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A method for determining the total alkaloid content of cloverleaf alkaloids in cloverleaf plants

A technology of harringtonine and harringtonine, which is applied in the field of alkaloid total alkali content testing, can solve the problems of lack of a method for measuring the total alkaloids of harringtonine alkaloids and the like, and achieves the advantages of shortened reaction time, high detection sensitivity and rapid experimentation. Effect

Active Publication Date: 2018-10-30
TROPICAL CORP STRAIN RESOURCE INST CHINESE ACAD OF TROPICAL AGRI SCI +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is still a lack of safe, fast and accurate assay methods for the total alkaloids of harrington alkaloids

Method used

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  • A method for determining the total alkaloid content of cloverleaf alkaloids in cloverleaf plants
  • A method for determining the total alkaloid content of cloverleaf alkaloids in cloverleaf plants
  • A method for determining the total alkaloid content of cloverleaf alkaloids in cloverleaf plants

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] The inventive method precision, stability, accuracy and recovery rate situation

[0041] (1) Stability test

[0042] (1) Take the leaves and twigs of Torreya hainanensis respectively, dry them at 105°C for 30 minutes, and then dry them at 80°C for 12 hours. After pulverizing the samples, filter them with a 40-mesh sieve to obtain dry powder.

[0043] (2) Weigh 3g of leaf and twig dry powder respectively, add 2mL of ammonia water (28%), add 30mL of chloroform, stir well with a vibrating agitator, soak overnight, centrifuge for 15min (3000r / min), take 10mL of chloroform layer solution, ventilate Chloroform was evaporated to dryness in the cupboard, the residue was dissolved in methanol, transferred to a 1mL volumetric flask to constant volume, and repeated three times.

[0044](3) Take 50 μL of leaf and twig sample solution of Torreya hainanensis and 100 μL of harringtonine standard solution (2 mg / mL, prepared with methanol, purchased from Shanghai Yuanye Biotechnology C...

Embodiment 2

[0064] Determination of the total alkaloid content of Herringbone alkaloids in leaves and shoots of Torreya hainanensis

[0065] (1) Take the leaves and twigs of Torreya hainanensis respectively, kill them in an oven at 105°C, then dry them at 80°C for 16 hours, grind them into fine powder, pass through a 40-mesh sieve, and obtain the crude drug;

[0066] (2) Weigh 3g of leaf and twig dry powder, add 2mL of ammonia water (28%), add 30mL of chloroform, stir evenly with a vibrating agitator, soak overnight, centrifuge for 15min (3000r / min), take 10mL of chloroform layer solution, and place in a fume hood Chloroform was evaporated to dryness, the residue was dissolved in methanol, quantitatively transferred to a 1mL small volumetric flask for later use, and repeated 3 times.

[0067] (3) Take 50 μL of leaf and twig sample solution, add freshly prepared chromotropic acid solution 300 μL (0.1g / ml, filter), add concentrated phosphoric acid 3mL, hydrogen peroxide 70 μL (0.025mol / L), ...

Embodiment 3

[0073] Determination of the total alkaloid content of Herringbone alkaloids in suspension culture cells of Torreya hainanensis

[0074] (1) Take 10 mL of suspension cultured cells and put them into a 50 mL centrifuge tube, centrifuge for 15 min (2000 rpm), remove the supernatant, and weigh 4 g of cells;

[0075] Among them, the reference for the cell culture process of Torreya hainanensis: Chen Lin, Yongcheng Li. Establishment of cell suspension culture system of Torreya hainanensis. Guangdong Agricultural Science, 2014, 24:52-57.

[0076] (2) Add 1 mL of ammonia water, add 15 mL of chloroform, shake and stir for 1 min, sonicate for 30 min (80 Hz, 30 min), soak overnight, filter, take 6 mL of filtrate, evaporate chloroform to dryness in a fume hood, dissolve the residue with methanol, and dilute to 1 mL Volumetric flask;

[0077] (3) Take 100 μL of the sample solution after constant volume, put it into a 50 mL beaker, evaporate methanol to dryness, add 300 μL of freshly prepa...

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Abstract

The invention discloses a method for determining the total alkali content of cephalotaxus alkaloids in a cephalotaxaceae plant. The method comprises the following steps that 1, a standard cephalotaxus alkaloid product is selected, a chromotropic acid solution, strong phosphoric acid and hydrogen peroxide are added, even shaking is performed, and then microwave heating is performed to obtain a standard production reaction solution; 2, the standard production reaction solution is cooled to reach room temperature, volume setting and color comparison are performed, a light absorption value obtained through color comparison is associated with the concentration of the standard cephalotaxus alkaloid solution in the step 1, and a standard light absorption value-concentration curve of the cephalotaxus alkaloids is established; 3, suspension culture cells of a cephalotaxaceae plant sample or the cephalotaxaceae plant are selected, and the total alkali of the cephalotaxus alkaloids of the cells is extracted; 4, the light absorption value is measured by referencing the steps 2 and 3, the measured light absorption value is substituted into the standard light absorption value-concentration curve to obtain the concentration of the cephalotaxaceae plant sample, and the total alkali content of the cephalotaxus alkaloids in the cephalotaxaceae plant sample is obtained through conversion. The method is simple, convenient, safe, quick and accurate.

Description

technical field [0001] The invention belongs to the technical field of testing the total alkaloid content of alkaloids, and in particular relates to a method for measuring the total alkaloid content of the cloverleaf alkaloids in the cloverleaf plants. Background technique [0002] Cephalotaxaceae (Cephalotaxaceae) only has one genus of Cephalotaxaceae, which has 7 species and 2 varieties, including C.oliveri, C.harringtonia, and C.harringtonia (C.Latifolia), rough torreya (C.sinensis), Hainan rough torreya (C.mannii), three-pointed fir (C.fortunei), alpine three-pointed fir (C.fortuneivar.alpine), Taiwan thick torreya (C. sinensis var.wilsoniana), Gongshan three pointed fir (C.lanceolata). The undergraduates are distributed in the south of East Asia and the Indochina Peninsula, and are mainly produced in China. Scattered in the east of the Hengduan Mountains, the Qinling Mountains to the Dabie Mountains, and the provinces and regions south of southern Jiangsu and Taiwan. ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/31
CPCG01N21/31
Inventor 丛汉卿江雪飞乔飞王峰王祝年
Owner TROPICAL CORP STRAIN RESOURCE INST CHINESE ACAD OF TROPICAL AGRI SCI
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