KR20090008728A - Quantitative analysis method of pelargonium sidoides syrup or solution - Google Patents

Abstract

A method for quantitative analysis of Pelargonium sidoides syrup or liquor is provided to identify the content of the extract undertaking the expression of drug effects in the Pelargonium sidoides syrup or liquor. The method for quantitative analysis of Pelargonium sidoides syrup or liquor comprises the steps of: (A) mixing 40-60 parts by weight of ethanol solution with 15-30 wt.%, 6-9 parts by weight of Folin & Ciocalteu's phenol reagent, 40-60 parts by weight of the aqueous solution of sodium carbonate with 10-20 wt.%, and 15-40 parts by weight of water based on 1 part by weight of the Pelargonium sidoides extract solution, and centrifuging the mixture to collect the supernatant as testing liquid; (B) mixing 8-12 parts by weight of ethanol solution with 15-30 wt.%, 1-2 parts by weight of Folin & Ciocalteu's phenol reagent, 8-12 parts by weight of the aqueous solution of sodium carbonate with 10-20 wt.%, and 3-8 parts by weight of water based on 1 part by weight of the aqueous solution of epicatechin ethanol, and centrifuging the mixture to collect the supernatant as standard liquid; (C) measuring the absorbance of the testing liquid and standard liquid by using water as control liquid; and calculating the total amount of phenol as epicatechin through the equation 1, wherein Et is the absorbance of the testing liquid, Es is the absorbance of the standard liquid, V is the volume of the testing liquid measured in ml unit, and S is the mass of the standard liquid measured in mg unit.

Description

Quantitative Analysis Method of Pelargonium sidoides Syrup or Solution

1 is a graph showing the relationship between the concentration and absorbance of the Pelargonium sidoides extract.

The present invention is used to treat Pelargonium sidoides ( Pelargonium) sidoides ) relates to quantitative analysis of syrups or solutions containing extracts.

Extracts derived from the roots of the species of pelargonium , such as Pelargonium sidoides and Pelargonium reniforme, are found in southern Africa. as a therapeutic agent for liver diseases, etc. (Watt, C. (1962) Medicinal and poisonous plants of southern and eastern Africa , Livingstone, Edinburgh, London, S. 449-455). In addition, it is known to be excellent for the treatment of respiratory diseases, especially tuberculosis.

Pelargonium sidoides extract, originally known as a traditional medicinal herb in South Africa, is known to be used in other regions, specifically in central Chile and Mexico. That is, in the region, the astringent function of the extract of Pelargonium sidoides was used to treat diarrhea and various gastrointestinal syndromes. These effects are known to be attributed to tannin, a constituent of the extract (Jose San Martin, A. (1983) Econom . Bot, 37 , 216-227. INI (1994) Atlas de las plantas de la medicina tradicional mexicans (Instituto Nacional Indigenista, Hrsg.) Vol II, Av.Revolucion no 1279, Col. Tiacopac, Mexiko, S. 667 und 950).

Furthermore, in the early 20th century in the United States, the root extract of Pelargonium sidoides is known as Umckaloabo in the upper airway (Kauffer, HJ (1915) Dental Cosmos 57 , 1366), such as rhinopharyngitis, tonsillitis It has been used in the treatment of tonsillitis, sinusitis, brochitis and the like.

This effect is known to be due to the antimicrobial effect of Pelargonium sidoides extract.

More specifically, coumarin experiments show that there is moderate cytotoxicity against GLC4, lung cancer cells, and COLO320 cells, and this cytotoxic effect is wholly dependent on the nature and position of the substituents in the aromatic ring. In the agar dilution test, experiments with coumarin and tannin compounds for antimicrobial action against various Gram-positive and Gram-negative bacteria showed moderate bacteriostatic effects. Minimal Inhibitory Concentration (MIC) values range from 200 to 2000 μg / ml (for coumarins) and 500 to 2000 μg / ml (for monomer tannin precursors), with highly antimicrobial coumarin trioxide, 4 Oxidized materials are representative. This clear antimicrobial activity is seen in the extract of Pelargonium sidoides, especially the extracts of roots and leaves, water is the largest antibacterial effect as a solvent. (Kayser, O. (1997) Phenolic constituents of Pelargonium sidiodes DC. and studies of the efficacy of the Umcka plant material ( Pelargonium sidiodes DC. and Pelargonium reniforme CURT .) Doctorial dissertion, Berlin Free University.)

