Method for detecting and separating lotus corniculatus glycoside in sedum spectabile
By extracting components from Sedum spectabile using ultrasound and utilizing the Fe3O4-PEI-CR binding reduction reaction, the problems of complex sample pretreatment and matrix interference in traditional methods were solved, enabling rapid and sensitive detection and separation of phytohexaenoic acid, ensuring detection accuracy and separation efficiency.
Patent Information
- Application Number
- CN202511841486.6
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-11-26
- Publication Date
- 2026-03-03
AI Technical Summary
Traditional methods for detecting basidiosin in Sedum spectabile involve complex sample pretreatment and are easily affected by complex matrices, lacking sensitive, rapid, green and environmentally friendly detection methods.
The effective components of Sedum spectabile were extracted using an ultrasonic method. Congo red was adsorbed using a composite material of iron(III) oxide/polyethyleneimine (Fe3O4-PEI). Combined with reducing agents such as calcium hypophosphate and nickel precursors, the presence of basidiosin was confirmed by pH changes through reduction and hydroxylamine hydrochloride reactions. Aldehyde compounds were then separated by electrostatic interactions.
It enables rapid, sensitive, and environmentally friendly detection and separation of basidiosin, ensuring detection accuracy and separation efficiency.
Abstract
Description
Technical Field
[0001] This invention belongs to the field of food technology, specifically relating to a method for the detection and separation of basidiosin from Sedum aizoon. Background Technology
[0002] Sedum erythrostictum H. ohba, also known as long-scented sedum, is a herbaceous plant belonging to the genus Sedum in the family Crassulaceae. It is an easy-to-cultivate, drought-tolerant, cold-hardy plant with a wide growing range. In-depth research on Sedum erythrostictum has revealed that its extracts contain a very complex, rich, and diverse range of active ingredients, including alkaloids, tannins, flavonoids, and their derivatives. Sedum erythrostictum possesses antioxidant, anti-glycation, and antibacterial bioactivities, showing significant efficacy in medical applications. Furthermore, Crassulaceae plants also contain a certain amount of cyanogenic glycosides, which are a class of secondary metabolites containing a cyanide group and are a type of natural plant toxin. The cyanogenic glycosides in Sedum erythrostictum exist in the form of arbutin. Due to its potential toxicity, arbutin requires rigorous detection and separation to ensure the safety and functionality of the product. Traditional testing methods use chromatography, but this method has drawbacks such as complex sample pretreatment, the need for derivatization, and susceptibility to interference in complex matrices. Therefore, it is extremely important to develop a sensitive, rapid, green and environmentally friendly detection method. Summary of the Invention
[0003] In order to improve the utilization of the potential nutritional, medicinal and commercial value of Sedum spectabile, this invention provides a method for the detection and separation of basidiosin.
[0004] The specific method is as follows:
[0005] (1) After pulverizing the dried Sedum spectabile, its effective components were extracted by ultrasonication. The Sedum spectabile sample was ultrasonicated twice with ethanol or methanol solution for 30 min each time. The filtrates were combined and then vacuum filtered to obtain the Sedum spectabile extract. The filtrate was concentrated by rotary evaporation and freeze-dried to obtain the Sedum spectabile extract.
[0006] (2) It is difficult for the cyanoglycoside ions of salvia miltiorrhiza to generate hydrocyanic acid. Therefore, the presence of the cyano group cannot be determined by simple picric acid paper reagent. In this invention, the cyano group of salvia miltiorrhiza is reduced to the aldehyde group by reducing agents such as calcium hypophosphite and nickel precursors, styrene aldehyde synthesis reaction (stannous chloride / HCl gas reduction), borohydride, triisopropoxyvanadium oxide / TMDS reduction system, etc.
