CN102095822A - Method for measuring organic acid in hawthorn tincture used in cigarettes - Google Patents

Abstract

The invention discloses a method for measuring organic acid in hawthorn tincture used in cigarettes, which is characterized by comprising the steps of: adopting methanol, water, glacial acetic acid and triethylamine as a quaternary mobile phase, isocratic-eluting, and detecting by adopting a high-performance liquid chromatography-diode array detection method. In the method for detecting the content of oleanolic acid and ursolic acid in hawthorn tincture, when the sample is pretreated, a hawthorn tincture sample only needs to be diluted with the methanol, and then filtered with a 0.45-Mum filter membrane for liquid chromatographic separation and analysis without interference of other impurities; in addition, the method has the advantages of being simple, convenient and fast, having wide linear range, good separation effect and accurate and reliable data, and the like.

Description

A kind of mensuration cigarette organic acid method in the hawthorn fruit tincture

Technical field

The present invention relates to chemical analysis detection technique field, be specifically related to a kind of mensuration cigarette organic acid method in the hawthorn fruit tincture.

Background technology

Hawthorn fruit tincture is the Fructus Crataegi with ethanol lixiviate maturation, heats continuous circulating reflux extraction, obtains behind the recovery solvent, and the aromatic odor of existing uniqueness is rich in multiple active component to the human body beneficial again, therefore is widely used in the cigarette blending as a kind of perfume material.But because each manufacturing enterprise's extracting technology exists than big-difference, the hawthorn fruit tincture quality that obtains is uneven.Therefore, for cigarette industry enterprise, it is necessary selecting for use suitable hawthorn fruit tincture chemical constitution index to control its quality.The index components of the hawthorn fruit tincture assay of bibliographical information is mainly volatile ingredient at present, but the most content of volatile ingredient are all less, are difficult to the index as the hawthorn fruit tincture quality control.And oleanolic acid is relative with ursolic acid content higher, so select oleanolic acid and ursolic acid as one of its quality control index.

By literature survey, oleanolic acid and ursolic acid content assay method mainly contain thin layer chromatography scanning, GC-MS(gas chromatography-mass spectrography) and high performance liquid chromatography etc.Thin layer chromatography scanning is subjected to environment temperature, humidity effect bigger, and two kinds of component separating are difficult, poor reproducibility; Shortcomings such as the gas chromatography-mass spectrography technology is applied to the separation determination of oleanolic acid and ursolic acid, need do derivatization treatment to oleanolic acid and ursolic acid, has complex operation, and analysis time is long.High performance liquid chromatography have separate fast, good selective.Yet, because oleanolic acid and ursolic acid isomers each other in the hawthorn fruit tincture is added hawthorn fruit tincture ingredient complexity, the comparatively difficulty that its compartment analysis is become.Therefore, can better control cigarette with the quality of hawthorn fruit tincture, be necessary to set up a kind of cigarette with two kinds of isomeride organic acid assay methods each other in the hawthorn fruit tincture for making cigarette industry enterprise.

Summary of the invention

A kind of mensuration cigarette that purpose of the present invention provides at analyzing in the hawthorn fruit tincture two kinds of problems that exist during the organic acid of isomeride each other in the above-mentioned prior art just is with organic acid method in the hawthorn fruit tincture.This method by isocratic elution, has realized the cigarette two kinds of oleanolic acid of isomers and separating and mensuration of ursolic acid each other in the hawthorn fruit tincture by adopting glacial acetic acid and triethylamine as the moving phase adjuvant simultaneously.

