CN110066305B - Mechanochemical extraction method for preparing crude naphthopyrone extract from berchemilla lineate - Google Patents
Mechanochemical extraction method for preparing crude naphthopyrone extract from berchemilla lineate Download PDFInfo
- Publication number
- CN110066305B CN110066305B CN201910323824.5A CN201910323824A CN110066305B CN 110066305 B CN110066305 B CN 110066305B CN 201910323824 A CN201910323824 A CN 201910323824A CN 110066305 B CN110066305 B CN 110066305B
- Authority
- CN
- China
- Prior art keywords
- extract
- crude
- extraction
- ball milling
- mechanochemical
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H1/00—Processes for the preparation of sugar derivatives
- C07H1/06—Separation; Purification
- C07H1/08—Separation; Purification from natural products
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H17/00—Compounds containing heterocyclic radicals directly attached to hetero atoms of saccharide radicals
- C07H17/04—Heterocyclic radicals containing only oxygen as ring hetero atoms
Abstract
The invention discloses a mechanochemical extraction method for preparing a crude naphthopyrone extract from berchemite, which comprises the following steps: drying, crushing and screening a raw material of the alclad gold, placing the obtained alclad gold powder into a ball mill for ball milling, mixing the ball-milled alclad gold powder with an extraction solvent, then placing the mixture into a water bath with the temperature of 20-50 ℃ for ultrasonic extraction for 15-60 min, then cooling to room temperature, filtering, and removing the solvent from the filtrate through rotary evaporation to obtain a tawny crude naphthopyrone extract; and after the crude extract is redissolved by methanol, standing, taking supernatant fluid to pass through a PTFE membrane with the size of 0.22 mu m, analyzing by using an ultra-high performance liquid chromatography, and calculating the yield of the crude extract according to the content of the main component of the crude extract of the naphthopyranone, namely rubrofusamycin-6-O-alpha-L-rhamnosyl- (1-6) -O-beta-D-glucopyranoside. The method has low cost, and the process of preparing and extracting the crude naphthopyrone extract is quick and efficient, and can provide technical basis for the comprehensive utilization of the berchemia lineata medicinal resource.
Description
Technical Field
The invention relates to the technical field of extraction of active ingredients of plant medicines, in particular to a mechanochemical extraction method for preparing a crude extract of naphthopyrone from berchemilla lineate.
Background
Iron coated gold (Berchemia lineate(L.)DC.) Is Rhamnaceae (Rhamnaceae) Dried root or stem of Berchemia leptostachya of Berchemia. The root of the plant is brown like iron, and the xylem is yellow like gold, so the plant is named as Tiebao gold. The plant of the source of the Baojin is mainly distributed in provinces such as Guangdong, Guangxi and Fujian in China, and is also found in Korea, Vietnam, India, Japan and other countries.
The berchemia lineata is an important national medicinal resource in China, can be used as a medicament for treating patient diseases (CN 102166275A), and can also be used for preparing health-care food together with bee pollen, honey and the like to improve the immunity of a human body (CN 102302105A).
The berchemia lineata has light, slightly bitter and cool nature, and has effects of dispelling pathogenic wind, removing dampness, relieving inflammation, relieving swelling, cooling blood, removing toxic substance, relieving rigidity of muscles, dredging collaterals, removing blood stasis, promoting blood circulation, warming stomach and invigorating spleen (determination of total flavone content in berchemia lineata Linn. Yao medicine berchemia lineata Linn (J)]. Chemical industry of Guangdong province240 plus 241, is a commonly used national medicine in the autonomous region of the Guangxi Zhuang nationality and the minority nationality region in the southwest of China, is recorded in the Lingnan medicine-gathering book at the earliest and is included in the quality standards of traditional Chinese medicines in Guangdong province, the quality standards of Fujian traditional Chinese medicines and the quality standards of traditional Chinese medicines in the Guangxi province. Research shows that the berchemia lineata contains naphthopyrones, flavonoids, phenols, anthraquinones, terpenoids, dimers and other compounds (Leclent, southern Fujian berchemia lineata and liver protection activity research [ D)]. HUAQIAO University2015, Shenyuxia, Tenghuali, Chenxiaolong, etc. St. St.John's wort root chemical composition research [ J]. Chinese herbal medicine2010(12) 1955-1957), in which naphthopyrones and flavonoids are the main active ingredients. Modern pharmacological studies have shown that naphthopyrones have antibacterial, anticancer, antimutagenic, hypolipidemic, and antioxidant effects (Gunn, Liuling, African fly, etc.; the naphtho-pyrones chemical components in Cassia tora and their biological activities have been studied [ J].The research of special products is carried out, 2017(3): 64-67.）。
the literature research shows that the common extraction method of the naphthopyrone compounds comprises the following steps: cold soaking method, heating reflux method and ultrasonic extraction method (Xuyilong, Tangliying, Zhouyidan, etc.. HPLC determination of 3 naphthopyranoside contents [ J ] in semen Cassiae]. Chinese experimental prescriptions magazine,2014(5): 54-56.). Wherein, the cold soaking method has simple process and low cost, but consumes time; the extraction solvent in the heating reflux method can be recycled during extraction, but the dosage of the solvent is more and the heating time is longer; the ultrasonic extraction method has the characteristics of simple and controllable operation, high extraction efficiency and the like. Currently, there are few studies on the extraction process of berchemia lineata in reported literature, wherein the ultrasonic extraction process of berchemia lineata is optimized by Zhudelwa et al through orthogonal experiments (Zhudelwa, Takayagming, Chenyonbin, etc.. the ultrasonic extraction process of berchemia lineata is preferred by the orthogonal experiments [ J]. Strait pharmacology2018(30): 43-45), but the process does not describe the extraction strategy of the naphthopyranone compounds in the berchemia lineate.
