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PHARMACOGNOSTICAL
Year : 2019  |  Volume : 40  |  Issue : 3  |  Page : 196-203  

Macro-micro-morphological diagnosis of leaves of two species of Cinnamomum (C. sulphuratum and C. verum) used as resource of bay leaf


1 Department of Pharmacognosy, Siddha Central Research Institute, (Central Council for Research in Siddha, Ministry of AYUSH, Government of India), Chennai, Tamil Nadu, India
2 Department of Director Incharge, Siddha Central Research Institute, (Central Council for Research in Siddha, Ministry of AYUSH, Government of India), Chennai, Tamil Nadu, India

Date of Submission14-Sep-2017
Date of Decision18-Oct-2017
Date of Acceptance13-Jun-2020
Date of Web Publication08-Aug-2020

Correspondence Address:
Dr. Sunil Kumar Koppala Narayana
Department of Pharmacognosy, Siddha Central Research Institute, (Central Council for Research in Siddha, Ministry of AYUSH, Government of India), Arumbakkam, Chennai - 600 106, Tamil Nadu
India
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/ayu.AYU_194_17

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   Abstract 


Background: Cinnamomum verum (true cinnamon or Tejpatra) is a well-known spice with immense medicinal properties. Its adulteration with leaf and bark of other species belonging to genus Cinnamomum is found to be a common practice in India. Aims: Cinnamomum sulphuratum is used as a substitute of C. verum owing to its apparent macroscopic similarities. Materials and Methods: Fresh leaves of C. verum and C. sulphuratum growing in South India were collected and studied to establish their macro–microscopic identity with pharmacognostical perspective. Results: Detailed microscopic evaluation by transverse section, maceration, and powder microscopy was conducted to delineate the two species. Detailed macroscopic identification served the purpose of identification of the entire drug on the spot, and microscopy has helped in the identification of fragmented and powdered form of the drugs. Conclusion: Further chemical and biological studies may be necessary to confirm whether these leaves can be used as a substitute or adulterant for other species Cinnamon.

Keywords: Cinnamomum sulphuratum, maceration, micrometry, powder microscopy, quantitative microscopy, Tejpatra


How to cite this article:
Narayana SK, Gopi DK, Rubeena M, Parameswaran SR. Macro-micro-morphological diagnosis of leaves of two species of Cinnamomum (C. sulphuratum and C. verum) used as resource of bay leaf. AYU 2019;40:196-203

How to cite this URL:
Narayana SK, Gopi DK, Rubeena M, Parameswaran SR. Macro-micro-morphological diagnosis of leaves of two species of Cinnamomum (C. sulphuratum and C. verum) used as resource of bay leaf. AYU [serial online] 2019 [cited 2020 Oct 1];40:196-203. Available from: http://www.ayujournal.org/text.asp?2019/40/3/196/291628




   Introduction Top


In India, significant quantities of medicinal plant resources are consumed by traditional healers and practitioners of Indian Systems of Medicine, which require continuous supply of medicinal plants for the manufacturing of medical formulations. Cinnamomum verum J. Presl (synonym Cinnamomum zeylanicum Blume) commonly known as the true cinnamon is one of the earliest known spices and medicinal plants.[1] It is traditionally used for bloating, nausea, flatulence, colic, and gastrointestinal tract spastic conditions and finds application in modern medicines too.[2] Almost every part of the cinnamon tree such as bark, leaves, flowers, fruits, and roots has some medicinal or culinary uses. Among the various species occurring in India, C. verum, Cinnamomum malabatrum, Cinnamomum sulphuratum, and Cinnamomum bejolghota are morphologically similar to one another. C. sulphuratum locally known as “Jangali dalchini” is one of the most traded medicinal plants sourced from tropical forests.[3] It is medium-to-large aromatic evergreen tree, distributed mainly in south[4] and northeastern parts[5] of India. On account of easy availability and similarity in flavor, different parts of C. sulphuratum are in use as substitutes for commercial cinnamon-derived spices.[6] In South India, it is distributed in Western Ghats regions of Tamil Nadu; Thiruvananthapuram and Wyanad in Kerala; and Coorg, South Kannada, Hassan, Mysore, Shimoga, and North Kannada districts in Karnataka.[7] The leaves and bark of the tree are aromatic; leaves are known by vernacular name Tejpatra by the Northeast Indian people and are used as a spice.[8] Medicinal uses of C. sulphuratum are similar to C. verum which include treating wounds, fever, intestinal worms, headache, and menstrual problems.[9] Leaves of the same genus are found to exhibit the number of similar common macro–microscopic characters and hence most likely to be used as an adulterant for official drug. In this article, a detailed macro–microscopic examination was carried out to differentiate these two species.


   Materials and Methods Top


Plant materials

The authentic leaf samples of C. verum were collected from botanical garden of Alva's Ayurveda Medical College, Moodbidri, Karnataka, and C. sulphuratum from wild population in Tala Kaveri of Kodagu (Coorg) district, Karnataka. Leaves were dried in the shade; a few preserved in formalin-ethyl alcohol-acetic acid solution, were used for histological studies. Powder of the dried leaves sieved through mesh 60 was stored in glass vials and used for microscopic evaluation.