On the other hand, Pelargonium sidoides extract is useful for the treatment of acute or chronic infectious diseases by affecting the immunomodulatory effect.

Specifically, Pelargonium sidoides extract has been reported to increase TNF-α, INF-β, nitric oxide (NO) synthesis (H. Kolodziej et al., Phytomedicine 10 (Suppl. 4) , 18-24 (2003)).

In addition, coumarin derived from Pelargonium sidoides extract has been shown to exhibit immunostimulatory effects in various experimental models. Specifically, in the intracellular Leishmania infection model, these substances showed moderately structure-dependent actions in relation to oxygen-dependent immune defense mechanisms, in particular intracellular infection via NO-mediated NO production mediated by simple coumarins. Moderate defensive stimulation of ROS increases with increasing oxygen production. In addition, gallic acid (gallic acid), which is another component of the extract, is an excellent derivative, and its activity is due to a high concentration of nitric oxide radicals. (Kayser, O. (1997) Phenolic constituents of Pelargonium sidiodes DC. and studies of the efficacy of the Umcka plant material ( Pelargonium sidiodes DC. and Pelargonium reniforme CURT .) Doctorial dissertion, Berlin Free University.)

These pelargonium sidoides extracts have been shown to be effective in acute or chronic infections, especially infections of the respiratory tract or ear throat, such as bronchitis, sinusitis, tonsillitis, nasopharyngitis.

On the other hand, syrups or solutions are easy to administer to children or the elderly who lack the ability to swallow tablets, have a rapid drug effect compared to tablets, and bioavailability that may be caused by differences in drug solubility in tablets when administered. It can also reduce the difference has the advantage that the drug can be kept constant. In particular, in the case of syrups, oral administration can significantly improve the bitter taste and unpleasant odor caused by the drug, and the patient's compliance with the drug can be improved, so that it can be easily administered to children or the elderly.

However, in the case of syrups or liquid preparations containing the Pelargonium sidoides extract, the content analysis method of the extract responsible for the expression of efficacy has not been established, which has hindered practical application.

The present invention has been made to solve the above problems, in the syrup or liquid formulation containing the Pelargonium sidoides extract, to provide a quantitative analysis method showing the content of the extract responsible for the expression of the drug do.

The quantitative analysis of Pelargonium sidoides extract of the present invention is characterized in that it comprises the following steps in order to achieve the object as described above:

(A) 40 to 60 parts by weight of 15 to 30% by weight of ethanol aqueous solution, 6 to 9 parts by weight of Floin Ciocalteus phenol reagent, 10 to 20% by weight, per 1 part by weight of Pelargonium sidoides extract 40 to 60 parts by weight of an aqueous sodium carbonate solution, and 15 to 40 parts by weight of water, and the sample solution preparation step of taking the supernatant as a sample solution by centrifuging the mixed solution,

(B) 8 to 12 parts by weight of 15 to 30% by weight of ethanol aqueous solution, 1 to 2 parts by weight of Floin Ciocalteus phenol reagent, 10 to 20% by weight, per 1 part by weight of epipicchin ethanol solution 8 to 12 parts by weight of an aqueous sodium carbonate solution, and 3 to 8 parts by weight of water, the standard solution preparation step of taking the supernatant as a standard solution by centrifuging the mixed solution, and

(C) measuring the absorbance of the sample solution and the standard solution using water as a control solution, and

(D) calculating the total amount of phenol as epicatechin by the following equation (1):

Where Et is the absorbance of the test liquid,

Es is the absorbance of the standard solution,

V is the volume of the sample solution measured in ml,

S is the mass of the standard solution measured in mg.

In addition, the mixture may further include the step of standing at room temperature for 10 to 30 minutes before centrifugation.

