A method for rapidly detecting L-carnitine and its kit

Abstract

The invention discloses a method and a kit for rapid and high efficient detection of content of L-carnitine. The method comprises the following steps: a microwell plate A which comprises freeze-drying acetyl coenzyme A and acetylcarnitine transferase, and a microwell plate B which is coated by a precipitating plate are prepared; a standard pore, a quality control pore and a sample pore are arranged on the microwell plate A, a chromogenic solution is firstly added into each pore, the standard pore, the quality control pore and a sample to be tested are respectively added into corresponding micropore for reaction, enzymatic chromogenic detection is carried out, and luminous absorption value (OD) is detected; referred to the microwell plate A, the same operation is carried out on the microwell plate B, after the reaction, the enzymatic chromogenic detection is carried out, and luminous absorption value (OD blank) is detected; based on difference of the luminous absorption values of twice enzymatic chromogenic detections, a standard curve is constructed in order to determine the content of L-carnitine in a sample to be tested. Preparation of reagents in the detection process is reduced, operation is simpler, enzyme activity is kept stable, the detection result is more reliable, and the method detection limit reaches 1mg / 100g.

Description

Technical field [0001] The invention relates to the field of food detection, in particular to a method for rapidly detecting L-carnitine and a kit thereof. Background technique [0002] L-carnitine, also known as L-carnitine, is an internal amino acid that promotes the conversion of fat into energy. Red meat is the main source of L-carnitine and has no toxic side effects to the human body. L-carnitine is also an essential nutrient for conditional babies It plays an important role in the metabolism of infants using fat as an energy source. L-carnitine is ubiquitous in infant foods, beverages, biscuits and some weight loss products. At present, the methods for determining L-carnitine in the literature include high performance liquid chromatography, capillary electrophoresis and mass spectrometry, but spectrophotometry is still the most important detection method. Spectrophotometric detection of L-carnitine mostly requires a chemical reaction. For example, the free L-carnitine in ...

AI Technical Summary

Problems solved by technology

The national standard method detects L-carnitine content at present, adopts ultraviolet spectrophotometer, mainly has the shortcoming of the following aspects: (1) acetyl-CoA and carnitine acetyltransferase are expensive, and detection cost is very big; (2) consumed The volume is large, and the traditional cuvette is used for testing, which requires a volume of 2-3mL, which is time-consuming in the colorimetric stage; (3) The enzyme used is stored in the form of an aqueous solution, and its validity period is limited, and the enzyme activity will be greatly affected. The degree of influence seriously affects the stability of the experiment; (4) the sample processing needs to weigh 5g, and the later sample processing consumes a large amount of reagents, and it cannot be transparent in the treatment of some substrates, which affects the subsequent detection effect

[0004] In the national standard GB 29989, the sample processing method is complex, time-consuming, and the amount of reagents is large, the detection process is cumbersome, and the repeatability is poor; while the existing L-carnitine detection kit reagent preparation and storage is cumbersome, and the reaction system is complex

Invention patent application CN 106198531A is actually a reduced version of the GB29989 detection system, changing the amount of reagents to achieve detection on the microplate, no substantial innovation in technology, still can not solve the problem of enzyme storage validity period and experimental stability

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