Phytochemical evaluation of the wild and cultivated varieties of Eranda Mula (Roots of Ricinus communis Linn.).

In Ayurveda, the roots of Eranda (Ricinus communis Linn.) are used in the treatment Amavata (rheumatism), Sotha (inflammation), Katisula (backache), Udararoga (disease of abdomen), Jwara (fever), etc, Due to high demand, root of the cultivated variety is mainly used in place of wild. But, a comparative phytochemical profile of both varieties is not available till date. Considering this, a preliminary study has been done to ensure basic phytochemical profile of both the varieties. Preliminary physicochemical parameters, phytochemical screening, quantitative estimation of alkaloid, high-performance thin layer chromatography (HPTLC), and heavy metal analysis were carried-out in the study. Analysis of physicochemical data reveals no significant difference in between both varieties of roots, while alkaloid was found to be more in cultivated variety (0.34%) than wild one (0.15%). Though, the analytical profiles are almost identical, except the quantity of alkaloid; inferences should be made through well designed pharmacological and clinical studies.

Introduction

Ricinus communis Linn (Euphorbiaceae) commonly known as Eranda in Ayurveda is a soft-wooded small tree wide spread throughout tropics and warm temperate regions of the world. In the Indian system of medicine, the leaf, root, and seed oil of this plant have been used for the treatment of inflammation and liver disorders.[1] In Ayurveda, the roots of Eranda are used in the treatment of Amavata (rheumatism), Sotha (inflammation), Katisula (backache), Udararoga (diseases of abdomen), Jwara (fever), etc.[2] Its roots have also been highlighted for its Vrishya (aphrodisiac) and Vatahara actions by Acharya Charaka.[3] This plant also possesses hepatoprotective,[45] anti-diabetic,[6] laxative,[7] anti-fertility,[8] anti-inflammatory and free radical scavenging activities.[9]

Due to high demand, roots of the cultivated variety are mainly used instead of wild. But, a comparative phytochemical profile of both varieties is not available till date. Hence, to ensure quality of both varieties, phytochemical evaluations of both the varieties was undertaken.

Materials and Methods

Collection of drug

Fresh roots of wild and cultivated varieties of R.communis Linn. after proper identification were collected from the adjacent area of Jamnagar, Gujarat, with the help of taxonomist. Specimen herbarium of both varieties for wild (No. 1490) and cultivated (No. 1491) were preserved in the Pharmacognosy Laboratory, IPGT and RA, Gujarat Ayurved University, Jamnagar for further reference. The obtained roots were shade dried and made into coarse powder with the help of mechanical grinder and preserved in a glass container for future studies [Figure 1].

Figure 1

Raw and crushed roots of wild and cultivated variety

Physicochemical study

Moisture content, ash values (total ash, acid insoluble ash), and extractive values (alcohol soluble extractive, water soluble extractive) were determined by following standard analytical procedures.[1011]

Preliminary phytochemical screening

Five grams coarse powder of the roots was subjected for extraction with methanol (100 ml), keeping it for overnight with initial occasional shaking up to 6 h. and then set aside. After 24 h, it was filtered and alcoholic extract was collected. Similarly, water extract was prepared. Both the extracts were evaporated to dryness. The dried extracts were weighed, and percentage yield was calculated. The extracts were used for preliminary phyto-chemical screening with a set of various chemical tests viz., Dragendorff's Mayer's, Hager's, and Wagner's tests for alkaloids; ferric chloride, lead acetate, potassium dichromate, and dilute iodine tests for tannins and phenolics; and foam test for saponin glycosides.[12]

High-performance thin layer chromatography

High-performance thin layer chromatography (HPTLC) was carried out by the standard methods.[13]

Chloroform extract of cultivated variety was labeled as track 1, while the wild variety as track 2. The solvent System used in the study was Toluene:Ethyl acetate:dimethylamine (7:2:2).

Chromatographic conditions

Application mode: Camag Linomate V; Development chamber: Camag Twin trough chamber; Plates: Pre-coated silica gel GF254 plates; Chamber saturation: 30 min; Development time: 30 min; Development distance: 7 cm; Scanner: Camag scanner II; Detection: Deuterium lamp and mercury lamp; Photo-documentation: Camag reprostar; Data system: Win cats software; Drying device: Oven and was visualized under 254 nm and 366 nm.

Quantitative estimation of alkaloid

The samples were estimated quantitatively for total alkaloid content by gravimetric method.[14]

Heavy-metal analysis

Heavy metal analysis of the root powder of both varieties, for arsenic, lead, mercury, and cadmium, by following standard procedure,[15] was carried out at Analytical Testing Laboratory, Konark Research Foundation, Daman.

Results and Discussion

Physico-chemical analysis

Results of physicochemical analysis, qualitative tests, and Rf values of HPTLC in wild and cultivated varieties of R.communis Linn. are mentioned in Tables 1–3 respectively. Alkaloid percentage was estimated as 0.34% in cultivated, and 0.15% in wild variety. Results of heavy metal analysis for arsenic, lead, mercury, and cadmium are mentioned in Table 4.

Table 1

Physicochemical analysis of Eranda Mula: Wild and cultivated varieties

Table 3

Rf values of chloroform extracts of wild and cultivated variety of Eranda Mula

Table 4

Heavy metal analysis of wild and cultivated variety of Eranda Mula

Analysis of physicochemical data [Table 1] reveals absence of foreign matter in both samples. Moisture content in wild variety (6.48% w/w) is less than cultivated variety (7.43% w/w). The difference in total ash, acid insoluble ash, water soluble extractive value and pH in between the samples is insignificant. Alcohol soluble extractive is comparatively higher in wild variety (22.56% w/v) than cultivated variety (17.92% w/v).

The root of cultivated and wild varieties of R.communis exhibits almost similar phytochemical profile indicating presence of alkaloids, cyanogenic glycosides, flavonoids, terpenoids, tannin, carbohydrates, and saponin [Table 2]. Quantitatively, tannin was found to be more in cultivated variety (0.34%) than the wild one (0.15%).

Table 2

Preliminary phytochemical profiles of Eranda Mula: Wild and cultivated varieties

Chloroform extract of wild variety showed 03 spots and cultivated variety showed 05 spots under 254 nm, among them Rf values 0.13 and 0.35 are similar in both samples. In 366 nm, chloroform extract of both varieties showed 05 spots, among them one Rf value 0.13 is similar in both samples. Other Rf values are nearer to each other, indicating that the components present in both varietiesmay be similar [Figure 2; Table 3]. In situ Ultra Violet spectral comparison graph also shows chemical similarity at 0.08 Rf, 0.15 Rf, and 0.36 Rf values [Figure 3].

Figure 2

Photograph of high-performance thin layer chromatography

Figure 3

In-situ UV spectral comparison grap

Heavy metal analysis of both varieties shows that the samples are free from heavy metal contamination and the observations are under the prescribed limits for heavy metals.[16] [Table 4].

Conclusion

Both cultivated and wild varieties have a chemical similarity except higher percentage of alkaloid in cultivated than wild variety. HPTLC finger printing shows similarities in wild and cultivated varieties. Heavy metals were within prescribed limit in both the varieties. Though, the analytical profiles are almost identical, except the quantity of alkaloid; inferences for clinical use should be made through well designed pharmacological and for clinical studies.