Pretreatment method for analyzing content of microcystin in seaweed

Abstract

The invention is applicable to the technical field of detection of the content of microcystic toxins in seaweeds, and provides a pretreatment method for analyzing the content of microcystic toxins in seaweeds, which comprises the following steps: S1, cleaning seaweeds with pure water, drying, crushing, grinding into powder, adding into a beaker, then adding an extracting solution, heating to 40-50 DEG C, and carrying out ultrasonic oscillation for 3-4 hours to obtain the extracting solution; s2, centrifuging the extracting solution, and taking supernate; s3, concentrating the supernate, pouring into an SPE column, adding an eluting agent, carrying out solid-phase extraction, collecting eluent, and concentrating for detection; preferably, the extracting solution in the step S1 is a mixed solution of methanol and pure water, and the mixing volume ratio of the methanol to the pure water is (65-75): 1. The pretreatment method for analyzing the content of the microcystic toxins in the seaweed, provided by the invention, is short in period, few in steps, simple to operate and high in accuracy of a detection result, and greatly facilitates rapid detection of the microcystic toxins.

Description

Technical Field

[0001] The invention relates to the technical field of microcystin content detection in seaweed, and in particular to a pretreatment method for analyzing the microcystin content in seaweed. Background Art

[0002] In the current field of marine environmental monitoring and food safety, the detection of microcystins in seaweed is particularly important. Microcystins are a class of toxic secondary metabolites produced by cyanobacteria (also known as blue-green algae), which pose a threat to aquatic life and human health.

[0003] Therefore, it is very important to detect the content of microcystin in seaweed. The detection methods of microcystin mainly include liquid chromatography-mass spectrometry (LC-MS / MS), enzyme-linked immunosorbent assay (ELISA), etc. Before detection and analysis, pretreatment is often required. The existing pretreatment steps are complicated, the operation is cumbersome, the cost is high, and the detection results are not accurate enough. Summary of the invention

[0004] The object of the present invention is to provide a pretreatment method for analyzing the content of microcystin in seaweed to solve the problems in the above-mentioned background technology.

[0005] To achieve the above object, the present invention provides the following technical solution: a pretreatment method for analyzing the content of microcystin in seaweed, comprising the following steps:

[0006] S1, wash the seaweed with pure water, dry, crush, grind into powder, add into a beaker, then add extract, heat to 40-50°C, and ultrasonically vibrate for 3-4h to obtain extract;

[0007] S2, centrifuging the extract and taking the supernatant;

[0008] S3, concentrate the supernatant, pour it into the SPE column, add the eluent, perform solid phase extraction, collect the eluate, concentrate it, and wait for detection.

[0009] Preferably, the drying temperature in S1 is 45°C to 50°C.

[0010] Preferably, the extract in S1 is a mixture of methanol and pure water, and the mixing volume ratio of methanol to pure water is 65 to 75:1.

[0011] Preferably, the centrifugation time in S2 is 8 to 10 min, the centrifugation temperature is 5 to 10° C., and the centrifugation speed is 10,000 r / min.

[0012] Preferably, the eluent in S3 is a methanol solution with a volume fraction of 20 to 30%.

[0013] Preferably, the elution rate in S3 is 3 to 6 mL / min.

[0014] The present invention has at least the following beneficial effects:

[0015] The present invention provides a pretreatment method for analyzing the content of microcystins in seaweed, which has the advantages of short cycle, few steps, simple operation, high accuracy of detection results, and greatly facilitates the rapid detection of microcystins. DETAILED DESCRIPTION

[0016] The technical solutions in the embodiments of the present invention are described clearly and completely below. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0017] Example 1

[0018] A pretreatment method for analyzing the content of microcystin in seaweed comprises the following steps:

[0019] S1, wash the seaweed with pure water, dry, crush, grind into powder, add into a beaker, then add extract, heat to 40°C, and ultrasonically oscillate for 3h to obtain extract;

[0020] The drying temperature is 45°C; the extract is a mixture of methanol and pure water, and the volume ratio of methanol to pure water is 65:1;

[0021] S2, centrifuging the extract for 8 min, at a temperature of 5°C and a speed of 10,000 r / min. After the centrifugation, taking the supernatant;

[0022] S3, concentrating the supernatant, pouring it into the SPE column, adding the eluent, performing solid phase extraction, collecting the eluate, concentrating it, and testing it;

[0023] The eluent is a methanol solution with a volume fraction of 20%, and the elution rate is 3 mL / min.

