INTRODUCTION:

Herbal remedies and natural plants are playing a paramount role in mankind health system and have been focused because of their capability to generate benefits especially in medical and pharmacological venue¹. Recently people were looking for the traditional health system like Unani, Ayurveda and Siddha for treatment of different diseases as the reported side effects with conventional drug therapy (allopathic drugs)². Phytochemical constituents were responsible for the medical power of the medicinal plants for performing their physiological actions ¹. Phytochemical constituents of medicinal plants used either for aliments curing or protection from various diseases³.

Plant material may vary in their phytochemical content due to their natural heterogeneity subsequently affecting their therapeutic activity according to the plant age, time of collection throughout the year, geographical places and environmental factors surrounding the plant⁴. There were an increasing demand for the documentation of plant standard parameters and standardization of plant material especially of plants belongs for same family⁵.

Medicinal herbs used for treatment of variant disease since ancient times⁶. The major cause of morbidity and mortality in the world is infectious diseases which cause a critical situations in health care system⁷. Many phytochemicals have important therapeutic activities such antibacterial, antifungal⁸, spasmolytic⁹, therefore the medicinal value which recorded for the corresponding plant due to presence of the phytochemical³.

Brassicaceae (old name: Cruciferae) or mustard family includes important traditional and industrial oil seed, condiment, vegetables and crop plant species [10]. Two botanically related species of Brassicaceae family¹¹were include in study Lepidium sativum Linn. (garden cress) and Nasturtium officinale R.Br. (watercress).

Lepidium staivum is fast growing annual herb, with many branches in upper part, major documented phytochemicals are sulphur glycosides, triterpenes, coumarins, flavonoids, sterols and various imidazole alkaloids. Different pharmacological have been recorded for the plant such as diuretic, demulcent, aperients, rubefacient, tonic, aphrodisiac, carminative, emmenagogue and galactagogue¹¹. Locally known as (Taratura) used for culinary purposes by native people of the region and medically Lepidium staivum seed used as antidiabetic herbal remedy.

Nasturtium officinale is perennial herb found in clumps in wet soils¹², important phutochemical constituents of water cress are lutein, zeaxanthin¹³. Reported therapeutic value for Nasturtium officinale are diabetes, diuretic¹⁴, antimicrobial¹⁵, anticarcinogenic and antiestrogenic¹⁶, finally used as dietary supplement and digestive aid¹⁷. Locally known as (Kuzara) used by local people for both culinary purposes and medically as an adjuvant of kidney stone removal.

The aim of present study was to compare the two botanically related species of Brassicaceae family Lepidium staivum and Nasturtium officinale grown naturally in mountain regions of Erbil city, Kurdistan Region sharing similar taste and flavor from physicochemical, phytochemical and biological view points, and emphasizing their botanical relation.

MATERIAL AND METHODS:

Plant material collection:

Arial parts of both Lepidium staivum and Nasturtium officinale were collected in mountain places of Erbil city, Kurdistan Region\Iraq, have been identified by Pharmacognosy Department, Pharmacy College\ Hawler Medical University, dried in shade. Dried plant material were kept under 25°C, until introduced for further study parts.

Physicochemical analysis:

Various physico-chemical parameters such as the percentage of foreign material, moisture content [loss on drying (LOD)], total ash, acid insoluble ash, water soluble ash were determined. Extractive value, color and consistency of plant material extracts obtained from different polarity solvents [petroleum ether (PE), chloroform (CH), ethanol (ETH), methanol (METH), and water (AQ)] for both plant materials were determined separately. All the parameters were taken in triplicate procedures and the obtained results was presented as percentage of mean ± standard deviation (mean ± SD)¹⁸^{, 19}.

Fluorescence analysis:

Plant materials extracts with different polarity solvents and powdered plant materials have been examined for the presence of fluorescence substances. Powdered plant materials were placed on a grease free microscopic slide and added 2 drops of freshly prepared reagent solutions (xylene, concentrated sulphuric acid, 1 M sodium hydroxide, picric acid, concentrated nitric acid, 10% nitric acid, 10% hydrogen peroxide, 10% potassium dichromate, iodine, aqueous ferric chloride, alcoholic ferric chloride, concentrated hydrochloric acid and acetic acid) mixed thoroughly and waiting for 1-2 minutes, then examine the mixture under visible day light, UV-light (254 nm) and UV light (366 nm), the observed color were recorded ^20,
21.

