Abstract: In this paper, the extraction and chromatographic analysis of effective chemical components of Solanum nigrum were discussed in detail.

Keywords: Solanum nigrum; Solanine; Ultrasonic wave; Chromatogram; Title of paper

1 materials and methods

1. 1 experimental instruments and materials

PHS-3C Precision Acidimeter * * * Beijing Precision Instrument Co., Ltd. * * *; DZF-6050 vacuum drying oven * * * Beijing Sanbo Technology Co., Ltd. * * *; SENCO R rotary evaporator * * * Beijing Bioinnovation Technology Co., Ltd. * * *; AY 120 electronic balance * * * Shimadzu Co., Ltd. * * *; High performance liquid chromatography * * * Shimadzu Co., Ltd. * * *; Methanol, ethanol and chloroform are analytically pure.

1.2 ultrasonic-assisted extraction

Weigh the Solanum nigrum L. sample, moisten it with V [HCl]: V * * * ethanol * * 1: 4 solution for 24 hours, then add ethanol to Solanum nigrum L. and ethanol according to 1g: 10ml, and perform ultrasonic extraction twice at room temperature. After filtration, the filtrates were combined. Distilling the filtrate under reduced pressure to obtain yellow-green solid. Dissolve the crude extract of steroidal alkaloids with 0.5mol/L HCl, extract with chloroform for three times, remove the chloroform layer, adjust the water phase to pH 1.5 with NaOH solution, extract with chloroform for three times, combine chloroform extracts, evaporate to dryness, dissolve with methanol and fix the volume to 50mL, and get the methanol solution of Solanum nigrum total alkaloids.

Preparation of 1.3 solanine

Quantitatively take out the methanol solution containing biological total alkali, evaporate under reduced pressure, dissolve the residue with 40 ml of V [HCl]: V * * * ethanol * * 1: 1, hydrolyze at 100℃, evaporate the ethanol in the solution after hydrolysis, and extract the residue with chloroform for three times, and take the acid water layer. After adjusting the pH to 1.5 with NaOH solution, extract with chloroform for three times, combine chloroform extracts, evaporate and dissolve with methanol, and fix the volume to 50 ml to obtain methanol solution of solanine.

1.4 chromatographic analysis

It is confirmed that the standard sample solanine has the maximum absorption peak at 2 10 hm by all-band scanning in ultraviolet region, so 2 10 hm is selected as the detection wavelength of solanine. Chromatographic conditions: acetonitrile: water 55: 45 * * * The water phase was adjusted to pH 2.7 * * with phosphoric acid, and the flow rate was 1.2mL/min. When the sample volume of solanine is 20ug/mL-200 ug /mL. 2***ug/mL*** has a good linear relationship with the peak area.

Two results

2. 1 extraction effect

Ultrasonic-assisted extraction of effective components from Chinese herbal medicines is rapid, simple and efficient. According to this principle, the experimental conditions selected in this paper are: the temperature is 25 degrees and the ethanol concentration is 95%. Under the optimum experimental conditions, the average yield of solanine in stems and leaves was 0.42%, and that in fruits was 3.22%. Three Solanum nigrum fruit samples were weighed and parallel extraction experiments were carried out to calculate the mass percentage of solanine products and the RSD value of the same batch of samples. The results showed that the average mass fraction of solanine in three kinds of Solanum nigrum samples was 2.53%, and the RSD was 4. 18%, which proved that the extraction process was stable and reliable.

2.2 Chromatographic effect

In this paper, the sample of steroid alkaloids hydrolyzed for 65438 0 h was selected as the research object. Without phosphoric acid, the peak shape of chromatographic peak is seriously tailed. With the increase of acidity, the peak shape began to sharpen. When pH is 2.7, steroid alkaloids and solanine are completely ionized. The peak shape is clear, and the resolution between the two chromatographic peaks is high.

3 discussion

Combining the viewpoints and means of modern medicine, it is of great significance to find and start new anticancer drugs with independent intellectual property rights in the treasure house of Chinese herbal medicine resources, especially in Chinese herbal medicines with definite curative effects, not only to promote human health, but also to promote the development of China's ethnic medicine industry and the modernization of Chinese medicine. Solanum nigrum is panicum miliaceum. The whole grass contains solanine, solanine, solanine and other steroidal alkaloids. Their aglycone is mainly solanine, and the content of alkaloids is the highest in fruits, the highest in immature fruits, and the content disappears when fruits mature.

Solanum nigrum L. is rich in medicinal and health-care components, and has many medicinal functions such as anti-inflammatory, antibacterial, antihypertensive and anticancer. But so far, the resource utilization rate of Solanum nigrum is still relatively low, and the report on its active components is even blank. In the last century, someone used the extraction technology of Solanum plants abroad to determine the content of solanine aglycone in Solanum nigrum by TLC. It was found that the content of solanine in Solanum nigrum fruit was significantly higher than that in whole grass, and even as high as 4.2% in immature olive. However, the extraction technology and rapid determination method of total alkaloids from Solanum nigrum have not been reported. The results of this study show that ultrasonic-assisted ethanol extraction of steroidal alkaloids from Solanum nigrum has the advantages of simple operation and short extraction time. At the same time, it has good repeatability, high stability and high recovery rate.

In recent years, high performance liquid chromatography (HPLC) has been widely used for the separation and identification of alkaloids with its absolute advantages. Especially, the application of RP-HPLC composed of nonpolar stationary phase and polar mobile phase accounts for about 90%, and the nonpolar stationary phase is mainly silanized reverse bond and silica gel-based phase. In this paper, the analytical conditions for the determination of steroidal alkaloids and their aglycone solanine by HPLC were studied and determined: the ultraviolet detection wavelength was 2 10hm. The mobile phase is acetonitrile: water with the ratio of 55: 45 * *, and the pH value of the water phase is adjusted to 2.7 * * with phosphoric acid. Ultraviolet, infrared and mass spectrometry analysis confirmed that it was the aglycone of steroidal alkaloids in Solanum nigrum.

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