[0024] Example 2

[0025] A pretreatment method for analyzing the content of microcystin in seaweed comprises the following steps:

[0026] S1, wash the seaweed with pure water, dry, crush, grind into powder, add to a beaker, then add the extract, heat to 45°C, and ultrasonically oscillate for 3.5 hours to obtain an extract;

[0027] The drying temperature is 45°C; the extract is a mixture of methanol and pure water, and the volume ratio of methanol to pure water is 70:1;

[0028] S2, centrifuging the extract for 9 minutes at a temperature of 8°C and a speed of 10,000 r / min. After the centrifugation, taking the supernatant;

[0029] S3, concentrating the supernatant, pouring it into the SPE column, adding the eluent, performing solid phase extraction, collecting the eluate, concentrating it, and testing it;

[0030] The eluent is a methanol solution with a volume fraction of 25%, and the elution rate is 5 mL / min.

[0031] Example 3

[0032] A pretreatment method for analyzing the content of microcystin in seaweed comprises the following steps:

[0033] S1, wash the seaweed with pure water, dry, crush, grind into powder, add to a beaker, then add the extract, heat to 50°C, and ultrasonically oscillate for 4 hours to obtain an extract;

[0034] The drying temperature is 50°C; the extract is a mixture of methanol and pure water, and the volume ratio of methanol to pure water is 75:1;

[0035] S2, centrifuging the extract for 10 min at a temperature of 10°C and a speed of 10,000 r / min. After the centrifugation, taking the supernatant;

[0036] S3, concentrating the supernatant, pouring it into the SPE column, adding the eluent, performing solid phase extraction, collecting the eluate, concentrating it, and testing it;

[0037] The eluent is a methanol solution with a volume fraction of 30%, and the elution rate is 6 mL / min.

[0038] The methods provided in the above examples 1-3 have a short cycle and fewer steps. According to experimental analysis, the recovery rates of MC-RR and MC-LR by this experimental method are 93.2% and 94.6%.

[0039] The above shows and describes the basic principles, main features and advantages of the present invention. It is obvious to those skilled in the art that the present invention is not limited to the details of the above exemplary embodiments, and that the present invention can be implemented in other specific forms without departing from the spirit or basic features of the present invention. Therefore, no matter from which point of view, the embodiments should be regarded as exemplary and non-restrictive. The scope of the present invention is defined by the appended claims rather than the above description, and it is intended that all changes falling within the meaning and scope of the equivalent elements of the claims are included in the present invention.

[0040] Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that various changes, modifications, substitutions and variations may be made to the embodiments without departing from the principles and spirit of the present invention, and that the scope of the present invention is defined by the appended claims and their equivalents.

Claims

1. A pretreatment method for analyzing the content of microcystin in seaweed, characterized in that: The following steps are involved: S1, wash the seaweed with pure water, dry, crush, grind into powder, add into a beaker, then add extract, heat to 40-50°C, and ultrasonically vibrate for 3-4h to obtain extract; S2, centrifuging the extract and taking the supernatant; S3, concentrate the supernatant, pour it into the SPE column, add the eluent, perform solid phase extraction, collect the eluate, concentrate it, and wait for detection.

2. A pretreatment method for analyzing the content of microcystin in seaweed according to claim 1, characterized in that: The drying temperature in S1 is 45°C to 50°C.

3. A pretreatment method for analyzing the content of microcystin in seaweed according to claim 1, characterized in that: The extract in S1 is a mixture of methanol and pure water, and the volume ratio of methanol to pure water is 65-75:

1.

4. A pretreatment method for analyzing the content of microcystin in seaweed according to claim 1, characterized in that: In the S2, the centrifugal time is 8 to 10 minutes, the centrifugal temperature is 5 to 10° C., and the centrifugal speed is 10,000 r / min.

5. A pretreatment method for analyzing the content of microcystin in seaweed according to claim 1, characterized in that: The eluent in S3 is a methanol solution with a volume fraction of 20-30%.

6. A pretreatment method for analyzing the content of microcystin in seaweed according to claim 1, characterized in that: The elution rate in S3 is 3-6 mL / min.