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Method for simultaneously and quantitatively detecting two components entering blood in trichosanthes kirilowii maxim, allium macrostemon and pinellia ternate and application

A quantitative detection method, the technology of Gualou Xiebai, is applied in the direction of measuring devices, material separation, and analysis of materials, so as to shorten the analysis time and improve the detection sensitivity.

Pending Publication Date: 2021-10-22
SHINEWAY PHARMA GRP LTD +1
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] Aiming at the defect that there is no quantitative detection method for the blood-entering components of Gualou Xiebai Pinellia at present, the present invention provides a simultaneous quantitative detection method and application of two blood-entering components in Gualou Xiebai Pinellia. The quantitative detection method can To provide a basis for further screening the quality markers and pharmacodynamic material basis of the classic famous prescription Gualou Xiebai Pinellia

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  • Method for simultaneously and quantitatively detecting two components entering blood in trichosanthes kirilowii maxim, allium macrostemon and pinellia ternate and application
  • Method for simultaneously and quantitatively detecting two components entering blood in trichosanthes kirilowii maxim, allium macrostemon and pinellia ternate and application
  • Method for simultaneously and quantitatively detecting two components entering blood in trichosanthes kirilowii maxim, allium macrostemon and pinellia ternate and application

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preparation example Construction

[0044] Preparation of internal standard solution: Weigh 5.00 mg of zeatin into a 10 mL volumetric flask, add methanol solution to the volume to obtain a 0.5 mg / mL internal standard stock solution, and dilute to obtain a 4 μg / mL internal standard solution for later use;

[0045]Preparation of plasma samples to be tested: Precisely draw 50 μL of rat plasma samples into 1.5 mL of EP, precisely add 10 μL of the above internal standard solution, vortex for 1 min to mix evenly, then add 140 μL of acetonitrile solution, vortex for 5 min, and mix thoroughly Evenly, centrifuge at 12000g for 10min, and the supernatant is the plasma sample to be tested;

[0046] Using UPLC-MS / MS to simultaneously detect the contents of guanosine and adenosine in the plasma samples to be tested;

[0047] Among them, mass spectrometry conditions: the ion source is ESI source, the capillary voltage is 3.2kv, the desolvation gas is nitrogen; the temperature of the desolvation gas is 500°C, the flow rate of t...

Embodiment 1

[0064] Example 1 Study on the Determination Method of Gualou Xiebai Banxia Drug Entering Blood Components

[0065] 1. Solution Preparation

[0066] 1.1 Preparation of mixed reference solution

[0067] Precisely weigh the reference substances trigonelline, uracil, thymidine, adenine, caffeic acid, uridine, guanine, guanosine, xanthine, adenosine, vanillic acid, 5-hydroxymethylfurfural, nicotinamide, Ding, quercetin, luteolin, protocatechuic acid, homoharringtonine, schiaftoside, cucurbitacin B, N-p-coumaroyl tyramide and ferulic acid 5.00mg each, placed in 10mL In the volumetric flask, add methanol to make the volume up to the mark, so that the final concentration is 0.5 mg / mL of each reference substance stock solution;

[0068] Then accurately measure each reference substance stock solution respectively, mix and dilute with methanol to prepare a mixed reference substance solution; the concentration of each reference substance in the mixed reference substance solution is as f...

Embodiment 2

[0099] Example 2 Methodological verification of the assay method for blood component content of Gualou Xiebai Pinellia jelly

[0100] Limit of detection and limit of quantitation (confirmation of prototype components in blood)

[0101] According to the signal-to-noise ratio of each reference substance in the mixed reference substance solution, calculate the required dilution factor, and properly dilute each reference substance stock solution to obtain the concentration of each reference substance in the mixed reference substance solution, and then follow the detection conditions under 2.1 Detect, determine the limit of quantification with the signal-to-noise ratio (S / N) greater than 10, determine the limit of detection with the signal-to-noise ratio (S / N) greater than 3, the detection limit and the limit of quantification of 22 kinds of components are indicated.

[0102] Table 5 Limits of quantification and detection of 22 components in rat plasma

[0103]

[0104]

[0...

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Abstract

The invention provides a method for simultaneously and quantitatively detecting two components entering blood in a trichosanthes kirilowii maxim, allium macrostemon and pinellia ternate preparation and application. The detection method comprises the following steps: detecting the contents of guanosine and adenosine in a to-be-detected plasma sample by adopting UPLC-MS / MS (Ultra Performance Liquid Chromatography-Mass Spectrometry / Mass Spectrometry), wherein chromatographic conditions are as follows: silica gel particles are used as a reversed-phase chromatographic column of a filler, and a mobile phase A is an aqueous solution containing 0.1% of formic acid, and a mobile phase B is acetonitrile; and performing gradient elution. According to the method, parameters in the UPLC-MS / MS method are limited and are matched with one another, so that the contents of two blood-entering components, namely guanosine and adenosine, in the trichosanthes kirilowii Maxim, allium macrostemon and pinellia ternate can be measured at the same time, and the method is simple, rapid, accurate, reliable, high in sensitivity, good in specificity and good in stability.

Description

technical field [0001] The invention relates to the technical field of drug metabolism in vivo, in particular to a simultaneous quantitative detection method and application of two blood-entering components in Gualou Xiebai Pinellia preparation. Background technique [0002] The classic famous prescription Gualou Xiebai Banxia Decoction comes from "Synopsis of the Golden Chamber". It is made from Gualou, Xiebai, Pinellia and rice wine. Tachycardia, intercostal neuralgia, breast hyperplasia, chronic obstructive pulmonary disease, traumatic pneumothorax, senile cough, chronic bronchial pneumonia, chronic cholecystitis, etc. [0003] At present, the research on Gualou Xiebai Banxia Decoction mainly focuses on the research on the chemical components, but the medicinal material basis of the efficacy of Gualou Xiebai Banxia Decoction is not very clear, and the detection method of the active ingredients in the blood is even blank . [0004] As we all know, the active ingredients ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02G01N30/72
CPCG01N30/02G01N30/72
Inventor 向铮求杰英刘眴蒂李丹侯金才
Owner SHINEWAY PHARMA GRP LTD
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