column: Welch Ultimate XB-NH2 4.6×250mm, 5μm

[0147] Mobile phase: acetonitrile-buffer (71:29), buffer: 0.6 g potassium dihydrogen phosphate and 0.279 g anhydrous disodium hydrogen phosphate, dissolve in water and dilute to 1000 ml; adjust the pH to 7.2 with 2 mol / L sodium hydroxide solution.

[0148] Column temperature: 35°C

[0149] Flow rate: 2.0ml / min

[0150] Injection volume: 10 μl

[0151] Detection wavelength: 210nm

[0152] Preparation of test solution: Take 20 mg of acarbose peak identification reference substance, add 1 ml of purified water to dissolve, and the acarbose peak identification solution is obtained.

[0153] According to the above chromatographic conditions, accurately measure 10 μL of the test solution and inject it into the liquid chromatograph, perform isocratic elution, and record the chromatogram.

[0154] Test results such as Figure 8 and as shown in Table 8.

[0155] Table 8

[0156]

[0157]

[0158] After the pH of the buffer in the mobile phase was adjusted to 7.2, impurity A became a single peak, and the separation degree between impurity A and acarbose was 1.7; the theoretical plate number of acarbose was 3984, which was greater than 2000 and qualified; the separation degrees of the remaining impurities and the main peak were all greater than 2.0, which were qualified. However, impurity C and acarbose could not achieve baseline separation, so the chromatographic column was further investigated to improve the separation degree between impurity C and acarbose.

[0159] Comparative Example 8

[0160] Chromatographic conditions

[0161] Chromatographic column: Waters Xbridge Amide 4.6mm×150mm, 3.5μm

[0162] Mobile phase: acetonitrile-buffer (75:25), buffer: 0.6 g potassium dihydrogen phosphate and 0.279 g anhydrous disodium hydrogen phosphate, dissolve in water and dilute to 1000 ml; adjust the pH to 7.2 with 2 mol / L sodium hydroxide solution.

[0163] Column temperature: 35°C

[0164] Flow rate: 1.0ml / min

[0165] Injection volume: 10 μl

[0166] Detection wavelength: 210nm

[0167] Preparation of test solution: Take 20 mg of acarbose peak identification reference substance, add 1 ml of purified water to dissolve, and the acarbose peak identification solution is obtained.

[0168] According to the above chromatographic conditions, accurately measure 10 μL of the test solution and inject it into the liquid chromatograph, perform isocratic elution, and record the chromatogram.

[0169] Test results such as Fig. 9 and as shown in Table 9.

[0170] Table 9

[0171]

[0172]

[0173] The peak-to-valley ratio of impurity A to acarbose under this method is 1.1, which is less than 2.0 and does not meet the requirements; the peak-to-valley ratio of impurity C to acarbose is 1.1, which is far inferior to the separation effect of the amino column; and due to the solvent effect, the peak shape of impurity D deteriorates; the separation degree of impurity G and impurity F is less than 1.5. In summary: the chromatographic conditions of this method need to be optimized.

[0174] Effect Example: Detection Method Verification of Example 1

[0175] 1.1 Preparation of stock solution

[0176]

[0177] 1.2 Accuracy solution preparation

[0178]

[0179]

[0180] 1.3 Measurement and calculation

[0181] Accurately measure 10 μl of each solution, inject it into the liquid chromatograph, and record the chromatogram. Calculate the recovery rate and RSD of the recovery rate of acarbose and acarbose impurities A, B, C, and D by the main component external standard method.

[0182]

[0183] 1.4 Verification Results

[0184] Results and conclusions of low concentration recovery rate of external standard method

[0185]

[0186] Results and conclusions of high concentration recovery rate of external standard method

[0187]

[0188] Conclusion: The recovery rate of 0.05% concentration should be between 70% and 130%, and the RSD should be no more than 10%; the recovery rate of 0.5% to 5.0% should be between 90% and 110%, and the RSD should be no more than 5%. From the above table results, it can be seen that the accuracy of impurities A, B, C, and D at low and high concentrations is within the range. The RSD of low concentration is less than 10%, and the RSD of high concentration is less than 2.5%, which meets the requirements.

[0189] 2. Limit of Quantitation

[0190] 2.1 Solution preparation

[0191] Each impurity stock solution was diluted to the negligible limit (0.05%) concentration.

[0192] 2.2 Measurement and calculation

[0193] Accurately measure 10 μl of each solution, inject it into the liquid chromatograph, and record the chromatogram. Calculate the S / N of the quantitative limit solution and the detection limit solution.

[0194] 2.3 Verification Results

[0195] Signal-to-noise ratio (S / N) of the quantitative limit solution

[0196] name S / N Impurity A 23 Impurity B 15 Impurity C 25 Impurity D 46

[0197] Conclusion: The signal-to-noise ratio (S / N) of the quantitative limit is not less than 10, which meets the requirements.

