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Determination method of content of metformin HCL (hydrochloride)

A technology of metformin hydrochloride and a determination method, which is applied in the directions of measuring devices, instruments, scientific instruments, etc., can solve problems such as the detection of metformin that has not been seen, and achieve the effects of improving bioavailability, high purity and strong activity

Inactive Publication Date: 2015-07-15
SHANGHAI JIAOTONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] At present, there is no detection of metformin in the mitochondria, and there are related literature reports on whether there will be metformin in the mitochondria

Method used

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  • Determination method of content of metformin HCL (hydrochloride)
  • Determination method of content of metformin HCL (hydrochloride)
  • Determination method of content of metformin HCL (hydrochloride)

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0075] Preparation of solid lipid nanoparticles

[0076] Take 300 μL of distilled water to form the inner aqueous phase. Weigh 50 mg of glyceryl monostearate and 15 mg of soybean lecithin, dissolve them in a mixed solvent of 0.5 mL of ethyl acetate and 0.5 mL of dichloromethane, fully dissolve, and form an organic phase, and prepare a concentration of 1.0% polyvinyl alcohol (PVA) as outer aqueous phase. The inner aqueous phase was added dropwise to the organic phase in an ultrasonic ice bath (power 100W, ultrasonic 0.5min), and then the colostrum was added dropwise into the outer aqueous phase with a syringe, and the probe was ultrasonically emulsified (power 100W, ultrasonic 3min) to obtain W / O / W complex milk. The organic solvent was removed by rotary evaporation to obtain the metformin hydrochloride lipid nanoparticle emulsion with blue opalescence. Centrifuge the nanoparticle emulsion at 12000rpm / min for 30min, take the precipitate, redissolve the precipitate with ultra...

Embodiment 2

[0094] Determination of Metformin Hydrochloride Content in and Out of Mitochondria

[0095] 1. Mitochondrial Sample Preparation

[0096] 1) Cell inoculation: select MCF-7 human breast cancer cells in the logarithmic growth phase 10 7 Each plate was cultured in RPMI-1640 medium containing 10% fetal bovine serum, and cultured in an incubator at 37°C, 5% CO 2 ;

[0097] 2) Preparation of sample solution: prepare metformin hydrochloride solutions with a concentration of 0.4mM, 2mM, and 10mM with the medium, filter and sterilize through a 0.22μm microporous membrane, and prepare and use immediately;

[0098] 3) Cell administration: After staying overnight, discard the extracellular fluid, add metformin hydrochloride solution with a concentration of 0.4mM, 2mM, and 10mM prepared with 1640 medium, and a blank control group, and stop culturing after 4 hours of drug action, 1000r / min Centrifuge for 10 minutes, discard the supernatant

[0099] 4) Digest the cells: wash three times w...

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Abstract

The invention relates to a detection method for metformin inside / outside cells and inside and outside mitochondria, aiming at detecting drug concentration of the metformin in cancer cells, detecting whether metformin can enter cancer cell mitochondria, dynamically observing variation conditions of drug concentrations inside and outside the cells and inside and outside the mitochondria, and verifying whether the metformin can enter the cancer cells and the mitochondria of the cancer cells. A determination method provided by the invention has great significances on studying a dynamic variation rule among the drug concentration, physiological parameters of the cell and environment physicochemical property, and lays certain foundation for studying a mechanism of the anti-tumor effect of the metformin.

Description

technical field [0001] The invention belongs to the field of measuring the content of metformin hydrochloride, in particular to a method for measuring the content of metformin hydrochloride inside / outside cells and mitochondria. Background technique [0002] The preeminence of metformin in the treatment of type 2 diabetes has attracted the attention of the world. In recent years, its indications have been further extended with the continuous development of various researches. The latest research shows that metformin can inhibit the proliferation of breast cancer cells, pancreatic cancer cells, melanoma cells and prostate cancer cells cultured in vitro. [0003] However, what is even more striking is the observation in clinical studies that metformin has a certain degree of protective effect on cancer patients. Patients using metformin have a reduced risk of breast cancer, liver cancer and other tumors, and tumor-related mortality is also reduced. This phenomenon has recentl...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N30/88
Inventor 沈琦易朝丽朱涛
Owner SHANGHAI JIAOTONG UNIV
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