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Method for simultaneously detecting additive, L-carnitine and D-carnitine

A feed additive, carnitine technology, applied in the field of photoelectric analysis, can solve the problem of consuming a lot of manpower and material resources, and achieve the effect of simplifying the test process, high separation efficiency, and good separation effect

Active Publication Date: 2021-12-17
SICHUAN NEW HOPE ANIMAL HUSBANDRY TECH +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0015] The main purpose of the present invention is that it is difficult to simultaneously take into account a variety of different detection and analysis objects for various detection methods in the prior art, especially for L-carnitine that needs derivatization treatment , consume a lot of manpower and material resources to carry out the deficiency of laboratory analysis, provide a kind of high performance liquid chromatography that can detect multiple components at the same time

Method used

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  • Method for simultaneously detecting additive, L-carnitine and D-carnitine
  • Method for simultaneously detecting additive, L-carnitine and D-carnitine
  • Method for simultaneously detecting additive, L-carnitine and D-carnitine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] In this study, high performance liquid chromatography and Chiral CD-Ph S5 chromatographic column were used to separate L-carnitine and D-carnitine, which can accurately determine the content of L-carnitine and D-carnitine in feed additives and premixes.

[0048] 1 Experimental part

[0049] 1.1 Instruments, reagents and test materials

[0050] LC-1260 infinityII high performance liquid chromatography-VWD ultraviolet detector (Agilent, USA); LE204E / 02 one hundred thousandth balance (Mettler Toledo Shanghai Co., Ltd.); LG-22 high-speed refrigerated centrifuge (Sichuan Shuke Instrument Co., Ltd.); KQ-300VDE (Kunshan Ultrasonic Instrument Co., Ltd.)

[0051] Levocarnitine (L-carnitine, namely L-carnitine), 99.9%, SIGMA-ALDRICH; Acetonitrile (chromatographically pure, sigma); Sodium perchlorate (analytical pure, Chengdu Kelong)

[0052] L-carnitine standard stock solution: Weigh 100mg of L-carnitine each, dissolve in methanol and dilute to 100mL brown volumetric flask resp...

Embodiment 2

[0067] On the basis of the experimental results of Example 1, the optimal chromatographic conditions and sample pretreatment process were selected, and the standard addition recovery and precision experiments were carried out.

[0068] Sample pretreatment process: Weigh 0.05g feed additive sample (accurate to 0.0001g), place it in a 100mL volumetric flask, add 2 / 3 volume (about 65mL) of ultrapure water for ultrasonic extraction for 15min, take it out and cool it to room temperature, The pure water is fixed to the mark, filtered through a 0.22μm water phase membrane, and tested on the machine.

[0069] Chromatographic conditions:

[0070] High performance liquid chromatography - UV detector: wavelength 220nm; chromatographic column: Chiral CD-Ph S5 (4.6mm i.d. × 250mm); mobile phase: acetonitrile: 0.25mol / L NaClO 4 Solution (v / v)=20:80, isocratic elution; flow rate: 0.5mL / min; column temperature: 25°C; injection volume: 10μL.

[0071] 2.3.2 Recovery rate and precision of stan...

Embodiment 3

[0083] The sample pretreatment method is as follows: Weigh 0.05g feed additive sample, place it in a 100mL volumetric flask, add 2 / 3 volume of ultrapure water for ultrasonic extraction for 15min, take it out and cool it to room temperature, dilute to the mark with ultrapure water, and pass 0.22 It is filtered by μm aqueous phase membrane and tested on the machine.

[0084] Chromatographic conditions: high performance liquid chromatography - UV detector: wavelength 220nm; chromatographic column: Chiral CD-Ph S5 (4.6mm i.d. × 250mm); mobile phase: acetonitrile: 0.25mol / L NaClO 4 Solution (v / v)=20:80, isocratic elution; flow rate: 0.5mL / min; column temperature: 25°C; injection volume: 10μL.

[0085] This example is similar to Example 2, the sample pretreatment method and chromatographic conditions are as above, and the influence of different pH values ​​on the test results is mainly compared.

[0086]During the experiment, the pH value of the mobile phase was adjusted with hydro...

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Abstract

The invention relates to a method for simultaneously detecting an additive, L-carnitine and D-carnitine, which comprises the following steps: preparing a standard reagent, carrying out detection analysis by adopting a high performance liquid chromatograph according to the following chromatographic conditions, and establishing a regression equation, the standard reagent comprising a levocarnitine standard solution and an L-carnitine standard solution, chromatographic conditions being as follows: a chromatographic column being a chiral chromatographic column, a mobile phase being an acetonitrile-sodium perchlorate solution, the column temperature being 25 DEG C, and the flow velocity being 0.5 mL / min; performing sample pretreatment and detection; to be specific, weighing a feed additive sample or premix, adding ultrapure water for ultrasonic extraction for 5-30 min, taking the feed additive sample or premix out and performing cooling to the room temperature, using ultrapure water for metering the volume to the scale, and filtering the feed additive sample or premix through a 0.22-micro water phase filter membrane and performing detection on a machine; and substituting a result obtained by detection into the regression equation, and calculating the concentration of the additive. According to the detection method disclosed by the invention, the L-carnitine detection analysis efficiency is higher, the whole detection can be completed in 15 minutes, and the detection efficiency is improved.

Description

technical field [0001] The invention relates to a chromatographic detection method, in particular to a high-performance liquid chromatographic detection method, which can realize simultaneous detection and analysis of various additives including L-carnitine and D-carnitine, and belongs to the technical field of photoelectric analysis. Background technique [0002] L-carnitine, also known as L-carnitine, is a kind of amino acid that promotes fat conversion, and plays an important role in the fat metabolism process of organisms. Domestic studies have shown that adding L-carnitine to pig feed can improve the anti-stress ability of piglets, improve the carcass quality of fattening pigs, and improve the reproductive performance of sows; adding L-carnitine to aquatic feed can improve the growth performance of aquatic animals; Adding L-carnitine in the feed can reduce the incidence of ascites in broilers, improve the hatching time of broilers and the hatching rate of fertilized egg...

Claims

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Application Information

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IPC IPC(8): G01N30/02G01N30/74
CPCG01N30/02G01N30/74
Inventor 梁玉树何开蓉乔煦玮李勇熊安琪刘尚群王力杜倩男刘珊李欣欣
Owner SICHUAN NEW HOPE ANIMAL HUSBANDRY TECH
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