KR960007789B1 - Analysing method for allicine in garlic oil - Google Patents

Abstract

Allicine is changed to two isomer 144-I(3-vinyl-1,2-dithi-5-ene), 144-II(3-vinyl-1,2-dithi-4-ene) at 150 deg. C or more of gas chromatography. So, we have to use di-n-propyl disulfide as internal standard. Allicine is changeable with the passage of time. So, have to use diallyl disulfide as standard. The method shows good reproducibility of assay.

Description

New Analysis of Allicin in Garlic Oil

1 is a gas chromatogram of diallyl disulfide standard of garlic oil of Example 1,

2 shows diallyl disulfide, di-n-propyl disulfide (internal standard), 3-vinyl-1,2-dity-5-ene (144-I) and 3-vinyl-1, of the garlic oil of Example 1; Gastrogram of 2-dithi-4-ene- (144-II),

3 is a gas chromatogram of diallyl disulfide of garlic oil of a commercial product (Euromed: trade name),

4 shows diallyl disulfide, di-n-propyl disulfide (internal standard), di-n-propyl disulfide (internal standard) of garlic oil of a commercial product (Euromed: trade name), 3-vinyl-1,2-di- Gastrogram of 4-ene (144-I) and 3-vinyl-1,2-diti-4-ene- (144-II).

The present invention relates to a new method for analyzing allicin in Garlic oil.

Garlic (Allium sativum L) has been widely used worldwide as a seasoning and folk medicine since ancient times. Garlic has the effect of smoothing the function of heart and blood flow, prevents blood clots and arteriosclerosis of coronary arteries, and is effective in treating diseases caused by infections and diseases caused by abnormal blood flow, lowers blood cholesterol levels and coagulates blood. Since there is a pharmacological action for improving blood circulation and improving peripheral blood circulation disorders such as delaying time, it is effective to prevent and treat various adult diseases.

A study of the chemical constituents in garlic, especially those with unusual odors, was the first in 1844 to announce that Bertheim first announced the existence of an allyl compound with the structure of C ₃ H ₅ (Ann. Chem. Pharm., 51,289, 1844) and in 1892 Semmler announced the presence of sulfur compounds such as diallyl disulfide and diallyl trisulfide among the aromatic components in garlic. (Arch. Pharm., 230, 434, 1892). Later, in 1944, Cabillito et al. Extracted and distilled garlic from Allicin and released it (J. Am. Chem. Soc., 66, 1944). : Diluted to 125,000 and bactericidal.

Broadnitz et al., Allylic, diallyl thiosulfide, have the structural formula CH ₂ = CHCH ₂ SOSCH ₂ CH = CH ₂ , which is a precursor of several sulfur compounds in garlic (Precursor). Allin in garlic, or S-Allylcysteine sulfoxide, becomes allicin by the action of an enzyme called Allinase when the garlic is ground. This allicin is diallyl disulfide. (Diallyl disulfide), diallyl sulfide and diallyl trisulfide such as sulfur compounds (Michei H. Bronditz, J. Agr. Food. Chem., Vol 19, No. 2 , 273, 1971).

Garlic contains not only allicin but also carbohydrates such as reducing sugars, sugars, starches, and textins, amino acids such as cysteine, histidine, and lysine and nicotine phase, vitamin C, vitamin A, thiamine, riboflavin, niacin, and biotin. The main component that represents is known to be a sulfur compound allicin.

The preparation for using garlic as a medicine can be seen that it was taken as juice or syrup because garlic juice and garlic syrup were loaded into the British Pharmacopoeia in 1949. Pharmaceutical Standards and Test Methods in accordance with 10) The first individual formulation contains garlic flow extract (0.01% allicin), but no commercialized garlic powder is commercially available.

Garlic oil containing 0.1% of allicin, or Garlic oil, is widely used as a medicine in Europe, especially Germany and Spain.In Korea, the garlic oil has been imported from Germany and Spain for about 10 years. After blending with vitamins and the like to produce a drug having the above-mentioned garlic.

Garlic oil being imported, ie garlic oil, does not mean garlic oil, but refers to oil containing fresh garlic components (l in knoblauchzwiebeln) (Von Holger Miething, Deutsche Apotheker Zritung, 125 Jahrg. Nr. 41. 10 10. 1985).

In the conventional garlic oil production method, the steam distillation method or the solvent extraction method were tested in order to extract the active ingredient in garlic, but all these methods need to be heated during processing, and since allicin is very unstable and volatile in heat, Low.

As a result of a long study, the inventors found that the active ingredient of allicin, garlic, is very affinity for oil, so that the garlic component is well-transformed into the oil part, and thus, the garlic component dissolved in the oil is very stable. He applied for a patent.

Example 1

1Kg of garlic was finely ground in a blender, left overnight, and 1.5 liters of vegetable oil was added thereto, stirred for about 2 hours using a stirrer, and centrifuged at 5,000 rpm for 30 minutes in a high-speed centrifuge to obtain about 0.9l of separated oil. As a result of analyzing allicin content, garlic oil containing garlic (0.107%) was obtained.

Example 2

1Kg of garlic was finely ground in a blender, left overnight, and 1.5 liters of cottonseed oil was added thereto, stirred for about 2 hours with a stirrer, and centrifuged at 5,000 rpm for 30 minutes in a high-speed centrifuge to obtain about 0.9l of separated oil. , As a result of analyzing allicin content, garlic oil (garlic oil) containing 0.106% of allicin was obtained.