In recent years, mechanochemical assisted extraction technology has been successfully applied to the extraction of various plant active ingredients by more and more researchers (CN 103006724A, CN 107684577A, CN 101638407 a). The method does not simply depend on the polarity of a solvent, changes the principle of the traditional extraction method, and mainly assists extraction through the following two ways: firstly, plant cells generate a wall-breaking effect through mechanical force, and the release of effective components is promoted; secondly, the structure of the extracted matter is changed, the dissolubility of the extracted matter is changed, and the extracted matter is reduced through a reverse reaction after extraction. The mechanochemical auxiliary extraction can also utilize the physical and chemical properties of the target compound to selectively add a solid reagent,different auxiliaries can lead to different physicochemical conditions during ball milling, thereby changing the dissolution rate of the target compound in the extraction solvent; however, the peracid and alkali conditions may change the structure of the target compound, thereby reducing the extraction rate, so that the ball milling extraction requires the optimization of the auxiliary agent (Wuffeiflu, Zhaozhi, Xuyongping, etc.. the microshearing-auxiliary agent interaction technique using sodium chloride as the auxiliary agent to assist the extraction of the essential oil of citrus peel [ J]. Food safety Full quality detection bulletin, 2016(8): 3246-3252.). The mechanochemical auxiliary extraction method not only improves the extraction rate of target components, but also shortens the extraction time and reduces the requirements on extraction conditions, and is a high-efficiency and environment-friendly extraction method.
Disclosure of Invention
The invention aims to prepare a crude extract of naphthopyrone from berchemite, and provides a high-efficiency mechanochemical-assisted ultrasonic extraction method, which takes a mixed solvent of ethanol and water as an extraction solvent and obtains a high extraction rate in a short time.
The mechanochemical extraction method for preparing the crude extract of the naphthopyrone from the berchemite, which is characterized by comprising the following steps:
1) drying, crushing and sieving the berchemia lineata raw material to obtain berchemia lineata powder for later use;
2) carrying out mechanochemical treatment on the iron-coated gold powder obtained in the step 1), wherein the treatment process comprises the following steps: placing the iron-coated gold powder into a ball mill for ball milling, wherein the ball milling rotation speed is 100-400 rpm, and the ball milling time is 5-20 min, so as to obtain ball-milled iron-coated gold powder;
3) mixing the ball-milled iron-coated gold powder obtained in the step 2) with an extraction solvent according to a material-liquid ratio of 1: 20-100, wherein the unit of the material-liquid ratio is g/mL, then placing the mixture in a water bath at 20-50 ℃ for ultrasonic extraction for 15-60 min, cooling to room temperature after the ultrasonic extraction is finished, filtering, and removing the solvent from the filtrate through rotary evaporation to obtain a tawny naphthopyranone crude extract;
4) re-dissolving the crude extract obtained in the step 3) with methanol, standing, taking supernatant to pass through a 0.22 mu m PTFE membrane, analyzing by using an ultra-high performance liquid chromatography, and calculating the yield of the crude extract according to the content of the main component of the crude extract of naphthopyranone, namely rubrofusarin-6-O-alpha-L-rhamnosyl- (1-6) -O-beta-D-glucopyranoside.
The mechanochemical extraction method for preparing the crude extract of the naphthopyrone from the berchemite, which is characterized in that in the step 1), the mesh number of the obtained berchemite powder is more than 80 meshes.
The mechanochemical extraction method for preparing the crude naphthopyranone extract from the berchemite, which is characterized in that in the step 2), the ball milling filling rate in a ball mill is 2-16%, and the preferable rate is 5.36%.
The mechanochemical extraction method for preparing the crude naphthopyranone extract from the berchemite, which is characterized in that in the step 2), the ball milling speed is 200 rpm, and the ball milling time is 15 min.
