Method for determining concentration of 5'-methoxyl-3',4'-methylenedioxyphenyl cinnamic acid isobutyl amide in plasma

Abstract

The invention discloses a met / hod for determining the concentration of 5'-methoxyl-3',4'-methylenedioxyphenyl cinnamic acid isobutyl amide in plasma. A liquid chromatography-mass system is adopted to determine. The method comprises the following steps: taking a to-be-determined sample first, adding a certain quantity of organic solvent for extraction, separating through a chromatographic column after pretreatment, and determining by using a mass spectrometry detector. The method disclosed by the invention has the advantages of rapidity, accuracy, high sensitivity, simplicity and convenience in operation and is suitable for determining the concentration of 5'-methoxyl-3',4'-methylenedioxyphenyl cinnamic acid isobutyl amide in plasma.

Description

technical field

[0001] The invention belongs to the technical field of drug analysis, and relates to a method for analyzing and measuring drugs in vivo, in particular to a method for measuring the concentration of 5'-methoxy-3',4'-methylenedioxyphenylcinnamic acid isobutylamide in plasma. Background technique

[0002] Piper Laetispicum (Piper Laetispicum C.DC.) is a plant belonging to the family Piperaceae. It is mainly produced in Hainan, Guangdong, Guangxi, Yunnan and other places. It is used for promoting blood circulation, reducing swelling and relieving pain. It is mainly used for bruises, blood stasis and swelling . Recent studies have found that betel has a strong antidepressant effect, and its activity is related to a series of amide compounds. Among them, 5'-methoxy-3',4'-methylenedioxyphenyl cinnamic acid isobutylamide (C15H19NO4) (K6) was the alkaloid with the highest content.

[0003] At present, there is no literature report on the determination method of K6 i...

Examples

Embodiment 1

[0023] Example 1: Determination of K6 concentration in human plasma.

[0024] 1. Experimental materials and instruments

[0025] K6 reference substance: provided by Suzhou Yihua Biomedical Technology Co., Ltd., batch number: 20131121; tinidazole reference substance: provided by China Institute for the Control of Pharmaceutical and Biological Products, batch number: 100336-200402; test water: ultrapure water; methanol: chromatography Pure (Merck Company).

[0026] API4000LC / MS / MS coupled instrument (Applied Biosystems, USA), chromatographic workstation: Analyst1.4.2; Mettler MT-5 electronic balance (Mettler, Switzerland); LEGEND MICRO17R refrigerated high-speed centrifuge (Thermo, USA) ; 80-2 centrifuge (Jiangsu Jintan Ronghua Instrument Manufacturing Co., Ltd.) Millipore Drict-Q5 ultra-pure water machine (Millipore, France). Centrifugal concentrator: Labconco CentriVap Console (Labconco, USA)

[0027] 2. Liquid conditions

[0028] 1. Liquid chromatography conditions

[00...

Embodiment 2

[0048] Embodiment 2: the mensuration of K6 concentration in human plasma

[0049] Referring to Example 1, 6 females took the extract of effective parts of betelwort (containing 20.8 mg of K6) on an empty stomach, and took it with 250 mL of warm boiled water; 4 mL of peripheral venous blood was collected 3 hours after administration, injected into a heparin tube, protected from light, and centrifuged (3000r·min -1 , 10 min) to prepare plasma. The treatment and determination conditions of the drug-containing plasma samples were the same, and the results showed that the K6 contents in the plasma of the 6 women who were given the extract of the effective part of the betelwort were 9.323, 12.54, 12.55, 13.09, 5.011, and 7.569 ng·mL -1 .

technical field

[0001] The invention belongs to the technical field of drug analysis, and relates to a method for analyzing and measuring drugs in vivo, in particular to a method for measuring the concentration of 5'-methoxy-3',4'-methylenedioxyphenylcinnamic acid isobutylamide in plasma. Background technique

[0002] Piper Laetispicum (Piper Laetispicum C.DC.) is a plant belonging to the family Piperaceae. It is mainly produced in Hainan, Guangdong, Guangxi, Yunnan and other places. It is used for promoting blood circulation, reducing swelling and relieving pain. It is mainly used for bruises, blood stasis and swelling . Recent studies have found that betel has a strong antidepressant effect, and its activity is related to a series of amide compounds. Among them, 5'-methoxy-3',4'-methylenedioxyphenyl cinnamic acid isobutylamide (C15H19NO4) (K6) was the alkaloid with the highest content.

[0003] At present, there is no literature report on the determination method of K6 i...

Examples

Embodiment 1

[0023] Example 1: Determination of K6 concentration in human plasma.

[0024] 1. Experimental materials and instruments

[0025] K6 reference substance: provided by Suzhou Yihua Biomedical Technology Co., Ltd., batch number: 20131121; tinidazole reference substance: provided by China Institute for the Control of Pharmaceutical and Biological Products, batch number: 100336-200402; test water: ultrapure water; methanol: chromatography Pure (Merck Company).

[0026] API4000LC / MS / MS coupled instrument (Applied Biosystems, USA), chromatographic workstation: Analyst1.4.2; Mettler MT-5 electronic balance (Mettler, Switzerland); LEGEND MICRO17R refrigerated high-speed centrifuge (Thermo, USA) ; 80-2 centrifuge (Jiangsu Jintan Ronghua Instrument Manufacturing Co., Ltd.) Millipore Drict-Q5 ultra-pure water machine (Millipore, France). Centrifugal concentrator: Labconco CentriVap Console (Labconco, USA)

[0027] 2. Liquid conditions

[0028] 1. Liquid chromatography conditions

[00...

Embodiment 2

[0048] Embodiment 2: the mensuration of K6 concentration in human plasma

[0049] Referring to Example 1, 6 females took the extract of effective parts of betelwort (containing 20.8 mg of K6) on an empty stomach, and took it with 250 mL of warm boiled water; 4 mL of peripheral venous blood was collected 3 hours after administration, injected into a heparin tube, protected from light, and centrifuged (3000r·min -1 , 10 min) to prepare plasma. The treatment and determination conditions of the drug-containing plasma samples were the same, and the results showed that the K6 contents in the plasma of the 6 women who were given the extract of the effective part of the betelwort were 9.323, 12.54, 12.55, 13.09, 5.011, and 7.569 ng·mL -1 .

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