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Determination method of content of 1-deoxynojirimycin in silkworm larvae

The technology of deoxynojirimycin and determination method is applied to the field of determination of 1-deoxynojirimycin content in silkworm larvae, can solve problems such as inability to meet high-level demands, achieve positive social and economic benefits, and high reproducibility , stable and good effect

Inactive Publication Date: 2013-05-08
ZHEJIANG UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are multiple (4) chiral centers in the DNJ molecule, and there are as many as 15 or more stereoisomers related to it, and its derivatives synthesized by artificial chemistry are quite different from natural DNJ in terms of pharmacological effects and toxic and side effects. Differences, unable to meet the high-level needs of today's society

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0016] Take silkworm larva body powder, add 0.05 mol / L HCl at a ratio of 1:700 (W:V), vortex at room temperature for 15 s and sonicate for 30 min, then centrifuge at 21690 g for 20 min, collect the supernatant and precipitate Extract again according to the above steps, combine the two supernatants, and constant volume to obtain the test solution. Take 10 μL of the test solution, mix it with 10 μL of 0.4 mol / L potassium borate buffer (pH8.5), add 20 μL of 5 mmol / L FMOC-Cl solution, mix quickly and react in a water bath at 20°C for 20 min , add 10 μL 0.1mol / L glycine solution to neutralize the remaining FMOC-Cl to terminate the reaction. The reaction solution was diluted with 950 μL of 0.1% (V / V) acetic acid solution to stabilize the generated DNJ-FMOC, and filtered through a nylon straw filter with a pore size of 0.2 μm to obtain the measurement solution. Inject 10 μL of the measuring solution into the chromatographic column with a sample injector into the DIAMONSIC C 18 The ...

Embodiment 2

[0018] Take silkworm larvae body powder, add 0.05 mol / L HCl at a ratio of 1:800 (W:V), vortex at room temperature for 20 s and sonicate for 25 min, then centrifuge at 21690 g for 20 min, collect the supernatant and sediment Extract again according to the above steps, combine the two supernatants, and constant volume to obtain the test solution. Take 10 μL of the test solution, mix it with 10 μL of 0.4 mol / L potassium borate buffer (pH8.5), add 20 μL of 5 mmol / L FMOC-Cl solution, mix quickly and react in a water bath at 20°C for 25 min , add 10 μL 0.1mol / L glycine solution to neutralize the remaining FMOC-Cl to terminate the reaction. The reaction solution was diluted with 950 μL of 0.1% (V / V) acetic acid solution to stabilize the generated DNJ-FMOC, and filtered through a nylon straw filter with a pore size of 0.2 μm to obtain the measurement solution. Inject 10 μL of the measurement solution into the chromatographic column with a sample injector as the DIAMONSIC C 18 The co...

Embodiment 3

[0020] Take silkworm larva body powder, add 0.05 mol / L HCl at a ratio of 1:750 (W:V), vortex at room temperature for 18 s and sonicate for 20 min, then centrifuge at 21690 g for 20 min, collect the supernatant and precipitate Extract again according to the above steps, combine the two supernatants, and constant volume to obtain the test solution. Take 10 μL of the test solution, mix it with 10 μL of 0.4 mol / L potassium borate buffer (pH8.5), add 20 μL of 5 mmol / L FMOC-Cl solution, mix quickly and react in a water bath at 20°C for 23 min , add 10 μL 0.1mol / L glycine solution to neutralize the remaining FMOC-Cl to terminate the reaction. The reaction solution was diluted with 950 μL of 0.1% (V / V) acetic acid solution to stabilize the generated DNJ-FMOC, and filtered through a nylon straw filter with a pore size of 0.2 μm to obtain the measurement solution. Inject 10 μL of the measurement solution into the chromatographic column with a sample injector as the DIAMONSIC C 18 The ...

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Abstract

The invention discloses a determination method of the content of 1-deoxynojirimycin in silkworm larvae. The determination method comprises the following steps of: taking 0.05mol / L HCl as a solvent and swirling for 15-20 seconds at a room temperature; extracting for two times by using a step of ultrasonic treatment for 20-30 minutes to effectively extract DNJ (Deoxynojirimycin) from the silkworm larvae; marking the DNJ for 20-25 minutes at 20 DEG C by utilizing 5mmol / L 9-fluorenyl methylclhlorofonmate (FMOC-C1) to form a DNJ-FMOC complex with the fluorescence; and detecting by utilizing a reverse phase high performance liquid chromatography provided with a fluorescence detector and calculating the content of the DNJ according to a peak area. The determination method of the content of the DNJ in the silkworm larvae provided with the invention has the advantages of high precision, repeatability and high sample recovery rate and good stability, can be used for detecting the content of the DNJ in a lot of silkworm larva products, and has active social benefits and economic benefits for propelling pharmaceutical development of the silkworm larvae.

Description

technical field [0001] The invention relates to a method for determining the content of 1-deoxynojirimycin in silkworm larvae. Background technique [0002] Diabetes mellitus is a common multiple chronic disease of endocrine and metabolic disorders, and its main clinical symptoms are hyperglycemia. With the improvement of social and economic conditions and the change of diet structure, the prevalence of diabetes has increased significantly in recent years, and it has become a serious public health problem in today's medicine, which has caused a great impact on the development of the national economy. At present, the drugs used clinically to treat diabetes mainly include insulins, sulfonylureas, biguanides and α-glucosidase inhibitors. Insulin, sulfonylureas and biguanides can cause liver damage, blood sugar Too low, edema, etc., side effects are relatively large, and α-glucosidase inhibitors are the key areas of current development. [0003] 1-deoxynojirimycin (1-deoxynoji...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02
Inventor 时连根张作法刘淑梅金卫明费建明李玉峰赵丽华施国方黄秀娣
Owner ZHEJIANG UNIV
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