TW201502513A - Method for identification of cinnamomum kanehirai hayata - Google Patents

Abstract

The invention provides a method for identifying if a sample derived from Cinnamomum kanehirai Hayata or Cinnamomum micranthum (Hayata) Hayata comprising detecting the contents of 4-terpineol, camphor and safrole in a volatile component of the sample. If the content of 4-terpineol is more than the total contents of camphor and safrole, the sample is derived from Cinnamomum kanehirai Hayata. If the content of safrole is more than the content of camphor, and the content of camphor is more than the content of 4-terpineol, the sample is derived from Cinnamomum micranthum (Hayata) Hayata.

Description

Method for identifying burdock

The present invention relates to a tree species technique. In particular, the present invention relates to a technique for identifying burdock using volatile components.

Cinnamomum kanehirai is a native tree species native to Taiwan. Because of its thick and solid tree shape, it is called "cow". Burdock is a large tree with a height of 30 meters and a diameter of 30 to 60 cm or more. The bark is brown, rough and often longitudinally split; the leaves are globose, apex blunt, pubescent outside, hairy at the edges; Oval, oblong, broadly elliptical or obovate, 9 to 11 cm long, 4 to 5 cm wide, apex acute or short-tailed, base acute, leathery, entire, smooth and shiny, with a contrasting back Light, not obvious three veins, leaf veins often have tufts; middle ribs and lateral veins are bulging on both sides of the surface; petiole slender, 0.8 to 1.2 cm long, smooth and hairless. Most flowers, scented, flower stalks 0.2 to 0.3 cm, arranged in terminal and axillary cymes, sometimes in panicles, corolla ca. 0.5 to 0.6 cm. The bracts have furry; the perianth tube is bell-shaped, the perianth is 6 pieces, approximately equal, long elliptical, 0.2 to 0.25 cm long, smooth outside, with furry inside, 9 stamens, first and second round Filaments 0.15 cm long, glandular, with villi at base; third filaments with glands, velum at base, approximate sessile glabrous; degenerate stamens arched, stalk velate, first and second anthers introverted; third round Anthers extroverted. The fruit is compressed in shape and has a length of 1 to 1.2 cm and a diameter of about 1.2 cm. The base is decorated with bell-shaped perianth.

Because of the parasitism of burdock, burdock has received great attention in recent years. . The wild burdock grows on the aging burdock tree, and the fruit body of the fungus grows from the inner surface of the trunk. The shape of the fruit body varies, and it has a plate shape, a bell shape, a horseshoe shape or a tower shape. It is bright red at the beginning and gradually changes. It is white, light reddish brown, light brown or light tan. According to the early legends of Taiwan, Niu Zhizhi was discovered by the aborigines. Because the aboriginal people like to drink alcohol, the proportion of suffering from liver disease is high. Inadvertently found anthrax in the mountains, eat raw or drink after cooking. The decoction found that although it tastes bitter, it has the effect of protecting the liver and eliminating fatigue, and it has been confirmed that Antrodia camphorata has an anti-cancer effect.

Because Antrodia camphorata is an extremely important research topic in the field of medical and health foods today, the value of the best host of the Antrodia camphorata can only be produced from Burdock, so the unique host of Burdock is also greatly increased. However, the identification of burdock does not have a set of scientifically standardized standard procedures. Most of them are judged by the appearance and aroma of the empirical value, which is easy to produce misjudgment, and generally no one can correctly identify the burdock.

Cinnamomum micranthum and Cinnamomum camphora are both genus of the genus Amaranthus, often confused with burdock.

Toona sinensis is a common plant on the ground. Its botany features are evergreen trees with a tree height of up to 40 meters; it emits a unique fragrance of eucalyptus. The bark is dark brown with longitudinal cracks. Oval or elliptic ovate-like leaves alternate, thin leathery, entire, smooth surface, slightly white powder on the back, glabrous, leaf margin slightly undulate, with three veins from the base, veins with obvious glands. Hermaphrodites, flowers bisexual, panicles axillary at apex, yellow-green flowers, flowering from April to May. Spherical berry, mature from October to November, from green to black purple at maturity; the skin is purple-black and shiny.

It is a large tree with smooth branches and a tall oval shape. Leaves leathery alternate, broadly ovate, ovate or elliptic, with smooth lower surface, distinct veins, large main veins, new leaf petiole red, old leaves green, stalk length 1.5 to 2 cm. Inflorescences cone-shaped, inflorescences axillate at the top of branches, yellow-green, flowers very small but numerous flowers, 6 perianth. The fruit is a long elliptical ball, 1.62 cm long and 1.17 cm wide. It is purple-black when mature, and the seed hypocotyl is often Raw, can provide raw materials for the production of camphor.

