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Method for determining kutkoside I content in biological sample

A technology for biological samples and picroside, which is applied in the preparation of test samples, measuring devices, instruments, etc., can solve the problems of complex pretreatment methods, poor reproducibility, low sensitivity, etc., and achieve shortened measurement time and high recovery rate. , the effect of good sensitivity

Inactive Publication Date: 2006-08-09
周亚伟
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Problems solved by technology

Though the chemical analysis method of this compound has been discussed at present, only had a report about the content determination of berberine-I in blood plasma in 1997 to the content analysis method of berberine-I in the biological sample (Journal Chromatography B, 698 (1997) 317-320), this method adopts the high-performance liquid phase ultraviolet detection method to measure the concentration of tuberculin-I in rabbit plasma after oral administration 100mg, plasma sample extracts protein through methanol precipitation, ethyl acetate After treatment, detection was carried out at a wavelength of 275nm. The lower limit of quantification of this method was 50ng / mL, and only the content of berberine-I in plasma samples 5 hours after administration was determined. The sensitivity was low, and the pretreatment method used in this report Complex, low recovery rate, poor reproducibility, can not be used for the determination of the content of berberine-I in organisms

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  • Method for determining kutkoside I content in biological sample
  • Method for determining kutkoside I content in biological sample
  • Method for determining kutkoside I content in biological sample

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051] Example 1 The determination of the blood concentration of berberine-I freeze-dried powder for injection in rats

[0052] 1. Instruments and materials

[0053] 1.1 Instrument Shimadzu SPD-10AT VP Double-pump liquid chromatography; guard column: Shim-packGVP-ODS (10mm×4.6mm, 5μm) (Shimadzu Corporation of Japan); N2000 chromatographic workstation

[0054]1.2 Materials Piroside I reference substance (provided by Peking University Shijia Research Center, batch number: 010114); internal standard reference substance: paeoniflorin (National Institute for the Control of Pharmaceutical and Biological Products, batch number: 0736-200219); pyroside I freeze-dried powder Injection (provided by Peking University Shijia Research Center, batch number: 020912 specification 60mg / branch, purity: 96.63%); acetonitrile (chromatographically pure, DIKMA company); methanol (chromatographically pure); ascorbic acid, carbon tetrachloride (all prepared by Beijing Chemical Reagent Provided by th...

Embodiment 2

[0078] Example 2 Pirucetin-I freeze-dried powder injection is measured in blood concentration in Beagle dogs

[0079] Experimental animals were non-rodent Beagle dogs. Follow the same steps as described in Example 1.

[0080] The results showed that the intra-day and inter-day precision of the method was 5.23% and 6.19%, respectively, and the recovery rate was 91.73%-95.87% in the determination of berberine-I in Beagle dog plasma. After the intravenous injection of 2.5mg / kg of pipericolin-I in dogs, the 2min average blood concentration of berberine-I was 17.05μg / mL.

Embodiment 3

[0081] The concentration determination of embodiment 3 berberine-I freeze-dried powder injection in rat tissue

[0082] After injecting berberine-I 7mg / kg into the tail vein of rats, take heart, liver, spleen, lung, kidney, brain, stomach, intestine, uterus, testis, fat, muscle and other tissues and organs, and prepare tissue homogenate with normal saline , Determination after protein precipitation with acetonitrile. The chromatographic conditions are the same as in Example 1. The results showed that: berberine-I could be detected in all tissues, and the average drug concentration in kidney tissue was the highest at 5 minutes after administration, which was 66.61 μg / mL; the concentration in brain tissue was the lowest, at 0.65 μg / mL.

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Abstract

Present invention provides a method for fast determining picroside-Icontent in biological sample utilizing high performance liquid chromatography. It contains adopting internal standard method to proceed quantify using octodecyl silicane bonding silicagel column as stationary phase, using acetonitrile-water as mobile phase, adopting gradient elution method. Said biological sample needs processed by antioxidant and determining after by organic solvent precipitation protein. Said method has simple operation, high sensitivity, and high recovery greater than 90 per cent etc advantages.

Description

technical field [0001] The invention belongs to the field of drug analysis, in particular to a method for determining the content of berberine-I in biological samples. Background technique [0002] As a traditional Chinese medicinal material, berberine has the effect of reducing steam and reducing fever. It was first recorded in "Xinxiu Materia Medica" in Tang Dynasty. Berberine-I is the main active ingredient of berberine, which belongs to iridoids. Though the chemical analysis method of this compound has been discussed at present, only had a report about the content determination of berberine-I in blood plasma in 1997 to the content analysis method of berberine-I in the biological sample (Journal Chromatography B, 698 (1997) 317-320), this method adopts the high-performance liquid phase ultraviolet detection method to measure the concentration of tuberculin-I in rabbit plasma after oral administration 100mg, plasma sample extracts protein through methanol precipitation, et...

Claims

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Application Information

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IPC IPC(8): G01N30/88G01N30/02G01N30/06G01N1/28G01N30/86
Inventor 周亚伟
Owner 周亚伟
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