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Method for detecting 9 kinds of mycotoxins in cassia seed medicinal material

A mycotoxin and detection method technology, which is applied in the field of detection of 9 kinds of mycotoxins in cassia medicinal materials, can solve the problems of pollution and non-detection of mycotoxins, etc.

Inactive Publication Date: 2018-06-01
TIANJIN TASLY PHARMA CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Cassia seeds are most susceptible to aflatoxin contamination during storage, but contamination of cassia seeds with other mycotoxins has not been reported in the existing literature
In the 2015 edition of "Chinese Pharmacopoeia", only the detection of aflatoxin is recorded, and it is limited to contain aflatoxin B per 1000g of cassia seeds 1 , no more than 5μg, aflatoxin G 2 , Aflatoxin G 1 , Aflatoxin B 2 and aflatoxin B 1 The total amount should not exceed 10 μg. Existing methods only detect aflatoxin-related toxins and do not detect other mycotoxins

Method used

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  • Method for detecting 9 kinds of mycotoxins in cassia seed medicinal material
  • Method for detecting 9 kinds of mycotoxins in cassia seed medicinal material
  • Method for detecting 9 kinds of mycotoxins in cassia seed medicinal material

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0069] Embodiment 1: the method for detecting 9 kinds of mycotoxins in Semen Cassiae

[0070] 1. Positive ion mode:

[0071] Chromatographic column: ACQUITY UPLC BEH C18 column (100mm×2.1mm, 1.7μm); column temperature: 40°C; flow rate: 0.3mL / min; injection volume: 3.5μL; mobile phase A: acetonitrile, mobile phase B: 0.1% (v / v) formic acid water; gradient elution program: 0~8.0min, 27%A~77%A; 8.0~9.0min, 77%A~100%A; 9.0~10min, 100%A~27%A .

[0072] Ion source: electrospray ion source (ESI); scan method: positive ion scan; detection method: multiple reaction monitoring (MRM) mode; ionization voltage (IS): 5500V; atomization temperature (TEM): 550 ° C; air curtain gas (CUR): 35L / min; spray gas (Gas1): 55L / min; auxiliary heating gas (Gas2): 55L / min; collision gas pressure (CAD): medium.

[0073] Negative ion mode:

[0074] Chromatographic column: ACQUITY UPLC BEH C18 column (100mm×2.1mm, 1.7μm); column temperature: 40°C; flow rate: 0.3mL / min; injection volume: 2.0-5.0μL; mobil...

Embodiment 2

[0089] Embodiment 2: method screening

[0090] 1. MRM-IDA-EPI method and spectral library establishment

[0091] Use MRM as a probe scan to trigger IDA to work. When the MRM signal threshold is greater than 1000cps set by IDA, the EPI scanning function is started at the same time. The scanning range of EPI is: 250~800Da; the scanning rate is: 10000Da / s. The mixed standard solution of 9 kinds of mycotoxins was scanned by EPI. The collision energy (Collision Energy Spread, CES) of the product ion of the compound was set to 15V. Collect 1 EPI spectrum, get 1 comprehensive EPI spectrum after averaging, input the EPI spectrum of each mycotoxin into the database, and fill in the chemical name, relative molecular weight, molecular formula, chemical structure and American Chemical Abstracts (Chemical Abstracts) of each compound at the same time. AbstractsService, CAS) registration number, to establish a standard spectrum library of 9 kinds of mycotoxins.

[0092] 2. Optimization o...

Embodiment 3

[0101] Example 3: Methodological Validation

[0102] 1. Linear

[0103] Prepare matrix matching standard working solution according to Example 1, after LC-MS / MS is measured, draw standard curve with each component peak area (y) to each component mass concentration (x, μ g / L), the results are shown in the table 2.

[0104] Table 2 Linearity, detection limit and quantification limit of 9 mycotoxins

[0105]

[0106]

[0107] Table 2 result shows: all show good linear relationship between response value and concentration, correlation coefficient (r 2 ) are greater than 0.993.

[0108] 2. Limit of detection and limit of quantitation

[0109] Take the lowest concentration point of the standard curve, quantitatively dilute it with a blank matrix and analyze it. The concentration when the signal-to-noise ratio (S / N) ≥ 10 is the LOQ, and the concentration when the signal-to-noise ratio (S / N) ≥ 3 is the LOD. The measurement results are shown in Table 2.

[0110] 3. Repeatab...

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Abstract

The invention relates to a method for detecting 9 kinds of mycotoxins in a cassia seed medicinal material. The method comprises the following steps: a step 1: preparation of a standard substance stocksolution; a step 2: preparation of a mixed standard substance stock solution; a step 3: preparation of a standard working solution; a step 4: preparation of a tested object solution; a step 5: preparation of a substrate matching standard solution; a step 6: determination.

Description

technical field [0001] The invention relates to a method for detecting the types and contents of fungi in Chinese medicinal materials, in particular to a method for detecting nine types of mycotoxins in cassia seed medicinal materials. Background technique [0002] Mycotoxins are toxic secondary metabolites produced by fungi under suitable environmental conditions, mainly including aflatoxins, ochratoxin A, zearalenone, trichothecenes, fumonisins, and aflatoxins Chlorotoxins and ergot alkaloids, these mycotoxins can widely contaminate crops, plants and their by-products. [0003] The introduction of common mycotoxins is as follows: Aflatoxin, discovered in 1960, is 10 times as toxic as potassium cyanide and 68 times as toxic as arsenic. The Cancer Research Institute of the World Health Organization has designated it as a class of carcinogens; fumonisins induce the occurrence of human esophageal cancer, liver cancer, and gastric cancer; 3-acetyl deoxynivalenol, deoxynivaleno...

Claims

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Application Information

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IPC IPC(8): G01N30/02
CPCG01N30/02
Inventor 佟玲李东翔李爽段玺玉
Owner TIANJIN TASLY PHARMA CO LTD
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