Preparation and application of test paper for testing royal jelly freshness

A technology for testing test strips and royal jelly, applied in biological testing, measuring devices, material inspection products, etc., can solve problems such as loss of functional components, judging the freshness and quality of royal jelly, and deterioration.

Inactive Publication Date: 2008-06-25
BEE RES INST CHINESE ACAD OF AGRI SCI +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0002] Since fresh royal jelly is usually stored at a temperature of -20°C or lower to maintain its active ingredients, if the storage temperature is improper, the fu...

Method used

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  • Preparation and application of test paper for testing royal jelly freshness
  • Preparation and application of test paper for testing royal jelly freshness
  • Preparation and application of test paper for testing royal jelly freshness

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0165] Two-dimensional electrophoresis separation and mass spectrometry analysis of royal jelly protein

[0166] The royal jelly under different storage conditions was extracted by phosphoric acid extraction buffer (PB) and protein sample dissolution buffer (LB), respectively, and trichloroacetic acid was added to the extract to precipitate the protein, and the whole protein of royal jelly under different storage conditions was obtained. The protein is redissolved and the concentration is detected, then two-dimensional electrophoresis (solid-phase pH gradient, 17cm gel strip, pH3-10) is performed according to a certain loading amount, and the electrophoretic spectrum ( figure 1 ) is analyzed with professional analysis software. The target protein spots with high expression abundance were digested for mass spectrometry analysis, and the mass spectrometry results were submitted to a professional database for sequence comparison for protein identification ( figure 2 ). The ana...

Embodiment 2

[0168] Chromatographic Separation and Mass Spectrometry Analysis of Royal Jelly Protein

[0169] Royal jelly stored under different storage conditions was precipitated with acetone and redissolved in alkaline buffer (pH8.2) to obtain the total protein of fresh royal jelly, royal jelly stored at -20°C for one year, royal jelly stored at 4°C for one year, and royal jelly stored at room temperature for one year , the above protein samples were subjected to molecular sieve chromatographic separation ( image 3 ), it is determined that there are significant differences in the components with molecular weights from 10K to 100K. The fourth fraction (Re=17.34min) was selected and subjected to trypsin digestion, sodium upgrading chromatographic separation and mass spectrometry sequence analysis (Nano LC MS / MS, Table 1), and it was confirmed that royal jelly protein 5 can only be stored in fresh royal jelly and at -20°C detected in royal jelly.

Embodiment 3

[0171] Detect target protein gene clone expression

[0172] The respective forward and reverse primers were synthesized according to the sequences of apal jelly protein 4, apal jelly protein 5 and glucose oxidase provided in the NCBI database, and PCR amplification was carried out using the DNA of the head of the honeybee worker bee as a template. Connect the amplified target fragment with an appropriate carrier (T plasmid, kit) to form a recombinant, and then introduce the recipient bacteria (E. Nucleotide sequence analysis. The recombinants confirmed by sequencing were recombined into expression vectors and induced to express them. After the expression product was isolated and purified, it was detected by mass spectrometry.

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Abstract

The invention relates to a royal jelly freshness detection test paper product and a preparation and application method thereof, which belong to the medicine, foodstuff and biotechnological field. As proved by proteomic technology recently, glucose oxidases, royal jelly proteins 4 and royal jelly proteins 5 in royal jelly can be taken as freshness protein labelings. The invention realizes detection of royal jelly quality with quick speed, low cost and high sensitivity through preparation of a nano colloidal gold detection test paper for detecting the proteins. The detection test paper can be used in the aspects like royal jelly raw material quality identification, product quality control and activity value determination during manufacturing, marketing and using processes, and also can be used for detecting royal jelly content in other relative products. Over 90 percent royal jelly in the world is made in China, and annual output value of the royal jelly is over 2 billion; moreover, with improvement of living standard of people, the royal jelly gradually becomes an important natural health product, thereby development of quick and easy variety detection methods is imperative.

Description

technical field [0001] The present invention relates to a detection device for detecting the freshness of royal jelly in the fields of medicine, food and biotechnology, its preparation and application method, more specifically, relates to a simple, quick and effective method for judging the freshness of royal jelly Detection test paper, its preparation and application method. Background technique [0002] Since fresh royal jelly is usually stored at a temperature of -20°C or lower to maintain its active ingredients, if the storage temperature is improper, the functional ingredients will be lost or deteriorated and lose commercial value, but there is no simple, fast and effective method to judge Freshness and quality of royal jelly. We recently proved that glucose oxidase, apal jelly protein 4 and apal jelly protein 5 in royal jelly can be used as protein markers of freshness by proteomics technology. [0003] Because nanoscale colloidal gold (Colloidal gold) has a high ele...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/52G01N33/577G01N33/558
Inventor 李建科潘映红国占宝冯毛张朝晖
Owner BEE RES INST CHINESE ACAD OF AGRI SCI
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