CN103884702A - Determination method for content of volatile oil in atractylodes macrocephala volatile oil/beta-cyclodextrin inclusion compound - Google Patents
Determination method for content of volatile oil in atractylodes macrocephala volatile oil/beta-cyclodextrin inclusion compound Download PDFInfo
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- inclusion compound
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Abstract
The invention relates to the field of quality control of an atractylodes macrocephala volatile oil inclusion compound and particularly relates to a determination method for the content of volatile oil in an atractylodes macrocephala volatile oil/beta-cyclodextrin inclusion compound. The determination method comprises the following steps: I, preparing a volatile oil ethanol de-clathration solution in the inclusion compound: precisely weighing the atractylodes macrocephala volatile oil/beta-cyclodextrin inclusion compound and adding absolute ethyl alcohol; treating by ultrasounds for 10 minutes and refrigerating for 24 hours; centrifuging for 10 minutes at rotary speed of 2500r/min; taking supernatant liquid, wherein the mass/volume ratio of the inclusion compound to the absolute ethyl alcohol is 3 to 1; and II, carrying out fluorescence analysis on the supernatant liquid. The determination method for the content of the volatile oil in the atractylodes macrocephala volatile oil/beta-cyclodextrin inclusion compound is simple and convenient to operate and rapid in analyzing, only needs few samples, is high in determination sensitivity and good in repeatability; a fluorescence analysis method for the volatile oil in the atractylodes macrocephala volatile oil/beta-cyclodextrin inclusion compound is developed before and after oxygenolysis so as to obtain the indexes including the inclusion rate, the oil rate and the like of the inclusion compound.
Description
Technical field
The present invention relates to Baizhu volatile oil inclusion compound field of quality control, specifically the assay method of volatile oil content in a kind of Baizhu volatile oil/Benexate Hydrochloride.
Background technology
What Baizhu volatile oil/cyclodextrin inclusion compound assay used traditionally is steam distillation.Although the method is widely applied, its heat time is long, large to instability composition influence in volatile oil, and the sample size needing is also large, tests length consuming time, operation relative complex, and experimental error is large.
Therefore, need badly a kind of simple to operation, analyze and fast, only need minute quantity sample, measure highly sensitive and reproducible assay method Baizhu volatile oil inclusion compound is realized to Quality Control.
Summary of the invention
The present invention is directed to Baizhu volatile oil and there is the volatile general character of volatile oil material, the assay method of volatile oil content in this kind of Baizhu volatile oil/Benexate Hydrochloride is provided.
The present invention is achieved by the following technical solutions: the assay method of volatile oil content in Baizhu volatile oil/Benexate Hydrochloride, described Baizhu volatile oil is volatile oil before oxygenolysis or the volatile oil after oxygenolysis, volatile oil after described oxygenolysis is: the volatile oil before oxygenolysis is after oxygenolysis, and wherein the resolution ratio of atractylone reaches 100% Baizhu volatile oil;
The step of this assay method is:
I. the preparation of the de-bag of volatile oil ethanol solution in inclusion compound: precision takes Baizhu volatile oil/Benexate Hydrochloride, adds absolute ethyl alcohol, ultrasonic 10min, refrigeration 24h; With 2500r/min rotating speed, centrifugal 10min, gets supernatant liquor subsequently; The mass volume ratio of inclusion compound and absolute ethyl alcohol is 3:1;
II. supernatant liquor is carried out to fluorescence analysis:
If a. Baizhu volatile oil is the volatile oil before oxygenolysis, fluorescence condition is: take 429nm as emission wavelength, take any wavelength within the scope of 340～409nm as excitation wavelength, slit width is 5nm, and fluorescence analysis obtains its fluorescence intensity F;
If 1. excitation wavelength is 367nm, its fluorescence intensity is F
,, try to achieve volatile oil content C in its supernatant liquor according to formula (1)
,, obtain volatile oil percentage composition in inclusion compound by calculating;
F
， = 44.23 C
， + 9.30 （1）
In formula (1): F
,for the fluorescence intensity of supernatant liquor;
C
,for the content (W/V) of volatile oil in supernatant liquor;
If 2. excitation wavelength is other wavelength, its fluorescence intensity is F
,,, reduce multiple B=F
,/ F
,,, try to achieve volatile oil content C in its supernatant liquor according to formula (2)
,,, obtain volatile oil percentage composition in inclusion compound by calculating;
F
，， = （44.23 C
，， + 9.30）/B （2）
In formula (2): F
,,for the fluorescence intensity of supernatant liquor;
C
,,for the content (W/V) of volatile oil in supernatant liquor;
If b. Baizhu volatile oil is the volatile oil after oxygenolysis, fluorescence condition is: take 485nm as emission wavelength, take any wavelength within the scope of 315～465nm as excitation wavelength, slit width is 5nm, and fluorescence analysis obtains its fluorescence intensity F;
If 1. excitation wavelength is 396nm, its fluorescence intensity is F
,, try to achieve volatile oil content C in its supernatant liquor according to formula (3)
,, obtain volatile oil percentage composition in inclusion compound by calculating;
F
，= 38.68 C
， + 7.01 （3）
In formula (3): F
,for the fluorescence intensity of supernatant liquor;
C
,for the content (W/V) of volatile oil in supernatant liquor;
If 2. excitation wavelength is other wavelength, its fluorescence intensity is F
,,, reduce multiple B=F
,/ F
,,, try to achieve volatile oil content C in its supernatant liquor according to formula (4)
,,, obtain volatile oil percentage composition in inclusion compound by calculating;
F
，，=（ 38.68 C
，， + 7.01 ）/B （4）
In formula (4): F
,,for the fluorescence intensity of supernatant liquor;
C
,,for the content (W/V) of volatile oil in supernatant liquor.
