INTRODUCTION:

Plant materials manifest a vital model of the global drug market today. Being natural often they are preferred because of their lower probabilities of adverse effects to the synthetic medicines over a large extent of population across the world. As per a recent evaluation of World Health Organization (WHO), most of the people across the world depends upon herbal medicines for their primary health care.

Medicinal characters are represented by the plants because of the presence of Andrographolide and Neoandrographolide (some forms of diterpenoids).

It has been reported about its medicinal uses. In the current period, the order of deposition of toxic industrial effluents including heavy metals present in the soil, air and water are gradually ascending because of rapid urbanization and industrialization the use of pesticides, insecticides, etc for the sake to fulfill the demand of population growth is a serious concern today. Plants often bear these metals in their all body parts by the processes like root uptake, foliar adsorption, deposition of specific elements in leaves, etc and then transmitted ultimately to those organisms including human who inject them. Hence it's important to advocate quantitative and qualitative estimation of heavy metals in herbal products before use for the sake to assure the safety of health and hygiene. So assessment of the heavy metals or other minerals is indispensable including their compositions in the medicinal plants to estimate the quantities of the heavy metals in the plant body. Andrographis paniculata is of interest to this study. It is a plant of all seasons and with green leaves of narrow oval shaped tapering to the points at each end and having heights of 60-70cm. It’s called King of Bitters because of its extreme bitter taste in every parts of its body. It’s also widely used in Ayurvedic medicines and Unani type of medicines.

Plant descriptions:

A. paniculata is an annual branched herb, stem tetra angular, slightly winged, solid, woody, glabrous, green color leaves lanceolate shape, simple 6-10cm (approximately) long and 3-5cm wider, having acute apex, margins are entire, wedge-shaped at base, herbaceous in nature, smooth on both surface, green in color.

(Fig. A: A. Paniculata plant in IMMT BBSR campus)

Vernicular Names:

Arabic: Quasabhuva, Assames: Kalmegh, Azerbaijani: Acilarsahi, AcilarXant, Bengali: Kalmegh, English: The Creat, King of Bitters, French: Chirette Verte, Hindi: Kirayat, Kalpanath, Japanese: Senshinren.

The safety of the herbal plants from natural contaminants like microbes and heavy metals present in the environment which in turn find their way into the herbal materials. Herbal materials mean either whole plant or parts of medicinal plants which include herbs, gums, resins, and dry powders of the herbs in the crude state¹.

Most of the reports have indicated that herbal medicines contain toxic heavy metal contaminants which can cause adverse effects like cancer, liver dysfunction, skin eruption, lung disease, cerebral haemorrhage, alopecia, etc.²

Metals such as Fe, Ca, Mn, Co, Cu, Zn, Ni, in trace amounts are essential for human life as growth supplements. However, large amount of heavy metal accumulation in human body get deposited and captured by proteins to form complexes, eventually leading to malfunctioning or lethal of the cells³. Non- essential metals such as Pb, Cd are toxic even in trace amounts while few metals like Cr and as are considered as potential contaminants⁴.

The aim of the study is to assess different metal contents of A. paniculata plant body from both urban and rural regions in Khordha district, Odisha.

The minerals play important role in promoting health care. Deposition of toxic industrial effluents/wastes including heavy metals in the soil, air and water is immensely ascending due to massive urbanization and industrialization as well. And the use of pesticides, insecticides, weedicides, etc, in agricultural field is a serious concern today. Plants often accumulate these metals in their all body parts caused by root uptake, foliar adsorption, etc and then transmitted ultimately to those organisms including human who uptake them. Hence quantitative and qualitative estimation of heavy metals of herbal products are unavoidable before they are used for the sake to assure the safety of health (Health is Wealth). So assessment of the heavy metals or other minerals is very much necessary along with the compositions in the medicinal or aromatic plants to estimate the level of heavy metal contaminants in the plant body.