Phytochemical analysis:

Qualitative phytochemical screening:

Variant solvent extracts such as [petroleum ether, chloroform, methanol, ethanol, water] of powdered plant materials have been introduced to different chemical tests for identification of phytochemical constituent using standard conventional protocols ^{22, 23}.

Quantitative phytochemical analysis:

Total alkaloid:

Ten gram of powdered plant materials were mixed with (200 ml) of 10% (v\v) alcoholic acetic acid. The mixture were allowed to stand for 4hr at room temperature, then filtrated and concentrated to about one quarter of it is original volume on water bath. Concentrated ammonium hydroxide solutions were added to the filtrate until precipitation have been stopped. The precipitate were collected and washed with dilute ammonia solution. The residue was expressed as total alkaloidal content, calculated as percentage using the plant material dry weight as reference ²⁴. Data were estimated from triplicate procedure works expressed as (mean ±SD).

Total Flavonoid:

Ten gram of powdered pant material has been extracted using (80% (v\v)) aqueous methanol repeatedly at room temperature. Filtrate of the repeated extraction process introduced to drying using water bath until constant weight has been obtained. The weight of dried powdered extract is the total flavonoid expressed as percentage using the dry weight plant material as reference ²⁴. Data were collected from triplicate procedure work expressed as (mean ±SD).

Biological analysis:

Plant extracts with higher number phytochemical constituents were chosen for biological analysis.

Plant extracts preparation:

Powdered plant material (for both plants separately) introduced to extraction using methanol and ethanol as solvent of extraction using ultra sonic extractor method described by Alpuli et al 2009 ²⁵. Extracts concentrated and dried under vacuum using rotary vapor machine, 100 mg\ml concentration of extracts were prepared and analyzed for their biological activity using dimethyl sulfoxide (DMSO) (10%) as diluent.

Inoculum preparation:

Five standard pathogenic bacteria were selected for the antibacterial evaluation: Pseudomonas aeruginosa (ATCC 27853), Escherichia coli (ATCC 35218), Staphylococcus aureus (ATCC 25923), Streptococcus pyogenes (ATCC 19615), and Streptococcus pneumoniae (ATCC 6303). All strains were maintained on nutrient and blood agar slant accordingly, and stored at 4 ˚C until used. McFarland standards 0.5 are used as a reference to adjust the turbidity of bacterial inoculums ²⁶.

Antimicrobial evaluation:

Agar well diffusion method used for evaluation of antibacterial (biological) activity of plant extracts as described by Sandhya et al 2012²⁷ with slight modification. The bacterial inoculum was uniformly spread using sterile cotton swab on a sterile Muller Hinton agar plate. About 0.1 ml of plant extracts was added to each of the wells (6 mm) diameter holes cut in the agar. Positive control [azithromycin antibiotic (1 mg\ml)] and negative control [DMSO (10%)] were incorporated in each plate. The plates were incubated for 24 h at 37˚C under aerobic conditions. Following incubation, the inhibition of the bacterial growth around the wells was measured in mm including the well diameter. Tests were performed in triplicate, the mean ± SD of the results were taken in consideration.

Statistical analysis:

All data were estimated from triplicate procedures expressed as mean ± SD. Comparison between mean were performed using t-test considering (p value ˂ 0.05) statistically significant using Graph Pad Prism 6 program.

RESULTS:

Physicochemical analysis:

The physicochemical parameters for Lepidium staivum and Nasturtium officinale aerial parts were determined shown in Table.1. The extractive value for different polarity solvents with estimation of physical characteristics including texture of the extract [such as sticky (S), oil form, paste form, non-sticky (NO)] and color of the extracts of both plants were presented in Table.2 and Table.3. Comparisons between data for extractive values were shown in (Figure.1.). Mean values for each parameter were compared for both plants considering (p value < 0.05) statistically significant.