Baizhu volatile oil/Benexate Hydrochloride described in the present invention can obtain by inclusion compound preparation methods such as polishing well known in the art or ultrasonic methods.
Volatile oil before described oxygenolysis is the volatile oil extracting in Rhizoma Atractylodis Macrocephalae.
The mass volume ratio of step I inclusion compound and absolute ethyl alcohol is 3:1(W/V), this ratio is in other ratios, and the volatile oil content data accuracy calculating according to its fluorescence intensity is high; If other ratios, in supernatant liquor, the content value of volatile oil may, not within the range of linearity of typical curve, if still calculate the content of volatile oil in inclusion compound according to each formula of the present invention, can impact the accuracy of result of calculation.
Step I of the present invention, according to β-CD(beta-schardinger dextrin-) be slightly soluble in absolute ethyl alcohol, Baizhu volatile oil is soluble in absolute ethyl alcohol, by the parameter in step I, the Baizhu volatile oil in Baizhu volatile oil/Benexate Hydrochloride is dissociated out.
Step II of the present invention has the feature of strong fluorescent emission according to volatile oil ethanolic solution, according to the volatile oil heterogeneity feature after volatile oil and oxygenolysis before oxygenolysis, different fluorescence analysis conditions is set, and the formula of concentrating on studies according to inventor, try to achieve volatile oil content in volatile oil ethanolic solution (supernatant liquor), and then obtain volatile oil content in inclusion compound.
For the volatile oil before oxygenolysis, inventor thinks that any wavelength within the scope of 340～409nm is that excitation wavelength all can adopt.Because emission wavelength is 429nm, if excitation wavelength is greater than 409nm, excitation wavelength and emission wavelength values be apart from too little, and both may phase mutual interference, affects the accuracy of fluorescence intensity level; If excitation wavelength is less than 340nm, excitation wavelength is not within the scope of whole excitation peak, and fluorescence intensity level is too little, do not reach the requirement of assay.
For the volatile oil after oxygenolysis, inventor thinks that any wavelength within the scope of 315～465nm is that excitation wavelength all can adopt, and the value reason of the excitation wavelength within the scope of this is the same.
The assay method of volatile oil content in Baizhu volatile oil/Benexate Hydrochloride of the present invention, simple to operation, analyze fast, only need minute quantity sample, mensuration sensitivity is high, reproducible, be slightly soluble in absolute ethyl alcohol according to β-CD, Baizhu volatile oil is soluble in absolute ethyl alcohol, and volatile oil ethanolic solution has the feature of strong fluorescent emission, carry out before oxygenolysis, volatile oil fluorescence analysis method in rear Baizhu volatile oil/Benexate Hydrochloride, thereby obtain the inclusion rate of inclusion compound, the indexs such as oil content, it is the valuable Baizhu volatile oil inclusion compound quality control method of a kind of science, the solution of this problem is expected to promote the R&D work of Baizhu volatile oil/Benexate Hydrochloride anti-cancer agent preparation.
Embodiment
embodiment 1
Instrument and condition:
Varian Cary Eclipse fluorospectrophotometer, SK250H Ultrasound Instrument; KDC-1044 low speed centrifuge; FA-1004 analytical balance, GM-0.33 II vacuum diaphragm pump, β-CD, absolute ethyl alcohol, the volatile oil/Benexate Hydrochloride before oxygenolysis.