The mechanochemical extraction method for preparing the crude extract of the naphthopyrone from the berchemilla lineata is characterized in that in the step 3), an extraction solvent is any one of petroleum ether, chloroform, ethyl acetate, methanol, ethanol and water, or a mixed solvent of ethanol and water, preferably a mixed solvent of ethanol and water.
The mechanochemical extraction method for preparing the crude naphthopyranone extract from the berchemia lineate is characterized in that the volume ratio of ethanol to water in the mixed solvent of ethanol and water is 20-80: 20-80, preferably 60: 40.
The mechanochemical extraction method for preparing the crude extract of the naphthopyrone from the berchemite, which is characterized in that in the step 3), the feed-liquid ratio of the berchemite powder to the extraction solvent is 1: 50; the ultrasonic extraction temperature is 40 deg.C, and the ultrasonic extraction time is 45 min.
The mechanochemical extraction method for preparing the crude extract of the naphthopyrone from the berchemite, which is characterized in that in the step 4), the process of analyzing by adopting the ultra-high performance liquid chromatography is as follows: the column was Waters Acquity BEH Shield RP C18 (100X 2.1 mm, 1.7 μm); the flow rate is 0.2 mL/min; the sample injection amount is 1.0 mu L; the column temperature is 40 ℃; the detection wavelength is 280 nm; the mobile phase A is 0.1% formic acid water solution, and the mobile phase B is acetonitrile; gradient elution conditions: 0-2 min, 10% B; 2-3 min, 10% -15% of B; 5-6 min, 15% -17% of B; 9-13 min, 17% -22% of B; 13-20 min, 22% -44% of B; 20-30 min, 44% -61% B; 32-32.5 min, 61% -69% B; 35-36 min, 10% B.
The invention has the following advantages:
(1) in order to improve the application value of the berchemia lineata, the invention adopts a mechanochemical auxiliary ultrasonic extraction technology to extract active ingredients in the berchemia lineata, and aims to preferably select a better extraction process for preparing the crude extract of the naphthopyrone from the berchemia lineata. The method adopts the mechanochemical combined ultrasonic technology to extract the naphthopyrones compounds in the berchemia lineate for the first time, and compared with the traditional ultrasonic method, the method obviously improves the extraction yield.
(2) The invention provides technical support for fully utilizing the medicinal value of the berchemia lineata as an important medicinal resource in China. The method has low cost and rapid and efficient extraction process, and can provide technical basis for the comprehensive utilization of the berchemia lineata medicinal resource.
(3) The invention has simple process, environmental protection and low cost.
(4) The crude extract of the naphthopyranone is prepared from berchemilla lineate, the main component of the crude extract of the naphthopyranone is rubromycin-6-O-alpha-L-rhamnosyl- (1-6) -O-beta-D-glucopyranoside, the content of the rubromycin-6-O-alpha-L-rhamnosyl- (1-6) -O-beta-D-glucopyranoside in the crude extract of the naphthopyranone can be quantitatively analyzed by ultra-high performance liquid chromatography, the extraction efficiency of the crude extract of the naphthopyranone prepared from the berchemilla lineate is evaluated according to the yield of the rubromycin-6-O-alpha-L-rhamnosyl- (1-6) -O-beta-D-glucopyranoside, the molecular structural formula of the rubrofusarin-6-O-alpha-L-rhamnosyl- (1-6) -O-beta-D-glucopyranoside is as follows:
drawings
FIG. 1 is a flow chart of the mechanochemical extraction process for the preparation of crude naphthopyrone extract from berchemilla lineate of example 1.
Detailed Description
The present invention is further illustrated by the following examples, which should not be construed as limiting the scope of the invention.
1. The specifications of the instruments and the medicines related to the invention are as follows:
PM200 planetary ball mill (Retsch, germany); waters Acquity UPLC ultra high performance liquid chromatograph (Waters, usa); XS205 dual range analytical balance (Mettler Toledo, switzerland); KH5200DE ultrasonic cleaner (kunshan ultrasonic instruments ltd); r-215 rotary evaporator (Buchi, Switzerland); barnstead TII ultrapure water system (Thermo Scientific, USA).
Water is ultrapure water (self-made in laboratories); the remaining reagents were analytical grade.
The samples of berchemia lineata used in the experiment were from Guangzhou, Guangxi river basin and Fujian Sanming, Guangdong.
2. The determination of rubrofusarin-6-O-alpha-L-rhamnosyl- (1-6) -O-beta-D-glucopyranoside in the crude naphthopyranone extract in berchemia lineate in the following examples is analyzed by ultra high performance liquid chromatography.