Table. 1: Physicochemical parameters for Lepidium staivum and Nasturtium officinale:

Physicochemical parameters	*Lepidium* *staivum* ¹	*Nasturtium officinale* ¹
Foreign material	0.021± 0.001%^*	0.011±0.001%^*
LOD	86.66 ± 1.1574 %	84.55±1.1574%
Total Ash value	19.677 ±1%	21.29±0.577%
Water soluble ash	5.484 ± 0.5%^*	10.161 ±0.577%^*
Acid insoluble ash	3.225 ± 001%	3.225 ± 001%%

1 stands to mean ± SD were expressed as percentage (w\w%), n=3

* stands for significant difference between mean values (p value ˂ 0.05)

Table. 2: Extractive value and physical characteristics of different polarity solvent extracts of Lepidium staivum:

Type of extract	Extractive value (w\w%)1	Physical characteristics
Type of extract	Extractive value (w\w%)1	Consistency2	Color
Petroleum ether	1.0 ±0.1	NS (oil form)	Yellow
Chloroform	8.0 ±0.1	S (paste form)	Dark green
Methanol	18.1±0.1	NS (paste form)	Dark green
Ethanol	10.0667±0.0577	NS (paste form)	Dark green
Water	18.1±0.1527	NS (powder form)	Yellow

1Extractive value were expressed as mean of percentage ± SD (w\w), n=3

2 S: stands for sticky texture, NS: stands for non sticky texture

Table. 3: Extractive value and physical characteristics of different polarity solvent of Nasturtium officinale:

Type of extract	Extractive value (w\w%)1	Physical characteristics
Type of extract	Extractive value (w\w%)1	Consistency2	Color
Petroleum ether	2.1±0.1	S (oil form)	Light green
Chloroform	1.967±0.0577	S (oil form)	Light green
Methanol	6.0 ±0.1	NS (paste form)	Dark green
Ethanol	3.697±0.1527	NS (paste form)	Dark green
Water	18.0 ±0.1527	NS (powder form)	Brown

1Extractive value were expressed as mean of percentage ± SD (w\w), n=3

2 S: stands for sticky texture, NS: stands for non sticky texture

Fluorescence analysis:

Both plant materials of Lepidium sativum and Nasturtium officinale have been examined under UV in both forms extract and dry powdered plant material expressed in Table 4 and Table 5 for Lepidium sativum and Table 6 and Table 7 for Nasturtium officinale.

Table.4: Fluorescence analysis of Lepidium sativum extracts:

Plant extract1	UV light (254nm)	UV light (366 nm)
PE	Black	Light grey
CH	Navy	Light purple
METH	Brownish black	Black
ETH	Green	Black
AQ	Light green	Light blue

1 PE: stands for petroleum ether, CH: stands for chloroform, METH: stands for methanol, ETH: stands for ethanol, AQ: stands for water.

Figure. 1: Comparison between extractive values using different solvent for Lepidium sativum and Nasturitium officinale plants

Table.5: Fluorescence analysis of Lepidium sativum dry powder plant:

Powder plant + reagent	Visible light	UV (254 nm)	UV (366 nm)
Powder + Xylene	Yellowish green	Violet	Greenish purple
Powder + Concentrated H2SO4	Brown	Dark grey	Green
Powder + 1 M NaOH	Yellowish green	Green	Grey
Powder + 10% HNO3	Very light blue	Light blue	Violet
Powder + 10% H2O2	Light yellow	Green	Light green
Powder + 10% K2Cr2O7	Orange	Brown	Reddish pink
Powder + I2	Light green	Greyish green	Light grey
Powder + Alcoholic FeCl3	Orange	Dark brown	Brown
Powder + Aqueous FeCl₃	Yellow	Dark green	Green
Powder + Concentrated HCl	Green	Light green	Fluorescent light green
Powder + Picric acid	Yellow	Green	Grey
Powder + Concentrated HNO₃	Light yellow brown	Light green	Light grey
Powder + Acetic acid	Yellow	Green	Light green

Table.6: Fluorescence analysis of Nasturtium officinale extracts:

Plant extract 1	UV light (254nm)	UV light (365 nm)
PE	Dark Blue	Light violet
CH	Violet	Light blue
METH	Dark green	Brown
ETH	Light green	Dark green
AQ	Dark green	Greenish black

1 PE: stands for petroleum ether, CH: stands for chloroform, METH: stands for methanol, ETH: stands for ethanol, AQ: stands for water.