It is very similar to the shape of the burdock. In the form, the fruit of the scorpion is elliptical, and the burdock is a flat inverted cone or a spherical shape. In addition, the aroma is light and the burdock is thick. However, in the case of wood, the characteristics of flowers, leaves and fruits that are not related to botany are available for identification, and the appearance and odor of wood alone are highly likely to be misjudged.

In view of this, it is necessary for the industry to develop a method that can be quickly, easily operated, and accurately identified.

The invention develops a technology for identifying burdock, which can be applied to identify whether wood is derived from burdock, does not require botany-related flower, leaf and fruit identification, and does not require experience to develop, and is objective, accurate, rapid, and easy to operate. Identification method.

The present invention provides a method for identifying a sample to be tested, which is derived from burdock or cockroach, comprising measuring terpineol (4-terpineol), camphor (camphor) and baicalein in the volatile component of the sample to be tested ( The content of safrole), wherein: if the content of terpineol is greater than the total content of camphorin and baicalein, the sample to be tested is derived from burdock; and if the content of baicalein is greater than the content of camphorin, and the content of camphorin is greater than pine oil The content of the alcohol is determined to be derived from strontium; and the sample to be tested is a stem sample.

Figure 1 shows the TIC plot of a burdock sample.

Figure 2 shows the TIC plot of the rinder samples in the 14 to 21 minute interval.

Figure 3 shows the TIC plot of the ruthenium sample.

Figure 4 shows a TIC plot of the ruthenium sample over a 14 to 21 minute interval.

Figure 5 shows the TIC plot of the sample of the camphor.

Figure 6 shows the TIC plot of the citron sample in the 14 to 21 minute interval.

Figure 7 shows the results of the peak of the calf sample retention time of 15.06 minutes compared with the database.

Figure 8 shows the results of the 16.59 minute peak and database comparison of the calf sample retention time.

Figure 9 shows the results of the peak of the calf sample retention time of 20.63 minutes and the database comparison.

Figure 10 shows the results of the 15.19 minute peak and database comparison of the retention time of the ruthenium sample.

Figure 11 shows the comparison of the peak retention time of the sputum sample with 20.83 minutes and the database.

Figure 12 shows the results of the 15.13 minute peak and database comparison of the retention time of the sample of Toona sinensis.

Figure 13 shows the results of the peak of the camphor sample retention time of 20.64 minutes and the database comparison.

The present invention provides a method for identifying a sample to be tested, which is derived from burdock or cockroach, comprising measuring terpineol (4-terpineol), camphor (camphor) and baicalein in the volatile component of the sample to be tested ( The content of safrole), wherein: if the content of terpineol is greater than the total content of camphorin and baicalein, the sample to be tested is derived from burdock; and if the content of baicalein is greater than the content of camphorin, and the content of camphorin is greater than pine oil The content of the alcohol is determined to be derived from strontium; and the sample to be tested is a stem sample.

In a preferred embodiment of the present invention, the method according to the present invention may further comprise: identifying a sample to be tested as a method derived from burdock, medlar or citron, wherein: if the content of camphorin is greater than that of baicalein The content, and the content of baicalein is greater than the content of terpineol, the sample to be tested is derived from camphor.

The "burdock" as used in the present invention is also known as yak and black cockroach. Plant genus Magnolia door (Magnoliophyta), Magnolia Gang (Magnoliopsida), laurales (Laurales), evergreen Lauraceae (Lauraceae), cinnamon (Cinnamomum) of large trees. Its scientific name is Cinnamomum kanehirai Hayata , Cinnamomum kanehirai Hay . or Cinnamomum kanehirai .

The "冇樟", also known as Shenshui, Shuiyu, and Astragalus, is an evergreen tree of Magnolia plant, Magnolia, Odonata, Polygonaceae, and Eucalyptus. Its scientific name is Cinnamomum micranthum (Hayata) Hayata or Cinnamomum micranthum .

The "fragrant scorpion" as used in the present invention is also known as eucalyptus, hibiscus, black scorpion (Sichuan), fragrant scorpion, Panyu, Xiangrui, eucalyptus, and eucalyptus, belonging to the genus Magnolia, Magnolia, Acarina, Polygonaceae , the evergreen big tree of the genus. Its scientific name is Cinnamomum camphora (L.) Presl or Cinnamomum camphora .

The sample to be tested according to the present invention is a stem sample. In a preferred embodiment of the present invention, the sample to be tested is a stem of a living plant, preferably a stem of a living plant. section. In this preferred embodiment, the method according to the invention can be used to identify whether a living plant is a calf.

