Method of determining volume of fraction of volatile compounds in ethereal oil of mint leaves

FIELD: agriculture.

SUBSTANCE: invention relates to the field of agriculture and food industry, as well as aromatherapy. In the method the volume of fraction of volatile compounds of ethereal oil of mint leaves is determined in a period of technical ripeness. The volume of fraction is calculated from the difference between the amount of ethereal oil of secretory glands, obtained based on the number and size of the glands per unit of area of mint plant leaves using light microscope with a video system Lumenera Infinity 2 followed by data processing using the program Infinity camera V:5.02, and the volume of ethereal oil obtained by the method of hydrodistillation.

EFFECT: method enables to determine quickly the content of fraction of volatile compounds in the ethereal oil and carry out the selection of breeding material.

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The invention relates to food industry, aromatherapy and production drugstore sheet, and for the evaluation of hybrids and varieties of mint on the number of volatile compounds in the essential oil of mint. Aromatic properties of essential oils of peppermint known for a long time. The main purpose of peppermint is obtaining essential oils to extract a menthol for the pharmaceutical industry, the second, not less important, the use of mint plants is the food industry, pharmacy sheet and aromatherapy, where important aroma, i.e. the presence of volatile compounds. Mint leaves are used as an integral part as the flavoring agent in a variety of drinks, including teas, and for the preparation of gathering medicinal plants as native essential oil peppermint contains the fraction of volatile compounds volatile when heated and creating a unique aroma. When this oil each of the numerous interspecific hybrids, intraspecific clones and varieties has a unique flavor that is defined by the composition and the proportion of volatile compounds.

Essential oil accumulates in the spherical outgrowths of the epidermis of the leaves of mint plants, called secretory glands. They represent a spherical tank, sostojashie is from the cavity, filled with oil, with a thin outer shell, and legs, with which a piece of iron attached to the surface of the sheet.

Known method of extracting essential oils "enfleurage", the principle of which is that the oil evaporating from the flowers, is absorbed by the sorbent solid fats, activated carbon). Of forming a fragrant mass essential oil is extracted by solvent.

The closest to the technical nature of the claimed method is a method of extraction of raw materials solvents in a column apparatus, obtaining essential oils with subsequent determination of its composition by combining methods of gas chromatography and mass spectrometry (Kheifets L.A., Dashurin V.M., 1994). The method allows to determine the composition of the components of essential oil and, in particular, volatile compounds. The disadvantages of this method are the duration of the analysis and the need for high-precision and expensive equipment.

The problem solved by the invention is the creation of a more simple way of determining the volume fraction of volatile compounds of the essential oil of interspecific hybrids, intraspecific clones and varieties of mint. The problem is solved by taking into account the number and morphological parameters secretory glands on the leaves of mint, selected in the phase of technical splost is, calculate the volume of essential oil glands per unit area of leaves (1000 cm2). At the same time in the mint leaves is determining the content of essential oil method hydrodistillation (method Clevenger, modified Loshkareva (GOST 24027.2-80)). The difference is the content of essential oil between the two methods is the fraction of volatile compounds Requires the presence of a light microscope with video Lumenera Infinity 2 or digital camera line to fix the zoom when taking pictures, or binocular microscope with ocular micrometer; the device for determination of essential oils.

Progress

We analyzed mint plants are selected 10-30 leaves 5-7 pairs 2-3 storey main flowering shoots in the phase of technical maturity, because in this phase ends with the formation of the essential oil glands and has the highest oil content.

Using video or digital camera attached to a light microscope, photographed at magnification ×500 10 plots each sheet with top and bottom sides, because the glands are often located on both sides of the sheet. On the images, using Infinity camera V:5.02 count the number of glands in the frame, determine the surface area of the sheet in the frame in cm, number of glands on 1 cm2the upper and lower sides of a sheet. Also is measure the diameter and calculate the average value of the diameter of metal.

You can carry out these calculations using scale bars, for which her photograph and take into account that 1mm=km.

In the absence of a light microscope similar measurements can be performed by using a stereomicroscope. The number of glands in the field of view, the diameter and area of the field of view is done using the ocular micrometer.

Using the data obtained, calculate the volume of essential oil glands per 1000 cm2the leaf area by the formulas:

V1000=Vtop+Vbottom;

Vthe top or bottom=4/3πR3×n×1000,

where V1000- the volume of essential oil glands per 1000 cm2the leaf area, cm3;

Vtop- the volume of essential oil glands on the upper side, cm3;

Vbottom- the volume of essential oil glands on the underside, cm3;

π=3,14;

R is the radius of the glands, cm;

n is the number of glands on 1 cm2the surface of the sheet pieces

Simultaneously analyzed in leaves of mint plants carry out the determination of the content of essential oil method hydrodistillation. For this 200-300g fresh leaves are placed in the unit for essential oil distillation, and according to GOST 24027.2-80 the essential oil and measure its volume. Fixed mass of 50 mint leaves, determined by their size using planimetry and calculate the mass of 1000 cm2the surface area of the leaves by the formula:

M1000=m50×1000s50,

where M1000- weight of 1000 cm2leaf surface, g;

m50- weight of 50 leaves, g;

s50- surface area of 50 leaves, cm2.

Counting the number of essential oil obtained by the method of hydrodistillation at 1000 cm2the surface area of the leaves is carried out according to the formula:

V*1000=M1000×V*nandinewith aKand,

where V*1000- the amount of essential oil obtained by the method of hydrodistillation at 1000 cm2the surface area of leaves, cm3;

V* is the volume of essential oil obtained by the method of hydrodistillation, cm3;

M1000- weight of 1000 cm2the surface of leaves,

Hanging - hanging fresh leaves taken for hydrodistillation, was the determination of the volume fraction of volatile compounds in leaves of mint is calculated by the formula:

VLF=V1000-V*1000,

where VLF- the volume fraction of volatile compounds in the essential mA the Le mint leaves, cm3;

V1000- the volume of essential oil (total volume of iron) at 1000 cm2the leaf area, cm3;

V*1000- the amount of essential oil obtained by the method of hydrodistillation at 1000 cm2the surface area of leaves, cm3.

