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Method for measuring danshensu, m-methyl-danshensu, protocatechualdehyde, and protocatechuic acid in human blood plasma

A technique for meta-methyl salvia and protocatechualdehyde, which is used in measuring devices, instruments, scientific instruments, etc.

Active Publication Date: 2015-08-19
TIANJIN TASLY PHARMA CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Danshensu and protocatechuic aldehyde are the main components of compound Danshen dripping pills. In recent years, the content determination methods of danshensu, protocatechuic aldehyde and protocatechuic acid in relevant biological samples (such as HPLC, LC-MS / MS, etc.) There are many reports, but most of them are methods for the determination of the above compounds alone

Method used

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  • Method for measuring danshensu, m-methyl-danshensu, protocatechualdehyde, and protocatechuic acid in human blood plasma
  • Method for measuring danshensu, m-methyl-danshensu, protocatechualdehyde, and protocatechuic acid in human blood plasma
  • Method for measuring danshensu, m-methyl-danshensu, protocatechualdehyde, and protocatechuic acid in human blood plasma

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0097] 1. Preparation of internal standard solution

[0098] Precisely measure 10.0mg of vanillic acid (internal standard) and put it in a 50mL volumetric flask, dissolve it with methanol and dilute to the mark, and shake well to obtain a 200μg / mL internal standard stock solution. Precisely measure 125μL of this internal standard stock solution in In a 5mL volumetric flask, dissolve with methanol and dilute to the mark to obtain a 5μg / mL internal standard solution.

[0099] 2. Plasma sample processing method

[0100] Take 200 μL of plasma sample, add 50 μL of internal standard solution, 50 μL of 50% methanol aqueous solution, 20 μL of 1mol / L hydrochloric acid solution, vortex evenly, add 1 mL of ethyl acetate, vortex for 30 s, centrifuge at 15000 rpm for 1 min, and take the supernatant ethyl acetate 900 μL of the ester layer was dried under nitrogen flow at 40°C, reconstituted with 120 μL of 50% aqueous methanol solution, centrifuged at 15,000 rpm for 3 min, and 10 μL of the ...

Embodiment 2

[0106] 1. Preparation of internal standard solution

[0107] Precisely measure 10.0mg of vanillic acid (internal standard) and put it in a 50mL volumetric flask, dissolve it with methanol and dilute to the mark, and shake well to obtain a 200μg / mL internal standard stock solution. Precisely measure 125μL of this internal standard stock solution in In a 5mL volumetric flask, dissolve with methanol and dilute to the mark to obtain a 5μg / mL internal standard solution.

[0108] 2. Plasma sample processing method

[0109] Take 200 μL of plasma sample, add 50 μL of internal standard solution, 50 μL of 50% methanol aqueous solution, 20 μL of 1mol / L hydrochloric acid solution, vortex evenly, add 1 mL of ethyl acetate, vortex for 30 s, centrifuge at 15000 rpm for 1 min, and take the supernatant ethyl acetate 900 μL of the ester layer was dried under nitrogen flow at 40°C, reconstituted with 120 μL of 50% aqueous methanol solution, centrifuged at 15,000 rpm for 3 min, and 10 μL of the ...

Embodiment 3

[0114] 1. Preparation of internal standard solution

[0115] Precisely measure 10.0mg of vanillic acid (internal standard) and put it in a 50mL volumetric flask, dissolve it with methanol and dilute to the mark, and shake well to obtain a 200μg / mL internal standard stock solution. Precisely measure 125μL of this internal standard stock solution in In a 5mL volumetric flask, dissolve with methanol and dilute to the mark to obtain a 5μg / mL internal standard solution.

[0116] 2. Plasma sample processing method

[0117] Take 200 μL of plasma sample, add 50 μL of internal standard solution, 50 μL of 50% methanol aqueous solution, 20 μL of 1mol / L hydrochloric acid solution, vortex evenly, add 1 mL of ethyl acetate, vortex for 30 s, centrifuge at 15000 rpm for 1 min, and take the supernatant ethyl acetate 900 μL of the ester layer was dried under nitrogen flow at 40°C, reconstituted with 120 μL of 50% aqueous methanol solution, centrifuged at 15,000 rpm for 3 min, and 10 μL of the ...

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Abstract

The invention relates to a method for measuring drug contents, particularly to a method for measuring danshensu, m-methyl-danshensu, protocatechualdehyde, and protocatechuic acid in the human blood plasma. The method comprises the following steps: 1) preparation of reference substance storing solutions: weighting reference substances of danshensu, m-methyl-danshensu, protocatechualdehyde, and protocatechuic acid, and dissolving the reference substances with methanol to obtain reference substance storing solutions of danshensu, m-methyl-danshensu, protocatechualdehyde, and protocatechuic acid; 2) preparation of an internal standard solution: dissolving an internal standard substance of vanillic acid with methanol to obtain the internal standard solution; 3) a treatment method of a blood plasma sample: taking the blood plasma sample, adding the internal standard solution, methanol, hydrochloric acid, and ethyl acetate into the blood plasma sample, performing uniform mixing, performing centrifugation of the obtained mixture to obtain a supernatant ethyl acetate layer, drying the ethyl acetate layer, re-dissolving the ethyl acetate layer with methanol, and performing centrifugation of the obtained solution to obtain supernatant liquid of the blood plasma sample; and 4) content measurement: by adopting LC-MS / MS chromatography, injecting the reference substance storing solutions and the supernatant liquid of the blood plasma sample into a chromatographic instrument to obtain a chromatogram, and calculating concentrations of danshensu, m-methyl-danshensu, protocatechualdehyde, and protocatechuic acid in the human blood plasma according to the areas of chromatographic peaks in the chromatogram.

Description

Technical field: [0001] The invention relates to a method for determining the content of medicines, in particular to a method for determining danshensu, m-methyldanshensu, protocatechualdehyde and protocatechuic acid in human blood plasma. Background technique: [0002] Compound Danshen Dripping Pills is a traditional Chinese medicine for promoting blood circulation, removing blood stasis, regulating qi and relieving pain developed by Tasly Company. It is used for chest tightness and angina pectoris. Hypoxia protects ischemic myocardium, 3 anti-platelet aggregation prevents thrombosis, 4 improves microcirculation. [0003] Salvia miltiorrhiza in Compound Danshen Dripping Pills is bitter in taste and slightly cold in nature. It has the effects of promoting blood circulation and removing blood stasis, nourishing blood and calming nerves, cooling blood and expelling carbuncle, detoxifying and promoting granulation. It is a commonly used drug for promoting blood circulation and ...

Claims

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Application Information

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IPC IPC(8): G01N30/88
Inventor 李伟罗瑞芝褚扬杨柳马晓慧郭治昕周水平孙鹤
Owner TIANJIN TASLY PHARMA CO LTD
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