In another preferred embodiment of the present invention, the sample to be tested is a stem of an inanimate plant, preferably a part of a stem of an abiotic plant, such as but not limited to wood, eucalyptus, wood chips, wood. Piece. In this preferred embodiment, the method according to the invention can be used to identify whether a piece of wood, eucalyptus, wood chips, wood blocks are derived from burdock.

In a preferred embodiment of the present invention, the sample to be tested is pulverized into small pieces for further analysis. Those skilled in the art to which the present invention pertains may determine the method of comminution, such as cutting, cutting, planing, boring, and cutting using a knife or an axe.

The term "volatile component" as used in the present invention means a component which is volatile at normal temperature and pressure contained in a sample to be tested, preferably at a temperature of about 20 to about 30 ° C and 760 mmHg, and the vapor pressure is greater than 0.1 mmHg of substance. The method of obtaining the volatile component can be a general method. In a specific embodiment of the present invention, the volatile component of the sample to be tested is solid phase micro-extraction, steam distillation, carbon dioxide supercritical extraction, extrusion extraction, organic solvent. Extraction, microwave extraction, oil adsorption, water diffusion extraction or water vapor-solvent co-extraction. The above-described methods and conditions for the extraction or adsorption of oils and fats are determinable by those of ordinary skill in the art to which the present invention pertains.

Preferably, the volatile component of the sample to be tested is obtained by solid phase microextraction of the sample to be tested. Solid phase microextraction is a recent extraction method that has been widely used in environmental detection. It is mainly used to adsorb volatile compounds by adsorbing fibers, which is simple and fast and does not require any organic solvent extraction. technology.

In another embodiment of the present invention, the steam distillation can be divided into three modes of water distillation, water distillation, and steam distillation, and the appropriate manner can be selected according to the materials. In the extraction operation, the distillation unit needs to be sealed and matched with heating and cooling efficiency to obtain the best extraction results. The utility model has the advantages of low equipment cost, easy operation and mass production; the disadvantage is long working time, energy consumption, frequent loss of volatile and heat sensitive components, and high boiling point components are not easy to be distilled, which easily affects the integrity of volatile components.

In another embodiment of the present invention, the supercritical fluid extraction coefficient of carbon dioxide is about 10 times to about 100 times higher than that of the liquid, and if the gas has no surface tension, it is easy to penetrate into the sample to be tested. The volatile components are dissolved. Carbon dioxide is the most commonly used supercritical fluid because of its critical point temperature, slightly above room temperature (31 ° C), critical point pressure of 72.8 Bar, and is non-toxic, easy to obtain and inexpensive. When supercritical fluid extraction is used to extract in the vicinity of the supercritical region, the extraction method includes (1) static extraction; (2) dynamic extraction; (3) static extraction in the front stage and dynamic extraction in the latter stage to improve Extraction rate. After the completion of the extraction, it is preferred to further reduce the pressure and recover carbon dioxide to obtain a volatile component.

In another embodiment of the present invention, the squeeze extraction system smashes the sample to be tested to allow volatile components to flow out, and secondly, it can be rinsed with water, filtered, precipitated, centrifuged, and separated. The advantage is that the volatile components are not oxidized; the disadvantage is that the recovery rate and purity are low, and the operation is cumbersome.

In another embodiment of the present invention, the organic solvent extraction is performed by immersing the sample to be tested with an organic solvent such as n-hexane, filtering, and finally recovering the solvent in the extract to obtain a volatile component. The advantages are high purity and selectivity, good separation effect, and heat The sensitizing component has less damage; the disadvantage is that a suitable solvent must be selected, and during the operation, there is a solvent escaping, and if the solvent remains in the volatile component, the quality will be affected.

In another embodiment of the present invention, microwave extraction of microwave refers to electromagnetic wave extraction using a frequency of about 300 MHz to about 300 GHz. The high frequency electromagnetic wave can rapidly rub and collide polar molecules, and the extract absorbs microwave radiation energy, resulting in intracellular The rapid rise in temperature also causes the internal pressure of the cells to increase sharply. When the expansion of the cell membrane and the cell wall is exceeded, the cells are broken and various components are extracted. The advantages are fast, energy-saving, high-efficiency, high extraction rate; the disadvantage is to pay attention to the impact of electromagnetic waves on health, so it must be prevented from leaking.