[0198] 3. Exclusivity

[0199] 3.1 Solution preparation

[0200] Blank solution: water

[0201] Blank excipient: Take about 1700 mg of blank excipient, accurately weigh, place in a 25 ml volumetric flask, add appropriate amount of diluent, shake for 10 minutes, dilute to the scale with diluent, shake well, centrifuge, and take the supernatant.

[0202] 3.2 Measurement and calculation

[0203] Blank solution and blank excipients do not interfere with the determination;

[0204] 4. Conclusion

[0205]

[0206]

Claims

1. A method for detecting acarbose-related substances, It is characterized in that The method comprises the following steps: using high performance liquid chromatography to gradiently elute acarbose to be tested in an amide chromatography column; The mobile phase was a mixed solution of acetonitrile-ammonium acetate buffer; In the mobile phase, the volume ratio of acetonitrile and ammonium acetate buffer is (76-80):(24-20); In the mobile phase, the concentration of ammonium acetate buffer is 8-12 mM / L; The acarbose test product contains acarbose and / or acarbose-related substances.

2. The method for detecting acarbose-related substances according to claim 1, It is characterized in that The detection method meets one or more of the following conditions: (1) The gradient elution is a two-stage elution; (2) In the mobile phase, the concentration of ammonium acetate buffer is 10 mM / L; and (3) The pH of the mobile phase is 8.0-10.

0.

3. The method for detecting acarbose-related substances according to claim 2, It is characterized in that The detection method meets one or both of the following conditions: (1) In the two-stage elution, in the first gradient, the volume ratio of acetonitrile to ammonium acetate buffer is 80:20; in the second gradient, the volume ratio of acetonitrile to ammonium acetate buffer is 76:24; (2) In the two-stage elution, the elution time in the first gradient is 35 min, and the elution time in the second gradient is 25 min; (3) the pH of the mobile phase is 9.0; and (4) The pH of the mobile phase is adjusted using an alkali metal base.

4. The method for detecting acarbose-related substances according to claim 3, It is characterized in that The conditions of the gradient elution are: And / or, the alkali metal base is sodium hydroxide.

5. The method for detecting acarbose-related substances according to claim 1, It is characterized in that The detection method meets one or more of the following conditions: (1) The bonded phase of the amide chromatographic column is a triple-bonded amide-bonded phase; (2) The length of the amide chromatography column is 150-250 mm; (3) The inner diameter of the amide chromatographic column is 1.0-4.6 mm; (4) The particle size of the filler in the amide chromatographic column is 3.5-5 μm.

6. The method for detecting acarbose-related substances according to claim 5, It is characterized in that The detection method meets one or more of the following conditions: (1) The length of the amide chromatographic column is 150 mm; (2) The inner diameter of the amide chromatography column is 4.6 mm; and (3) The filler particle size in the amide chromatographic column is 3.5 μm.

7. The method for detecting acarbose-related substances according to claim 6, It is characterized in that The detection method meets one or more of the following conditions: (1) The amide chromatography column is XbridgeAmide, 4.6mm×150mm, 3.5μm; (2) The column temperature of the amide chromatography column is 50-60°C; (3) The flow rate of the mobile phase is 1.0 ml / min-2.0 ml / min; (4) The injection volume of the HPLC method is 5 μL-10 μL; (5) The detection wavelength of the high performance liquid chromatography is 200nm-260nm; and (6) The acarbose sample to be tested also contains inorganic impurities, residual solvents or pharmaceutical excipients.

8. The method for detecting acarbose-related substances according to claim 7, It is characterized in that The detection method meets one or more of the following conditions: (1) The column temperature of the amide chromatographic column is 55°C; (2) The flow rate of the mobile phase is 1.0 ml / min; (3) The injection volume of the HPLC method is 5 μL; and (4) The detection wavelength of the high performance liquid chromatography is 210 nm.

9. The method for detecting acarbose-related substances according to claim 8, It is characterized in that The inner diameter of the amide chromatographic column is 4.6 mm; the filler particle size of the amide chromatographic column is 3.5 μm; the length of the amide chromatographic column is 150-250 mm; the pH of the mobile phase is 9.0; the mobile phase is a mixed solution of acetonitrile-ammonium acetate buffer; the volume ratio of the acetonitrile and the ammonium acetate buffer is (76-80):(24-20); the concentration of the ammonium acetate buffer is 10 mM / L.

10. The method for detecting acarbose-related substances according to claim 1, It is characterized in that The detection method meets one or both of the following conditions: (1) The acarbose-related substance is one or more of the following structures: and (2) The acarbose test product is an acarbose bulk drug or an acarbose preparation.

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