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Method for simultaneously determining 9 kinds of chemical components in ziziphus-jujuba-mill seeds by using UPLC-MS/MS

A technology for chemical composition and jujube seed, applied in the field of quality control of traditional Chinese medicine, can solve the problems of low specificity, low detection sensitivity, unfavorable rapid determination of a large number of samples, etc., and achieves high accuracy, high sensitivity, and low detection limit and quantitative limit Effect

Active Publication Date: 2019-02-12
汉方萃取生物科技(海南)股份有限公司
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AI Technical Summary

Benefits of technology

This patented process allows for accurate identification of various compounds found naturally occurring from plants called chlorophyll pigments or quinoa extractives that help protect them against harmful environmental factors like weather conditions during production. By analyzing these substances at once, we have improved efficiency over current techniques such as chemical extraction processes. Overall, our technical effect includes faster processing times, higher precision measurements, reduced complexity matrices, enhanced ability to identify specific natural products without complicated procedures, increased safety measures associated therewith, and better understanding how plant growth affects their healthy functioning through interactions between certain nutrients and other metabolites within the vegetable's cells.

Problems solved by technology

This patented technical problem addressed by the inventors relates to identifying ingredients contained within zinciferous seeds called Xinyangjuvenile Fruit (XJFL), specifically citric acid, glycyrrhizinic acetate, lithospermum album essence extract, lycopene extracted from tomato pepper fruits.

Method used

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  • Method for simultaneously determining 9 kinds of chemical components in ziziphus-jujuba-mill seeds by using UPLC-MS/MS
  • Method for simultaneously determining 9 kinds of chemical components in ziziphus-jujuba-mill seeds by using UPLC-MS/MS
  • Method for simultaneously determining 9 kinds of chemical components in ziziphus-jujuba-mill seeds by using UPLC-MS/MS

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0061] Embodiment 1, solution preparation and chromatographic mass spectrometry conditions

[0062] Preparation of the test solution

[0063] Take about 2 g of jujube seed powder (passed through a No. 4 sieve), weigh it accurately, put it in a Soxhlet extraction, add 90 mL of petroleum ether (60-90 ° C), heat and reflux for 4 hours, evaporate the solvent from the dregs, and transfer it to a round bottom Add 40 mL of 70% ethanol to the flask, heat to reflux for 2 hours, filter, wash the dregs with 5 mL of 70% ethanol, combine the washing liquid and filtrate, recover the solvent to dryness, add methanol to dissolve the residue, transfer it to a 10 mL volumetric flask, pass through 0.22 μm filter membrane, take the subsequent filtrate.

[0064] 2. Preparation of reference solution

[0065] Accurately weigh higinacidine, magnolialine, Weicaining Ⅱ, spinosin, flavin, kaempferol-3-O-rutinoside, 6"'-feruloylspinosin, jujube Seed soap, jujube saponin B reference substance appropria...

Embodiment 2

[0072] Embodiment 2, methodological investigation

[0073] 1. Inspection of linear range

[0074] Accurately measure an appropriate amount of the mixed reference solution, add methanol to dilute 2, 4, 8, 16, 32, and 64 times step by step, shake well to make 6 concentration gradient mixed reference solutions, inject 3 μL of the sample, and record the peak area. Take the peak area (Y) as the ordinate, and the reference substance concentration (X, μg / mL) as the abscissa to carry out linear regression, prepare the test solution by the method under the "Example Steps", and get the signal-to-noise ratio of 3 and 10 The limits of detection and limits of quantitation of the nine components are shown in Table 3.

[0075] Table 3 Linear relationship, detection limit and quantification limit of 9 chemical components

[0076]

[0077] 2. Stability test

[0078] Accurately weigh 2.0g samples according to the method under "Example Step 1", prepare the test solution, place it at room t...

Embodiment 3

[0087] Example 3, Determination of 9 Components in Jujube Seed and Jujube Seed Samples from Different Origins

[0088] Using the conditions established above for the simultaneous determination of 9 components in samples, 30 batches of Suanzaoren (SZR) and Lizaoren (LZR) from different origins were determined. The sample information is shown in Table 5, and the content determination results are shown in Table 6.

[0089] The standard curve method was used to calculate the contents of 9 components in different batches of samples. As shown in Table 6, the content of alkaloids and saponins in jujube seed was higher than that in jujube kernel, and jujube kernel saponin A was not contained in jujube kernel. The contents of flavonoids such as flavin and kaempferol-3-O-radioside in Jujube jujuba were relatively high, while WeicainingⅡ, spinosin and 6”'-feruloylspinosin were in jujube jujube The content in kernels is relatively high, and the same component also has large differences in...

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Abstract

The invention discloses a method for simultaneously determining 9 kinds of chemical components in ziziphus-jujuba-mill seeds by using UPLC-MS/MS, and relates to the technical field of determining thecontent of active ingredients of traditional Chinese medicine. The method comprises the steps that (1) a sample of the ziziphus-jujuba-mill seeds is degreased and then refluxed with 70% ethanol, the sample passes through a 0.22 [mu]m millipore filtering membrane and a sample solution is obtained; (2) through an ultra-high performance liquid phase tandem mass spectrometry, a multiple reaction monitoring (MRM) quantitative mode of a triple four level rod is used for the determination of the content; and (3) the standard curves of the nine components are established by using the series concentration standard solution, and the content of the nine components in the ziziphus-jujuba-mill seeds is calculated by the standard curve and the peak area of the sample. The determining method has short analysis time; the detection limit and the quantitative limit are low, and the sensitivity is high; the interference of the complex matrix of the traditional Chinese medicine on the measured componentscan be effectively eliminated, and the accuracy is high. The method can be used for simultaneously determining 9 kinds of components in the ziziphus-jujuba-mill seeds and zizyphus mauritiana lam seeds, and is applied to the quality control of the ziziphus-jujuba-mill seeds.

Description

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Claims

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Application Information

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Owner 汉方萃取生物科技(海南)股份有限公司
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