The previous studies on this plant are discussed consisting of pharmacological activities of the plant like hepato protective activity^5,6, immunological potential⁷, anti-cancer potential⁸, cytotoxicity against cancer cells, anti-inflammatory and anti-angiogenic activities⁹, anti- hyperglycemic and renal protective activities, anti-malarial activities^10,11,12, cardiovascular activities¹³, anti-fertility effect¹⁴, anti-venom activities. Although they have lots of pharmacological application, their various agricultural, medical and technological applications demonstrate their specific wide distribution in the environment; imparts a focus over their potential impacts on human health and the sustainable environment as well. Their toxicity depends on several factors including the doses, route of exposure and chemical species, as well as the age, gender, genetics status of exposed individuals. Because of their high degree of toxicity, As ^15-18, Cd^19-21, Cr^22,23, Pb^24,25 and Hg are classified as human carcinogens according to the U.S. Environmental Protection Agency, and the International Agency for Research on Cancer (IARC).

This study deals with the characterization of metals content present in A. paniculata plant collected from two regions which are analyzed by using the following physical and chemical methods.

MATERIALS AND METHODS:

At the very beginning the plant samples i.e. Andrographis paniculata, were collected from two places of Khordha region v.i.z: CSIR-IMMT campus, Bhubaneswar and Bolgarh region along with soil sample. Then these plants were washed with tap water and dried in sunlight up to one week and then dried in oven for 24h to have moisture-free plant samples. Then all the parts like roots, stems and leaves were separated well for individual estimation purposes. Further all the three parts are individually made to powdered form. Further ashing was carried out using muffle furnace (M/s Thereek Furnace) at 500℃ for 6 h each.

Then these are taken out of the furnace and kept in a desiccators for drying. The ashes were individually taken in beakers for acid digestion for 1hr at 80°C maintained using magnetic stirrer. The ashes were oxidized with Aqua-regia (mixture solution of HNO₃ and HCl of 1:3 ratio). Each sample was filtered using Whatmann Filterpaper (42) and the filtrate was made up to 25mL volume (in 25mL volumetric flask) by addition of de-ionized water to it. Then these solutions of root, stem and leaf were taken for estimation of concentration of metals (Zn, Mg, Mn, Co, Cu, Cr,) and Heavy metals like Ni, Cd, Pb, As by Atomic Absorption Spectroscopy and ICP- OES. Other metals like Ca, Na, K are estimated by using Flame Photometry.

All the experiments are duplicated i.e. two sample solutions of each of the ashes were taken for precision and accuracy of the results.

Plant Samples collected from IMMT, BBSR campus

Table-1: Plant Samples taken from IMMT, BBSR campus with various quantities

Samples	Dried weight of powdered samples(g)	Weight of the ash samples(g)
L₁	0.7655	0.1835
L₂	0.6679	0.2323
S₁	3.0000	2.1898
S₂	3.0000	2.1898
R₁	1.0000	0.6000
R₂	1.0000	0.4000

Plant samples collected from Bolgarh, Khordha:

Table-2: Plant samples collected from Bolgarh with various quantities

Sample	Dried weight of powdered samples (g)	Weight of the ash samples (g)
L₃	2.0762	0.6000
L₄	2.0762	0.4000
S₃	0.7358	0.3000
S₄	0.7358	0.2000
R₃	2.5385	2.2961
R₄	2.5385	2.2961

Now all the digested samples are individually filtered by using Whatmann Filter paper (42) and the filtrates are transferred into 25 mL volumetric flask and volumes were made up to the mark. Now these homogeneous sample solutions of roots, stems, leaves are ready for AAS, ICP-OES and Flame Photometry. It is important to note that the original solutions are then made diluted separately from the original solution as per the require ppm or mg/L for analysis purposes.

Table-3: Plant samples collected from IMMT, BBSR campus

Sample ID	Original sample (mg/L(ppm) in 100g)
Sample ID	Pb	Cd	Cr	Ni	Co	Cu	Mn	Zn	Fe	Mg
L₁	2.874	0.190	0.749	2.656	1.607	0.664	3.337	1.280	21.389	Not detected
S₁	0.101	0.013	0.780	0.251	0.092	7.247	2.226	1.940	NA	Not detected
R₁	0.504	0.091	2.433	0.775	0.412	4.391	38.541	13.958	1032.916	0.750
R₂	0.225	0.087	2.631	1.118	0.231	3.975	31.875	8.125	2281.875	0.637

Analysis results of Table-2:

