INTRODUCTION:

Medicinal plants are the richest bio resource of drugs for traditional systems of medicine, modern medicines, Nutraceuticals, food supplements, folk medicines, pharmaceutical intermediates and chemical entities for synthetic drugs. Cinnamon is a common spice used by different cultures around the world for several centuries. It is obtained from the inner bark of trees from the genus Cinnamomum, a tropical evergreen plant that has two main varieties; Cinnamomum zeylanicum Blume (Cinnamomum verum/true cinnamon) and Cinnamon cassia (also known as Cinnamomum aromaticum/ Chinese cinnamon). Cinnamomum zeylanicumBlume cultivated in south India, butoriginates from the island Sri Lanka, southeast of India [1, 2]. It is also known as Dalchini (Hindi) and Twak (Sanskrit).

Cinnamon is obtained by drying the central part of the bark and is marketed as quills or powder. In Ayurvedic medicine Cinnamon is considered a remedy for respiratory, digestive and gynaecological ailments [3]. Twak used as a one of the ingredient in Sitopaladi Churna, Caturaja Churna etc[4]. Almost every part of the cinnamon tree including the bark, leaves, flowers, fruits and roots, has some medicinal or culinary use. Three of the main components of the essential oils obtained from the bark of Cinnamomum zeylanicum are trans-cinnamaldehyde, eugenol and linalool, which represent 82.5% of the total composition [5]. Trans cinnamaldehyde, accounts for approximately 49.9–62.8% of the total amount of bark oil [6,7]. Cinnamaldehyde and eugenol are the major components of Cinnamomum zeylanicum [8].

Cinnamomum cassia, called Chinese cassia or Chinese cinnamon, is an evergreen tree originating in southern China, and widely cultivated there and elsewhere in southern and eastern Asia (India, Indonesia, Laos, Malaysia, Taiwan, Thailand, and Vietnam) [9]. It is one of several species of Cinnamomum used primarily for their aromatic bark, which is used as a spice. In the United States, Chinese cassia is the most common type of cinnamon used. The buds are also used as a spice, especially in India, and were once used by the ancient Romans. Cassia bark is used as a flavouring agent for confectionery, desserts, pastries, and meat; it is specified in many curry recipes, where Ceylon cinnamon is less suitable. Cassia is sometimes added to Cinnamomum zeylanicum, but it is a much thicker, coarser product. Cassia is sold as pieces of bark or as neat quills or sticks. Cassia sticks can be distinguished from Cinnamomum zeylanicum sticks. The Cinnamomum zeylanicum sticks have many thin layers and can easily be made into powder using a coffee or spice grinder, whereas cassia sticks are extremely hard and are usually made up of one thick layer. One important difference between Cinnamomum cassia Blume and Cinnamomum zeylanicum is their coumarin (1, 2-benzopyrone) content [10]. The levels of coumarins in Cinnamomum cassia appear to be very high and pose health risks if consumed regularly in higher quantities. Due to a blood-thinning component called coumarin, which could damage the liver if taken in huge amounts, European health agencies have warned against consuming high amounts of cassia. Other bioactive compounds found in the bark, powder and essential oils of C. cassia are cinnamaldehyde and styrene. In high doses these substances can also be toxic for humans. According to the German Federal Institute for Risk Assessment (BfR), 1 kg of Cinnamomum cassia powder contains approximately 2.1-4.4 g of coumarin, which means 1 teaspoon of Cinnamomum cassia powder would contain around 5.8-12.1 mg of coumarin [11]. Cinnamomum cassia has shown various pharmacological activities like anti-inflammatory, antioxidant, anticancer activities, etc. It has been shown to possess many pharmacological actions for the treatment of various diseases such as cancer, diabetes; bacterial infections, etc. [12]. In-vitro and in-vivo studies in animals and humans have demonstrated numerous beneficial health effects of Cinnamomum zeylanicum, such as anti-inflammatory properties, anti-microbial activity, reducing cardiovascular disease, boosting cognitive function and reducing risk of colonic cancer [13,14].

This study aimed to revealed specific identities for crude drug taken which will be useful in identification and control to adulterations of the raw drug.

