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Sample rapid pretreatment method for determining melatonin in milk, serum and products thereof

A technology of melatonin and dairy products, which is applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of cumbersome pretreatment steps, not suitable for general promotion, and large consumption of consumables, etc., to achieve a straight baseline and low cost , the effect of simple operation

Pending Publication Date: 2020-11-10
CHINA AGRI UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] Some methods for detecting melatonin content in milk, serum and related products have been disclosed in the prior art, but the pretreatment method steps are cumbersome, the consumption of consumables is large, and the experiment time is long; at the same time, some detection methods need to use liquid phase Chromatography-mass spectrometry method, high consumption cost, not suitable for general promotion

Method used

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  • Sample rapid pretreatment method for determining melatonin in milk, serum and products thereof
  • Sample rapid pretreatment method for determining melatonin in milk, serum and products thereof
  • Sample rapid pretreatment method for determining melatonin in milk, serum and products thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] 1. Experimental samples

[0025] The collected milk is stored in centrifuge tubes, transported at low temperature, and samples are stored at -80°C.

[0026] 2. Melatonin standard preparation

[0027] Weigh 0.05g of melatonin (brand name: sigma), first dissolve it with 5mL of methanol, then dilute with methanol to 4, 10, 20, 50, 100ng / mL (ie ppb), store in a brown sample bottle, - Store at 20°C.

[0028] 3. Sample pretreatment

[0029] Take out the milk sample to be tested, dissolve it, blow it evenly, and store it at 4°C for later use; take 200 μL of milk, add 200, 400, 600 and 800 μL of methanol respectively, and dilute the milk to 2, 3, 4 and 5 times the volume of the original sample; Vortex for 30s; 4°C, 14000r / min, centrifuge for 20min; finally draw the supernatant with a 1mL syringe, filter with a 2μm filter, draw 350μL of the liquid and store it in a brown sample bottle for detection.

[0030] 4. Detection by high performance liquid chromatography

[0031] 12...

Embodiment 2

[0058] 1. Experimental sample processing

[0059] The sheep's blood was collected by phlebotomy. After the blood was allowed to stand at room temperature for 30 minutes, it was centrifuged at 5000r / min for 10 minutes, and the serum was drawn into a clean centrifuge tube, numbered and labeled, stored at -80°C, and transported on ice for short distances.

[0060] 2. Melatonin standard preparation

[0061] Weigh 0.05g of melatonin (brand name: sigma), first dissolve it with 5mL of methanol, and then dilute with methanol to 2, 4, 10, 20, 50, 100ng / mL (ie ppb) to make a standard product, store in brown Store in the sample bottle at -20°C, and the standard is valid within one month after preparation.

[0062] 3. Sample pretreatment

[0063] Take out the sample to be tested, dissolve, and pipette evenly; take 200 μL of the sample to be tested, add 200, 400, 600 and 800 μL of methanol respectively, and dilute the sample to be tested to 2, 3, 4 and 5 times the volume of the original...

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Abstract

The invention provides a rapid sample pretreatment method for determining melatonin in milk, serum and products thereof. According to the method, a sample is diluted by using methanol, and the melatonin content is determined by using high performance liquid chromatography, so that the purpose of rapidly and accurately determining melatonin in milk, serum and products thereof is achieved. Accordingto the method disclosed by the invention, the effect is optimal when milk is diluted to 5 times of the volume of an original sample, and the melatonin recovery rate is 91.5-134.7%; better recovery rate can be obtained when the sheep serum is diluted to 2 times, 3 times, 4 times and 5 times of the original volume, and the recovery rate is 81.02%-97.44%. The method has the advantages of low cost, simple steps and time saving, and provides a beneficial reference for further development of research related to melatonin in milk and serum.

Description

technical field [0001] The invention belongs to the field of agricultural animal husbandry and veterinary medicine, and the research objects are cattle, sheep, pigs and other livestock milk, serum and products thereof, in particular to a sample pretreatment method for rapidly and effectively measuring melatonin content therein. Background technique [0002] Melatonin (Melatonin, MT) is a hormone secreted by the pineal gland. Current studies have shown that melatonin is involved in the regulation of biological rhythms, sleep and immune cells, and is an important reproductive hormone. Accurate determination of melatonin content in milk, cow, sheep, pig and other livestock milk and milk products, serum and its products provides important reference significance for the detection of livestock growth status and the promotion and application of melatonin. [0003] Some methods for detecting melatonin content in milk, serum and related products have been disclosed in the prior art, ...

Claims

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Application Information

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IPC IPC(8): G01N30/02G01N30/06G01N30/34G01N30/36G01N30/74G01N30/86
CPCG01N30/02G01N30/06G01N30/34G01N30/36G01N30/74G01N30/8679
Inventor 刘国世王静姚松阳马慧张鲁常毅姬鹏云
Owner CHINA AGRI UNIV
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