The yield of the main components is calculated by adopting an external standard method by taking rubrofusarin-6-O-alpha-L-rhamnosyl- (1-6) -O-beta-D-glucopyranoside (the purity is more than or equal to 99.0%) which is self-made by a laboratory as a reference substance. Taking the peak area y as a vertical coordinate and the mass concentration x (mg/mL) of the reference substance as a horizontal coordinate, drawing a standard curve, wherein a regression equation is as follows:
y = 8×106 x - 29136，R2=0.9991
yield of rubrofusarin-6-O-alpha-L-rhamnosyl- (1-6) -O-beta-D-glucopyranoside in crude extract of naphthopyranoneY) The calculation formula is as follows:
yield of the productY =rubrofusarin-6-O-alpha-L-rhamnosyl- (1-6) -O-beta-D-glucopyranoside mass (mg)/weight of the raw material of the iron-coated gold (g) × 100%.
Example 1
Crushing the berchemia lineata of Guangdong Guangzhou origin and sieving the crushed berchemia lineata with a 80-mesh sieve to obtain the berchemia lineata powder with the mesh number larger than 80 meshes. 0.30 g of the iron-coated gold powder is added into a stainless steel ball milling tank, 10 stainless steel balls with the diameter of 8 mm are added (the ball milling filling rate is 5.4%), the ball milling time is 15min, and the ball milling rotating speed is 200 rpm.
Weighing 0.10 g of ball-milled powder and an ethanol aqueous solution (V ethanol: V water = 60/40) according to a feed-liquid ratio of 1: 50 (the feed-liquid ratio is g/mL), placing the mixture in a water bath at 40 ℃ for ultrasonic extraction for 45 min (the ultrasonic frequency is 100 HZ), filtering, and removing the solvent from the filtrate through rotary evaporation to obtain 11 mg of crude naphthopyrone extract.
Dissolving 10.59 mg of the crude naphthopyrone extract with 1 mL of methanol, taking supernatant, filtering with a 0.22 mu m PTFE membrane, and performing sample introduction and ultra high performance liquid chromatography analysis, wherein the chromatographic conditions are as follows: the column was Waters Acquity BEH Shield RP C18 (100X 2.1 mm, 1.7 μm); the flow rate is 0.2 mL/min; the sample injection amount is 1.0 mu L; the column temperature is 40 ℃; the detection wavelength is 280 nm; the mobile phase A is 0.1% formic acid water solution, the mobile phase B is acetonitrile, and the gradient elution conditions are as follows: 0-2 min, 10% B; 2-3 min, 10% -15% of B; 5-6 min, 15% -17% of B; 9-13 min, 17% -22% of B; 13-20 min, 22% -44% of B; 20-30 min, 44% -61% B; 32-32.5 min, 61% -69% B; 35-36 min, 10% B. The yield of rubrofusarin-6-O-alpha-L-rhamnosyl- (1-6) -O-beta-D-glucopyranoside in the crude extract was calculated to be 143.1% (mg/g).
Example 1A schematic of a mechanochemical extraction process for the preparation of crude naphthopyrone extract from berchemilla lineate is shown in FIG. 1.
Example 2
Crushing the berchemia lineata of Guangdong Guangzhou origin and sieving the crushed berchemia lineata with a 80-mesh sieve to obtain the berchemia lineata powder with the mesh number larger than 80 meshes. 0.31 g of the iron-coated gold powder is added into a stainless steel ball milling tank, 15 stainless steel balls with the diameter of 8 mm are added (the ball milling filling rate is 8.0 percent), the ball milling time is 15min, and the ball milling rotating speed is 200 rpm.
0.10 g of the ball-milled powder was weighed, and the remaining subsequent ultrasonic extraction conditions and ultra-high performance liquid chromatography conditions were the same as in example 1, to obtain 11 mg of a crude naphthopyrone extract. The yield of rubrofusarin-6-O-alpha-L-rhamnosyl- (1-6) -O-beta-D-glucopyranoside in the crude extract was calculated to be 141.9% (mg/g).
Comparing example 2 with example 1, it is found that when 15 stainless steel balls were added (the ball-milling packing ratio reached 8.0%), the yield did not increase any more, but rather, the yield tended to decrease slightly. The main reason is that the excessive filling rate reduces the space in the ball milling tank, limits the movement of ball milling beads, reduces mechanical acting force and leads to the reduction of yield.
Example 3
Crushing the berchemia lineata of Guangdong Guangzhou origin and sieving the crushed berchemia lineata with a 80-mesh sieve to obtain the berchemia lineata powder with the mesh number larger than 80 meshes. 0.30 g of the iron-coated gold powder is added into a stainless steel ball milling tank, 20 stainless steel balls with the diameter of 8 mm are added (the ball milling filling rate is 10.7%), the ball milling time is 15min, and the ball milling rotating speed is 200 rpm.