Table.7: Fluorescence analysis of Nasturtium officinale dry powder plant:

Powder plant + reagent	Visible light	UV (254 nm)	UV (366 nm)
Powder + Xylene	Light green	violet	Light bluish purple
Powder + Concentrated H2SO4	Brown	Grey	Grayish blue
Powder + 1 M NaOH	Light yellow	Green	Light green
Powder + 10% HNO3	Light orange	Grey	Grayish blue
Powder + 10% H2O2	Very light green	Grey	Blue
Powder + 10% K2Cr2O7	Orange	Reddish brown	Brown
Powder + I2	Light yellow	Light blue	Blue
Powder + Alcoholic FeCl3	Orange	Brown	Light brown
Powder + Aqueous FeCl3	Yellow	Green	Greyish green
Powder + Concentrated HCl	Yellow	Green	Light green
Powder + Picric acid	Yellow	Olive green	Yellowish green
Powder + Conce. HNO3	Yellow	Light violet	Blue
Powder + Acetic acid	Yellowish brown	Green	Light brown (caramel)

Table. 8: Phytochemical screening of variant polarity solvent extracts of Lepidium sativum:

Phytochemical	Performed chemical test	Solvent extracts¹
Phytochemical	Performed chemical test	PE	CH	METH	ETH	AQ
Alkaloid	Dragendorf test	-	+	+	+	+
Anthraquinone glycoside	Borntragers test	-	-	-	-	-
Cardioactive glycoside	Keller kiliani test	-	-	-	-	-
Flavonoid	Alkaline test	-	-	+	+	+
Terpenoid	Salkowski test	-	-	-	-	-
Saponin	Forth test	-	-	-	-	-
Tannin	Braymer^,s test	-	-	-	-	-
Phlobatannin	Phlobatannin test	-	-	-	-	-
Phytosterol	Lieberman Burchard test	-	-	+	+	-
Phytosterol	H₂SO₄ test	-	-	+	+	-
Phenol	Lieberman test	-	-	+	+	+
Quinone	Quinone test	-	-	-	-	-
Glycoside	Fehling test	-	-	-	-	-
Carbohydrate	Molisch test	+	-	+	+	+

1 PE: stands for petroleum ether, CH: stands for chloroform, METH: stands for methanol, ETH: stands for ethanol, AQ: stands for water.

2 (-) stands for absence, (+) stands for presence.

Table. 9: Phytochemical screening of variant polarity solvent extracts of Nastrutium officinale:

Phytochemical	Performed chemical test	Solvent extracts¹
Phytochemical	Performed chemical test	PE	CH	METH	ETH	AQ
Alkaloid	Dragendorf test	+	+	+	+	+
Anthraquinone glycoside	Borntragers test	-	-	-	-	-
Cardioactive glycoside	Keller kiliani test	-	-	-	-	-
Flavonoid	Alkaline test	-	-	+	+	+
Terpenoid	Salkowski test	-	-	-	-	-
Saponin	Forth test	-	-	-	-	-
Tannin	Braymer^,s test	-	-	-	-	-
Phlobatannin	Phlobatannin test	-	-	-	-	-
Phytosterol	Lieberman Burchard test	+	+	+	+	-
Phytosterol	H₂SO₄ test	+	+	+	+	-
Phenol	Lieberman test	-	-	+	+	-
Quinone	Quinone test	-	-	-	-	-
Glycoside	Fehling test	-	-	-	-	-
Carbohydrate	Molisch test	-	-	+	+	+

1 PE: stands for petroleum ether, CH: stands for chloroform, METH: stands for methanol, ETH: stands for ethanol, AQ: stands for water.

2 (-) stands for absence, (+) stands for presence.

Figure. 3:Muller Hinton agar plate shows the antibacterial activity of methanolic extract of Nastrutium officinale against P. aeruginosa bacteria, A: stands for methanolic extract, B:stands for ethanolic extract, C: stands for positive control [azithromycine antibiotic], D: stands for negative control [DMSO]

Biological analysis:

Plant extracts with larger number of phytochemical constituents were introduced for biological analysis. All strains showed resistance for the extracts at tested extract concentration except P. aeruginosa exhibits sensitivity against methanolic extract (METH) of Nastrutium officinalis, results were shown in Table.10. expressed as inhibition zone ± SD measured in millimeter (mm) and (Figure.3.).

