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UHPLC-MS/MS (Ultra High Performance Liquid Chromatography-Mass Spectrometry-Mass Spectrometry) analytical method for measuring rat curcumin plasma concentration

A technology of plasma concentration and analysis method, which is applied in the field of UHPLC-MS/MS analysis for measuring the plasma concentration of rat curcumin, can solve the problem that the lower limit of quantification cannot meet the detection requirements, the operation of measuring the plasma concentration of rat curcumin is cumbersome, and the injection time Long and other problems, to achieve the effect of simplifying the sample pretreatment process, meeting the detection requirements, high accuracy and precision

Inactive Publication Date: 2017-06-13
GUANGZHOU ZHONGDA NANSHA TECH INNOVATION IND PARK +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This prior art has the following disadvantages: the use of HPLC to measure the plasma concentration of curcumin in rats is cumbersome, the sample injection time is long, and the lower limit of quantification cannot meet the detection requirements, etc.

Method used

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  • UHPLC-MS/MS (Ultra High Performance Liquid Chromatography-Mass Spectrometry-Mass Spectrometry) analytical method for measuring rat curcumin plasma concentration
  • UHPLC-MS/MS (Ultra High Performance Liquid Chromatography-Mass Spectrometry-Mass Spectrometry) analytical method for measuring rat curcumin plasma concentration
  • UHPLC-MS/MS (Ultra High Performance Liquid Chromatography-Mass Spectrometry-Mass Spectrometry) analytical method for measuring rat curcumin plasma concentration

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Embodiment 1 establishes the standard curve of measuring curcumin concentration in blood plasma

[0046] 1. Preparation of standard working solution:

[0047](1) Preparation of curcumin standard working solution: Accurately weigh 3.47 mg of curcumin reference substance, dissolve in 1.735 mL of acetonitrile solution and shake well to obtain a standard stock solution of 1 mg / mL (1000 ng / µL), and then use Methanol / water=1 / 1 diluent solution gradient dilution to prepare curcumin standard solutions with concentrations of 100, 10, 7.5, 5, 2.5, 0.5, 0.25, 0.05, 0.02, 0.01 ng / µL.

[0048] (2) Preparation of internal standard tinidazole (Tinidazole) standard solution: Accurately weigh 10.03 mg of tinidazole standard product, dissolve in 10 mL of 90% methanol aqueous solution and shake well to obtain 1 mg / mL (1000 ng / µL) The stock solution was then diluted with 90% aqueous methanol to obtain a 0.2 µg / mL standard stock solution.

[0049] 2. Establish a standard curve

[0050] T...

Embodiment 2

[0061] Example 2 Detection of spiked samples

[0062] 1. Pretreatment of spiked plasma samples

[0063] (1) About 300 µL of blood was collected from the orbital venous plexus of blank rats. The blood samples were placed in heparinized EP tubes, and the whole blood was centrifuged at 4000 rpm for 5 min to absorb the upper plasma, that is, the blank rat plasma, which was divided into 100 µL and stored at -80°C.

[0064] Curcumin was added to blank rat plasma to obtain spiked plasma samples with curcumin concentrations of 1, 2, 100, and 8000 ng / mL.

[0065] (2) Take 100 µL of the spiked plasma sample, and after processing according to the sample pretreatment in Example 1, take 80 µL of the supernatant and transfer it into the sample bottle.

[0066] 2. Detection of spiked plasma samples

[0067] 10 µL was injected for UPLC-MS / MS analysis, and the conditions were the same as in Example 1. The plasma sample concentration calculated according to the standard curve is 1.0±0.1, 2....

Embodiment 3

[0069] Example 3 Sample detection

[0070] 1. Study on the absolute bioavailability of curcumin preparations after single gavage in SD rats. Plasma sample pretreatment

[0071] (1) After a single gavage and intravenous injection of SD rats, about 300 µL of blood was collected from the orbital venous plexus according to the sampling time plan of the experimental design. Blood samples were placed in heparinized EP tubes, and the whole blood was centrifuged at 4000 rpm for 5 min to absorb the upper layer of plasma, which was divided into 100 µL and stored at -80°C until assayed.

[0072] (2) 100 µL of rat plasma sample was pretreated with the sample pretreatment method described in Example 1, and 80 µL of the supernatant was taken and transferred into an injection bottle.

[0073] 2. Study on absolute bioavailability of curcumin preparations by single oral gavage in SD rats Plasma sample detection

[0074] 10 µL was injected for UPLC-MS / MS analysis, the conditions were the same...

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Abstract

The invention discloses a UHPLC-MS / MS (Ultra High Performance Liquid Chromatography-Mass Spectrometry-Mass Spectrometry) analytical method for measuring rat curcumin plasma concentration. The method comprises the following steps: (1) preparing a curcumin standard solution and an internal standard tinidazole standard solution, and establishing a standard curve; and (2) adding an internal standard into to-be-measured plasma, performing pretreatment, performing LC-MS / MS analysis, and calculating the concentration of curcumin in the to-be-measured plasms by utilizing the standard curve. According to the method disclosed by the invention, the chromatographic conditions and internal standard substances are reasonably selected, the sensitivity is improved, the analysis time is shortened, and the sample pretreatment process is simplified. Meanwhile, the method disclosed by the invention has high accuracy and precision degree, and the detection requirements of non-clinical pharmacokinetic study are met.

Description

technical field [0001] The invention belongs to the technical field of pharmacokinetic analysis. More specifically, it relates to a UHPLC-MS / MS analysis method for determining the plasma concentration of curcumin in rats. Background technique [0002] Curcumin is a phenolic pigment extracted from the rhizomes of some plants in Zingiberaceae and Araceae. Among them, turmeric contains about 3% to 6%, which is a rare pigment with diketone in the plant kingdom. class of compounds. Curcumin is an orange-yellow crystalline powder with a slightly bitter taste and is insoluble in water. Curcumin has the effects of lowering blood fat, anti-tumor, anti-inflammation, choleretic, and anti-oxidation, and is a new anti-cancer drug with good application prospects. [0003] Pharmacokinetic evaluation is to quantitatively study the absorption, distribution, metabolism and excretion of drugs in vivo, and to use mathematical principles and methods to explain the changes in blood drug concen...

Claims

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Application Information

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IPC IPC(8): G01N30/02G01N30/72G01N30/88
CPCG01N30/02G01N30/7266G01N30/88G01N2030/884
Inventor 温鼎声姜福林陈江英钟国平黄民
Owner GUANGZHOU ZHONGDA NANSHA TECH INNOVATION IND PARK
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