The contents of fractions of volatile compounds in the percent of essential oil of mint leaves is determined by the formula:

Vl.f×100V1000,

where VLF- the volume fraction of volatile compounds in the essential oil

mint leaves, cm3;

V1000- the volume of essential oil (total volume of iron) at 1000 cm2

the leaf area, cm3.

Example 1. The calculation of VLFfor varieties of mint old breeding, zoned for Ukraine and southern regions of Russia, grown on the experimental area of the laboratory in the Main Botanical garden of Russian Academy of Sciences. In the analyzed plants selected 30 leaves 5-7 pairs 2-3 storey main flowering shoots in the phase of technical maturity of each variety counted the number of secretory glands, their diameter and V1000. At the same time the samples were taken from fresh leaves of each variety and the content of essential oil methods hydrodistillation. Determine the volume fraction of volatile compounds and calculated their essential oil content varieties of mint old selection.(table 1).

Table 1.
The determination of the volume fraction of volatile compounds in the essential oil of peppermint.
gradethe amount of essential oils, cm3The contents of fractions of volatile compounds in the essential oil of peppermint, %
the total amount of iron,
V1000
the amount of oil after hydrodistillation, V*1000the amount of volatile compounds, VLF
1. Krasnodar 20,509of 0.1330,37673,9
2. Kuban 60,6460,2120,43467,2
3. Simferopol 2000,6640,1800,48472,9
4. Muscovite0,6050,2240,381630
5. Uichanco0,766of 0.3370,42956,1
6. Michalska0,5000,1660,33466,8
7. Medicines 40,5830,2800,30352,0

As can be seen from the tables, in the composition of essential oil of varieties of mint old selection based on M.piperita, the content of the volatile fraction of connections exceeds 50%, and in varieties Krasnodar 2 and Simferopol 200 exceeds 70%. These varieties more efficient use of fresh - food industry or aromatherapy, and for drugstore sheet. When used to obtain essential oils should apply the methods of maceration, as when using the method of hydrodistillation fraction of volatile compounds are lost, leading to loss of essential oil and deterioration of its quality.

Example 2. The calculation of VLFfor interspecific hybrids, intraspecific mint clones grown on the experimental area of the laboratory in the Main Botanical garden of Russian Academy of Sciences.

In the analyzed plants from the gap 30 leaves 5-7 pairs 2-3 storey main flowering shoots in the phase of technical maturity of each variety counted the number of secretory glands, their diameter and V1000- At the same time the samples were taken from fresh leaves of each variety and the content of essential oil methods hydrodistillation. Determine the volume fraction of volatile compounds and calculated their essential oil content varieties of mint old breeding (table 2).

Table 2.
The determination of the volume fraction of volatile compounds in the essential oil of peppermint.
gradethe amount of essential oils, cm3The contents of fractions of volatile compounds in the essential oil of peppermint, %
the total amount of iron, V1000the amount of oil after hydrodistillation, V*1000the amount of volatile compounds, VLF
Intraspecific tetraploid clone of M. arvensis,0,5730,313is 0.260to 45.4
Intraspecific clone of M. arvensis (NV-74)0,6700,1510,51977,5
Intraspecific clone of M. arvensis (17-Taiwan)0,5020,170of 0.33266,2
Intraspecific clone of M. arvensis (18-Japan)0,5410,1910,35064,7
Intraspecific clone of M. arvensis (24-Syktyvkar)0,0940,0250,06973,5
Intraspecific clone of M. arvensis (19-Crimea)0,4600,0720,38884,4
Interspecific hybrid M. piperita and M. crispa
Intraspecific clone of M. crispa (Crimea)
0,6110,1650,44673,0
0,7450,100to 0.64586,6

As can be seen from the tables, in the composition of the essential oil most intraspecific clones mint M arvensis, the content of the fraction of volatile compounds is from 65 to 85% of these samples can be used in fresh appearance is in the food industry, pharmacy list and in aromatherapy. Only tetraploid clone of M. arvensis content fraction of volatile compounds was less than 50%, so it can be recommended for the production of essential oils by the method of hydrodistillation.

Thus, our proposed method for determining volume fraction of volatile compounds in the essential oil of mint leaf allows you to quickly assess the content of the fraction of volatile compounds in the essential oil and filter material for food industry, pharmacy production sheet and aromatherapy.

Sources of information

1. The Kheifets L.A., Dashurin V.M. fragrances and other products for perfumes. - M.: Chemistry, 1994. - P.185.

2. GOST 24027.2-80. Raw medicinal plant. Methods for the determination of moisture, ash content, extractive and tannins, essential oils.

1. The method for determining the volume fraction of volatile compounds of essential oil of mint leaf in the period of technical maturity, characterized in that calculated by the difference between the volume of essential oil secretory glands, obtained on the basis of the number and size of glands per unit area of leaves of mint plants using a light microscope with video Lumenera Infinity 2 with subsequent data processing using Infinity camera V:5.02 and the amount of essential oil obtained by the method of hydrotest is LaCie.

2. The method according to claim 1, characterized in that the amount of essential oil secretory glands obtained using a light microscope with a digital camera and a scale ruler.

3. The method according to claim 1, characterized in that the amount of essential oil secretory glands obtained using the binocular microscope with ocular micrometer.



 

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