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HPLC (High Performance Liquid Chromatography) analysis method for determining content of curcumin and paclitaxel in biological sample

An analytical method and technology for biological samples, applied in the field of HPLC for determining the content of curcumin and paclitaxel in biological samples, can solve the problems of cumbersome sample pretreatment and determination operations, difficulty in realizing combined use of CU and PTX, and high requirements for instruments and equipment, and achieve The effect of reduced time, high sensitivity, simplified pretreatment steps

Active Publication Date: 2018-12-28
SOUTHWEST MEDICAL UNIVERISTY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The prior art of pharmacokinetic research on CU and PTX still has the following disadvantages: the existing method can only measure the content of CU and PTX biological samples alone, and it is difficult to realize the combination of CU and PTX; while the UPLC-MS / MS method is used to simultaneously measure CU and When it comes to PTX content, the sample pretreatment and determination operations are cumbersome, time-consuming, and require high equipment

Method used

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  • HPLC (High Performance Liquid Chromatography) analysis method for determining content of curcumin and paclitaxel in biological sample
  • HPLC (High Performance Liquid Chromatography) analysis method for determining content of curcumin and paclitaxel in biological sample
  • HPLC (High Performance Liquid Chromatography) analysis method for determining content of curcumin and paclitaxel in biological sample

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Embodiment 1: establish and measure the standard curve of CU, PTX concentration in CU single product, PTX single product and CU joint PTX compound product in rat plasma

[0046] 1. Standard solution preparation:

[0047] (1) Preparation of CU and PTX standard solutions: Accurately weigh 4.00 mg of CU and PTX reference substances, place them in 50 mL volumetric flasks, add methanol to dissolve, shake and shake well, and set the volume to 50 mL to obtain 80 μg / mL of CU and PTX The stock solution was diluted with methanol to prepare CU and PTX standard solutions with concentrations of 0.02, 0.04, 0.08, 0.16, 0.32, 0.80, 1.60, 3.20, 4.80, 6.60, and 8.00 μg / mL.

[0048] (2) Preparation of CU and PTX compound control solution: Take the newly prepared CU and PTX reference substance stock solution, dilute with methanol, and prepare concentrations of 0.02, 0.04, 0.08, 0.16, 0.32, 0.80, 1.60, 3.20, 4.80, respectively. 6.60, 8.00μg / mL CU and PTX composite reference substance solu...

Embodiment 2

[0054] Embodiment 2: sample detection

[0055] 1. Detection of spiked rat plasma samples

[0056] (1) Pretreatment of blank rat plasma and preparation of test solution

[0057] About 500 μL of blood was collected from the heart of a blank rat. The blood sample was placed in a heparinized EP tube, and the whole blood was centrifuged at 54,000 rpm for 3 minutes to absorb the upper layer of plasma, that is, the blank rat plasma, for use.

[0058] Add CU, PTX, and CU and PTX compound control solution to blank rat plasma to obtain CU concentrations of 0.1124, 0.9050, and 3.062 μg / mL; PTX concentrations of 0.1005, 0.4726, and 1.4204 μg / mL; Spiked plasma samples with concentrations of 0.1872, 0.7027, 3.1453 μg / mL, PTX concentrations of 0.1097, 0.3086, 1.2386 μg / mL.

[0059] Take 100 μL of the spiked plasma sample, and after processing according to the plasma sample pretreatment method in Example 1, take 160 μL of the supernatant and transfer it into an injection bottle.

[0060] ...

Embodiment 3

[0071] Embodiment 3: establish and measure the standard curve of CU, PTX concentration in CU single product, PTX single product and CU joint PTX compound product in rabbit plasma

[0072] 1. Standard solution preparation:

[0073] (1) Preparation of CU and PTX standard solutions: Accurately weigh 4.00 mg of CU and PTX reference substances, place them in 50 mL volumetric flasks, add methanol to dissolve, shake and shake well, and set the volume to 50 mL to obtain 80 μg / mL of CU and PTX The stock solution was diluted with methanol to prepare CU and PTX standard solutions with concentrations of 0.008, 0.016, 0.032, 0.08, 0.16, 0.8, 5, 10, 15, 20, 25, and 30 μg / mL, respectively.

[0074] (2) Preparation of CU and PTX composite standard solution: take the newly prepared CU and PTX reference substance stock solution, dilute with methanol, and prepare concentrations of 0.008, 0.016, 0.032, 0.08, 0.16, 0.64, 0.8, 5, 10, 15 , 20, 25, 30 μg / mL CU and PTX composite standard solution.

...

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Abstract

The invention relates to the field of pharmacokinetic analysis, in particular to an HPLC (High Performance Liquid Chromatography) method for determining the content of curcumin and paclitaxel in a biological sample. The method comprises a standard curve establishment step and a sample detection step in sequence. The HPLC analysis method is characterized in that a Luna@5[mu]m C18 (4.6mmx250mm) chromatographic analysis column and a gradient elution variable-wavelength method are adopted. Through adoption of the method, simultaneous determination of the content of curcumin and paclitaxel in the biological sample is realized by only HPLC; the efficiency of pharmacokinetic study of curcumin and paclitaxel combination is improved rapidly; the development process of curcumin and paclitaxel combination can be promoted rapidly; and the time, manpower and economic cost is greatly lowered for research and development enterprises and scientific research institutions.

Description

technical field [0001] The invention relates to the field of pharmacokinetic analysis, in particular to an HPLC method for determining the contents of curcumin and paclitaxel in biological samples. Background technique [0002] Curcumin (CU) is a monomer isolated and purified from traditional Chinese medicine. It is a rare pigment with diketones in the plant kingdom. [0003] It is a diketone compound, an orange-yellow crystalline powder, insoluble in water. It has anti-inflammatory, anti-tumor, anti-aging and other pharmacological effects, high efficiency and low toxicity, and has a good prospect as a new generation of anti-tumor drugs. It mainly realizes its anti-tumor effect by blocking the signal pathway conduction NF-κB, STAT3 and MAPK, regulating the expression of related tumor genes and proteins BcL-2, p53, etc., regulating the mediation of adhesion molecules, and inhibiting tumor angiogenesis. [0004] Paclitaxel (PTX) is a natural diterpenoid with anticancer activ...

Claims

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Application Information

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IPC IPC(8): G01N30/02
CPCG01N30/02
Inventor 赵领魏郁梦皮超李兰梅郭璞袁纪元黄绍秋赵仕杰邹永根杨红茹
Owner SOUTHWEST MEDICAL UNIVERISTY
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