[0007] (3) Congo red (CR) is adsorbed onto the composite material Fe3O4-PEI / polyethyleneimine (Fe3O4-PEI). The Congo red changes color due to changes in the solution's pH. Adding the aforementioned reduced aldehyde compound and hydroxylamine hydrochloride to the Fe3O4-PEI-CR solution causes a nucleophilic addition reaction between the aldehyde group and hydroxylamine hydrochloride, ultimately resulting in dehydration to aldoxime, water, and hydrochloric acid. The hydrochloric acid alters the pH, causing a color change in the Fe3O4-PEI-CR solution, confirming the presence of aldehydes, specifically holologous glycosides. Simultaneously, Fe... 3+ and Fe 2 + It can form chemical bonds with functional groups such as hydroxyl and oxime groups in the aldoxime molecule or be adsorbed through electrostatic interactions, thus further separating the aldoxime compound. Detailed Implementation
[0008] Example 1
[0009] (1) Weigh 20.0g of Sedum spectabile sample, sonicate twice with 200mL of methanol aqueous solution (95:5, V / V) for 30min each time, combine the filtrates, vacuum filter and rotary evaporate, and freeze dry to obtain Sedum spectabile extract powder for later use.
[0010] (2) Magnetic Fe3O4 was prepared by coprecipitation. 20 mL of ultrapure water was added to a 50 mL three-necked flask and stirred. Inert gas was passed through to remove oxygen for about 30 min. 4 mmol FeCl3·6H2O and 2.4 mmol FeSO4·7H2O were added and stirred for another 10 min (stirring speed 500 r / min). The solution turned orange-red. The solution was heated to 60 °C in a water bath and 2.5 mL of concentrated ammonia was slowly added dropwise (to be added in about 1 min). The pH of the solution was between 9 and 10. The solution quickly changed from orange-red to black. The reaction was continued for another 30 min and then heated to 80 °C and stirring was stopped. The solution was left to stand for 30 min to obtain a black solution. The solution was collected under an external magnetic field and washed 3 times. The solution was then dried under vacuum at 80 °C. The preparation of magnetic Fe3O4-supported amino-adsorbed Congo red (Fe3O4-PEI-CR) was first carried out by dispersing 0.5 g of prepared Fe3O4 in 100 mL of water and sonicating for 30 min. Then, under stirring, 5 mL of 5 g / L polyethyleneimine (PEI) aqueous solution and an equal volume of 1 mg / mL Congo red solution were added, and the mixture was stirred for at least 2 h (temperature controlled at 0 °C). The solution was then transferred to an 80 °C constant temperature water bath and reacted for 6 h to obtain the Fe3O4-PEI-CR solution.
[0011] (3) Selecting compounds of calcium hypophosphate and nickel as reducing agents, under alkaline conditions, Ni(OAc)₂·4H₂O, alkaline solution Ca(OAc)₂·H₂O or K₂CO₃, and Ca(H₂PO₂)₂ are mixed evenly in a molar ratio of 2:4:4, and then dissolved in a mixture of ethanol and water to obtain the desired reducing agent mixture. Calcium hypophosphate will reduce Ni... 2+ Reduced to Ni 0 Ni 0 The reduction of cyanide to aldehyde is crucial. Adding 2.0g of Sedum spectabile extract to the reducing agent mixture ultimately yields an aldehyde compound solution.
[0012] (4) In Fe3O4-PEI-CR solution, the above-mentioned reduced aldehyde compound solution and hydroxylamine hydrochloride (NH2OH.HCl) are added dropwise. The two react to produce aldoxime, water and hydrochloric acid, thereby changing the... p H value. The solution color changes from red to blue, confirming the presence of aldehyde compounds, while the magnetic composite material separates the aldoxime. The test was performed three times to ensure accuracy.
Claims
1. A method for detecting basidiosin in Sedum spectabile, characterized in that, After reducing the cyano group of arbutin to an aldehyde group, it reacts with hydroxylamine hydrochloride to generate hydrochloric acid and an aldoxime compound. Changes in pH cause discoloration of the Fe3O4-PEI-CR magnetic composite solution, confirming the presence of aldehydes, specifically arbutin. Simultaneously, the presence of Fe in the magnetic composite material is detected. 3+ and Fe 2+ Aldehyde compounds can be further separated by forming chemical bonds or electrostatic interactions with functional groups such as hydroxyl and oxime groups in the oxime molecule.