Table-4: Plant sample collected from Bolgarh, Khordha

Sample ID	Metal content (mg/L(ppm) in 100g)
Sample ID	Pb	Cd	Cr	Ni	Co	Cu	Mn	Zn	Fe	Mg
L₃	0.820	0.075	2.064	54.79	0.291	16.870	0.791	6.104	443.541	441.666
L₄	0.781	0.093	1.841	56.562	0.393	18.850	0.825	5.943	547.500	639.375
S₃	2.991	0.116	0.826	75.416	0.458	5.408	1.808	1.358	246.666	773.330
S₄	3.30	0.225	0.531	81.875	0.287	4.312	2.850	3.137	324.375	1100.620
R₃ & R₄	0.166	0.013	1.535	15.241	0.940	0.274	5.372	1.149	Not studied	Not studied

ICP-OES Analysis Data: (iCAP7600Duo)

Table-5: Plant samples collected from IMMT, BBSR campus

Sample ID	Metal content(mg/L(ppm) in 100g)
Sample ID	Fe	Cu	Co	Mn	Ni	Cd	Zn	Pb	Cr	V
R₁	1029.583	4.570	0.233	27.787	0.641	0.054	10.216	0.484	5.558	1.962
R₂	Not detected	4.130	0.212	25.787	0.725	0.050	11.037	0.319	5.356	1.893
S₁	130.00	0.645	0.034	2.887	0.180	0.018	1.380	0.344	3.297	0.342
S₂	154.580	0.651	0.039	2.930	0.192	0.018	1.365	0.365	3.350	0.347
L₁	4871.394	4.404	0.462	53.551	1.022	0.032	27.324	0.630	3.540	3.508
L₂	Not detected	4.294	0.495	54.928	0.979	0.021	27.023	0.609	3.637	3.544

Table-6: Plant sample collected from Bolgarh, Khordha campus

Sample ID	Metal content(mg/L(ppm) in 100g)
Sample ID	Fe	Cu	Co	Mn	Ni	Cd	Zn	Pb	Cr	V
R₃	256.794	0.540	0.152	5.084	0.193	0.002	1.793	0.505	4.967	0.450
R₄	252.384	0.529	0.146	4.913	0.175	0.001	1.771	0.478	4.868	0.424
S₃	339.166	55.358	5.308	44.449	0.850	0.016	68.850	12.183	6.575	4.466
S₄	378.750	128.50	5.887	54.137	1.250	0.016	166.25	22.500	11.250	4.525
L₃	470.000	5.225	16.170	59.308	0.750	0.004	23.879	21.640	10.479	4.929
L₄	477.500	5.418	18.643	64.493	1.050	0.018	25.887	30.956	12.181	5.112

Flame Photometry Data: (Flame photometry 128)

Table-7: Plant sample collected from IMMT-BBSR campus

Sample ID	*Metal content(mg/L(ppm) in 100g)*
Sample ID	Ca	Na	K
R₁	2553.229	16.075	1738.333
S₂	796.876	145.561	1386.542
L₂	1554.1541	1140.766	13845.2432

Table-8: Plant sample collected from Bolgarh, Khordha

Sample ID	*Metal content(mg/L(ppm) in 100g)*
Sample ID	Ca	Na	K
R₃	283.633	115.957	773.049
S₃	8038.616	92.191	26881.666
L₄	12717.956	75.468	19861.250

OBSERVATIONS:

We have also viewed the metals concentration in A. paniculata plant studied by BCSIR, Chittagong cultivation area in Bangladesh²⁶ (a review collected) and the data is shown in figure-1 and another study was carried out in Chennai, India²⁷ as shown in figure-2.

Fig.-1: AAS data of BCSIR, Bangladesh

Fig.-2: AAS data of Chennai, India

The Atomic Absorption Spectroscopy (AAS) characterization (Table-3) of the plant sample collected from IMMT, BBSR region suggests that leaves are richer in Fe, Mn, Ni, Pd metals and stems are rich in Fe metal. This advocated that urban area A. paniculata plants are rich in trace metas like Fe, Cu, along with other toxic metals with exceptional extent.

As per Table-4 of AAS data, it represents that plant (A. paiculata) sample collected from Bolgarh region, Khordha, leaves are richer in Mg, Fe, Ni, Stems are also rich in Mg, Fe and Ni and roots are rich in Mn and Fe. Consequently here it is cleared that rural area A. paniculata plants are composed of Mg, Fe along with toxic metals with less extent as compared to urban area. In short, AAS data proves that urbanization and industrialization (environmental conditions) have a major impact on metals deposition in the concerned plants.