0.10 g of the ball-milled powder was weighed, and the remaining subsequent ultrasonic extraction conditions and ultra-high performance liquid chromatography conditions were the same as in example 1, to obtain 10 mg of a crude naphthopyrone extract. The yield of rubrofusarin-6-O-alpha-L-rhamnosyl- (1-6) -O-beta-D-glucopyranoside in the crude extract was calculated to be 138.5% (mg/g).
Example 4
Crushing the berchemia lineata of Guangdong Guangzhou origin and sieving the crushed berchemia lineata with a 80-mesh sieve to obtain the berchemia lineata powder with the mesh number larger than 80 meshes. 0.30 g of the iron-coated gold powder is added into a stainless steel ball milling tank, 5 stainless steel balls with the diameter of 8 mm are added (the ball milling filling rate is 2.7%), the ball milling time is 15min, and the ball milling rotating speed is 200 rpm.
0.10 g of the ball-milled powder was weighed, and the remaining subsequent ultrasonic extraction conditions and ultra-high performance liquid chromatography conditions were the same as in example 1, to obtain 10 mg of a crude naphthopyrone extract. The yield of rubrofusarin-6-O-alpha-L-rhamnosyl- (1-6) -O-beta-D-glucopyranoside in the crude extract was calculated to be 115.8% (mg/g).
Example 5
Crushing the berchemia lineata of Guangdong Guangzhou origin and sieving the crushed berchemia lineata with a 80-mesh sieve to obtain the berchemia lineata powder with the mesh number larger than 80 meshes. 0.31 g of the iron-coated gold powder is added into a stainless steel ball milling tank, 10 stainless steel balls with the diameter of 8 mm are added (the ball milling filling rate is 5.4%), the ball milling time is 20 min, and the ball milling rotating speed is 200 rpm.
0.10 g of the ball-milled powder was weighed, and the remaining subsequent ultrasonic extraction conditions and ultra-high performance liquid chromatography conditions were the same as in example 1, to obtain 10 mg of a crude naphthopyrone extract. The yield of rubrofusarin-6-O-alpha-L-rhamnosyl- (1-6) -O-beta-D-glucopyranoside in the crude extract was calculated to be 125.4% (mg/g).
Comparing example 5 with example 1, it is understood that when the ball milling time is increased to 20 min, the yield does not increase any more but tends to decrease. The main reasons are that the mechanical force of the ball mill is too long, and the heat in the ball mill is accumulated, so that the effective components are damaged, and the yield is reduced.
Example 6
Crushing the berchemia lineata of Guangdong Guangzhou origin and sieving the crushed berchemia lineata with a 80-mesh sieve to obtain the berchemia lineata powder with the mesh number larger than 80 meshes. 0.31 g of the iron-coated gold powder is added into a stainless steel ball milling tank, 10 stainless steel balls with the diameter of 8 mm are added (the ball milling filling rate is 5.4%), the ball milling time is 5min, and the ball milling rotating speed is 200 rpm.
0.10 g of the ball-milled powder was weighed, and the remaining subsequent ultrasonic extraction conditions and ultra-high performance liquid chromatography conditions were the same as in example 1, to obtain 10 mg of a crude naphthopyrone extract. The yield of rubrofusarin-6-O-alpha-L-rhamnosyl- (1-6) -O-beta-D-glucopyranoside in the crude extract was calculated to be 118.5% (mg/g).
Example 7
Crushing the berchemia lineata of Guangdong Guangzhou origin and sieving the crushed berchemia lineata with a 80-mesh sieve to obtain the berchemia lineata powder with the mesh number larger than 80 meshes. 0.30 g of the iron-coated gold powder is added into a stainless steel ball milling tank, 10 stainless steel balls with the diameter of 8 mm are added (the ball milling filling rate is 5.4%), the ball milling time is 15min, and the ball milling rotating speed is 300 rpm.
0.10 g of the ball-milled powder was weighed, and the remaining subsequent ultrasonic extraction conditions and ultra-high performance liquid chromatography conditions were the same as in example 1, to obtain 10 mg of a crude naphthopyrone extract. The yield of rubrofusarin-6-O-alpha-L-rhamnosyl- (1-6) -O-beta-D-glucopyranoside in the crude extract was calculated to be 131.1% (mg/g).
As is clear from comparison between example 7 and example 1, when the rotation speed reached 300 rpm, the yield did not increase but slightly decreased. The main reason is that high energy is generated by excessively high rotating speed, so that effective components are damaged, and the yield is reduced.
Example 8
Crushing berchemia lineata in the Guangxi river basin as a production place, and sieving the crushed berchemia lineata with a 80-mesh sieve to obtain berchemia lineata powder with the mesh number larger than 80 meshes. 0.30 g of the iron-coated gold powder is added into a stainless steel ball milling tank, 10 stainless steel balls with the diameter of 8 mm are added (the ball milling filling rate is 5.4%), the ball milling time is 15min, and the ball milling rotating speed is 200 rpm.