As a result of extracting the garlic component as other cooking oil, the same results as in Example 1 or 2 were obtained. This result can be predicted even if the composition of each cooking oil is triglyceride of unsaturated fatty acid of about the same.

Previous analysis of allicin

As for the analysis of allicin, various methods have been attempted to analyze allicin directly or its degradation products by measuring absorbance, quantifying total sulfur, and using a device such as liquid chromatography or gas chromatography. These various attempts have been repeated because of the instability of allicin because each assay is difficult to apply universally to garlic oil and to obtain reproducible results.

According to Broadnizi, garlic extracts and allicin compounds were almost decomposed after 20 hours at 20 ^° C., changing to diallyl disulfide 66%, diallyl sulfide 14% and diallyl trisulfide 9%.

The inventors also synthesized allicin by the method of Herwig Jansen et al. (Planta Medica 559, 1987) and analyzed immediately after the synthesis and after 16 hours at room temperature, the amount of diallyl disulfide was significantly increased. .

Some have synthesized allicin, purified by liquid chromatography, and then adsorbed onto dry silica gel and stored in a dryer at -24 ^o C. These studies indicate that 100% pure allicin standards are not available and that allicin, which is adsorbed and diluted on silica gel, cannot guarantee stability in distribution and is difficult to store in general laboratories.

Thus, the inventors gave the principle to the Broadnichi test (.g Agr. Food. Chem., 19,723, 1971) and did not directly use allicin as a standard, but di-n-propyl disulfide. As a result of analysis using disulfide) as an internal standard solution, the present inventors have found a surprising fact that allicin can be easily analyzed.

In other words, an object of the present invention is to easily quantify and analyze allicin by using di-n-propyl disulfide as an internal standard instead of using allicin directly as a standard and using diallyl disulfide, which is a major degradation product of allicin, as a standard. It provides a new method of analysis of allicin.

This principle is based on the dehydration of allicin at temperatures above 150 ^o C, resulting in isomers 144-I (3-Vinyl-1,2-dithi-5-ene) and 144-II (3-Vinyl-1, 2-dithi-4-ene), which is converted to 144-I and 144-II by the high temperature of gas chromatography when gastrograph is applied to gas chromatography, and this component is detected by the detector of gas chromatography. . To quantify these two components, di-n-propyl disulfide, a sulfur compound that is not contained in garlic and has similar reactivity in gas chromatography, was used as an internal standard solution. Phosphorous diallyl disulfide was used as a standard, and then quantified and combined.

Test Example

Allicin was quantified by the following method.

Internal standard solution: 30 mg of di-n-propyl disulfide was precisely taken and dissolved in acetonitrile to obtain 500 ml.

Sample solution: 1.5 mg of allicin was precisely weighed, placed in a centrifuge tube, 5 ml of internal standard solution was added to the solution, left for 3 minutes in an ultrasonic bath, centrifuged, and the supernatant was taken as a sample solution.

Diallyl disulfide standard solution: 10 mg of diallyl disulfide was precisely taken and dissolved in acetonitrile to make 100 ml.

Operation: The test solution and the standard solution were tested according to the gas chromatography method under the following conditions.

Analysis condition

Gas Chromatography: Hewlett-Packard 5890 A, HP3394 A Integrator

Column: Ultra I

25m × 0.32mm × 0.52μm

Temperature: Injection 250 ^o C

Column 120 ^o C

Detector 300 ^o C

Detector: Hydrogen Ionization Detector

Mobile phase: Helium (He)

Flow rate: 25ml / min

Injection volume: 3μl

formula :

At: peak area of the sample solution

As: Internal standard liquid or standard liquid peak area

Cs: Standard solution concentration (mg / ml)

Sa: amount of sample taken (mg)

Result: Allicin content in garlic oil

Example 1

One). 144-I and 144-II

2). Diallyl disulfide

* Imports (Euromed, Spain)

One). 144-I and 144-II

2). Diallyl disulfide

1 is a gas chromatogram of diallyl disulfide standard of garlic oil prepared in Example 1, and FIG. 2 is diallyl disulfide, di-n-propyl disulfide (internal standard) of garlic oil of Example 1, 3-vinyl Gas chromatogram of -1,2-dithi-5-ene (144-I) and 3-vinyl-1,2-dithi-4-ene (144-II), and FIG. 3 is a commercially available product (Euromed: trade name). Is a gas chromatogram of diallyl disulfide standard of garlic oil, and FIG. 4 shows diallyl disulfide, di-n-propyl disulfide (internal standard) of garlic oil of a commercially available product (Euromed: trade name), 3-vinyl-1,2 -Gastrotogram of dithi-5-ene (144-I) and 3-vinyl-1,2-diti-4-ene (144-II).

As apparent from the above experimental results, the above analysis method is a very accurate and convenient method for analyzing allicin.

Claims (1)

In the analysis method of allicin in garlic oil, the content of allicin in garlic oil was obtained by using di-n-propyl disulfide as an internal standard and gastrographing with diallyl disulfide, a major degradation product due to changes in allicin, as a standard product. Method for the analysis of allicin in garlic oil, characterized by measuring the.