Simultaneous quantitative measuring method of 9 in-blood constituents of spine date seed water extract

A technology for quantitative determination and blood components, applied in the direction of measuring devices, material separation, and analysis of materials, can solve the problems of long analysis time, low analysis sensitivity, and unfavorable pharmacokinetic research of traditional Chinese medicine, so as to improve detection sensitivity and shorten analysis time. the effect of time

Active Publication Date: 2019-01-18
汉方萃取生物科技(海南)股份有限公司
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Problems solved by technology

[0004] The present invention is in order to solve the low analysis sensitivity of HPLC-UV in the quantitative analysis of the components in the blood of Jujube Seed at the present stage, it is not suitable for analyzing the trace components in the blood plasma, and the analysis time is too long; although LC-MS / MS has high measurement sensitivity, it only Applied to the determination of the pharmacokinetics of a single component, which is not conducive to the study of the pharmacokinetics of traditional Chinese medicine, a method for the simultaneous quantitative determination of 9 blood components in the water extract of Suanzaoren is provided, and Application of Oral Administration in the Pharmacokinetics of Normal and PCPA Insomnia Model Rats

Method used

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  • Simultaneous quantitative measuring method of 9 in-blood constituents of spine date seed water extract
  • Simultaneous quantitative measuring method of 9 in-blood constituents of spine date seed water extract
  • Simultaneous quantitative measuring method of 9 in-blood constituents of spine date seed water extract

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Embodiment 1

[0027] Example 1: A method for the simultaneous quantitative determination of 9 blood components in the water extract of Suanzaoren, using the UHPLC-QExactive Orbitrap HR / MS method to quantitatively determine 9 active ingredients in the water extract of Suanzaoren: black medicine Alkaline, magnolialine, weixining, spinosin, flavin, kaempferol-3- O -Rutinoside, 6 ''' - Feruloyl spinosin, jujube saponin A and jujube saponin B; the specific steps are as follows:

[0028] (1) Preparation of a single stock solution of a reference substance: Accurately weigh the reference substances respectively, add the initial mobile phase of the chromatography analyzed by UHPLC-Q ExactiveOrbitrap HR / MS, and prepare 2.032 mg / mL of aconitine and 1.130 mg / mL of magnolialine The stock solution of single reference substance was prepared by adding methanol to Weicaining 2.184 mg / mL, spinosin 2.016 mg / mL, flavin 1.972 mg / mL, kaempferol-3- O - Rutinoside 2.058 mg / mL, 6 ''' - A single reference substan...

Embodiment 2

[0038] Embodiment 2: methodological investigation

[0039] 1. Specificity test: the blank plasma, blank plasma plus working solution, and drug-containing plasma from 6 rats from different sources were processed and analyzed according to the conditions in Example 1. Compare the chromatograms of the three, and observe whether there is interference in the blank plasma at the peak positions of the components to be tested and the internal standard. The result is as figure 1 As shown, the retention times of the components to be tested and the internal standard were: 3.23 min for hidradoline, 4.12 min for magnolialine, 2.85 min for weicaining, 6.13 min for spinosin, 6.87 min for flavin, and 6.87 min for kaempferol-3 - O - Rutinoside 7.37 min, 6 ''' - Feruloyl spinosin 8.14 min, jujuboside A 10.76 min, juboside B 11.14 min, palmatine hydrochloride 10.22 min, daidzin 4.60 min, astragaloside 4 11.10 min. There was no interference in the blank plasma corresponding to the peak positi...

Embodiment 3

[0052] Example 3: Pharmacokinetic determination of 9 components in rat plasma after oral administration of jujube seed aqueous extract in normal and PCPA insomnia model rats

[0053] (1) Preparation of Suanzaoren water extract: take about 500 g of Suanzaoren decoction pieces, crush (60% pass through No. 1 Pharmacopoeia sieve), accurately weigh, add 10 times the amount of water, soak for 30 min, heat to boiling, and reflux extraction 2 h, filter, add 8 times the amount of water to the filter residue, heat and reflux for extraction for 2 h, filter, and combine the filtrates; concentrate under reduced pressure to 0.5 g / mL (crude drug amount), freeze-dry, and dissolve in water when used.

[0054] (2) Animal experiments and collection of plasma samples: healthy male SD rats, weighing 280±20 g, provided by Beijing Weitong Lihua Experimental Animal Technology Co., Ltd. The rats were randomly divided into two groups, the normal group and the model group, with 6 rats in each group. Th...

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Abstract

The invention belongs to the technical field of research of internal metabolism of Chinese patent medicine, and provides a simultaneous quantitative measuring method of 9 in-blood constituents of a spine date seed water extract in order to solve problems that the HPLC-UV sensitivity is low and the time is over-long in a conventional quantitative analysis of the in-blood constituents of a spine data seed so that the analysis is not suitable for analysis of micro-constituents in blood plasma, and LC-MS / MS is only used for measurement of pharmacokinetics with a single constituent and does not facilitate the research of traditional Chinese medicine pharmacokinetics. Nine in-blood constituents of the spine data seed water extract are simultaneously and quantitatively measured by employing a UHPLC-Q Exactive Orbitrap HR / MS method, the detection sensitivity is improved, and the analysis time is reduced; simultaneous quantitative analysis of three polar compounds including flavone, saponin andalkaloids is conducted; and an analysis technical guarantee is provided for the research of the pharmacokinetics of the spine date seed in rats of normal and PCPA insomnia models, and references areprovided for simultaneous quantitative analysis of multiple constituents of other oral traditional Chinese medicine.

Description

technical field [0001] The invention belongs to the technical field of in vivo metabolism research of Chinese patent medicines, and in particular relates to a simultaneous quantitative determination method for nine blood-entering components in water extracts of jujube seeds. Background technique [0002] Ziziphus jujube is a plant of Rhamnaceae Ziziphus jujuba Mill. var. spinosa (Bunge) The dry and mature seeds of Huex H. F. Chou have the effects of nourishing the heart and nourishing the liver, calming the mind, calming the mind, suppressing sweat, and promoting body fluid. It is the first choice for clinical treatment of insomnia. Modern pharmacological studies have shown that total saponins, total flavonoids, and total alkaloids in Suanzaoren are the main active ingredients for sedation and hypnosis. [0003] As we all know, the absorption of active ingredients of traditional Chinese medicine into the blood is the premise of its curative effect, and multi-component, ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02G01N30/06
CPCG01N30/02G01N30/06
Inventor 闫艳张敏杜晨晖张福生秦雪梅
Owner 汉方萃取生物科技(海南)股份有限公司
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