Instrumentation and techniques

Detailed macroscopy of the leaves of C. verum and C. sulphuratum was studied with respect to the description provided in floras.[10],[11] Leaf petiole and lamina were transversely cut and slides were prepared[12] as per the standard procedures for histological examinations.[12] Photomicrographs were taken using Leica microscope attached with Canon digital camera. Isolation of the tissues was carried out by Schultz's maceration process or by boiling with 5% KOH.[13] Measurements of the various elements[14] were taken with the help of stage and ocular micrometers. The microscopic diagnostic characters of the powder were studied by clearing the powder with chloral hydrate. Cell contents were tested with usual reagents.[15] Microscopic characters were drawn with Prism Type Camera Lucida.


   Results Top


Macroscopy

Morphological descriptions outlined in floras were used as a guideline to propose diagnostic differentiating macrocharacters of the two drugs. The macroscopic and sensory characters of the two species were compared, and the differences were observed in size of leaf blade and petiole (C. verum is bigger than C. sulphuratum). Petiole is not hairy in C. verum when compared to C. sulphuratum and is lamina glabrous and smooth in C. verum while coriaceous and tomentose in C. sulphuratum which also shows algal growth on the surface. The lower surface of the leaves of C. verum is paler compared to C. sulphuratum. The secondary veins were distinct in C. verum while faint in C. sulphuratum. Leaves of C. verum are sweet, pungent, and aromatic while of C. sulphuratum are astringent and fragrant [Table 1] and [Figure 1]a, [Figure 1]b.
Table 1: Comparative macroscopic and organoleptic characters of leaves of Cinnamomum spp.

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Figure 1: (a) Macroscopic characters of the leaves of Cinnamomum verum, (b) macroscopic characters of the leaves of Cinnamomum sulphuratum, (c) Detailed transverse section of petiole of Cinnamomum verum, (d) detailed transverse section of petiole of Cinnamomum sulphuratum, (e) detailed transverse section of the leaf passing through midrib of Cinnamomum verum, (f) detailed transverse section of the leaf passing through midrib of Cinnamomum sulphuratum

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Microscopy

Transverse section of the petiole

C. verum is cordate with a small projection within the depressed upper surface (circular to almost elliptical without any projections in C. sulphuratum); below the epidermis, cavities containing volatile oils are present in C. verum (absent in C. sulphuratum); rosette crystals are present in C. sulphuratum (absent in C. verum); stomata anomocytic cells are present in C. verum (paracytic in C. sulphuratum); and isolated sclerenchyma cells are seen on both sides of the vascular bundles in C. verum (in C. sulphuratum, it is present on the outer side) [Table 2] and [Figure 1]c, [Figure 1]d.
Table 2: Comparative microscopic characters of the transverse section of petiole of Cinnamomum spp.

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Transverse section of the leaf passing through midrib

Both upper and lower surfaces are broad and conical in C. verum, while the upper region is semicircular and the lower is broad in C. sulphuratum; hypodermal sclerenchymatous patch is 10–12 cells wide in C. verum while 8–10 cells wide in C. sulphuratum; lower cortex is made up of collenchymatous cells of 2–3 layers in C. verum while 6–8 layers in C. sulphuratum. Xylem in C. verum is elongated, roughly ellipsoidal with 28–30 radial rows of vessels while in C. sulphuratum is concavo convex in outline with 20 radial rows of xylem vessels.

Lamina of both species have thick cuticle covering the upper epidermis and thin cuticle covering the lower cells, being nonlignified, nonpapillose in C. verum while being lignified and highly papillose in C. sulphuratum. Single-layered palisade cells are present in both; the number of layers of spongy mesophyll varies from 4 to 5 layers in C. verum while 7 to 8 layers in C. sulphuratum [Table 3] and [Figure 1]e, [Figure 1]f.
Table 3: Comparative microscopic characters of the transverse section of lamina passing through the midrib of Cinnamomum spp.

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Microscopy of the macerated leaf

Leaf on maceration with KOH yielded information on different tissue systems such as type of hairs, xylem vessels, tracheids, stone cell fibers, crystals of calcium oxalate from the petiole and lamina. Brief differentiations are depicted [Figure 2]A, [Figure 2]B, and [Figure 3]A, [Figure 3]B. The inference is found to be more informative in differentiating the two species.
Figure 2: (A) Elements of maceration of Cinnamomum verum petiole (a) epidermis, (b) stone cell with wide lumen, (c) sclereisidal fiber, (d) prismatic crystals of calcium oxalate, (e) vessels with wide simple pits, (f) stone cell with narrow lumen, (g) spiral vessel, (h) mucilage and volatile oil cells, (i) sclereids. (B) Elements of maceration of Cinnamomum sulphuratum petiole (a) epidermis, (b) rosette crystals, (c) tablet-shaped crystals, (d) spiral vessel, (e) highly pitted vessel, (f) fiber with narrow end, (g) sclereids, (h) stone cells, (i) sclereids and oleoresin cells