In another embodiment of the present invention, the oil adsorption is suitable for extracting a heat sensitive component, including a cold suction method and a hot suction method, and the cold suction method first applies a grease such as lard, tallow or a mixture thereof to a glass plate. First, the sample to be tested is placed thereon, the volatile components are adsorbed, the oil is collected, and the volatile components in the oil are extracted by ethanol, and finally the ethanol is recovered; the hot suction method is to apply oil, such as lard or The butter is heated and melted, and the sample to be tested is immersed therein to absorb the aroma thereof, and the adsorbed and cooled fat is treated with ethanol. The advantage is that the ingredients are pure; the disadvantage is that it is very time consuming and labor intensive.

In another embodiment of the present invention, the water diffusion extraction also utilizes steam distillation, but the flow of the vapor is opposite to the steam distillation method, but spreads from the top to the bottom and passes through the sample to be tested by gravity. The extracted volatile components are of high quality and quantity, and energy saving is an advantage.

In another embodiment of the present invention, the water vapor-solvent co-extraction first puts the material into a container, adds an appropriate amount of water, and then adds another container to a suitable solvent, and then simultaneously heats both, and the water vapor, The volatile component and the evaporated solvent are introduced into a common condenser tube, that is, the extraction is carried out, and the volatile component is dissolved in the solvent. Since the solvent containing the volatile component and the water have different specific gravity, it is divided into two layers, and then The extracted solvent is collected, and finally the solvent is recovered to obtain the extracted volatile components. The advantage is that the solvent extraction method saves a lot of solvent, and the extraction effect of the trace components is better.

The method for analyzing the volatile components in the sample to be tested according to the present invention may be a general method, and it is preferred to simultaneously detect the type and content of the volatile component. In a specific embodiment of the present invention, the volatile component of the sample to be tested is analyzed by gas chromatography mass spectrometry, by thin layer chromatography or by gas chromatography ion flame detection. The method and conditions of operation of the foregoing analysis are those of ordinary skill in the art to which the invention pertains.

In a specific embodiment of the present invention, gas chromatography uses gas as the mobile phase, and the carrier gas itself has no separation effect. Generally, the commonly used gases are N ₂ , He, H ₂ . The main separation function of gas chromatography comes from the stationary phase (column material), and the specific temperature rise conditions can be used to effectively separate specific analytes. The common ion source of mass spectrometry is an electron impact mass spectrometer. When the sample enters the ion source, free electrons are generated by filament heating, and then accelerated, and then collided with the gasified molecules, so that the analyte becomes ion fragments and enters the mass spectrometer. After entering the mass spectrum, it is detected in the form of mass-to-charge ratio (m/z). Since different materials have different fragments, the distribution of the fragments can be used and compared with the database or standard. Therefore, the sample with high volatility is separated by gas chromatography, and then the molecular weight is measured by a mass spectrometer. Accurate qualitative and quantitative purposes can be achieved.

In a specific embodiment of the present invention, the thin layer chromatography analysis is performed by uniformly coating an adsorbent such as tannin or alumina on an aluminum sheet, a film or a glass plate to form a chromatography plate as a stationary phase. The volatile components are spotted on the stationary phase and separated by a different developing solvent (mobile phase). Through the capillary action, the moving phase moves from bottom to top, and the solubility of the mobile phase differs due to the different adsorption forces of the different substances and the stationary phase. Therefore, when the moving phase rises, the weakly moving movement is faster, and the strong adsorption force is slower. Because the adsorption power of different substances is different, the separation can be achieved on the chromatography plate. Qualitative, semi-quantitative analysis can be achieved with appropriate developers or coloration in different spectra.

In a specific embodiment of the present invention, gas chromatography in gas chromatography ion flame detector analysis, as described above, the ion flame detector mainly uses flame combustion to free organic matter, because it becomes conductive after being released. Ion, the detector can receive the signal that the current becomes stronger. And the current intensity is proportional to the carbon number of the organic matter, so the FID is suitable for a substance containing a CH-group, such as a fatty acid.

Preferably, the volatile component of the sample to be tested is analyzed by gas chromatography mass spectrometry. Gas chromatography-mass spectrometry is a method for simultaneously characterizing and quantifying volatile components, and the operation is fast, and the detection result can be obtained in a short time.

In a preferred embodiment of the present invention, when using gas chromatography mass spectrometry, a TIC pattern in the interval of 14 to 21 minutes can be selected, wherein the camphorin peaks at about 14.9 to about 15.4 minutes, and terpineol is about A peak appears from about 16.4 to about 16.9 minutes, and a peak appears in about 20.4 to about 20.9 minutes.

In a preferred embodiment of the present invention, the method according to the present invention comprises the steps of: (a) providing the sample to be tested; (b) solid phase micro-extracting the sample to be tested to obtain the volatile component; c) gas chromatography mass spectrometry analysis of the content of terpineol, camphorin and baicalein in the volatile component; and (d) comparing the content of terpineol, camphorin and baicalein in the volatile component.