Purpose of the present invention can realize by following technique measures:

Method of the present invention adopts quaternary moving phase by to the simple pre-treatment of hawthorn fruit tincture sample, carries out under following liquid phase chromatogram condition;

Specifically, said method comprising the steps of:

A, specimen preparation: accurately take by weighing the hawthorn fruit tincture sample (being accurate to 0.001g) of 3.0g, place the 10mL volumetric flask, be diluted to scale 10mL, shake up with methyl alcohol, through 0.45 μ m membrane filtration, stand-by then;

B, sample that step a is prepared are with liquid chromatography-diode array detector analysis, and analysis condition is as follows:

Chromatographic column: be 4.6mm * 250mm, the Waters SymmetryC of 5 μ m ₁₈Post;

Moving phase: be to be that methanol-water-glacial acetic acid-triethylamine of 87:13:0.04:0.02 is prepared from by weight ratio;

Flow velocity: 1.0mL/min;

Column temperature: 30 ℃;

Detect wavelength: 210nm;

Sample size: 10 μ L;

Through isocratic elution, realize two kinds of organic acids separation determination simultaneously.

Described organic acid is oleanolic acid and ursolic acid.

Beneficial effect of the present invention is as follows:

The organic acid analysis method is compared in the hawthorn fruit tincture of the present invention and prior art, this method only needs the hawthorn fruit tincture sample is diluted with methyl alcohol, then through 0.45 μ m membrane filtration, can feed liquor phase chromatographic resolution analyze, do not have the interference of other impurity, have easyly, quick, the range of linearity is wide, good separating effect, data wait advantage accurately and reliably.In addition, because oleanolic acid and ursolic acid belong to pentacyclic triterpenoid together, be isomers, structure polarity is similar, and the two is difficult to separate.It is the quaternary moving phase high performance liquid chromatography-photodiode array detection method of moving phase adjuvant that glacial acetic acid and triethylamine are adopted in this experiment, oleanolic acid in the hawthorn fruit tincture and ursolic acid the two effective separation can have been confirmed that this method is simple and feasible.

 

Description of drawings

Accompanying drawing is oleanolic acid and a ursolic acid chromatogram in the hawthorn fruit tincture.

Among the figure: 1. oleanolic acid 2. ursolic acid

Embodiment

The present invention is further described below with reference to embodiment:

Embodiment 1

Accurately take by weighing the hawthorn fruit tincture sample (being accurate to 0.001g) of 3.0g, place the 10mL volumetric flask, be diluted to scale 10mL, shake up with methyl alcohol, through 0.45 μ m membrane filtration, stand-by then;

The liquid-phase chromatographic analysis condition:

Chromatographic column: waters Symmetry C18 post (4.6mm * 250mm * 5 μ m)

Flow velocity: 1.0mL/min

Column temperature: 30 ℃

Detect wavelength: 210 nm

Sample size: 10 μ L

Moving phase: methanol-water-glacial acetic acid-triethylamine (87:13:0.04:0.02)

Under this chromatographic condition, the chromatogram of hawthorn fruit tincture sample as shown in Figure 1.

Get 3 parts in embodiment 1 hawthorn fruit tincture sample, record oleanolic acid and content of ursolic acid and be respectively 87.28 μ g/g and 117.17 μ g/g, its RSD is respectively 2.0% and 4.0%, and the result shows that this method has reappearance preferably to oleanolic acid and content of ursolic acid mensuration.

Claims (1)

1. measure cigarette organic acid method in the hawthorn fruit tincture for one kind, it is characterized in that: this method comprises the steps:

A, specimen preparation: accurately take by weighing the hawthorn fruit tincture sample of 3.0g, place the 10mL volumetric flask, be diluted to scale 10mL, shake up with methyl alcohol, through 0.45 μ m membrane filtration, stand-by then;

B, sample that step a is prepared are with liquid chromatography-diode array detector analysis, and analysis condition is as follows:

Chromatographic column: be 4.6mm * 250mm, the Waters SymmetryC of 5 μ m ₁₈Post;

Moving phase: be to be that methanol-water-glacial acetic acid-triethylamine of 87:13:0.04:0.02 is prepared from by weight ratio;

Flow velocity: 1.0mL/min;

Column temperature: 30 ℃;

Detect wavelength: 210nm;

Sample size: 10 μ L;

Behind isocratic elution, realize two kinds of organic acids of oleanolic acid and ursolic acid separation determination simultaneously.

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