Method development for the estimation of Coumarin content in different Cinnamomum species

Abstract:

Both Cinnamomum zeylanicum (Ceylon Cinnamon, True Cinnamon) and Cinnamomum cassia (Cassia, Chinese Cinnamon) are visibly almost similar and widely used for culinary purpose due to its aroma and flavor. However Cassia is known to contain high amount of coumarin which is known to have toxic effects on the liver, kidneys and lungs. A rapid and simple High Performance Liquid Chromatography (HPLC) method has been developed for the quantification of coumarin in Cinnamomum zeylanicum and Cinnamomum cassia samples. Analysis was performed using C18 column (250mm x 4.6 mm; i.d. 5 µm) and UV detector. Samples were extracted by 90% methanol. The calibration plot was linear over the range studied (1.0-20 mg/L) with a correlation of 0.99. The method was also validated for the linearity, range, precision, recovery and detection limits. Thus, the method is suitable for routine analysis of coumarin in Cinnamomum zeylanicum and Cinnamomum cassia samples.

Keyword:

Cinnamomum zeylanicum, Cinnamomum cassia, HPLC, Coumarin

Introduction:

Cinnamon is one of the oldest known spices. The names “cinnamon” and “cassia” cause considerable confusion, as they are often used interchangeably. There are four main economically important species of cinnamon in the genus Cinnamomum. The first one is Cinnamomum verum, translation “true cinnamon”. It is also called Sri Lankan or Ceylon cinnamon. Sri Lanka is the only regular supplier of true cinnamon bark and leaf oils. C. verum’s older botanical name, Cinnamomum zeylanicum, is derived from Sri Lanka’s older name, Ceylon. The other three main species of cinnamon are Cinnamomum cassia (C. aromaticaum, also called Chinese cinnamon), Cinnamomum burmannii (also called Korintje, Java, or Indonesian cinnamon), and Cinnamomum loureiroi (also known as Vietnamese or Saigon cinnamon).

The dried inner bark of the cinnamon plant has been used as a flavoring agent in foods, beverages, chewing gums, etc. C. zeylanicum’s flavour is often considered to be the most delicate and complex of the major species of cinnamons.

>Whilst both species of Cinnamons, namely, C. Zeylanicum and C. Cassia, are widely used is regular culinary purposes, each also has a content of the potentially dangerous compound, coumarin.

Coumarin is a colorless crystalline solid with a sweet odor resembling the scent of vanilla and a bitter taste. Coumarin is naturally occurring in Cinnamomum plants. Although coumarin is safe in small doses, studies have demonstrated high intakes to have toxic effects on the liver, kidneys and lungs. Some evidence even suggests that it causes cancer.

>Compared to C. zeylanicum though, C. cassia holds the highest content of coumarin. The present study is designed for routine estimation of the same.

Coumarin - Wikipedia

Coumarin

Materials and Method:

5 different samples of C. zeylanicum and C. Cassia were extracted by 100 ml 90% methanol. The solutions were filtered and subjected to HPLC analyses. Quantification was done from the standard curve of Coumarin.

Results and Discussions:

The method validation for coumarin was carried out by HPLC. The validation data are incorporated in table 1. Estimation of coumarin content was carried out in five different samples of C. zeylanicum and C. cassia. The results were incorporated in table 2, 3; figure 1.

Table 1: validation data of coumarin

Parameters	Results
Linearity range	1.0-20.0 mg/L
Slope	79617
Intercept	1279
Regression Equation	y = 79617x + 1279
Correlation Coefficient	0.993
LOD	0.006
LOQ	0.02

Table 2: Coumarin content of C. zeylanicum

*C. zeylanicaum* samples	Coumarin content (mg/kg)
Sample 1	9.8
Sample 2	10.0
Sample 3	12.6
Sample 4	8.6
Sample 5	14.7

Table 3: Coumarin content of C. cassia

*C. cassia* samples	Coumarin content (mg/kg)
Sample 1	410.7
Sample 2	657.4
Sample 3	529.6
Sample 4	796.1
Sample 5	605.5

Fig 1: Coumarin content of C. zeylanicum and C. cassia samples

Conclusion:

In this proposed method the linearity was observed in the concentration range of 1.0-20.0 mg/L for coumarin with co-efficient of correlation, r² = 0.993. The result of the analysis of Cinnamomum samples by the method was found to be highly reproducible and reliable. The matrix and other ingredients present in the samples did not interfere with determination of coumarin. So, the developed HPLC method is simple, precise and accurate and can be used for routine quantification of coumarin in C. zeylanicum and C. cassia samples.

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Method development for the estimation of Coumarin content in different Cinnamomum species

Testimonial

Mitra S.K. have added value to our business process by effective inspection, strict quality control and timely response. It is a pleasant experience to work with “Team Mitra S.K.”. We hope the same service level will continue in the future too....

Sirsendu Mukherjee

Mitra SK Inspection Gulf LLC has been appointed by us and the services provided by them has been up to the mark and as per our expectations....

Ajay Mathur

Amar Singh Gohil

The Inspection services provided by Mitra S. K. Inspection Gulf has been prompt and accurate and has been in the line with our expectations....

Naser Bustami

We would like to express our high appreciation of the business association our company had with Mitra S. K. as a testing and inspection house for their quality of service, good documentation and reporting during the job ....

R. Jeyakumar

Tulus Situmeang

Their job was excellent and showed Mitra S.K. differential from others....

Geraldo Pimenta

Konstantin Gertsen

We’ve been pleased with the improvement we have seen in CIQ results, and Mitra S.K’s reports have certainly helped us to resolve any disputes with our customers in a more timely manner….....

Brian Mcdonald

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