In addition, the centrifugation is preferably performed for 5 to 20 minutes.

In addition, the absorbance is preferably measured at 720 nm.

In addition, the 15 to 30% by weight of the ethanol aqueous solution is preferably prepared by diluting 95 to 96% by weight of ethanol aqueous solution and water in a volume ratio of 1: 2 to 5.

Hereinafter, preferred embodiments of the present invention will be described in detail. In addition, in the following description, many specific details such as specific components are disclosed, which are provided to help a more general understanding of the present invention, and it is common in the art that the present invention may be practiced without these specific details. It is self-evident to those who have knowledge of the world. In describing the present invention, when it is determined that a detailed description of a related known function or configuration may unnecessarily obscure the subject matter of the present invention, the detailed description thereof will be omitted.

The quantitative analysis of the Pelargonium sidoides extract of the present invention begins with the preparation of a sample solution containing the extract of Pelargonium sidoides and a standard solution as a base to be compared thereto.

First, all the ferrago sidoides extract to be analyzed is included as long as it is a substance containing the extract of the ferrago sidoides irrespective of its formulation, and in the present invention, a syrup or liquid containing the extract is particularly preferable. Do.

40 to 60 parts by weight of 15 to 30% by weight of an ethanol aqueous solution, 6 to 9 parts by weight of a Floin Ciocalteus phenol reagent, and 10 to 20% by weight of an aqueous sodium carbonate solution of 40 to 60 parts by weight of the extract Parts and 15 to 40 parts by weight of water are mixed.

Here, the 15 to 30% by weight of ethanol aqueous solution is preferably prepared by diluting 95 to 96% by weight of ethanol aqueous solution and water in a volume ratio of 1: 2 to 5.

The mixed solution is then centrifuged and the supernatant taken as the sample solution.

At this time, the centrifugation is preferably performed for 5 to 20 minutes.

In particular, in the present invention, it is more preferable to further include the step of leaving the mixture at room temperature for 10 to 30 minutes before centrifugation in terms of improving the reproducibility of the analysis results.

The epicatechin standard solution is prepared separately from the sample solution. First, 8 to 12 parts by weight of 15 to 30% by weight of an ethanol aqueous solution of 15 to 30% by weight of the epicatechin ethanol solution of moderately diluted epicatechin with 15 to 30% by weight of ethanol solution, 1 to 2 parts by weight of a polline sycartheus phenol reagent , 8 to 12 parts by weight of an aqueous 10-20% by weight sodium carbonate solution, and 3 to 8 parts by weight of water.

Thereafter, the stationary step and the centrifugation step are the same as in the sample preparation step.

Then, the absorbance is measured by using the prepared sample solution and the standard solution as the control solution, the measurement wavelength is most preferably carried out at 720 nm.

Finally, the content of Pelargonium sidoides extract is determined by calculating the total amount of phenol as epicatechin by the following equation:

[Equation 1]

Where Et is the absorbance of the test liquid,

Es is the absorbance of the standard solution,

V is the volume of the sample solution measured in ml,

S is the mass of the standard solution measured in mg.

EMBODIMENT OF THE INVENTION Hereinafter, the Example of this invention is described.

Example

Example  One: Sample  pharmacy

A mixture of 200 μl of syrup containing Pelargonium sidoides extract and 96% by weight of ethanol solution was mixed with distilled water and 1: 3. The flask was charged with a volume of distilled water, and distilled water was added to make the whole amount 25 ml. The mixture was left at room temperature for 20 minutes, then centrifuged for 10 minutes, and the supernatant was taken as a sample solution.

Example  2: Standard Solution

10 mg of epicatechin was collected and placed in a 20 ml flask, and 96% by weight of an ethanol solution was mixed with distilled water and diluted 1: 3 to dilute ethanol dilutions to make the total amount 20 ml. 1 ml of the solution was taken and the distillation of the ethanol was added again to make 10 ml of the total amount, where 1 ml was taken. 1 ml of the last solution taken, 10 ml of the ethanol dilution solution, 1.5 ml of a Pauline Siaceticus phenol reagent, and 10 ml of an aqueous sodium carbonate solution were rinsed into a 25 ml flask, and distilled water was added to make 25 ml. The mixture was left at room temperature for 20 minutes, then centrifuged for 10 minutes, and the supernatant was taken as a standard solution.

