CN105929086A

CN105929086A - Method for extracting total anthraquinone from rheum officinale

Info

Publication number: CN105929086A
Application number: CN201610234890.1A
Authority: CN
Inventors: 陈学松; 廖强; 韦日伟; 陈佳丽; 王�华; 姚泳成
Original assignee: Wuzhou Institutes for Food and Drug Control
Current assignee: Wuzhou Institutes for Food and Drug Control
Priority date: 2016-04-15
Filing date: 2016-04-15
Publication date: 2016-09-07

Abstract

The invention discloses a method for extracting total anthraquinone from rheum officinale. The method comprises the following steps: step one, grinding rheum officinale and sieving; step two, weighing 0.15 part by weight of rheum officinale powder, evenly mixing rheum officinale powder with a proper amount of diatomite, and saving the mixture for later use; step three, adding a little amount of quartz sand into an extraction tank provided with a filter membrane, evenly mixing, adding a proper amount of diatomite into the extraction tank until diatomite reaches the opening of the extraction tank; covering the extraction tank by a cover, carrying out static extraction; and obtaining the liquid extract after extraction; wherein the extraction conditions are as follows: the extracting solvent is acetic acid-methanol (1%), the extraction temperature is 100 DEG C, the static extraction lasts for 5 minutes, the flush volume is 100%, and the static circulation is performed for three times. The method has the advantages that the operation is simple, the extraction time is short, and the requirements on the skills of operators are lowered effectively.

Description

A kind of extracting method of All Kind of Anthraquinone In Rhubarb

Technical field

The present invention relates to test in laboratory field, the extracting method of a kind of All Kind of Anthraquinone In Rhubarb.

Background technology

Pharmacopeia in 2010 have recorded the method extracting All Kind of Anthraquinone In Rhubarb: takes this product powder (crossing No. four sieves) about 0.15g, accurate title Fixed, put in tool plug conical flask, accurate addition methanol 25ml, weighed weight, it is heated to reflux 1 hour, lets cool, more weighed weight, Supply the weight of less loss with methanol, shake up, filter.Precision measures subsequent filtrate 5ml, puts in flask, flings to solvent, adds 8% salt Acid solution 10ml, supersound process 2 minutes, then add chloroform 10ml, it is heated to reflux 1 hour, lets cool, put in separatory funnel, With a small amount of chloroform washing container, being incorporated in separatory funnel, divide and take chloroform layer, acid solution uses chloroform extraction 3 times again, Every time 10ml, merges chloroform liquid, decompression and solvent recovery as, residue adds methanol makes dissolving, is transferred in 10ml measuring bottle, Add methanol to scale, shake up, filter, take subsequent filtrate, to obtain final product.Algoscopy precision respectively draws reference substance solution and test sample The each 10 μ l of solution, inject chromatograph of liquid, measure, to obtain final product.This product is pressed dry product and is calculated, containing general anthraquinone with aloe-emodin (C15H10O5), chrysophanic acid (C15H8O6), rheum emodin (C15H10O5), chrysophanol (C15H10O4) and physcione (C16H12O5) total amount meter, must not be less than 1.5%.Chromatographic condition is: chromatographic condition and system suitability are with octadecane Base silane bonded silica gel is filler；With methanol 0.1% phosphoric acid solution (85:15) for flowing phase；Detection wavelength is 254.nm.Reason Opinion plate number is calculated by rheum emodin peak should be not less than 3000.

The method operating process of pharmacopeia is complicated, and the operant skill of operator is required height

Summary of the invention

It is an object of the invention to provide the extracting method of a kind of All Kind of Anthraquinone In Rhubarb, the method is simple to operate, and extraction efficiency is high.

