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Content determination method of sodium taurochenodeoxycholate in bile

A technology of sodium taurochenodeoxycholate and a detection method, applied in the field of medicine, can solve the problems of low sensitivity, poor repeatability, unsuitable detection of sodium taurochenodeoxycholate, etc., and achieves good reproducibility, Accurate and practical effects

Inactive Publication Date: 2014-03-26
SHANGHAI NATURE STANDARD R&D & BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The existing detection method has low sensitivity and poor repeatability, and is not suitable for detecting sodium taurochenodeoxycholate in bile

Method used

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  • Content determination method of sodium taurochenodeoxycholate in bile
  • Content determination method of sodium taurochenodeoxycholate in bile
  • Content determination method of sodium taurochenodeoxycholate in bile

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Chromatographic conditions and system adaptability: Agilent ZORBAX Extend C18 (4.6×250mm, 5μm) is used for the chromatographic column; the mobile phase is gradient elution of acetonitrile-0.03% triethylamine aqueous solution, 0-30min15% acetonitrile-90% acetonitrile; flow rate 1ml / min; ELSD nebulization temperature 105°C, gas flow rate 3L / min; column temperature 25°C, injection volume 20μl;

[0025] Preparation of the test solution: Take about 0.5g of bile, weigh it accurately, put it in a 25ml volumetric flask, add an appropriate amount of ethanol, ultrasonically treat it (220W, 40kHz) for 20min, let it cool, dilute to the mark with ethanol, and shake well. Filter with a 0.45μm microporous membrane, that is, too.

[0026] Preparation of standard solution: Accurately weigh 12.3mg of sodium taurochenodeoxycholate reference substance, place it in a 25ml volumetric flask, dilute to the mark with methanol, and shake well; that is ready.

[0027] Determination method: Prec...

Embodiment 2

[0030] Chromatographic conditions and system adaptability: the chromatographic column uses Symmetrix ODS-R (4.6 × 250mm, 5 μm); the mobile phase is acetonitrile-0.03% triethylamine aqueous solution gradient elution, 0 ~ 30min15% acetonitrile ~ 70% acetonitrile; flow rate 1ml / min; ELSD atomization temperature is 105°C, gas flow rate is 3L / min; column temperature is room temperature; injection volume is 10μl;

[0031] Preparation of the test solution: Take about 0.5g of bile, weigh it accurately, put it in a 25ml volumetric flask, add an appropriate amount of ethanol, ultrasonically treat it (220W, 40kHz) for 20min, let it cool, dilute to the mark with ethanol, and shake well. Filter with a 0.45μm microporous membrane, that is, too.

[0032] Preparation of the reference substance solution: Accurately weigh 11.8 mg of the reference substance sodium taurochenodeoxycholate, place it in a 25ml volumetric flask, dilute to the mark with methanol, and shake well; that is.

[0033] De...

Embodiment 3

[0036] Chromatographic conditions and system adaptability: Agilent ZORBAX SB-C18 (4.6×250mm, 5μm) is used for the chromatographic column; the mobile phase is gradient elution of acetonitrile-0.03% triethylamine aqueous solution, 0-40min15% acetonitrile-90% acetonitrile; flow rate 1ml / min; ELSD nebulization temperature 105°C, gas flow rate 3L / min; column temperature 25°C, injection volume 5μl;

[0037] Preparation of the test solution: Take about 0.5g of bile, weigh it accurately, put it in a 25ml volumetric flask, add an appropriate amount of ethanol, ultrasonically treat it (220W, 40kHz) for 20min, let it cool, dilute to the mark with ethanol, and shake well. Filter with a 0.45μm microporous membrane, that is, too.

[0038] Preparation of reference solution: Accurately weigh 10.5 mg of sodium taurochenodeoxycholate reference substance, place it in a 25ml volumetric flask, dilute to the mark with methanol, and shake well; that is.

[0039] Determination method: Precisely dra...

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Abstract

The invention discloses a method for detecting sodium taurochenodeoxycholate content in bile by using a reversed-phase high-performance liquid chromatography-evaporative light scattering detector (ELSD) method. The method is characterized by comprising the following steps: ultrasonically extracting sodium taurochenodeoxycholate in bile by using ethanol so as to prepare a test solution; absorbing the test solution to inject into a high performance liquid chromatograph; determining by adopting the reversed-phase high-performance liquid chromatography-ELSD method; carrying out gradient elution by utilizing a C18 column of taking octadecylsilane chemically bonded silica as a filler, and adopting an acetonitrile-triethylamine solution as a flow phase, wherein the flow velocity is 1ml / min, and the column temperature is 25 DEG C; detecting by using an ELSD, carrying out qualitative analysis according to the retention time and the chromatogram of the sample; and carrying out quantitative analysis according to the peak area. The method has the advantages of simplicity and convenience in operation, high accuracy, and good repeatability, and can be applied to detection of sodium taurochenodeoxycholate in the bile.

Description

technical field [0001] The invention relates to a method for detecting the content of sodium taurochenodeoxycholate (Sodium taurochenodeoxycholate) from bile, which belongs to the technical field of medicine. Background technique [0002] Bile salt is the main active ingredient of bile, which mainly has the functions of clearing away heat and detoxifying, reducing phlegm and promoting dampness. [0003] Sodium taurochenodeoxycholate, molecular formula: C 26 h 44 NNaO 6 S, molecular weight 521.68, CAS number 6009-98-9, its structure is shown in the figure: [0004] [0005] The existing detection method has low sensitivity and poor repeatability, and is not suitable for detecting sodium taurochenodeoxycholate in bile. Therefore, it is necessary to explore an easier and more accurate assay method. Contents of the invention [0006] The purpose of the present invention is to provide an easy-to-operate, high-accuracy method for determining the content of sodium tauroch...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/89
Inventor 费文静钱勇丁慧诸晨许纪峰谢天培
Owner SHANGHAI NATURE STANDARD R&D & BIOTECH
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