0.10 g of the ball-milled powder was weighed, and the remaining subsequent ultrasonic extraction conditions and ultra-high performance liquid chromatography conditions were the same as in example 1, to obtain 15 mg of a crude naphthopyrone extract. The yield of rubrofusarin-6-O-alpha-L-rhamnosyl- (1-6) -O-beta-D-glucopyranoside in the crude extract was calculated to be 416.9% (mg/g).
Example 9
Crushing the berchemia lineate whose producing area is Fujian Sanming, and sieving the crushed berchemia lineate with a 80-mesh sieve to obtain the berchemia lineate powder with the mesh number larger than 80 meshes. 0.30 g of the iron-coated gold powder is added into a stainless steel ball milling tank, 10 stainless steel balls with the diameter of 8 mm are added (the ball milling filling rate is 5.4%), the ball milling time is 15min, and the ball milling rotating speed is 200 rpm.
0.10 g of the ball-milled powder was weighed, and the remaining subsequent ultrasonic extraction conditions and the conditions of the ultra-high performance liquid chromatography were the same as in example 1, to obtain 13 mg of a crude naphthopyrone extract. The yield of rubrofusarin-6-O-alpha-L-rhamnosyl- (1-6) -O-beta-D-glucopyranoside in the crude extract was calculated to be 280.8% (mg/g).
Comparing example 1, example 8 and example 9, it can be seen that the content of naphthopyrone substances in the raw materials of the berchemite in different producing areas is very different.
Example 10
Crushing the berchemia lineata of Guangdong Guangzhou origin and sieving the crushed berchemia lineata with a 80-mesh sieve to obtain the berchemia lineata powder with the mesh number larger than 80 meshes. 0.32 g of the iron-coated gold powder is added into a stainless steel ball milling tank, 10 stainless steel ball milling beads with the diameter of 8 mm (the ball milling filling rate is 5.4%) and 450 mg of an auxiliary agent (the auxiliary agent is sodium hydroxide) are added, the ball milling time is 15min, and the ball milling rotating speed is 200 rpm.
0.10 g of the ball-milled powder was weighed, and the remaining subsequent ultrasonic extraction conditions and ultra-high performance liquid chromatography conditions were the same as in example 1, to obtain 10 mg of a crude naphthopyrone extract. The yield of rubrofusarin-6-O-alpha-L-rhamnosyl- (1-6) -O-beta-D-glucopyranoside in the crude extract was calculated to be 42.2% (mg/g).
Example 11
Crushing the berchemia lineata of Guangdong Guangzhou origin and sieving the crushed berchemia lineata with a 80-mesh sieve to obtain the berchemia lineata powder with the mesh number larger than 80 meshes. 0.31 g of the iron-coated gold powder is added into a stainless steel ball milling tank, 10 stainless steel ball milling beads with the diameter of 8 mm (the ball milling filling rate is 5.4%) and 450 mg of an auxiliary agent (the auxiliary agent is anhydrous sodium carbonate) are added, the ball milling time is 15min, and the ball milling rotating speed is 200 rpm.
0.10 g of the ball-milled powder was weighed, and the remaining subsequent ultrasonic extraction conditions and ultra-high performance liquid chromatography conditions were the same as in example 1, to obtain 10 mg of a crude naphthopyrone extract. The yield of rubrofusarin-6-O-alpha-L-rhamnosyl- (1-6) -O-beta-D-glucopyranoside in the crude extract was calculated to be 49.4% (mg/g).
Example 12
Crushing the berchemia lineata of Guangdong Guangzhou origin and sieving the crushed berchemia lineata with a 80-mesh sieve to obtain the berchemia lineata powder with the mesh number larger than 80 meshes. 0.32 g of the iron-coated gold powder is added into a stainless steel ball milling tank, 10 stainless steel balls with the diameter of 8 mm (the ball milling filling rate is 5.4%) and 450 mg of an auxiliary agent (the auxiliary agent is sodium bicarbonate) are added, the ball milling time is 15min, and the ball milling rotating speed is 200 rpm.
0.10 g of the ball-milled powder was weighed, and the remaining subsequent ultrasonic extraction conditions and ultra-high performance liquid chromatography conditions were the same as in example 1, to obtain 10 mg of a crude naphthopyrone extract. The yield of rubrofusarin-6-O-alpha-L-rhamnosyl- (1-6) -O-beta-D-glucopyranoside in the crude extract was calculated to be 76.8% (mg/g).
