Method for fast screening and preparing spice

A spice, fast technology, applied in the direction of biochemical equipment and methods, microbial determination/inspection, etc., can solve the problem of spice toxicity, chronic toxicity, not well studied, and safety, etc., to reduce development costs and development costs. The effect of shortening the test cycle and improving the detection efficiency

Active Publication Date: 2012-10-17
YUNNAN RES INST OF TOBACCO SCI
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  • Summary
  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

People use these substances in large quantities in food without considering their safety, and...

Method used

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  • Method for fast screening and preparing spice
  • Method for fast screening and preparing spice
  • Method for fast screening and preparing spice

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] 1 Experimental part

[0021] 1.1 Instruments and materials

[0022] Series II CO2 incubator (Thermo Forma Company); inverted microscope (Olympus Company); microplate reader (Bio-Rad Company); 96-well cell culture plate, cell culture flask (Corning Company).

[0023] 1.2 Reagents

[0024] DMEM medium, fetal bovine serum (Gibco); pH 7.2 phosphate buffer (CMF-PBS), trypsin, trypan blue, penicillin-streptomycin (Solarbio); neutral red dye (Sigma) ; MTT (Sigma, USA); Dimethyl sulfoxide (DMSO, cell culture grade, Biomol); Formaldehyde (AR grade); Absolute ethanol (AR grade); Glacial acetic acid (AR grade); Lauryl Sodium sulfate (SDS, AR grade).

[0025] 1.3 Preparation method of test plant extract sample

[0026] Weigh 500 g of dried jujube samples, crush them, and reflux extract three times with ethanol solvent 3 times that of the samples. The extract was concentrated under reduced pressure by a rotary evaporator to obtain jujube extract. Evaporate the solvent from the...

Embodiment 2

[0051] Weigh 1000 g of the dried Herba sativa sample, pulverize it, and extract it with 95% ethanol solvent 3 times that of the sample under reflux. The extract was concentrated by a rotary evaporator under reduced pressure to obtain Lingxiangcao extract (Lingxiangcao total extract, LXC-0).

[0052] According to the experimental part of Example 1, the neutrophil toxicity test was carried out on the extract of Lingxiangcao.

[0053] The result shows that the IC of the cell lethal dose of Lingxiang herbal extract 50 It was 56.32 (μg / mL). When the dose of Lingxiangcao extract was 50 μg / mL, its cytotoxicity grade was grade 2, which was mild cytotoxicity; when the dose was 100 μg / mL, its cytotoxicity grade was grade 3, which was moderate cytotoxicity. In order to reduce the cytotoxicity of the herb extract and improve the safety of herb extract. Through different extraction and separation techniques, the total extract of natural fragrance raw materials is divided into different ...

Embodiment 3

[0059] Different extraction techniques were used to prepare ling vanilla extracts.

[0060] Weighed 500 g of the dried Herba spirulina sample, crushed it, extracted three times with petroleum ether solvent 3 times that of the sample, and concentrated the extract under reduced pressure in a rotary evaporator to obtain the petroleum ether extract of Herb spirulina. According to the experimental part of Example 1, the neutral erythrocyte toxicity test was carried out on the extract of spirit vanilla petroleum ether. The results show that the IC of the half lethal dose of the spirit vanilla petroleum ether extract 50 It was 328.11 (μg / mL). Compared with Ling vanilla crude paste (ethanol extract), the safety of the product has been improved.

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Abstract

The invention relates to a method for fast screening and preparing spice, particularly relates to a method for fast screening and preparing safe spice based on a cell toxicity testing technology and an extraction separation technology and belongs to the field of spice research and development. The method disclosed by the invention comprises the steps of: carrying out in-vitro cell toxicity test on the initial extractum of the spice by a neutral red uptake assay or an MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) method on the basis of combining the existing in-vitro cell toxicity testing technology with the extraction separation technology; separating the initial extractum of the spice into different extracts by the existing extraction separation technology, carrying out in-vitro cell toxicity test on each extract, and finding out the main source of cell toxicity; and removing the poisonous part and preparing the safe spice by the existing extraction separation technology. The method has the characteristics that the usage of animals can be effectively reduced; manpower and material resources are saved; pain of a living animal is reduced; the test period is shortened; and the detection efficiency is improved, and the development test cost of the spice is reduced.

Description

Technical field: [0001] The invention relates to a method for quickly screening and preparing spices, in particular to a method for rapidly screening and preparing safe spices based on cytotoxicity detection technology and extraction and separation technology. It belongs to the research and development field of spices. Background technique: [0002] There are more than 1,700 kinds of edible spices, which are the largest family of food additives. Among them, about 1,300 are natural substances extracted from spice plants that are commonly eaten by humans, and have a safe edible history in special or common areas. Because these substances are used in small amounts and are highly volatile, it is difficult to cause damage to the human body, so most of the natural food flavors are considered safe (GRAS substances). People use these substances in food in large quantities without considering their safety, and in fact, the toxicity, especially the chronic toxicity, of this kind of s...

Claims

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Application Information

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IPC IPC(8): C12Q1/02
Inventor 杨叶昆李雪梅米其利夭建华魏玉玲徐济仓陈建华
Owner YUNNAN RES INST OF TOBACCO SCI
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