Emission wavelength: 429nm;
Excitation wavelength is respectively: 340nm, 367nm, 409nm;
Slit width is 5nm;
F
， = 44.23 C
， + 9.30 （1）
F
，， = （44.23 C
，， + 9.30）/B （2）。
Experimental procedure:
I. the preparation of the de-bag of volatile oil ethanol solution in inclusion compound:
Precision takes the volatile oil/Benexate Hydrochloride 30mg before oxygenolysis, adds 10mL absolute ethyl alcohol, ultrasonic 10min, refrigeration 24h; With 2500r/min rotating speed, centrifugal 10min, gets supernatant liquor subsequently;
II. supernatant liquor is carried out to fluorescence analysis:
In the time that excitation wavelength is 367nm, fluorescence analysis obtains its fluorescence intensity 31.136, and according to formula (1), in supernatant liquor, volatile oil content is 0.4937mg/mL, volatile oil before the oxygenolysis that contains 4.937mg in 10mL volatile oil ethanolic solution, in inclusion compound, volatile oil content is 16.45%;
In the time that excitation wavelength is 340nm, fluorescence analysis obtains its fluorescence intensity 6.742, B=31.136/6.742=4.62,
F
，， = （44.23 C
，， + 9.30）/B=9.57 C
，，+2.01 （2）
According to formula (2), in supernatant liquor, volatile oil content is 0.4947mg/mL, the volatile oil before the oxygenolysis that contains 4.947mg in 10mL volatile oil ethanolic solution, and in inclusion compound, volatile oil content is 16.49%;
In the time that excitation wavelength is 409nm, fluorescence analysis obtains its fluorescence intensity 16.755, B=31.136/16.755=1.87,
F
，， = （44.23 C
，， + 9.30）/B=23.65 C
，，+4.97 （2）
According to formula (2), in supernatant liquor, volatile oil content is 0.4983mg/mL, the volatile oil before the oxygenolysis that contains 4.983mg in 10mL volatile oil ethanolic solution, and in inclusion compound, volatile oil content is 16.61%.
In inclusion compound, the mean value of volatile oil content is 16.51%, RSD=0.504%.
embodiment 2
Instrument and condition:
Varian Cary Eclipse fluorospectrophotometer, SK250H Ultrasound Instrument; KDC-1044 low speed centrifuge; FA-1004 analytical balance, GM-0.33 II vacuum diaphragm pump, β-CD, absolute ethyl alcohol, the volatile oil/Benexate Hydrochloride after oxygenolysis.
Emission wavelength: 485nm;
Excitation wavelength is respectively: 315nm, 396nm, 465nm;
Slit width is 5nm;
F
，= 38.68 C
， + 7.01 （3）
F
，，=（ 38.68 C
，， + 7.01 ）/B （4）。
Experimental procedure:
I. the preparation of the de-bag of volatile oil ethanol solution in inclusion compound:
Precision takes the volatile oil/Benexate Hydrochloride 30mg after oxygenolysis, adds 10mL absolute ethyl alcohol, ultrasonic 10min, refrigeration 24h; With 2500r/min rotating speed, centrifugal 10min, gets supernatant liquor subsequently;
II. supernatant liquor is carried out to fluorescence analysis:
In the time that excitation wavelength is 396nm, fluorescence analysis obtains its fluorescence intensity 31.918, and according to formula (3), in supernatant liquor, volatile oil content is 0.6440mg/mL, volatile oil after the oxygenolysis that contains 6.440mg in 10mL volatile oil ethanolic solution, in inclusion compound, volatile oil content is 21.46%;
In the time that excitation wavelength is 315nm, fluorescence analysis obtains its fluorescence intensity 0.602, B=31.918/0.602=53.02,
F
，，=（ 38.68 C
，， + 7.01 ）/B =0.73 C
，，+0.13 （4）
According to formula (2), in supernatant liquor, volatile oil content is 0.6466mg/mL, the volatile oil before the oxygenolysis that contains 6.466mg in 10mL volatile oil ethanolic solution, and in inclusion compound, volatile oil content is 21.55%;
In the time that excitation wavelength is 465nm, fluorescence analysis obtains its fluorescence intensity 2.573, B=31.918/2.573=12.40,
F
，，=（ 38.68 C
，， + 7.01 ）/B =3.12C
，，+0.57 （4）
According to formula (2), in supernatant liquor, volatile oil content is 0.6420mg/mL, the volatile oil before the oxygenolysis that contains 6.420mg in 10mL volatile oil ethanolic solution, and in inclusion compound, volatile oil content is 21.40%.
In inclusion compound, the mean value of volatile oil content is 21.47%, RSD=0.352%.
