High efficiency liquid chromatography method for detecting carnitine content in fish plasma

Abstract

The invention discloses a high efficiency liquid chromatography method for detecting carnitine content in fish plasma. A p-BPB (para-bromophenacyl bromide) derivatization reagent with a strong ultraviolet absorption property and carnitine are subjected to a pre-column derivatization reaction, so as to reduce the polarity, increase the ultraviolet absorption and enhance the detection sensitivity of the method; separation and detection are performed under the combination of normal phase high-efficiency liquid chromatography. A reliable analysis method is provided for the detection of carnitine in aquatic products as well as the pharmacokinetic study of the carnitine in aquatic animal bodies, and a basic theory foundation is provided for rational drug use in the aquatic industry. In a word, the method has important and profound significance for the content analysis of carnitine-type compounds in the aquatic products as well as evaluation of food nutrition and safety and meanwhile, the method is a necessary premise and condition for research and development of carnitine.

Description

technical field [0001] The invention belongs to the technical field of carnitine content detection, in particular to a high-performance liquid chromatography method for detecting carnitine content in fish plasma. Background technique [0002] A large number of studies have shown that L-carnitine is easy to use. Adding L-carnitine in feed can improve the growth of aquatic animals, resist fatty liver, improve fish quality, and reduce the toxic effects of pesticides, drugs, ammonia and other toxic substances on aquatic animals. And other effects, it has been paid attention to as a new type of feed additive, and related research and utilization have also been carried out in the pharmaceutical and food industries. [0003] The price of L-carnitine is relatively high. If the amount is used too much, it is not economical, but if the amount is too small, the desired effect cannot be achieved. Therefore, it is necessary to determine the dosage of L-carnitine according to the animal ...

AI Technical Summary

Problems solved by technology

According to the derivatization reagent used, there are two kinds of pre-column derivatization high-performance liquid chromatography methods: (1) pre-column UV derivatization high-performance liquid chromatography: p-bromophenylacetyl bromide (p-BPB) is the most commonly used method. The selected derivatization reagent has strong ultraviolet absorption at a wavelength of 260nm, is cheap and easy to buy, and Poorthuis et al. have detected the carnitine content in the urine of patients with congenital metabolic defects by this method, but according to further The research found that the derivative product generated by the derivatization reaction between the derivatization reagent p-BPB and carnitine is very low, and contains more impurities, which is difficult to meet the requirements of blood sample detection

In order to speed up the reaction between carnitine and derivatization reagents, Minkler P et al. selected p-bromophenylacetyl trifluoromethanesulfonate as the derivatization reagent under the catalysis of diisopropylethylamine, which can be derivatized with carnitine in about 10 minutes. complete, but the disadvantage is that p-bromophenylacetyl trifluoromethanesulfonate is expensive, difficult to purify and buy, and the recovery rate of this method is mostly lower than 80%.

In 2002, Rosenthal reported that using tetrabutylammonium hydroxide as a catalyst, p-BPB and carnitine can obtain derivatives with high reaction yields. This method is suitable for the determination of free and total carnitine content in blood, and the recovery rate is 97%. , which improves the p-BPB carnitine derivatization method to a certain extent, but the sample reaction time is relatively long, and it needs to go through reaction steps such as shaking, centrifugation, and heating, and it takes at least 3 hours to complete a sample pretreatment

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