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Figure 3: (A) Histology of Cinnamomum verum leaves: (a) transverse section of petiole, (b) transverse section of lamina showing vascular strands, (c) epidermis of petiole in surface view, (d) transverse section of petiole showing group of stone cells, (e) upper epidermis showing cicatrix, (f) lower epidermis showing stomata and cicatrix. (B) Histology of Cinnamomum sulphuratum leaves: (a) transverse section of petiole, (b) transverse section of lamina showing vascular strands, (c) epidermis of petiole in surface view, (d) transverse section of petiole showing group of stone cells, (e) upper epidermis showing cicatrix, (f) lower epidermis showing stomata, papillae, and cicatrix

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Micrometry

Micrometric measurements were taken for various tissues in the macerated leaf [Table 4] and [Figure 4]a, [Figure 4]b.
Table 4: Micrometry of leaves of Cinnamomum spp.

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Figure 4: (a) Microscopy of macerated leaf of Cinnamomum verum, (b) microscopy of macerated leaf of Cinnamomum sulphuratum

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Quantitative microscopy

Values for different leaf constants such as upper epidermal cells, vein islet number, and stomatal number are documented in [Table 5].
Table 5: Quantitative microscopy of leaves of Cinnamomum verum and Cinnamomum sulphuratum

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Powder

Pearl glands were observed in C. verum (absent in C. sulphuratum); fragments of pitted and reticulate vessels present in C. verum (absent in C. sulphuratum); tracheidal elements in C. sulphuratum possessed wide lumen (narrow lumen in C. verum); stone cells with wide lumen are present in C. verum (found rarely in C. sulphuratum); oval-shaped stone cells are extant in both the species, but the boat-shaped one is present exclusively in C. sulphuratum; stone cells with three-sided thickening was seen in C. verum (absent in C. sulphuratum); prismatic crystals of calcium oxalate predominantly were present in C. verum (absent in C. sulphuratum) [Figure 5]a and [Figure 5]b.
Figure 5: (a) Powder microscopy of leaf of Cinnamomum verum, (b) powder microscopy of leaf of Cinnamomum sulphuratum

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   Discussion Top


The morphological similarities, wider distribution, and resemblance in fragrance of the leaves of various species of Cinnamomum have created a great difficulty in differentiating and lead to the adulteration and substitution of herbal samples. C. verum has been used in many Ayurvedic and Siddha formulations. Due to the similarities with C. verum, C. sulphuratum is used widely as the substitute. Anatomical descriptions of leaf of Cinnamomum species have been provided earlier,[16],[17] but detailed anatomical studies of leaf have been published for only a restricted number of Cinnamomum species.[18],[19],[20],[21],[22],[23],[24],[25],[26]

Even though penninerved and triplinerved leaves have been reported in Cinnamomum,[27] the two species under the study possessed triplinerved leaves. C. verum and C. sulphuratum had thick epidermis with lignin deposition in contrast to the suberin deposition in Cinnamomum tamala.[26] The stomata observed in C. verum were anomocytic while it was paracytic in C. sulphuratum. Earlier anomocytic stomata have been reported in C. malabatrum and paracytic in C. tamala[25] and Cinnamomum camphora.[27] In C. verum and C. sulphuratum, volatile oil cells were present in mesophyll cells but completely absent in Cinnamomum magnifolium.[23] In Cinnamomum amoenum, three-layered palisade parenchyma was observed,[23] while in C. verum and C. sulphuratum, it is single layered. The variation in the number of layers of spongy parenchyma has been encountered before in C. malabatrum (5–6) and C. tamala (10–12).[20],[25]

Macroscopic and organoleptic features depicting the differences in petiole, leaf, smell, and taste can help to a certain extent in delimiting the species, but the detailed and exhaustive microscopic evaluation which displayed the differences in hypodermal sclerenchymatous layers, shape and number of xylem elements, differences in the number of phloem patches, and the presence of isolated sclerenchyma cells surrounding the vascular bundles has proved to be a reliable tool in authenticating the two species.


   Conclusion Top


The lack of botanical identity of herbal drugs may lead to adulteration and substitution. When the concerned herbal material is not present in sufficient quantities, there is a tendency to substitute it with available samples. The differences exhibited in macro–microscopy of the species may lead to the differences in their medicinal values too. Hence, the usage of allied species may not ensure the exact bioactivities for which detailed pharmacological studies may be required. The detailed macro–microscopic profiles laid down for C. verum and C. sulphuratum leaves will thus serve as diagnostic tools for their differentiation.

Acknowledgment

Authors are grateful to Prof. R. Ramasamy, Director General, CCRS, for support. Deep gratitude is extended to Dr. Prof. Malati G. Chauhan and Sri A. P. G. Pillai for their guidance. Dr. Remashree, CMPR, Arya Vaidya Sala, Kottakkal, are acknowledged for providing photomicrography facility.

Financial support and sponsorship

Nil.

Conflicts of interest

There are no conflicts of interest.



 
   References Top

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    Figures

  [Figure 1], [Figure 2], [Figure 3], [Figure 4], [Figure 5]
 
 
    Tables

  [Table 1], [Table 2], [Table 3], [Table 4], [Table 5]



 

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