MATERIALS AND METHODS:

Plant material:

Cinnamomum zeylanicum and Cinnamomum cassia samples were collected from the local market of Chennai, Tamilnadu and authenticated by Department of Botany at CSMRADDI, Chennai. Powdered samples were used for Physico chemical parameters like total ash, acid insoluble ash, extractive values and HPTLC fingerprint. Analysis studies were carried out as per the API, WHO and AOAC guidelines [15-17].

Preliminary Phytochemical Analysis:

Preliminary phytochemical results showed the presence or absence of certain phytochemicals in the Cinnamomum zeylanicum and Cinnamomum cassia Samples. The tests performed using alcoholic extract was subjected to various qualitative tests for the identification of phytoconstituents present. Preliminary phytochemical test revealed the presence of coumarin, flavonoids, glycoside, tannin, sugar terpenoids and absence of alkaloids. [18-19].

Physicochemical Parameters:

Different physicochemical parameters like total ash, acid insoluble ash, water soluble ash, ethanol soluble extractive value and water soluble extractive value [15-17].

HPTLC fingerprinting profile:

The HPTLC method can be used for phytochemical profiling of plants and quantification of compounds present in plants. 2 g of the sample was soaked with 20 ml ethanol overnight, boiled for 10 minutes, concentrated and made up to 10 ml volumetric flask. The test samples solution were applied 10 µl and 12 µl on Tracks-1and 2 and 7µl, 10 µl and 12 µl on Track- 3, 4 and 5forCinnamomum zeylanicumand Cinnamomum cassia respectively on E. Merck aluminium plate pre-coated with Silica gel 60F₂₅₄ of 0.2 mm thickness using CAMAG Linomate IV applicator. The plate was developed in the solvent system of Toluene: Ethylacetate: Formic acid (9: 0.9: 0.1) and dried. The plate was observed through CAMAG TLC Visualizer under UV at 254 nm and 366 nm and photos were documented. Finally the plate was dipped in Vanillin-Sulphuric acidreagentand heated in hot air oven at 105 ^oC untill the colour of the spots were appeared and photo was documentation under white light. Before derivatization the was scanned using CAMAG TLC Scanner with WINCATS 4.05 version software at a wavelength of UV 254 and 366 nm using deuterium and mercury lamps respectively [20-23]. The R_fvalues and fingerprint data were recorded by WIN CATS software.

RESULTS AND DISCUSSION:

Physico-chemical analysis:

Physico-chemical parameters such as water soluble extractives, alcohol soluble extractive, total ash content and acid insoluble ash of the Cinnamomum zeylanicum (CZ) and Cinnamomum cassia (CC) samples were determined. Total ash value of plant material indicated the amount of minerals and earthy materials attached to the plant material. Analytical results showed total ash values content of CZ and CC were 4.02 and 3.1 % w/w respectively. The amount of acid insoluble siliceous matter present in the plant samples were 0.26 and 0.23 % w/w respectively. The water-soluble extractive value was indicating the presence of sugar, acids and inorganic compounds. The water soluble extractive value in the plant samples were 6.89(CZ) and 4.71(CC) % w/w and alcohol soluble extractive value were 5.83 (CZ) and 4.83(CC) % w/w. The alcohol soluble extractive values indicated the presence of polar constituents like phenols, alkaloids, steroids, glycosides, flavonoids and secondary metabolites in the plant samples.

Preliminary Phytochemical Analysis:

Preliminary phytochemical screening of Cinnamomum zeylanicum and Cinnamomum cassia revealed the presence of phytochemicals like alkaloids, saponins, flavonoids, tannins and terpenoids. Steroid was found to be absent in Cinnamomum cassia and Glycosides in Cinnamomum zeylanicum.

HPTLC fingerprinting profile:

Qualitative phytochemical analysis provides the information regarding the presence of primary and secondary metabolites of the raw drug for identity and purity. HPTLC fingerprinting is proved to be a linier, precise, accurate method for herbal identification and can be used further in authentication and characterization of the important medicinal plant. The HPTLC method can be used for phytochemical profiling of plants and quantification of compounds present in plants.

The test samples solution were applied 10 µl and 12 µl on Tracks-1and 2 and 7µl, 10 µl and 12 µl on Track- 3, 4 and 5forCinnamomum zeylanicumand Cinnamomum cassia respectively on TLC plate. The plate was developed in the solvent system of Toluene: Ethylacetate: Formic acid (9: 0.9: 0.1) and dried. The plate was visualized under UV 254 nm and 366 nm and after that plate was dipped in Vanillin-Sulphuric acidreagent.

