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Content determination method of sanguisorbin I

A technology of Burnet saponins and a determination method, which are applied in the directions of measuring devices, pharmaceutical formulations, medical preparations containing active ingredients, etc., can solve the problems of long peak time, inability to effectively separate saponins, and low analysis efficiency, and achieve The effect of short peak time

Inactive Publication Date: 2012-10-10
CHENGDU DIAO PHARMA GROUP
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Problems solved by technology

However, it has been found in practice that the existing detection methods for the content of tannin and / or gallic acid are not suitable for the detection of Burnet saponin I
[0007] Zou Shengqin et al disclosed the high-performance liquid chromatography determination method of ursolic acid and oleanolic acid content in Burnet [Zou Shengqin, Chen Wu. High-performance liquid chromatography determination of ursolic acid and oleanolic acid content in Burnet Zhen Guoyi Guoyao, 2006, 17(8): 1373~1374]; Xie Junbo et al disclosed the method for the determination of three kinds of triterpenes and their saponins in Sijiqing by HPLC-ELSD method, including Sangui saponin I [Xie Junbo, Bi Zhiming , Li Ping. Determination of triterpenes and saponins in Sijiqing by HPLC-ELSD method], although Sijiqing also contains Burnet saponin I, but because Sijiqing and Burnet are different plants, except Burnet I , the other chemical components contained in the two are different, so the method suitable for the determination of the content of Burnet saponin I in Sijiqing is not suitable for the content determination of Burnet Burnet medicinal material, Burnet extract or Burnet preparation; Ming et al. disclosed the HPLC fingerprints of Sangui saponins and flavonoids, wherein the HPLC chromatographic conditions of Sangui saponins are: chromatographic column: Shim-pack CLC ODS column (6.0mm×150mm, 5μm), mobile phase : Acetonitrile-water (30:70) detection wavelength: 206nm, flow rate: 1.0ml / min, column temperature: 25°C
[Sha Ming, Zhang Dongfang, Meng Xiansheng, etc. Study on the quality evaluation of traditional Chinese medicine Burnet by DNA fingerprinting and HPLC fingerprinting, Chinese Journal of Pharmaceutical Sciences, 2002, 37(11): 815-818], but the peak eluting time using this method is too long (20min), the analysis efficiency is low, and the saponin I of Burnet eucalyptus and its similar structure saponins cannot be effectively separated
The analysis methods reported above all have the technical defects that the peak elution time is as long as 20 minutes, the analysis efficiency is low, and the saponin I of Burnet eucalyptus and its similar structure saponins cannot be effectively separated.
[0008] Therefore, so far, there is no method capable of quickly and accurately measuring the content of Burnet eucalyptus saponin I in Burnet burnet medicinal materials and its extracts or preparations at home and abroad.

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  • Content determination method of sanguisorbin I
  • Content determination method of sanguisorbin I
  • Content determination method of sanguisorbin I

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0068] Example 1 Selection of high performance liquid chromatography column, mobile phase and elution program

[0069] Sanyu saponins belong to triterpene saponins, which have relatively high polarity. Experiments show that they are retained on C18 chromatographic columns, so C18 analytical columns are used.

[0070] The choice of mobile phase: Sanguirin I is a neutral compound, and there is no need to add acid-base additives or ion pair reagents in the mobile phase. Therefore, the mobile phase systems such as methanol-water and acetonitrile-water are selected for comparison and screening. In UV detection, the acetonitrile-water system has a straighter baseline and sharper peaks than the methanol-water system. However, after multiple adjustments, the separation chromatographic peak is still better than methanol-water. There are few systems, and the impurity peak immediately before the saponin I can not be detected at all whether using isocratic elution or gradient elution (see Figu...

Embodiment 2

[0076] Example 2 The effect of column particle size on the separation effect of Sanguis saponins I reference substance

[0077] (1) Sample solution preparation: Take an appropriate amount of the crude Sanguisaponin I, dissolve it with methanol and dilute it to a solution containing 0.5mg per 1ml to obtain;

[0078] (2) Chromatographic conditions: use methanol-water system as mobile phase gradient elution, the elution procedure is 0~7min 60%→64% methanol aqueous solution, 7~12min 64% methanol aqueous solution, 12~20min 64%→80 % Methanol aqueous solution, 20~27min 80% methanol aqueous solution, flow rate 1.0ml / min, column temperature 25℃, detection wavelength 210nm, ELSD: 85℃, 2.8L / min, gain1; YMC-Pack proC was investigated 18 Column (150×4.6mm, 3.5μm) (UV detection) and SynergiHydro-RP C 18 The separation effect of the column (150×4.6mm, 4μm) (ELSD detection) on Sanguisorba saponins I is shown in Figure 2.

[0079] The results in Fig. 2 show that the main peak of Sanguirin I and the i...

Embodiment 3

[0080] Example 3 Isocratic elution of methanol-water system for separation of Sanguisorba saponins I from Sanguisorba sanguis medicinal materials and total Sanguis saponins

[0081] (1) Preparation of medicinal material sample solution: Take about 0.4g of Sanguiran medicinal material powder (passed through No. 4 sieve), accurately weigh it, place it in a 100ml measuring flask, add 80ml of 60% ethanol aqueous solution, shake well, ultrasound 15min, and place until Set the volume to the mark at room temperature, shake well, filter and take the filtrate for sample analysis or centrifuge to get the supernatant.

[0082] (2) Preparation of the sample solution of the total glycosides of Sanguinis: take an appropriate amount of the crude powder of Sanguirell medicinal materials, add 10 times the volume of water to decoct, each time for 1 hour, cook three times, filter, combine the filtrate, apply the macroporous adsorption resin, and wash thoroughly with distilled water , Then eluted with...

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Abstract

The invention discloses a content determination method of sanguisorbin I. The method comprises the following steps of: (1) respectively dissolving a standard product of sanguisorbin I and a sample containing sanguisorbin I into an ethanol aqueous solution to respectively prepare a standard solution and a sample solution; and (2) respectively injecting the standard solution and the sample solution which have equal volumes into a high-efficiency liquid-phase chromatograph, and then measuring and calculating the peak area of the standard solution and the sample solution to acquire the content of sanguisorbin I in the sample. The invention provides a rapid and accurate method for measuring the content of sanguisorbin I in a medical herb of garden burnet and extractives or preparations thereof.

Description

[0001] The present invention is a divisional application, the original application number is 201010170728.0, the original application date is April 30, 2010, and the original name is the content determination method of Sanguis saponins I. Technical field [0002] The invention relates to a method for determining the content of Sanguis saponins I in traditional Chinese medicine Sanyu medicinal materials, extracts and preparations thereof, and belongs to the technical field of traditional Chinese medicine. Background technique [0003] Chinese medicine Sanguisorba officinalis L. or Sanguisorba officinalis L. var. longifolia (Bert.) Yüet Li is the dried root of Sanguisorba officinalis L. [0004] The 2005 edition of the Chinese Pharmacopoeia contains this variety, which records the method for determining the content of tannin. The content of the specific content determination item is: "Take 0.4g of this product powder (passed through the fourth sieve), accurately weigh it, and determine...

Claims

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Application Information

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IPC IPC(8): G01N30/34G01N30/02A61K36/739A61P7/00A61K125/00
Inventor 姬建新黎秀丽刘志勇白红艳邹文俊王军姚华
Owner CHENGDU DIAO PHARMA GROUP
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