DISCUSSION:

Brassicaceae plant family include many important plant species were used as crop and vegetable, and considered as a source for a large number of bioactive compounds ²⁸. Two botanically related species Lepidium sativum and Nastrutium officinale of Brassicaceae plant family were chosen for comparison from different points of view as an interest to their traditional usage in the Kurdistan Region for both culinary and medicinal purposes. Evaluated physicochemical parameters showed that Lepidium sativum expressed significant higher percentage of foreign material and moisture content (p < 0.05) than it is corresponding related species Nasturtium officinale, while the later one showed significant larger total ash value (p < 0.05), [similar finding were recorded by E. D’agaro, 2006 ²⁹ and Pradhan et al, 2015 ³⁰]. Total ash value is an indication for the physiological matrix of plant and quantity of inorganic minerals ³¹, the results confirmed that the Nasturtium officinale had greater physiological matrix and mineral content than the compared one. An approximately duplicated value for water soluble ash value were estimated for Nasturtium officinale. Both species have exhibited similar value of acid soluble ash which reveals the lowest quantity of environmental contamination for the plants. Different solvent polarity extractive value have been estimated for both plants, results showed significant highest extractive value (p ˂ 0.0001) were obtained for both plants using water as solvent of extraction. Lepidium sativum exhibited an approximately similar extractive value of water and methanol solvent of extraction. Generally the extractive values obtained from Lepidium sativum plant was significantly greater (P < 0.0001) than Nasturtium officinale except petroleum ether extractive value for the later one was higher (P < 0.0001). High extractive values is an indication of absence of exhausted material in sample specially water solvent ³². Another physicochemical parameter used for evaluation of crude drugs is fluorescence analysis, different colors were obtained on addition of variant reagents for both plants, some reagents give similar colors or near colors such alcoholic ferric chloride, potassium dichromate which revealed the botanical relation of plant materials and similar constituents. The botanical relation further have been underlined from results of qualitative analysis of both plants. The qualitative phytochemical analysis of plant materials showed the presence of five out of thirteen screened phytochemicals in the solvents of ethanol and methanol, other solvent extracts exhibited the presence of some phytochemical constituents. Identical phytochemical constituents have been detected in both plants such as alkaloids, flavonoids ^3,33, phenols³, phytosterols ³⁴, and carbohydrates ^34,35, which confirmed the botanical relation of both species of Brassicaceae family. Quantitative phytochemical analysis showed variation in the concentration of the present phytochemical constituents Lepidium sativum showed significantly higher alkaloidal content (6.6 ± 0.001%) (P value < 0.0001) while Nasturtium officinale exhibited significant greater flavonoidal content (7.633±0.115%) (P value < 0.0001).

Biological analysis were based on phytochemical quality of the extracts. Extracts containing the greater number of phytochemical constituents were introduced for antibacterial evaluation against five standard bacterial strains, all bacterial species were showed resistance to plant extracts except P. aeruginosa expressed sensitivity against methanolic extract of Nasturtium officinale, antibacterial plant activity may be due higher flavonoidal content of the plant species in correspondence to it is related species (Lepidium sativum) since flavonoids were known for their antibacterial activity ³⁶.

CONCLUSION:

Present study was concluded that Nasturtium officinale was the biologically active species with greater physiological matrix can be used as source of new antimicrobial agents. In an attempt to emphasize the botanical relation of two Brassicaceae plant species we concluded that, Lepidium sativum and Nasturtium officinale showed similarity in qualitative phytochemical evaluation of crude drugs and fluorescence analysis (in some aspects), which highlighted the relation of two species botanically.

ACKNOWLEDGEMENTS:

The author is gratefully acknowledging the financial support of Pharmacognosy Department, Pharmacy College, Hawler Medical University.

REFERENCES:

1. Akinmoladun AC, et al. Pytochemical constituents and antioxidant activity of extract from the leaves of the Ocimum graticcimum. Science Research Essay. (2) ; 2007: 163-166.

2. Darshan S and Ved DK. A balanced perspective for management of Indian medicinal plants. Indian For. (129); 2003: 275-288.