Example  3: Absorbance Measurement (1)

In the sample solution of Example 1 and the standard solution of Example 2, absorbance (Agilent. Model 8453) was measured at a total length of 10 mm and a wavelength of 720 nm using water as a control solution. The measurement was repeated five times and the results are shown in Table 1.

Exam number Absorbance One 0.61215 2 0.61744 3 0.61690 4 0.59956 5 0.61438 6 0.61800

As a result of the measurement, a high level of reproducibility was shown with a relative standard deviation of 1.14.

Example  4: absorbance measurement (2)

The Example 3 was repeated, but the amount of syrup containing the Pelargonium sidoides extract of Example 1 was measured by varying 50, 80, 100, 150, 200 μl. The results are shown in Table 2 and FIG.

Sample concentration (μl / ml) Absorbance 2 0.16457 3.2 0.26766 4 0.33972 6 0.50115 8 0.65023

As a result of the measurement, a high linearity of the correlation coefficient 0.99973 can be confirmed, and at least in the above concentration range, a proportional relationship between concentration and absorbance can be used.

Although the above has been illustrated and described with respect to the preferred embodiments of the present invention, the present invention is not limited to the specific embodiments described above, those skilled in the art without departing from the gist of the present invention various modifications Of course, implementation is possible. Therefore, the scope of the present invention should not be construed as being limited to the above embodiments, but should be defined by the claims below and equivalents thereof.

Due to the quantitative analysis of the syrup or solution containing the Pelargonium sidoides extract of the present invention, it is possible to ensure the quality control and pharmacological effect expression by measuring the content of the Pelargonium sidoides extract in the syrup or solution, The analysis method has the advantage of excellent reproducibility and high reliability through relatively simple operation.

Claims (5)

Quantitative analysis of Pelargonium sidoides extract, comprising the following steps: (A) 40 to 60 parts by weight of 15 to 30% by weight of ethanol aqueous solution, 6 to 9 parts by weight of Floin Ciocalteus phenol reagent, 10 to 20% by weight, per 1 part by weight of Pelargonium sidoides extract 40 to 60 parts by weight of an aqueous sodium carbonate solution, and 15 to 40 parts by weight of water, and the sample solution preparation step of taking the supernatant as a sample solution by centrifuging the mixed solution, (B) 8 to 12 parts by weight of 15 to 30% by weight of ethanol aqueous solution, 1 to 2 parts by weight of Floin Ciocalteus phenol reagent, 10 to 20% by weight, per 1 part by weight of epipicchin ethanol solution 8 to 12 parts by weight of an aqueous sodium carbonate solution, and 3 to 8 parts by weight of water, the standard solution preparation step of taking the supernatant as a standard solution by centrifuging the mixed solution, and (C) measuring the absorbance of the sample solution and the standard solution using water as a control solution, and (D) calculating the total amount of phenol as epicatechin by the following equation (1): [Equation 1]

Where Et is the absorbance of the test liquid, Es is the absorbance of the standard solution, V is the volume of the sample solution measured in ml, S is the mass of the standard solution measured in mg. The method of claim 1, Quantitative analysis of Pelargonium sidoides extract, further comprising the step of standing at room temperature for 10 to 30 minutes before centrifuging the mixture. The method according to claim 1 or 2, The centrifugation is quantitative analysis method of Pelargonium sidoides extract, characterized in that performed for 5 to 20 minutes. The method according to claim 1 or 2, The absorbance is quantitative analysis of Pelargonium sidoides extract, characterized in that measured at 720 nm. The method according to claim 1 or 2, The ethanol aqueous solution of 15 to 30% by weight is prepared by diluting 95 to 96% by weight of ethanol aqueous solution and water in a volume ratio of 1: 2 to 5 quantitative analysis of Pelargonium sidoides extract.

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