Examples 13 to 22
The procedure of example 12 was repeated except that: the results of the determination of whether to add the auxiliary agent, the type of the added auxiliary agent, the ball milling time, the ball milling rotating speed, the ball milling filling rate, the change of specific conditions and the yield of the rubrofusarin-6-O-alpha-L-rhamnosyl- (1-6) -O-beta-D-glucopyranoside are shown in Table 1.
TABLE 1 Effect of mechanochemical parameters on the yield of rubrofusamycin-6-O-alpha-L-rhamnosyl- (1-6) -O-beta-D-glucopyranoside
From examples 1-22, it can be seen that the production location of the berchemie and the ball milling conditions had an effect on the extraction yield of rubrofusarin-6-O-alpha-L-rhamnosyl- (1-6) -O-beta-D-glucopyranoside. Under the condition of adding alkaline and strong acid auxiliary agents, the extraction rate of the rubrofusarin-6-O-alpha-L-rhamnosyl- (1-6) -O-beta-D-glucopyranoside is greatly reduced, and the main reason is that the mechanical force action in alkaline or strong acid environment influences the structural stability of the rubrofusarin-6-O-alpha-L-rhamnosyl- (1-6) -O-beta-D-glucopyranoside. A
The statements in this specification merely set forth a list of implementations of the inventive concept and the scope of the present invention should not be construed as limited to the particular forms set forth in the examples.
Claims (9)
1. A mechanochemical extraction method for preparing a crude naphthopyrone extract from berchemilla lineate is characterized by comprising the following steps:
1) drying, crushing and sieving the berchemia lineata raw material to obtain berchemia lineata powder for later use;
2) carrying out mechanochemical treatment on the iron-coated gold powder obtained in the step 1), wherein the treatment process comprises the following steps: placing the iron-coated gold powder into a ball mill for ball milling, wherein the ball milling rotation speed is 100-300 rpm, the ball milling filling rate in the ball mill is 2-8%, and the ball milling time is 5-20 min, so that ball-milled iron-coated gold powder is obtained;
3) mixing the ball-milled iron-coated gold powder obtained in the step 2) with an extraction solvent according to a material-liquid ratio of 1: 20-100, wherein the unit of the material-liquid ratio is g/mL, then placing the mixture in a water bath at 20-50 ℃ for ultrasonic extraction for 15-60 min, cooling to room temperature after the ultrasonic extraction is finished, filtering, and removing the solvent from the filtrate through rotary evaporation to obtain a tawny naphthopyranone crude extract;
4) re-dissolving the crude extract obtained in the step 3) with methanol, standing, taking supernatant to pass through a 0.22 mu m PTFE membrane, analyzing by using an ultra-high performance liquid chromatography, and calculating the yield of the crude extract according to the content of the main component of the crude extract of naphthopyranone, namely rubrofusarin-6-O-alpha-L-rhamnosyl- (1-6) -O-beta-D-glucopyranoside.
2. The mechanochemical extraction process for preparing crude naphthopyranone extract from berchemilla lineata as claimed in claim 1, wherein in step 1), the obtained berchemilla lineata powder has a mesh size of 80 meshes or more.
3. The mechanochemical extraction process for preparing crude naphthopyranone extract from berchemite according to claim 1, wherein in step 2), the ball mill packing rate in the ball mill is 5.36%.
4. The mechanochemical extraction process for preparing crude naphthopyranone extract from berchemie lineate as claimed in claim 1, wherein in step 2), the ball milling speed is 200 rpm and the ball milling time is 15 min.
5. The mechanochemical extraction method for preparing crude naphthopyranone extract from berchemilla lineate as claimed in claim 1, wherein in step 3), the extraction solvent is any one of petroleum ether, chloroform, ethyl acetate, methanol, ethanol and water, or a mixed solvent of ethanol and water.
6. The mechanochemical extraction method for preparing crude naphthopyranone extract from berchemilla lineate as claimed in claim 5, wherein the extraction solvent is a mixed solvent of ethanol and water, and the volume ratio of ethanol to water in the mixed solvent of ethanol and water is 20-80: 20-80.
7. The mechanochemical extraction process for preparing crude naphthopyranone extract from berchemilla lineate as claimed in claim 6, wherein the volume ratio of ethanol to water in the mixed solvent of ethanol and water is 60: 40.
8. The mechanochemical extraction method for preparing crude naphthopyranone extract from berchemia lineate as described in claim 1, wherein in step 3), the feed-to-liquid ratio of berchemia lineate powder to extraction solvent is 1: 50; the ultrasonic extraction temperature is 40 deg.C, and the ultrasonic extraction time is 45 min.