Claims (1)
1. the assay method of volatile oil content in Baizhu volatile oil/Benexate Hydrochloride, it is characterized in that, described Baizhu volatile oil is volatile oil before oxygenolysis or the volatile oil after oxygenolysis, volatile oil after described oxygenolysis is: the volatile oil before oxygenolysis is after oxygenolysis, and wherein the resolution ratio of atractylone reaches 100% Baizhu volatile oil;
The step of this assay method is:
I. the preparation of the de-bag of volatile oil ethanol solution in inclusion compound: precision takes Baizhu volatile oil/Benexate Hydrochloride, adds absolute ethyl alcohol, ultrasonic 10min, refrigeration 24h; With 2500r/min rotating speed, centrifugal 10min, gets supernatant liquor subsequently; The mass volume ratio of inclusion compound and absolute ethyl alcohol is 3:1;
II. supernatant liquor is carried out to fluorescence analysis:
If a. Baizhu volatile oil is the volatile oil before oxygenolysis, fluorescence condition is: take 429nm as emission wavelength, take any wavelength within the scope of 340～409nm as excitation wavelength, slit width is 5nm, and fluorescence analysis obtains its fluorescence intensity F;
If 1. excitation wavelength is 367nm, its fluorescence intensity is F
,, try to achieve volatile oil content C in its supernatant liquor according to formula (1)
,, obtain volatile oil percentage composition in inclusion compound by calculating;
F
， = 44.23 C
， + 9.30 （1）
In formula (1): F
,for the fluorescence intensity of supernatant liquor;
C
,for the content (W/V) of volatile oil in supernatant liquor;
If 2. excitation wavelength is other wavelength, its fluorescence intensity is F
,,, reduce multiple B=F
,/ F
,,, try to achieve volatile oil content C in its supernatant liquor according to formula (2)
,,, obtain volatile oil percentage composition in inclusion compound by calculating;
F
，， = （44.23 C
，， + 9.30）/B （2）
In formula (2): F
,,for the fluorescence intensity of supernatant liquor;
C
,,for the content (W/V) of volatile oil in supernatant liquor;
If b. Baizhu volatile oil is the volatile oil after oxygenolysis, fluorescence condition is: take 485nm as emission wavelength, take any wavelength within the scope of 315～465nm as excitation wavelength, slit width is 5nm, and fluorescence analysis obtains its fluorescence intensity F;
If 1. excitation wavelength is 396nm, its fluorescence intensity is F
,, try to achieve volatile oil content C in its supernatant liquor according to formula (3)
,, obtain volatile oil percentage composition in inclusion compound by calculating;
F
，= 38.68 C
， + 7.01 （3）
In formula (3): F
,for the fluorescence intensity of supernatant liquor;
C
,for the content (W/V) of volatile oil in supernatant liquor;
If 2. excitation wavelength is other wavelength, its fluorescence intensity is F
,,, reduce multiple B=F
,/ F
,,, try to achieve volatile oil content C in its supernatant liquor according to formula (4)
,,, obtain volatile oil percentage composition in inclusion compound by calculating;
F
，，=（ 38.68 C
，， + 7.01 ）/B （4）
In formula (4): F
,,for the fluorescence intensity of supernatant liquor;
C
,,for the content (W/V) of volatile oil in supernatant liquor.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
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|---|---|---|---|
| CN201410144714.XA CN103884702B (en) | 2014-04-12 | 2014-04-12 | The assay method of volatile oil content in Baizhu volatile oil/Benexate Hydrochloride |
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2014
- 2014-04-12 CN CN201410144714.XA patent/CN103884702B/en not_active Expired - Fee Related
Patent Citations (6)
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| CN101825572A (en) * | 2010-06-11 | 2010-09-08 | 重庆大学 | Method for differentiating Chinese spirits with different flavor types with fluorescein |
| US20120043479A1 (en) * | 2010-08-17 | 2012-02-23 | Dow Agrosciences Llc. | Normalization of Biomolecules |
| WO2013088883A1 (en) * | 2011-12-15 | 2013-06-20 | コニカミノルタ株式会社 | Device for chromatographic analysis and method for chromatographic analysis |
| CN102706844A (en) * | 2012-06-07 | 2012-10-03 | 苏州市中心血站 | Method for detecting content of methylene blue in blood plasma |
| CN103149187A (en) * | 2013-02-21 | 2013-06-12 | 江南大学 | Fluorescent method for rapidly determining content of aliphatic acid |
| CN103396890A (en) * | 2013-07-26 | 2013-11-20 | 山西省肿瘤医院 | Method for preparing stable bighead atractylodes rhizome volatile oil |
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| 阎克里 等: "白术挥发油提取方法研究", 《中国药物与临床》 * |
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