Sea pumpkin saponin content determination method

A technology of sea cucumber saponins and determination methods, applied in the directions of measuring devices, instruments, scientific instruments, etc., can solve the problems of sample loss, measurement result errors, large differences in sea cucumber saponins, etc., and achieve stable peak area, short detection time, and high sensitivity. Effect

Inactive Publication Date: 2008-07-09
OCEAN UNIV OF CHINA
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

In addition, sea cucumber also contains a large amount of complex components such as protein, polysaccharide, fat, trace elements, etc., which will inevitably bring many difficulties to the determination of saponin content in sea cucumber
[0003] The patent application No. 200510014471.9 discloses a method for the determination of bitter melon saponins. This method is not very selective for the target measured substance, and the impurity components in the sample are likely to cause large measurement errors, and usually require more complicated sample pretreatment. process, while macroporous resin, silica gel and other column chromatography processes will inevitably cause the loss of samples, which will easily bring further errors to the measurement results
The patent application number 2005100

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0009] (1) Weigh 500.8 mg of commercially available dry sea cucumber A (Apostichopus japonicus), crush it, add 1 mL of 2 mol / L trifluoroacetic acid aqueous solution, and hydrolyze it at 120 ° C for 2 hours. The aqueous solution was neutralized, filtered, and the volume was adjusted to 5mL as the test solution.

[0010] (2) Weigh quinolose and make a solution of 1 mmol / L with water as a stock solution. Further diluted successively to 0.01mmol / L, 0.05mmol / L, 0.1mmol / L, 0.2mmol / L, 0.4mmol / L, 0.6mmol / L, 0.8mmol / L, 1.0mmol / L, prepared (8 concentrations Gradient standard aqueous solution.

[0011] (3) Take 250 μL of the standard aqueous solution of the above concentration respectively, add 250 μL of a methanol solution of 1-phenyl-3-methyl-5-pyrazolone with a concentration of 0.5 mol / L and 250 μL of 0.3 mol / L hydrogen Sodium oxide aqueous solution, derivatized at 70°C for 30 minutes, cooled for 10 minutes, neutralized with 250 μL 0.3mol / L hydrochloric acid aqueous solution; added ...

Embodiment 2

[0014] (1) Weigh 75.4 mg of commercially available dried sea cucumber B (sea cucumber B. dermatitis), crush it, add 0.2 mL of 2 mol / L trifluoroacetic acid aqueous solution, and hydrolyze it at 120 ° C for 2 hours, and neutralize the hydrolyzed solution with 5 mol / L sodium hydroxide solution , and dilute to 10mL as the test solution.

[0015] (2) Obtain working curve y=10219x+9.0095 (R 2 =0.9995), wherein x is the concentration of quinolose standard solution, and y is the peak area of ​​quinolose.

[0016] (3) Draw 250 μ L of the sample test solution of sea cucumber B, carry out derivation reaction and liquid chromatographic analysis according to the same method as Example 1, record the quinolose peak area as 2784.27, bring into the working curve to calculate the quinolose in the sample The content of sea cucumber B is 0.591%, multiplied by the conversion coefficient of sea cucumber B (leathered sea cucumber), the saponin content in sea cucumber B can be obtained. Sea cucumbe...

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Abstract

The invention discloses a method for determining holothurin content, which is characterized in that the method comprises the following steps of: hydrolyzing a sample with a trifluoroacetic acid aqueous solution, neutralizing the hydrolysate, filtering, adding water to fixed volume, adding 1-pheny-3-methy-5-pyrazolone derivative agent to carry out derivation reaction, determining the content of quinose in the derivatives by high-performance liquid chromatographic analysis, and calculating the holothurin content in the sample by multiplying the content of quinose by a conversion coefficient. The invention has the advantages of simple operation, high sensitivity, less sample consumption and good stability, and is suitable for content determination of glycosides of various sea cucumbers.

Description

technical field [0001] The invention relates to a method for determining biologically active substances, in particular to a method for determining the content of sea cucumber saponins. Background technique [0002] Sea cucumber saponin is an important biologically active substance in sea cucumber, and there are more than 100 kinds of sea cucumber saponins that have been discovered so far. The structure of saponins in sea cucumbers is complex and varies with different species of sea cucumbers and growth environments. Different species of sea cucumbers have different structures of saponins, and the same species of sea cucumber usually contains several or even dozens of saponins with similar structures and subtle differences. In addition, sea cucumber also contains a large amount of complex components such as protein, polysaccharide, fat, and trace elements, which will inevitably bring many difficulties to the determination of saponin content in sea cucumber. [0003] The pat...

Claims

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Application Information

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IPC IPC(8): G01N30/02
Inventor 董平薛长湖于林芳李兆杰徐杰
Owner OCEAN UNIV OF CHINA
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