Solid phase microextraction and gas chromatography mass spectrometry can be used to quickly identify burdock in one hour.

The invention is illustrated by the following examples, which are not intended to limit the invention.

Sample: Wood that has been identified as burdock, medlar, and citron.

Equipment: Gas Chromatograph: Trace GC Ultra, thermo

Mass spectrometer: ITQ 900, thermo

Chromatography column: VF-5ms 30 m* 0.25 mm ID* 0.25 μm, Varian

Solid phase micro extraction unit: 65μm PDMS/DVB

Sample processing and analysis: the wood was pulverized, and the pulverized wood chips were placed in a septum capped serum bottle, and then a solid phase micro-extraction device was inserted into the well and adsorbed in a 60 ° C water bath for 5 minutes, followed by adsorption. The solid phase micro-extraction apparatus was injected into the gas chromatograph, and the analysis conditions used were as follows: split ratio: 1:10

Use carrier gas: He(6N)

Injection chamber temperature: 250 ° C

Ion source: 200 ° C

Conditions: temperature rise conditions as shown in Table 1 and carrier gas conditions shown in Table 2

The measured gas chromatographic TIC diagrams of burdock, sorghum and citron are shown in Figures 1 to 6, wherein Figures 2, 4 and 6 are TIC diagrams for the 14 to 24 minute interval, indicating terpineol, Camphorin and The location and content of baicalein.

The results of the corresponding peak m/z and database comparison in the sample are shown in Figures 7 to 13, so it is determined that the peak of the residence time of 15.6 minutes in the burdock sample is camphorin, the peak of the residence time of 16.59 minutes is terpineol and the residence time. The peak of 20.63 minutes is baicalein; the peak of residence time in the sample is 15.19 minutes, the peak of camphorin and retention time is 20.83 minutes, and the peak of retention time of 15.13 minutes in camphor sample is peak of camphorin and retention time of 20.64 minutes. It is jaundice.

The contents of terpineol, camphorin and baicalein in Figures 2, 4 and 6 are shown in Table 3.

The above-described embodiments are merely illustrative of the principles and effects of the invention, and are not intended to limit the invention. Modifications and variations of the embodiments described above will be apparent to those skilled in the art without departing from the spirit of the invention. The scope of the invention should be as set forth in the appended claims.

Claims (10)

A method for identifying a sample to be tested is derived from burdock or cockroach, comprising measuring the content of terpineol (4-terpineol), camphor (camphor) and safrole in the volatile component of the sample to be tested. Wherein: if the content of resveratrol is greater than the total content of camphorin and baicalein, the sample to be tested is derived from burdock; and if the content of baicalein is greater than the content of camphorin and the content of camphorin is greater than the content of terpineol, Then, the sample to be tested is derived from strontium; and the sample to be tested is a stem sample. According to the method of claim 1, the method further comprises identifying a sample to be tested as a method derived from burdock, medlar or citron, wherein: if the content of camphorin is greater than the content of baicalein, and the content of baicalein is greater than pine oil The content of the alcohol, the sample to be tested is derived from the camphor. According to the method of claim 1 or 2, wherein the calf is Cinnamomum kanehirai Hayata , Cinnamomum kanehirai Hay ., Cinnamomum kanehirai or Cinnamomum micranthum . The method of claim 1 or 2, wherein the lanthanum is Cinnamomum micranthum (Hayata) Hayata or Cinnamomum micranthum . According to the method of claim 2, wherein the citron is Cinnamomum camphora (L.) Presl or Cinnamomum camphora . The method of claim 1 or 2, wherein the sample to be tested is a stem of a living plant. The method of claim 1 or 2, wherein the sample to be tested is a stem of an inanimate plant. According to the method of claim 1 or 2, wherein the volatile component of the sample to be tested is The sample to be tested is solid phase micro-extraction, steam distillation, carbon dioxide supercritical extraction, extrusion extraction, organic solvent extraction, microwave extraction, oil adsorption, water diffusion extraction or steam-solvent co-extraction. And got it. The method of claim 1 or 2, wherein the volatile component of the sample to be tested is analyzed by gas chromatography mass spectrometry, by thin layer chromatography or by gas chromatography ion flame detection. The method of claim 1 or 2, comprising the steps of: (a) providing the sample to be tested; (b) solid phase microextraction of the sample to be tested to obtain the volatile component; (c) a gas phase layer The content of terpineol, camphorin and baicalein in the volatile component was analyzed by mass spectrometry; and (d) the content of terpineol, camphorin and baicalein in the volatile component was compared.