Establishment method of inducing callus cultured Allium sativum L. enzymatic hydrolysate HPLC fingerprint

An enzymatic hydrolysis product and callus technology, which is applied in the field of establishment of HPLC fingerprints of garlic enzymatic hydrolysis products of inducing callus culture, and achieves the effects of simple and easy chromatographic conditions, little interference of chromatographic signals, and accurate internal quality and quality.

Inactive Publication Date: 2015-12-16
TIANJIN NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0009] The technical problem to be solved in the present invention is to provide a method for establishing the HPLC fingerprint of induced callus cultured garlic in view of the deficiencies of the existing quality management detection methods for induced callus cultured garlic

Method used

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  • Establishment method of inducing callus cultured Allium sativum L. enzymatic hydrolysate HPLC fingerprint
  • Establishment method of inducing callus cultured Allium sativum L. enzymatic hydrolysate HPLC fingerprint
  • Establishment method of inducing callus cultured Allium sativum L. enzymatic hydrolysate HPLC fingerprint

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Effect test

Embodiment 1

[0027] Embodiment 1: the method for establishing the HPLC fingerprint of the garlic enzymatic hydrolyzate of induced callus culture, the method steps are as follows:

[0028] (1) Samples, kits and instruments: Agilent 1200 high-performance liquid chromatography was used to induce callus to culture garlic, and the reagents were all chromatographically or analytically pure;

[0029] (2) Preparation of the test solution of garlic enzymatic hydrolysis product: Weigh 20g of garlic bulbs, crush and grind, add 25-30ml of distilled water, enzymatically hydrolyze in a warm bath at 50-55℃ for 1h, add methanol to dilute to 50ml, centrifuge at 5000×g for 15min , and the supernatant was filtered through a 0.45 μm microporous membrane;

[0030] (3) Preparation of the test solution of garlic enzymatic hydrolysis product: Activate the C18 solid-phase extraction column with 5ml methanol and 10ml ultrapure water, and equilibrate for 15min; Rinse and dry under low vacuum; then elute with 5ml of...

Embodiment 2

[0036] The establishment method of the HPLC standard fingerprint of the garlic enzymatic hydrolyzate of induced callus culture, the method steps are as follows:

[0037] (1) Samples, kits and instruments: A total of 10 batches of garlic were induced for callus culture, using Agilent 1200 high-performance liquid chromatography, and all reagents were chromatographically or analytically pure;

[0038] (2) Preparation of the test solution of garlic enzymatic hydrolysis product: crush and grind 20g of garlic bulbs, add 25-30ml of distilled water, enzymolyze in a warm bath at 50-55℃ for 1h, add methanol to 50ml, centrifuge at 5000×g for 15min, and take The supernatant was filtered through a 0.45 μm microporous membrane;

[0039] (3) Preparation of the test solution of garlic enzymatic hydrolysis product: Activate the C18 solid-phase extraction column with 5ml methanol and 10ml ultrapure water, and equilibrate for 15min; Rinse and dry under low vacuum; then elute with 5ml of methano...

Embodiment 3

[0047] Utilize to carry out chromatographic analysis to the HPLC standard fingerprint of induced callus culture garlic enzymatic hydrolyzate and its parent and other varieties garlic, the method steps are as follows:

[0048] (1) Samples, kits and instruments: parent Baodi "six petals red" garlic, Xinjiang white garlic, using Agilent 1200 high-performance liquid chromatography, reagents are chromatographically pure or analytically pure;

[0049] (2) Preparation of the test solution of garlic enzymatic hydrolysis products: Weigh 20g of garlic bulbs, crush and grind them, add 25-30ml of distilled water, enzymatically hydrolyze in a warm bath at 50-55℃ for 1h, add methanol to 50ml, and centrifuge at 5000×g After 15 minutes, the supernatant was filtered through a 0.45 μm microporous membrane;

[0050] (3) Preparation of the test solution of garlic enzymatic hydrolysis product: Activate the C18 solid-phase extraction column with 5ml methanol and 10ml ultrapure water, and equilibrat...

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Abstract

The invention discloses an establishment method of inducing callus cultured Allium sativum L. enzymatic hydrolysate HPLC fingerprints. The establishment method of the inducing callus cultured Allium sativum L. enzymatic hydrolysate HPLC fingerprints specifically comprises the steps of preparing inducing callus cultured Allium sativum L. bulbs into solution for test products through steps such as solid-phase extraction, and performing high-performance chromatographic analysis and detection under certain conditions to obtain HPLC fingerprints of inducing callus cultured Allium sativum L. enzymatic hydrolysate. By analyzing the HPLC fingerprints of 10 batches of samples, characteristic common peaks are determined and standard fingerprints are obtained. The standard fingerprints can provide a reliable basis for variety identification and production quality control of inducing callus cultured Allium sativum L. The establishment method of the inducing callus cultured Allium sativum L. enzymatic hydrolysate HPLC fingerprints has the advantages that the operation is convenient, the stability is high, the specificity is strong, the repeatability is good, the characteristic peaks are more, the analysis efficiency is high, the truth identification of the inducing callus cultured Allium sativum L. can be realized by adopting the invention, and the interior product quality can be comprehensively, accurately and effectively evaluated.

Description

[0001] The invention was funded by the "Key Project of Tianjin Agricultural Commission (201302030) and the Application Development Research Fund of Tianjin Normal University". technical field [0002] The invention belongs to a method for constructing plant fingerprints for induced callus culture, in particular to a method for constructing high-performance liquid chromatography (HPLC) fingerprints for induced callus cultured garlic and standard fingerprints thereof. Background technique [0003] garlic( Allium sativum L. ) Liliaceae Allium, biennial herbaceous plant, food and medicine, is widely planted in the world. At present, my country is the largest garlic producer, with 692,000 hectares of garlic planted in China, accounting for about 1 / 2 of the world's planting area. The total output of garlic in my country accounts for more than 3 / 4 of the world's total output, with an output of 12.088 million tons. As an indispensable vegetable and condiment in people's life, garli...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02G01N30/06
Inventor 王振英任春雪尚云涛范宝莉刘晓颖党光彭永康陈宏王中良
Owner TIANJIN NORMAL UNIVERSITY
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