9. The mechanochemical extraction process for preparing crude naphthopyranone extract from berchemie lineate as claimed in claim 1, wherein in step 4), the analysis process by ultra performance liquid chromatography is as follows: the column was Waters Acquity BEH Shield RP C18 (100X 2.1 mm, 1.7 μm); the flow rate is 0.2 mL/min; the sample injection amount is 1.0 mu L; the column temperature is 40 ℃; the detection wavelength is 280 nm; the mobile phase A is 0.1% formic acid water solution, and the mobile phase B is acetonitrile; gradient elution conditions: 0-2 min, 10% B; 2-3 min, 10% -15% of B; 5-6 min, 15% -17% of B; 9-13 min, 17% -22% of B; 13-20 min, 22% -44% of B; 20-30 min, 44% -61% B; 32-32.5 min, 61% -69% B; 35-36 min, 10% B.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910323824.5A CN110066305B (en) | 2019-04-22 | 2019-04-22 | Mechanochemical extraction method for preparing crude naphthopyrone extract from berchemilla lineate |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910323824.5A CN110066305B (en) | 2019-04-22 | 2019-04-22 | Mechanochemical extraction method for preparing crude naphthopyrone extract from berchemilla lineate |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN110066305A CN110066305A (en) | 2019-07-30 |
| CN110066305B true CN110066305B (en) | 2020-12-01 |
Family
ID=67368426
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201910323824.5A Active CN110066305B (en) | 2019-04-22 | 2019-04-22 | Mechanochemical extraction method for preparing crude naphthopyrone extract from berchemilla lineate |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN110066305B (en) |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110003290B (en) * | 2019-04-22 | 2021-01-26 | 浙江工业大学 | Mechanochemical extraction method for preparing hydrolysable tannin crude extract from water chestnut shells |
| CN113082082A (en) * | 2021-04-27 | 2021-07-09 | 浙江工业大学 | Mechanochemical extraction method of mulberry leaf extract with alpha-glucosidase inhibitory activity |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102093449B (en) * | 2011-01-06 | 2014-01-22 | 刘斌 | Method for separating and preparing fusarubin gentiobioside from cassia seeds |
-
2019
- 2019-04-22 CN CN201910323824.5A patent/CN110066305B/en active Active
Also Published As
| Publication number | Publication date |
|---|---|
| CN110066305A (en) | 2019-07-30 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Wang et al. | Ultrasound-assisted extraction of geniposide from Gardenia jasminoides | |
| CN104906153A (en) | Technological method for efficiently extracting ginkgo flavone | |
| CN1164582C (en) | Process for preparing danshen salviandic acid | |
| CN102127140A (en) | Method for combinedly extracting Morroniside, dogwood polysaccharide and ursolic acid from dogwood | |
| CN110066305B (en) | Mechanochemical extraction method for preparing crude naphthopyrone extract from berchemilla lineate | |
| CN102178741A (en) | Guava leaf extract with function of reducing blood sugar as well as preparation method and application thereof | |
| CN110618211A (en) | Method for extracting scutellaria chemical components by using eutectic solvent | |
| CN115721671A (en) | Method for extracting and separating effective components of liquorice root in full-industrial-chain cooperation manner | |
| CN115894515A (en) | Method for integrated extraction, enrichment, separation and purification of effective components of ginkgo leaves in full industrial chain | |
| CN103224491A (en) | Method for extracting high-purity puerarin by using water as solvent | |
| CN104725450A (en) | Method for extracting high-purity oleuropein from jasminum grandiflorum | |
| CN106176844A (en) | The Folium Hibisci method for preparing extractive that a kind of antioxidation is strong | |
| CN103180334B (en) | Prepare the method for lactone glucoside of Radix Paeoniae and peoniflorin | |
| CN101530607B (en) | Method for extracting gingerol type material from ginger with high efficiency | |
| Liu et al. | Ultrasonic extraction and purification of scutellarin from Erigerontis Herba using deep eutectic solvent | |
| CN104069191B (en) | A kind of extraction process of water polygonum flaccidum general flavone | |
| Wu et al. | Preparation of paeoniflorin from the stems and leaves of Paeonia lactiflora Pall.‘Zhongjiang’through green efficient microwave-assisted extraction and subcritical water extraction | |
| CN103566195A (en) | Method for extracting alkaloid from dendrobium candidum | |
| CN101712731A (en) | Preparation and application of emodin molecular imprinted polymer | |
| CN102614231A (en) | Method for preparing cynomorium songaricum terpene and cynomorium songaricum polysaccharose from cynomorium songaricum | |
| CN108392506A (en) | The extracting method and pharmaceutical composition of a kind of wilsonii activity extract and its application | |
| CN101773528B (en) | Preparation method of total terpene lactones with low phenolic acid | |
| CN104491048A (en) | Loquat leaf total sesquiterpene extract as well as preparation method and application thereof | |
| CN110003290B (en) | Mechanochemical extraction method for preparing hydrolysable tannin crude extract from water chestnut shells | |
| CN1687092A (en) | General glycoside extractive of xanthium and preparation method |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |