CN105954378A - Method for determining total anthraquinone in Chinese rhubarb - Google Patents

Method for determining total anthraquinone in Chinese rhubarb Download PDF

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Publication number
CN105954378A
CN105954378A CN201610254539.9A CN201610254539A CN105954378A CN 105954378 A CN105954378 A CN 105954378A CN 201610254539 A CN201610254539 A CN 201610254539A CN 105954378 A CN105954378 A CN 105954378A
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China
Prior art keywords
extraction
rhubarb
instrument
methanol
anthraquinone
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CN201610254539.9A
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Chinese (zh)
Inventor
陈学松
廖强
韦日伟
陈佳丽
王�华
姚泳成
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Wuzhou Institutes for Food and Drug Control
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Wuzhou Institutes for Food and Drug Control
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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02—Column chromatography
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02—Column chromatography
    • G01N30/04—Preparation or injection of sample to be analysed
    • G01N30/06—Preparation
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02—Column chromatography
    • G01N2030/022—Column chromatography characterised by the kind of separation mechanism
    • G01N2030/027—Liquid chromatography
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02—Column chromatography
    • G01N30/04—Preparation or injection of sample to be analysed
    • G01N30/06—Preparation
    • G01N2030/062—Preparation extracting sample from raw material

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  • Physics & Mathematics (AREA)
  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Analytical Chemistry (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • General Physics & Mathematics (AREA)
  • Immunology (AREA)
  • Pathology (AREA)
  • Other Investigation Or Analysis Of Materials By Electrical Means (AREA)

Abstract

The invention discloses a method for determining total anthraquinone in Chinese rhubarb. The method comprises the following steps: step 1, slicing Chinese rhubarb, adding Chinese rhubarb slices into a pulverizer and adding nanometer quartz sand at the same time so as to obtain Chinese rhubarb powder, wherein a weight ratio of nanometer quartz sand to Chinese rhubarb is 0.1-0.2: 1; step 2, mixing the Chinese rhubarb powder with diatomite, adding the obtained mixture into an extraction pool in which quartz sand is laid, and carrying out extraction by using an ASE rapid extraction instrument, wherein extraction parameters are that an extraction solvent is a mixed solution of ethanol and methanol in a weight ratio of 0.3-0.4: 0.6-0.7, extraction temperature is 80 to 90 DEG C, static extraction time is 5 min, a flushing volume is 100%, one static cycle is carried out, static extraction pressure is 1500 psi and purging time in static extraction is 100 s; step 3, subjecting extract obtained in the previous step to centrifugation so as to obtain supernatant; and step 4, analyzing the supernatant by using liquid chromatography. The method for determining total anthraquinone in Chinese rhubarb is thorough in extraction and high in measurement precision.

Description

A kind of method measuring All Kind of Anthraquinone In Rhubarb
Technical field
The present invention relates to test in laboratory field, a kind of method measuring 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 Weighed, put in tool plug conical flask, accurate addition methanol 25ml, weighed weight, it is heated to reflux 1 hour, lets cool, then claim Determine weight, supply the weight of less loss with methanol, shake up, filter.Precision measures subsequent filtrate 5ml, puts in flask, flings to molten Agent, adds 8% hydrochloric acid solution 10ml, supersound process 2 minutes, then adds chloroform 10ml, be heated to reflux 1 hour, put Cold, put in separatory funnel, with a small amount of chloroform washing container, be incorporated in separatory funnel, divide and take chloroform layer, acid solution Again with chloroform extraction 3 times, each 10ml, merge chloroform liquid, decompression and solvent recovery as, residue adds methanol Make dissolving, be transferred in 10ml measuring bottle, add methanol to scale, shake up, filter, take subsequent filtrate, to obtain final product.Algoscopy is divided Accurate reference substance solution and each 10 μ l of need testing solution of drawing, injection chromatograph of liquid, measure, to obtain final product.This product is by dry Dry product calculate, containing general anthraquinone with aloe-emodin (C15H10O5), chrysophanic acid (C15H8O6), rheum emodin (C15H10O5), Radix Et Rhizoma Rhei Phenol (C15H10O4) and physcione (C16H12O5) total amount meter, must not be less than 1.5%.
In order to improve, pharmacopeia is tested during detection relatively complicated, high to the operant skill requirement of operator problem, I Be attempted with ASE rapid extraction-ASE and extract, extractant is 1% acetic acid and methanol aqueous solution；Extraction efficiency is higher.
Therefore, ASE rapid extraction-ASE is the development trend to Chinese crude drug detection method fast and accurately, extraction quick to ASE Follow the example of to optimize further and can improve accuracy of detection and detection speed.
Summary of the invention
It is an object of the invention to provide a kind of method measuring All Kind of Anthraquinone In Rhubarb, the method is extracted more thoroughly, certainty of measurement High.
The technical scheme that the present invention provides is: a kind of method measuring All Kind of Anthraquinone In Rhubarb, comprises the following steps:
Step 1: join in pulverizer after Radix Et Rhizoma Rhei is cut into slices, be simultaneously introduced nano quartz sand, nano quartz sand and Radix Et Rhizoma Rhei Weight ratio is 0.1-0.2:1, obtains Radix Et Rhizoma Rhei powder；
Step 2: use ASE quickly to extract by joining to be equipped with in the abstraction pool of quartz sand after Radix Et Rhizoma Rhei powder and kieselguhr mixing Take instrument extraction；Extraction parameters is as follows: extractant be weight ratio be that the mixing of the ethanol of 0.3-0.4:0.6-0.7 and methanol is molten Liquid；Extraction temperature 80-90 DEG C；Static extracting time 5min；Flush volume 100%；Quiet cycle number of times 1 time；Static The pressure of extraction is 1500psi；The purge time of described static extracting is 100s；
Step 3: after extract centrifugation, obtain supernatant；
Step 4: use liquid chromatography to be analyzed supernatant；
Wherein, the analysis condition of liquid chromatography is:
A) instrument: agilent1260 Ultra Performance Liquid Chromatography instrument
B) chromatographic column specification: C181.8 μm 4.6 × 100mm
C) column temperature: 40 DEG C
D) flow velocity: 0.5mL/min
E) flowing phase: 0-6min:0.1% phosphoric acid-methanol, 0.1% phosphoric acid solution-methanol volume ratio is 40:60；
6min:0.1% phosphoric acid-methanol, 0.1% phosphoric acid solution-methanol volume ratio is 10:90；
F) detection wavelength: 254nm.
In the above-mentioned method measuring All Kind of Anthraquinone In Rhubarb, step 3 is particularly as follows: by extract at the bar of 15000r/min Under part, centrifugal 3min, takes supernatant.
In the above-mentioned method measuring All Kind of Anthraquinone In Rhubarb, described instrument is agilent1260 Ultra Performance Liquid Chromatography instrument.
In the above-mentioned method measuring All Kind of Anthraquinone In Rhubarb, described chromatographic column is AgilentZORBAXExtend-C181.8μm4.6×100mm。
In the above-mentioned method measuring All Kind of Anthraquinone In Rhubarb, the instrument that described ASE extraction is used is ASE350 Accelerate solvent extraction instrument.
