Method for detecting biopterin in marine water body

A detection method, biopterin technology, applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of difficult to use detection methods, low content of biopterin, etc., to improve detection sensitivity, high measurement accuracy, and improve detection accuracy Effect

Active Publication Date: 2020-08-07
XIAMEN UNIV
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Problems solved by technology

Previous studies have found that biopterin-α-glucoside has a significant effect on resisting the harm of ultraviolet radiation during the growth of cyanobacteria. In order to reveal the regulation process of biopterin in the life activities of marine plankton and strengthen the understanding of marine chemistry, it is necessary to To provide a detection method for biopterin in microscopic organisms in marine water, but one of the current difficulties is that the content of biopterin in marine water is extremely low, and conventional detection methods are difficult to use

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  • Method for detecting biopterin in marine water body
  • Method for detecting biopterin in marine water body
  • Method for detecting biopterin in marine water body

Examples

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Embodiment 1

[0042] A method for analyzing and measuring biopterin in marine micro organisms, the specific steps are as follows:

[0043] 1) Pretreatment

[0044]①Collect water samples, use a graduated cylinder to accurately measure 150mL, and filter with a 0.22μm polycarbonate membrane. After filtration, take the filter membrane and place it in a 25mL centrifuge tube, and store it in a freezer at -80°C in the dark.

[0045] ②When in use, take out the preserved filter membrane to a centrifuge tube, add cell lysate, vortex for 30 minutes, ultrasonically break for 5 minutes, and then ultrasonic constant temperature water bath for 30 minutes, so that the cells are completely broken, the organic matter in the particles is dissolved, and the dissolution solution is obtained; among them, the cells The components of the lysate were 5% methanol and 95% chromatographic grade pure water, adjusted to pH = 3.5 with hydrochloric acid; after refrigeration at low temperature (4°C), the amount of sample a...

Embodiment 2

[0078] Optimization of Detection Conditions

[0079] 1. Sample pH

[0080] Weigh a number of standard samples with known biopterin concentrations of 1 μg / L, 10 μg / L and 38 μg / L, respectively, add hydrochloric acid and sodium hydroxide to adjust the pH of the sample solution to 3, 4, 5, 6, 7, 8, 9, after constant volume, extract 2 mL of supernatant after ultrasonication and centrifugation, after filtering through a 0.22 μm aqueous phase membrane, adopt step 2) reversed-phase high performance liquid chromatography in Example 1, and get the sample to be tested on the machine concentration.

[0081] The result is as Figure 5 and Figure 6 as shown, Figure 5 It is the detection result of samples with different concentrations under different pH conditions of samples in the condition optimization experiment of another embodiment of the present invention. Figure 6 It is the recovery rate result of samples with different concentrations under different pH conditions in the condi...

Embodiment 3

[0090] The present embodiment is based on the detection of biopterin in marine water body through field research.

[0091] 1. Collection and preservation of marine microbiological samples

[0092] In April 2017, the natural water bodies of the Jiulong River Estuary and Xiamen Bay were investigated. The voyages of the Kowloon Estuary and Xiamen Bay used GO-FLO water collectors to collect surface water at a water depth of 0.5-1.0 meters. Accurately measure 150mL of water sample with a graduated cylinder, first pre-filter it with a 10μm sieve, and then filter it through a 0.22μm polycarbonate membrane.

[0093] 2. Pretreatment of particulate matter samples

[0094] (1) Broken: Take the filter membrane to a 10mL centrifuge tube, add 4mL of cell lysate (5% methanol, 95% pure water, hydrochloric acid to adjust to pH=3.5), shake for 5min, cool in crushed ice for 30 seconds, and repeat Six times, after the oscillation, each sample was ultrasonically crushed for five minutes, and ea...

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Abstract

The invention provides a method for detecting biopterin in a marine water body. The method comprises the following steps of: performing pretreatment, specifically, filtering a water sample, discardinga filtrate, taking particulate matters, adding a cell lysis solution crush the particulate matters to obtain a dissolved liquid, adjusting the pH value of the dissolved liquid, and performing extraction to obtain a water phase containing biopterin; and 2) detecting the water phase containing the biopterin by using reversed-phase high performance liquid chromatography. According to the reversed-phase high performance liquid chromatography, isocratic elution is adopted, and mobile phases are methanol and water. According to the method for detecting the content of the biopterin in the marine water body provided by the embodiment of the invention, the pretreatment under specific conditions is carried out; detection sensitivity is effectively improved; the detection limits of the method are further reduced; the method can adapt to the current situation that the content of the biopterin in the marine water body is low; the method has high accuracy; and the method has important significancein revealing the regulation and control process of biopterin in marine plankton, especially the life activity of microorganisms, and strengthening the understanding of marine chemistry.

Description

technical field [0001] The invention relates to the technical field of biological detection, in particular to a method for detecting biopterin in marine water. Background technique [0002] Biopterin (BTP) is a metabolite of guanosine triphosphate (GTP), and GTP cyclohydrolase I, which controls the conversion of GTP into pterin, is found in both prokaryotes and eukaryotes. In nature, biopterin can be synthesized from bacteria to higher animals and plants (except insects). As an endogenous coenzyme factor of more than 70 kinds of enzymes, biopterin participates in the life activities of animals, plants, bacteria and fungi. Biopterins include nitrogenase, nitric oxide synthase, nitrate reductase, sulfite oxidase, carbon monoxide oxidoreductase, phenylalanine hydroxylase, tyrosine hydroxylase, tryptophan hydroxylase Enzymes and essential coenzyme factors for anabolic processes such as folic acid and vitamins have important physiological functions and are crucial in the marine...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/06G01N30/74G01N30/89
CPCG01N30/06G01N30/74G01N30/89
Inventor 梅康石梦秋王德利
Owner XIAMEN UNIV
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