The technical scheme that the present invention provides is: the extracting method of a kind of All Kind of Anthraquinone In Rhubarb, comprises the following steps:

Step 1: after Radix Et Rhizoma Rhei is pulverized, sieve；

Step 2: weigh 0.15 weight portion Radix Et Rhizoma Rhei powder, mix homogeneously stand-by with proper amount of silicon diatomaceous earth；

Step 3: be initially charged a small amount of quartz sand in the abstraction pool putting filter membrane in advance well, add appropriate diatom after mix homogeneously Native to the Chi Kou reaching abstraction pool；The lid covering abstraction pool carries out static extracting；Extraction is extracted liquid after terminating；

Extraction conditions is: extractant is 1% acetic acid and methanol aqueous solution；Extraction temperature 100 DEG C；Static extracting time 5min； Flush volume 100%；Quiet cycle number of times 3 times；Wherein, acetic acid and methanol aqueous solution are 99 unit bodies hydrops, 0.5 unit bodies Long-pending acetic acid and the carbinol mixture of 0.5 unit volume.

In the extracting method of above-mentioned All Kind of Anthraquinone In Rhubarb, the volume of described abstraction pool is 10ml.

In the extracting method of above-mentioned All Kind of Anthraquinone In Rhubarb, the pressure of described static extracting is 1500psi.

In the extracting method of above-mentioned All Kind of Anthraquinone In Rhubarb, the purge time of described static extracting is 100s.

In the extracting method of above-mentioned All Kind of Anthraquinone In Rhubarb, step 2 is particularly as follows: by extract under conditions of 15000r/min Centrifugal 3min, takes supernatant.

In the extracting method of above-mentioned All Kind of Anthraquinone In Rhubarb, the instrument that described ASE extraction is used is that ASE350 is quick Solvent extraction instrument

The present invention is after using technique scheme, and it has the beneficial effect that

The extracting method of the present invention is simple to operate, and the used time is short, effectively reduces the requirement of the operant skill to technical staff.

Accompanying drawing explanation

Fig. 1 is the rectangular plots of aloe-emodin chromatography validation verification in the present invention；

Fig. 2 is the rectangular plots of chrysophanic acid chromatography validation verification in the present invention；

Fig. 3 is the rectangular plots of rheum emodin chromatography validation verification in the present invention；

Fig. 4 is the rectangular plots of chrysophanol chromatography validation verification in the present invention；

Fig. 5 is the rectangular plots of physcione chromatography validation verification in the present invention；

Detailed description of the invention

Below in conjunction with detailed description of the invention, technical scheme is described in further detail, but does not constitute the present invention Any restriction.

Embodiment 1:

Size-reduced for Radix Et Rhizoma Rhei machine is pulverized, excessively No. four sieves, about 0.15g, accurately weighed, mix homogeneously with proper amount of silicon diatomaceous earth, stand-by, First add a small amount of quartz sand at abstraction pool, then sample is moved into puts filter membrane in advance well10ml abstraction pool adds Proper amount of silicon diatomaceous earth, shaking is allowed to Chi Kou in the same horizontal line, tighten abstraction pool upper cover gently.After extraction terminates, extraction Liquid shifts in 50ml volumetric flask, and with methanol dilution to scale, under 15000r/min, centrifugal 3min, takes supernatant, enters LC measures.

Extraction conditions (being shown in Table 1) is:

Table 1

Acetic acid and methanol aqueous solution are 99 unit bodies hydrops, the acetic acid of 0.5 unit volume and the carbinol mixture of 0.5 unit volume

Analysis method is LC liquid chromatography, liquid-phase chromatographic analysis condition:

A) instrument: agilent1260 Ultra Performance Liquid Chromatography instrument

B) chromatographic column specification: Agilent ZORBAX Extend-C18 1.8 μm 4.6 × 100mm, i.e. carbon 18 chromatographic column, Diameter 4.6mm, length 100mm, particle diameter 1.8 microns；

C) column temperature: 40 DEG C

D) flow velocity: 0.5mL/min

E) flowing phase: 0-6min:0.1% phosphoric acid-methanol (volume ratio 40:60)

6min:0.1% phosphoric acid-methanol (volume ratio 10:90), flowing before i.e. the 6th minute is 40:60 for volume ratio mutually The mixture of 0.1% phosphoric acid-methanol, when the 6th minute, flowing mixing for 0.1% phosphoric acid-methanol that volume ratio is 10:90 mutually Compound.

F) detection wavelength: 254nm

Chromatography validation verification:

Take aloe-emodin, chrysophanic acid, rheum emodin, chrysophanol, reference substance in right amount, accurately weighed, add methanol and make every 1ml Solution containing 16ug, physcione 8ug.Obtain.