The present invention is after using technique scheme, and it has the beneficial effect that
The present invention, during detection, first passes through smashing and is then passed through ASE rapid extraction-ASE and extracts, it is possible to increase Separation accuracy, particularly in crushing process, uses nano quartz sand can improve cytoclastic effect, because nanometer stone Sand can stick to the surface of cell, and in telling crushing process, the nano quartz sand sticking to cell surface can and mutually be touched The cell friction hit and and Disintegrating knife friction, raising cell pulverization effect.
Meanwhile, extract uses ethanol methyl alcohol mixed liquor, i.e. can realize extracting effect the most thoroughly by a quiet cycle Really.
Use liquid phase chromatography analytical method, the content of general anthraquinone in Radix Et Rhizoma Rhei can be measured the most accurately.
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 this Bright any restriction.
Embodiment 1:
Join in pulverizer after 1g Radix Et Rhizoma Rhei is cut into slices, be simultaneously introduced the weight of nano quartz sand, nano quartz sand and Radix Et Rhizoma Rhei Ratio is 0.15:1, obtains Radix Et Rhizoma Rhei powder；0.1725g Radix Et Rhizoma Rhei powder is mixed homogeneously with proper amount of silicon diatomaceous earth, stand-by, first at abstraction pool Add a small amount of quartz sand, 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, extract is turned Moving in 50ml volumetric flask, 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
Analysis method is LC liquid chromatography, liquid-phase chromatographic analysis condition:
A) instrument: agilent1260 Ultra Performance Liquid Chromatography instrument
B) chromatographic column specification: AgilentZORBAXExtend-C181.8 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: (volume ratio 40:60, under the conditions of the volume ratio in the present embodiment is 20 DEG C to 0-6min:0.1% phosphoric acid-methanol Volume ratio)
6min:0.1% phosphoric acid-methanol (volume ratio 10:90), flowing before i.e. the 6th minute is the 0.1% of 40:60 for volume ratio mutually The mixture of phosphoric acid-methanol, when the 6th minute, flowing was mutually for the mixture of 0.1% phosphoric acid-methanol that volume ratio is 10:90.
F) detection wavelength: 254nm.
Comparative example 1
Join in pulverizer after 1g Radix Et Rhizoma Rhei is cut into slices, obtain Radix Et Rhizoma Rhei powder；Accurately weighed powder 0.15g, with proper amount of silicon diatomaceous earth Mix homogeneously, stand-by, first add a small amount of quartz sand at abstraction pool, then sample is moved into puts filter membrane in advance wellAdding proper amount of silicon diatomaceous earth in 10ml abstraction pool, shaking is allowed to Chi Kou in the same horizontal line, tighten extraction gently Pond upper cover.After extraction terminates, extract is shifted in 50ml volumetric flask, with methanol dilution to scale, at 15000r/min Under be centrifuged 3min, take supernatant, enter LC and measure.
Extraction conditions (being shown in Table 2) is:
Table 2
Close with pharmacopeia
Abstraction pool volume (ml) 10
Extractant 1% acetic acid and methanol aqueous solution
Pressure (psi) 1500
Temperature (DEG C) 100
The static extracting time (min) 5
Quiet cycle number of times 3
Flush volume (%) 100
Purge time (s) 100
Analysis method is LC liquid chromatography, liquid-phase chromatographic analysis condition:
A) instrument: agilent1260 Ultra Performance Liquid Chromatography instrument
B) chromatographic column specification: AgilentZORBAXExtend-C181.8 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 the 0.1% of 40:60 for volume ratio mutually The mixture of phosphoric acid-methanol, when the 6th minute, flowing was mutually for the mixture of 0.1% phosphoric acid-methanol that volume ratio is 10:90.
F) detection wavelength: 254nm.
The degree of accuracy test of the extracting process of embodiment 1
Using the assay method of embodiment 1, the assay method of comparative example 1 and official method to extract Radix Et Rhizoma Rhei, Radix Et Rhizoma Rhei is 3 batches, lot number is respectively 150912,1511012,20150701；
Concrete test result see table 3；
Table 3
The above-described presently preferred embodiments of the present invention that is only, all made in the range of the spirit and principles in the present invention any repair Change, equivalent and improvement etc., should be included within the scope of the present invention.

