Pioglitazone detection device and application thereof

A technology of pioglitazone and detection device, which is applied in the direction of measurement device, analysis by chemical reaction of materials, instruments, etc., can solve the problems of poor specificity, expensive equipment, poor sensitivity, etc., and achieves convenient use, economical speed, and production. easy effect

Pending Publication Date: 2019-11-05
广东省食品检验所
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

LC and LC-MS / MS detection have the advantages of high precision, high separation efficiency, good selectivity, low false positive rate, good reproducibility, etc., but the operation is complicated, the equipment is expensive, the operator has high technical requirements, and it cannot be displayed immediately Results and other issues, not suitable for rapid online detection and monitoring
TLC, NIR and RS are easy to operate and fast in analysis, but their sensitivity is poor, there is often interference between multiple components, the specificity is not good, and Chinese patent medicines and complex food matrices are prone to false positives, generally need to use dedicated Further validation with more robust methods

Method used

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  • Pioglitazone detection device and application thereof
  • Pioglitazone detection device and application thereof
  • Pioglitazone detection device and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] The preparation of embodiment 1 pioglitazone hapten

[0044] 3.53 g of pioglitazone, 1.52 g of ethyl bromoacetate and 0.1 g of sodium carbonate were sequentially added into a 100 mL three-necked flask, and reacted overnight in 45 mL of DMF solution at 90°C. Reaction solution is rotary steamed, added water, adjusted to slightly acidic with dilute hydrochloric acid, extracted 2 to 3 times with ethyl acetate, combined organic phase, evaporated to dryness, mixed sample, purified by column to obtain pioglitazone intermediate in Chinese patent medicine and health food .

[0045]Dissolve the pioglitazone intermediate in 10mL of methanol, then add 20mL of 10% sodium hydroxide solution, stir and react at 60°C for 5h, add water, adjust the solution to slightly acidic with dilute hydrochloric acid, extract with ethyl acetate 2 to 3 times, combine the organic phases, Evaporate to dryness, mix the sample, and purify through the column to obtain the pioglitazone hapten.

[0046] Pi...

Embodiment 2

[0048] The preparation of embodiment 2 pioglitazone conjugated antigen

[0049] Dissolve 0.1 mmol of the pioglitazone hapten prepared in Example 1 in 2 mL of DMF, add 0.2 mmol of DCC and 0.15 mmol of NHS with stirring, and react overnight with magnetic stirring at 4°C. After centrifugation, the supernatant is liquid A. Weigh 140 mg of hemocyanin (KLH) and dissolve it in 10 mL of PBS (pH 8.0) with a concentration of 0.1 mol / L, add 1 mL of DMF, stir and dissolve to prepare liquid B, and under magnetic stirring, liquid A is gradually dropped into liquid B, 4 React at ℃ for 12 hours, centrifuge the reaction solution, take the supernatant, dialyze with normal saline at 4℃ for 3 days, change the dialysate 3 times a day, and obtain pioglitazone-conjugated antigen. The obtained conjugated antigen was dispensed into 0.5mL centrifuge tubes at a concentration of 1 mg / mL, and stored frozen in a -20°C refrigerator; the pioglitazone conjugated antigen had the structure shown in formula (II)...

Embodiment 3

[0051] The preparation of embodiment 3 pioglitazone monoclonal antibody

[0052] The pioglitazone-coupled antigen prepared in Example 2 was used to immunize four 6-week-old BALB / C mice. After boosting the immunization three times, blood was collected to measure the titer. immunized mice. Three days later, the mice were killed by decapitation, the spleen was taken under aseptic conditions to prepare splenocytes, mixed with vigorously growing mouse myeloma cells in a 50mL centrifuge tube at a ratio of 8:1, and 30mL serum-free IPMI1640 medium was added, 1100r / Centrifuge for 5 min to discard the supernatant, shake the cell mass gently, and place in a 37°C water bath. Slowly add 1mL of 50% PEG-4000 to the cells, drop it within 1min, and gently stir the sediment at the bottom. After standing for 1min, slowly and uniformly add 1mL of serum-free medium along the tube wall for the first 30s, then add 2mL for the last 30s, and then Quickly add 27mL to terminate the fusion process, ce...

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Abstract

The invention relates to a pioglitazone detection device, and belongs to the technical field of safety detection for traditional Chinese medicine patent medicines and health products. The detection device comprises a test strip and a reaction cup; the test strip comprises a detection line and quality control line, the detection line comprises a pioglitazone coupling antigen, and the quality control line comprises an anti-pioglitazone antibody; and the reaction cup comprises a pioglitazone antibody labeled with colloidal gold. The detection device has the advantages of high sensitivity and specificity and rapid and simple use, can meet requirements for detecting the residual quantity of pioglitazone, and is also characterized by being convenient to use, easy to manufacture, low in cost, andeconomic and rapid.

Description

technical field [0001] The invention belongs to the technical field of safety detection of Chinese patent medicines and health products, and in particular relates to a pioglitazone detection device and its application. Background technique [0002] Thiazolidinedione compounds (TZDs) are a class of commonly used hypoglycemic drugs that can improve insulin resistance and increase the sensitivity of human tissues to insulin. They are clinically used to treat type 2 diabetes and can significantly improve insulin resistance and related metabolism. disorder. Hypoglycemic drugs are prescription drugs and have strict dosages according to their indications, while illegally added hypoglycemic drugs have risks such as dosage, type, drug interaction, and unknown contraindications, and are potentially harmful. With the increasing prevalence of diabetes year by year, propaganda points such as "pure natural", "traditional Chinese medicine conditioning" and "old traditional Chinese medicin...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577G01N33/558G01N33/531G01N21/78
CPCG01N21/78G01N33/531G01N33/558G01N33/577
Inventor 刘海虹雷毅汪廷彩钟海鹰蔡若纯申超群邓皇翼
Owner 广东省食品检验所
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