In this study discussed about two tracks of similar concentration applied of test samples of Cinnamomum zeylanicum and Cinnamomum cassia on track 2 and 5 respectively.

It was observed a significant variation with respect to Cinnamomum zeylanicum and Cinnamomum cassia on track 2 and track 5 under UV 254 that spot with Rf value 0.05 and 0.51 is missing in track 2 (CZ) ) in comparission with CC. Simillarly a spot with Rf value 0.31 is missing in track 5 (CC) in comparission with CZ.

It was also observed on track 2 and track 5 under UV 366 that spot with Rf value 0.31, 0.38 and 0.71 is missing in track 2 (CZ) ) in comparission with CC. Simillarly a spot with Rf value 0.40 is missing in track 5 (CC) in comparission with (CZ).

After derivatisation under white light it was observed on track 2 and track 5 that spot with Rf value 0.73 is missing in track 2 (CZ) in comparission with (CC). Simillarly a spot with Rf value 0.62 and 0.95 is missing in track 5 (CC) in comparission with (CZ).

The chromatograms were scanned by densitometer at 254 nm and 366 nm before derivatization. The R_fvalues and fingerprint data were recorded by WIN CATS software. The HPTLC chromatogram, peaks, R_fvalues and area obtained for tracks after scanning at UV 254 nm and 366 nm are depicted in respective Figures 2 to 6 and Table 1 to 4. The chromatograms of Cinnamomum zeylanicum and Cinnamomum cassia at UV 254 and 366 nm revealed that all sample constituents were clearly separated without any tailing and diffuseness.

HPTLC fingerprinting analysis various peaks were detected and it was found that some compounds appear in CZ fingerprints profile but some peask not appear in CC fingerprints .The Ethanol extract of Cinnamomum zeylanicum at 254 nm and 366 nmexemplified 5 and 3 peaks respectively at the Rf values 0.24, 0.30, 0.57, 0.70 and 0.93 and 0.38, 0.66 and 0.79 were found to be more predominant as the intensity of area was ranging from 881.8 AU to 21778.8AU. The Ethanol extract of Cinnamomum cassia at UV 254 nm and 366 nm exemplified 3 and 1 peaks respectively at the Rf values 0.25, 0.55 and 0.71 and 0.80 were found to be more predominant as the intensity of area was ranging from 832.9 AU to 6127.4AU. The remaining components are less in quantity as the area of all spots.

Thus, the developed chromatogram will be specific with selected solvent system Toluene: Ethylacetate: Formic acid (9: 0.9: 0.1), R_fvalue and serve the better tool for standardization of the drug.

Figure 2. 3D view of all tracks of Densitometric chromatogram of Cinnamomum zeylanicum and Cinnamomum cassia of Ethanolic extract at 254 nm.

CONCLUSION:

Samples were studied and described preliminary phytochemical analysis, physicochemical parameters and HPTLC fingerprint studies. These parameters will be useful in authentification and identifying the adulterants and /or substituent’s for quality control of raw drugs. Sample exhibits a set of diagnostic characters, which will help to identify the drug in dried condition. It has been concluded from this study of various species, such as estimation of physicochemical parameters like water soluble extract, alcohol soluble extract, total ash, acid insoluble ash and high performance thin layer chromatography profile (HPTLC) is highly essential for raw drugs or plant parts used for the preparation of compound formulation drugs. The results obtained from qualitative evaluation of HPTLC finger print profiles could be useful in the authentication and quality control of the raw drug.

In this study steroid was found to be absent in Cinnamomum cassia (CC)and Glycosides in Cinnamomum zeylanicum (CZ).HPTLC fingerprint profiles discussed about two tracks of similar concentration applied of test samples of CZ and CC. It was observed a significant variation with respect to CZ showed 8 spots and CC showed 7 spotsunder UV 254 nm. Under UV 366 nm CZ showed 6 spots and CCshowed 5 spots.This could help to distinguish between the two different species of this herbal medicine.

ACKNOWLEDGEMENT:

The authors are very grateful to Director General and Deputy Director General, CCRAS, New Delhi for providing encouragement and facilities for carrying out this work.

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Received on 28.02.2018 Modified on 22.03.2018

Asian J. Research Chem. 2018; 11(2):344-350.

DOI:10.5958/0974-4150.2018.00062.7