Comparison of metals content of A. paniculata from the two regions analyzed by AAS

Fig.-3.0: Data of leaves sample of BBSR

Fig.-3.1: Data of leaves samples of Bolgarh

Fig.-4.0: Data of stems sample of BBSR

Fig.-4.1: Data of stems sample of Bolgarh

Fig.-5.0: Data of roots sample of BBSR

Fig.-5.1: Data of roots sample of Bolgarh

By considering ICP-OES data, it has been obtained that all the parts of plants collected from IMMT, BBSR campus and Bolgarh, khordha are rich in iron metals compared to other metals exist. The comparisons of the metal concentrations are given as;

Fig.-6.0: Data of roots sample of BBSR

Fig.-6.1: Data of roots sample of Bolgarh

Fig.-7.0: Data of stems sample of BBSR

Fig.-7.1: Data of stems sample of Bolgarh

Fig.-8.0: Data of leaves sample of BBSR

Fig.-8.1: Data of leaves sample of Bolgarh)

Further for Na, Ca and K, Flame photometry was used and the observations were studied. The A.paniculta plant samples were collected from IMMT-BBSR whose leaves are rich in Potassium (K) i.e. 13845.243 ppm/100g as compared to Na and Ca. Similarly stems are rich in K i.e. 1386.542ppm/100g, roots are rich in Ca i.e. 2553.229 ppm/100g comparison with K and Na. This indicates that the plant as a whole rich in Potassium (K) as compared to Na and Ca. A. paniculata samples collected from Khordha region, leaves are rich in K i.e. 19861.250 ppm/100g, stems are rich in K i.e. 26881.66 ppm/100g and roots are rich in K i.e.773.049ppm/100g. This indicates the plant from Khordha region is also rich in K as compared to both Na and Ca, the flame photometry data of both the regions are compared as shown in the below pie chart;

Fig.-9.0: Data of root sample of BBSR

Fig.-9.1: Data of root sample of Bolgargh

Fig.-10.0: Data of stem sample of BBSR

Fig.-10.1: Data of stem sample of Bolgarh

Fig.-11.0: Data of leaf sample of BBSR

Fig.-11.1: Data of leaf sample of Bolgarh

As per some FESEM microstructural images of leaves, the micro-structures represent some slab like structures which are interlinked together forming a flower like shape which may look like micro-structure of CaPO₄ on which the work is under process for clarifications.

CONCLUSIONS:

Commercially available raw materials of the herbal products are used in home remedies for common ailments worldwide. In this investigation, greater level of Ca, K, Mg, and Fe were detected when compared to other metals using AAS, ICP-OCP, and FP. As per the literature study, consumption of such herbal plants or medicinal plants might help to get rid of various types of diseases. Thus our findings are indicating that the pharmaceutical or medicinal plants or their parts which have been used for the treatment of different diseases or as synthetic medicines must be checked for heavy metals (Pd, Zn, Cd, Ni) contamination for the sake of making it safe for human consumption.

It should be collected from the areas that are not contaminated with heavy metals for local or pharmaceutical purposes. This study gives a brief knowledge including metal contents in A.paniculata plant which is very useful for mankind. Last but not the least this work is continuing to know about mineral compositions by micro-structural and XRF studies.

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Received on 11.06.2022 Modified on 24.06.2022

Res. J. Pharmacognosy and Phytochem. 2022; 14(4):257-264.

DOI: 10.52711/0975-4385.2022.00045

Vernicular Names:

Arabic: Quasabhuva, Assames: Kalmegh, Azerbaijani: Acilarsahi, AcilarXant, Bengali: Kalmegh, English: The Creat, King of Bitters, French: Chirette Verte, Hindi: Kirayat, Kalpanath, Japanese: Senshinren.

MATERIALS AND METHODS:

Plant samples collected from Bolgarh, Khordha:

Sample

Dried weight of powdered samples (g)

Weight of the ash samples (g)

L3

2.0762

0.6000

L4

2.0762

0.4000

S3

0.7358

0.3000

S4