Claims (5)

1. the method measuring All Kind of Anthraquinone In Rhubarb, it is characterised in that comprise the following steps:
Step 1: join in pulverizer after Radix Et Rhizoma Rhei is cut into slices, be simultaneously introduced nano quartz sand, nano quartz sand and Radix Et Rhizoma Rhei Weight ratio is 0.1-0.2:1, obtains Radix Et Rhizoma Rhei powder；
Step 2: join after Radix Et Rhizoma Rhei powder and kieselguhr are mixed and be equipped with in the abstraction pool of quartz sand employing ASE Rapid Extraction Instrument extracts；Extraction parameters is as follows: extractant be weight ratio be ethanol and the mixed solution of methanol of 0.3-0.4:0.6-0.7； Extraction temperature 80-90 DEG C；Static extracting time 5min；Flush volume 100%；Quiet cycle number of times 1 time；Static extracting Pressure be 1500psi；The purge time of described static extracting is 100s；
Step 3: after extract centrifugation, obtain supernatant；
Step 4: use liquid chromatography to be analyzed supernatant；
Wherein, the analysis condition of liquid chromatography is:
A) instrument: agilent1260 Ultra Performance Liquid Chromatography instrument Y002
B) chromatographic column specification: C181.8 μm 4.6 × 100mm
C) column temperature: 40 DEG C
D) flow velocity: 0.5mL/min
E) flowing phase: 0-6min:0.1% phosphoric acid-methanol, 0.1% phosphoric acid solution-methanol volume ratio is 40:60；
6min:0.1% phosphoric acid-methanol, 0.1% phosphoric acid solution-methanol volume ratio is 10:90；
F) detection wavelength: 254nm.
The method of mensuration All Kind of Anthraquinone In Rhubarb the most according to claim 1, it is characterised in that step 3 particularly as follows: By extract centrifugal 3min under conditions of 15000r/min, take supernatant.
The method of mensuration All Kind of Anthraquinone In Rhubarb the most according to claim 1, it is characterised in that described instrument is Agilent1260 Ultra Performance Liquid Chromatography instrument.
The method of mensuration All Kind of Anthraquinone In Rhubarb the most according to claim 3, it is characterised in that described chromatographic column is AgilentZORBAXExtend-C181.8μm4.6×100mm。
The method of mensuration All Kind of Anthraquinone In Rhubarb the most according to claim 1, it is characterised in that described ASE extraction The instrument that method is used is ASE350 Accelerate solvent extraction instrument.
CN201610254539.9A 2016-04-22 2016-04-22 Method for determining total anthraquinone in Chinese rhubarb Pending CN105954378A (en)

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Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
RU2001396C1 (en) * 1991-05-06 1993-10-15 Государственный научно-исследовательский институт химии и технологии элементоорганических соединений Method of analysis of antraquinones
CN101603953A (en) * 2009-07-20 2009-12-16 韩桂茹 The Chrysophanol of polytype and archen method for quantitatively determining in rheum officinale and the compound preparation thereof
CN104257768A (en) * 2014-09-28 2015-01-07 江西百神药业股份有限公司 Application of total anthraquinone with various ingredients in stable and uniform proportion and composition of total anthraquinone to treatment of pancreatitis

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
RU2001396C1 (en) * 1991-05-06 1993-10-15 Государственный научно-исследовательский институт химии и технологии элементоорганических соединений Method of analysis of antraquinones
CN101603953A (en) * 2009-07-20 2009-12-16 韩桂茹 The Chrysophanol of polytype and archen method for quantitatively determining in rheum officinale and the compound preparation thereof
CN104257768A (en) * 2014-09-28 2015-01-07 江西百神药业股份有限公司 Application of total anthraquinone with various ingredients in stable and uniform proportion and composition of total anthraquinone to treatment of pancreatitis

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
XINGQIANG WU 等: "A Novel Selective Accelerated Solvent Extraction for Effective Separation and Rapid Simultaneous Determination of Six Anthraquinones in Tartary Buckwheat and Its Products by UPLC–DAD", 《FOOD ANAL. METHODS》 *
叶碧莎 等: "保健食品中总蒽醌的测定", 《中国卫生检验杂志》 *
李鹏: "加速溶剂提取技术在中药质量控制中的研究", 《中国优秀博硕士学位论文全文数据库（硕士） 医药卫生科技辑》 *
江连洲 等: "《酶在大豆制品中的应用》", 31 August 2015, 北京：中国轻工业出版社 *

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Application publication date: 20160921