Method for ionic liquid extraction/HPLC (high performance liquid chromatography) detection of nuciferine in lotus leaves

A technology of ionic liquid and nuciferine, which is applied in the direction of measuring devices, material separation, instruments, etc., can solve the problems of target product decomposition and extraction rate reduction, and achieve the effect of good repeatability and simple method

Inactive Publication Date: 2016-11-23
ZHEJIANG OCEAN UNIV
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Problems solved by technology

[0005] The traditional method uses organic reagents as solvents to extract the nuciferine in the lotus leaf by heating reflux, Soxhlet extraction, ultrasonic extraction, etc. The traditional method requires a higher temperature and a long extraction time to achieve the purpose of extracting nuciferin , while higher temperature or long-term extraction may lead to the decomposition of the target product, reducing the extraction rate

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  • Method for ionic liquid extraction/HPLC (high performance liquid chromatography) detection of nuciferine in lotus leaves
  • Method for ionic liquid extraction/HPLC (high performance liquid chromatography) detection of nuciferine in lotus leaves
  • Method for ionic liquid extraction/HPLC (high performance liquid chromatography) detection of nuciferine in lotus leaves

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Experimental program
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Effect test

Embodiment 1

[0030] Embodiment 1: detection condition

[0031] 1-1. HPLC chromatographic conditions

[0032] HPLC chromatographic conditions are: adopt Kromasil ODS-C 18 Column (4mm×250mm, 5um), mobile phase is acetonitrile-water-triethylamine-glacial acetic acid (56:44:1:0.5); flow rate: 1ml×min -1 ; Detection wavelength: 270nm; Column temperature: 25°C.

[0033] 1-2. Preparation of standard curve

[0034] Precisely measure 10 mg of Nuciferine standard substance (National Institute for Drug Control), set the volume in a 10ml volumetric flask for later use, and configure the concentrations of 0.005mg / ml, 0.01mg / ml, 0.03mg / ml, and 0.05mg / ml respectively , 0.07mg / ml nuciferine standard solution, adopt HPLC method to measure the content of nuciferine in lotus leaf. Take the peak area as the ordinate, and the concentration of nuciferine as the abscissa to make a standard curve, and the concentration of the nuciferine standard substance and its corresponding peak area are shown in Table 1. ...

Embodiment 2

[0037] Embodiment 2: sample extraction

[0038]The dried lotus leaf sample (Zhilin Pharmacy, Zhoushan, Zhejiang) was pulverized and passed through a 20-mesh sieve for subsequent use; 1.0 g of the lotus leaf powder sample was accurately weighed and placed in a round flask, then the ionic liquid solution was added, and a stirrer was placed in the Microwave-assisted extraction at 60 °C.

[0039] 2-1. Selection of ionic liquid types

[0040] Using V 离子液体 / V 甲醇 (Volume ratio 1:5) of [Bmin]Br, [Bmim]BF 4 、[Bmim]PF 6 、[Hmim]PF 6 、[Dmim]PF 6 Six kinds of ionic liquid solutions were used as extraction solutions. The solid-to-liquid ratio of the lotus leaf powder sample to the above-mentioned ionic liquid solutions was 1:30, and microwave-assisted extraction was performed at a microwave power of 400w for 10 minutes. The sealing areas corresponding to each ionic liquid solution are shown in Table 1, and the effects of different extraction solutions on the extraction of nuciferine ...

Embodiment 3

[0069] Example 3: Recovery and precision

[0070] Under the optimal extraction conditions mentioned above, the nuciferine in the lotus leaf was extracted, kept at room temperature for one week, and the day-to-day precision and intra-day precision experiments were carried out. The test was performed every two days within 6 days, and the measurement was repeated 3 times; within 13 hours, the measurement was performed every 4 hours, and the measurement was repeated 3 times. The experimental results show that the standard deviation of the peak area determined within 2 days is less than 3.5%, and the standard deviation of the peak area determined within 4 hours is less than 1.8%, indicating that the precision is good, reliable and stable. The results are shown in Table 8, the recoveries of the three levels ranged from 92.3% to 102.7%, and the relative standard deviation was less than 11%. The results show that the method of the present invention using ionic liquid extraction / HPLC ...

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Abstract

The invention relates to a method for ionic liquid extraction/HPLC (high performance liquid chromatography) detection of nuciferine in lotus leaves. The method comprises steps as follows: (1) sample extraction: a lotus leaf powder sample is weighed and put in a container provided with a stirring bar, an ionic liquid solution in the solid-to-liquid ratio being 1: (20-60) is added, and extraction is performed under the microwave assistance at the microwave power being 200-600 w for 2-12 min; (2) detection: HPLC is adopted for detection under the HPLC chromatographic conditions that a Kromasil ODS-C18 column is adopted and has the specifications being 4 mm*250 mm, 5 mu m; a flowing phase is acetonitrile-water-triethylamine-glacial acetic acid; the flowing speed is 0.5-1 ml/min; the detection wavelength is 270 nm; the column temperature is 25 DEG C. The method is simple, convenient, rapid, accurate, high in repeatability and suitable for measuring the content of nuciferine in the lotus leaves.

Description

technical field [0001] The invention relates to the field of detection of active components of traditional Chinese medicinal materials, in particular to a method for detecting nuciferine in lotus leaves by using ionic liquid extraction / HPLC. Background technique [0002] The lotus leaf is the leaf of the perennial aquatic herbaceous Nymphaeaceae plant, the upper surface is dark green or yellow-green, rough; the lower surface is light grayish brown, smoother, with 21 to 22 thick veins, which shoot out from the center to the surroundings, with protrusions in the center Ye Xixia Qianji. my country is rich in lotus leaf resources, which are distributed in all places except Qinghai and Tibet, and are most widely distributed in the three river basins of the Yangtze River, Yellow River and Pearl River. Traditional Chinese medicine believes that the lotus leaf is bitter and astringent in nature, and it belongs to the liver, spleen, stomach, and heart meridian. Diarrhea, hot blood ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02G01N30/14C07D221/18
Inventor 欧阳小琨杨立业李庆龙
Owner ZHEJIANG OCEAN UNIV
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