Method for detecting content of lycorine

Abstract

The invention discloses a method for detecting the content of lycorine. The method comprises the following steps: (1) preparing a sample solution to be detected; (2) using liquid chromatography-tandem secondary mass spectrometry to detect the sample solution to be detected; (3) calculating the content according to a standard curve. The method provided by the invention can rapidly extract and accurately, qualitatively and quantitatively analyze the alkaloid substance-lycorine in lycoris plants, greatly shortens the analysis time, and improves the analytical accuracy; after the method is adopted, the detection result can be obtained within the program time of 6.0min, retention time of the lycorine is within 3min, the recovery rate of the lycorine is 98.7-99.8%, and the relative standard deviation (RSD) is 0.7-5.1%; the method is free from other characteristic peak interference, has the separation degree of complete separation, and greatly increases the analysis speed and accuracy; an effective and accurate analytical method is provided for the study of the accumulation law and anabolism regulation of the lycorine in the lycoris plants.

Description

technical field [0001] The invention belongs to the field of detection of alkaloid substances, in particular to a method for detection of lycorine content. Background technique [0002] Lycoris spp. is a bulbous herb of Amaryllidaceae (Amarylllidaceae), many of which are unique ornamental and medicinal flowers in China, rich in a variety of unique alkaloid components of Lycoris spp. So far, more than 500 alkaloid compounds with different structures have been isolated from Lycoris plants. Phytochemical studies have shown that these compounds have various pharmacological functions and have high medicinal value. Among them, lycorine has important value in antibacterial, antimalarial, tumor treatment, antiviral and cardiovascular protection (Li et al., 2005). So far, wild lycoris resources are still the most important source of medicinal lycorine. Therefore, accurate qualitative and quantitative analysis of lycorine content in lycoris is one of the important contents of breedin...

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Problems solved by technology

[0004] Li Mingkai (Establishment of Analytical Method for Lycorisine Alkaloids and Research on Biosynthesis, Anhui Agricultural University, 2012: 1-69) realized the preliminary separation of lycorine and other alkaloids by using thin-layer chromatography separation technology, and found that Two-way thin-layer chromatography is more suitable for the separation of many different alkaloids than one-way chromatography. For comparison, to achieve preliminary qualitative identification of alkaloid species in the sample, but not for accurate structural and quantitative analysis

[0005] Quan et al. (Quan et al., Photosynthetic characteristics of Lycoris aurea and monthly dynamics of alkaloid contents in its bulbs, African Journal of Biotechnology, 11(15): 3686-3691, 2012) utilized high-performance liquid chromatography detection technology, through samples and known The comparison of the retention time and the peak area of ​​the alkaloid standard substance under the same conditions carried out the quantitative research of galantamine and lycorine in Lycoris hudixiao, and its single extraction of Lycoris bulb material needs fresh sample 20g ( water content of about 90%), equivalent to 2g of Lycoris bulb dry sample, need to consume 110ml of methanol, 10ml of hydrochloric acid, 15ml of chloroform, which consumes a lot of plant samples and reagents, and at the same time, it only adopts the method of HPLC, according to the retention time of the substance. Content detection cannot be accurately qualitative for the substance to be tested, so the quantitative result of the substance is unreliable, and the structure, qualitative and precise quantitative analysis cannot be carried out

[0006] Sahar et al. (Sahar et al., 4'-O-Methylnorbelladine feeding enhances galanthamine and lycorine production by Leucojum aestivum L.shoot cultures, Engineering in Life Sciences, 00: 1-6, 2015) adopted the method of HPLC-ESI-MS, for Quantitative analysis of lycorine in Xia Xue Pian lotus buds. The elution time of lycorine is 25.5 minutes. The HPLC analysis time is relatively long, which takes about 36 minutes. The content is detected according to the retention time and parent ion information. This method Compared with Quan et al.'s method, the accuracy has been improved, but the obvious defect is that there may be many kinds of material components with the same elution time and precursor ion size, so there are still defects in qualitative and quantitative accuracy.

[0007] Moreover, in the prior art, there are also problems of long extraction time and cumbersome operation. For example, in the method of Quan et al., the main extraction steps are drying, filtration, methanol extraction for 3 hours, filtration, evaporation, pH adjustment, and chloroform extraction for 3 times. , evaporation, methanol reconstitution, filtration

In the method of Sahar et al. (Sahar et al., 2015), the main extraction steps are: methanol extraction for 24 hours, concentration, sulfuric acid acidification, chloroform extraction, evaporation, reconstitution, and filtration, all of which bring about rapid quantitative analysis of lycorine. inconvenient

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