Take aloe-emodin, chrysophanic acid, rheum emodin, chrysophanol, physcione reference substance in right amount, accurately weighed, put brown amount In Ping, add methanol make aloe-emodin 0.01295mg/ml, chrysophanic acid 0.01422mg/ml, rheum emodin 0.01021mg/ml, Chrysophanol 0.01215mg/ml, the solution of physcione 0.005385mg/ml, then respectively precision draw this solution 0.5 μ l, 0.8 μ l, 1 μ l, 1.5 μ l, 2ul enter LC and measure, the results detailed in Table 2-1, table 2-2, table 2-3, table 2-4, table 2-5； Accompanying drawing 1, accompanying drawing 2, accompanying drawing 3, accompanying drawing 4, accompanying drawing 5 respectively with table 2-1, table 2-2, table 2-3, table 2-4,2-5 pair, table Should, accompanying drawing 1, accompanying drawing 2, accompanying drawing 3, accompanying drawing 4, the abscissa indicated concentration of accompanying drawing 5, vertical coordinate represents peak area.

Table 2-1 aloe-emodin linear test result

Table 2-2 chrysophanic acid linear test result

Table 2-3 rheum emodin linear test result

Table 2-4 chrysophanol linear test result

Table 2-5 chrysophanol linear test result

The repeatability checking of the extracting process of 1.1 embodiments 1:

Take sample (lot number: the 20150701) 0.15g of identical lot number, totally 5 parts, accurately weighed, extract by ASE extracting method Need testing solution, sample size is 1 μ L, with above-mentioned chromatographic condition parallel test, record aloe-emodin in sample, chrysophanic acid, Rheum emodin, chrysophanol, the content of physcione are shown in Table three, and RSD is 1.02%, and test shows that ASE extracting method repeatability is good Good, the results detailed in Table 3

Table 3

The degree of accuracy test of the extracting process of 1.2 embodiments 1

Using the ASE extracting process of embodiment 1 and the extracting process of pharmacopeia to extract Radix Et Rhizoma Rhei, Radix Et Rhizoma Rhei is 3 batches, and lot number divides It is not 150912,1511012,20150701；

The ASE extracting process of embodiment 1 to using the equipment parallel testing twice of two set same sizes with a batch of Radix Et Rhizoma Rhei, point Do not represent for ASE1 and ASE2.

Concrete test result see table 4；

Table 4

Above-described be only presently preferred embodiments of the present invention, all made in the range of the spirit and principles in the present invention any amendment, Equivalent and improvement etc., should be included within the scope of the present invention.

Claims

1. the extracting method of an All Kind of Anthraquinone In Rhubarb, it is characterised in that comprise the following steps:

Step 1: after Radix Et Rhizoma Rhei is pulverized, sieve；

Extraction conditions is: extractant be volume ratio be 1% acetic acid and methanol aqueous solution；Extraction temperature 100 DEG C；During static extracting Between 5min；Flush volume 100%；Quiet cycle number of times 3 times, wherein, acetic acid and methanol aqueous solution be 99 unit bodies hydrops, The acetic acid of 0.5 unit volume and the carbinol mixture of 0.5 unit volume.

The extracting method of All Kind of Anthraquinone In Rhubarb the most according to claim 2, it is characterised in that the volume of described abstraction pool For 10ml.

The extracting method of All Kind of Anthraquinone In Rhubarb the most according to claim 2, it is characterised in that the pressure of described static extracting Power is 1500psi.

The extracting method of All Kind of Anthraquinone In Rhubarb the most according to claim 2, it is characterised in that blowing of described static extracting Flyback time is 100s.

The extracting method of All Kind of Anthraquinone In Rhubarb the most according to claim 1, it is characterised in that step 2 will be particularly as follows: will extract Take liquid centrifugal 3min under conditions of 15000r/min, take supernatant.

The extracting method of All Kind of Anthraquinone In Rhubarb the most according to claim 1, it is characterised in that described ASE extraction The instrument used is ASE350 Accelerate solvent extraction instrument.