3. Iqbal H, et al. Phytochemicals screening and antimicrobial activities of selected medicinal plants of Khyber pakhtunkhwa Pakistan. Journal of Pharmacy and Pharmacology. (6); 2011: 746-750.

4. Bigoniya P, Singh CS and Shukla A. Pharmacognostical and physicochemical standardization of ethnopharmacologically important seeds of Lepidium sativum Linn. and Wrightia tinctoria R. Indian journal of natural products and resources. 2(4); 2011: 464-471.

5. Dahanukar SA, Kulkarni RA and Rege NN. Pharmacology of medicinal plants and natural products. Indian Journals of Pharmacology. 32; 2000: 81-18.

6. Dragland S, et al. Several culinary and medicinal herbs are important sources ofdietary antioxidants. Nutrition. 133; 2003: 1286-1290.

7. Laredo JV, Agut M and Calvo-Torras MA. Antimicrobial activity of essences from Labiatae. International Microbiology. 82; 1995: 171–162.

8. Lemos TLG, et al. Antimicrobial activity of essential oils of Brazilian plants. Phytoptherapy Research. 4; 1990: 82-84.

9. Benoitvical F, et al. Antiplasmodial activity of Colchlospermum planchonii and C. tinctorium tubercle essential oils. Journal of Essential Oil Research. 13; 2001: 65-67.

10. Warwick SI. Brassicaceae in Agriculture. In Genetics and Genomics of the Brassicaceae, Plant 33 Genetics and Genomics: Crops and Models 9. Edited by Schmidt R and Bancroft I. Springer Science+Business Media LLC, New York. 2011. pp. 34-35.

11. Rahul KS, Kapil V and Hansraj M. Evaluation of antifungal effect of ethanolic extract of Lepidium sativum seed. International Journal of Phytopharmacology. 3(2); 2012: 117-120.

12. Washington Department of Ecology. Aquatic Plant Identification: Manual for Washington’s Freshwater Plants, Nasturtium officinale, watercress. Retrieved 2009.

13. Anonymous. US Department of Agriculture, Agricultural Research Service. USDA National Nutrient Database for Standard Reference, Release 16; 2003.

14. Miraldi E, Ferri S and Mostaghimi V. Botanical drugs and preparations in the traditional medicine of west Azarbaijan (Iran)'. Journal Ethnopharmacology. 75; 2001:77-87.

15. Ozen T. Investigation of antioxidant properties of Nasturtium officinale (Watercress) leaf extracts. Acta Poloniae Pharmaceutica Drug Research. 66 (2); 2009: 187-93.

16. Tamayo C, et al. The chemistry and biological activity of herbs used in Flor-Essence herbal tonic and Essiac. Phytotherapy Research. 14; 2000: 1-4.

17. Sezik E, et al. Composition and antimicrobial activity of Juniperus excelsa essential oil. Journal of Ethnopharmacology. 75; 2000:115-195.

18. 18.Govt. of India. Ministry of Health and Family Welfare India Pharmacopeia, Controller of Publications. New Delhi; 1996: A53-55.

19. WHO. Quality Control Methods for Medicinal Plant Materials. Geneva: 1998. p. 10-31.

20. Gupta MK, Sharma PK, Ansari SH and Lagarkha R. Pharmacognostical evaluation of Grewia asiatica fruits. International Journal of Plant Science. 1 (2); 2006: 249-251.

21. Kokashi CJ, Kokashi RJ and Sharma M. Fluorescence of powdered vegetable drugs in ultra- violet radiation. Journal of American Pharmacy Association. 47; 1958: 715-717.

22. Kokate CK. Practical Pharmacognosy, 4th ed. Vallabh Prakashan, Delhi. 1997. pp. 107 -111.

23. Khandelwal KR. Practical Pharmacognosy Techniques and Experiments, 15th ed. Nirali Prakashan, Pune. 2006. pp.150–163.

24. Krishnaiah D, Devi T, Bono A and Sarbatly R. Studies on phytochemical constituents of six Malaysian medicinal plants. Journal of Medicinal Plants Research. 3(2); 2009: 67-72.

25. Alupuli A, Calinescu I and Lavric V. Ultrasonic vs. microwave extraction intensification of active principles from medicinal plants. AIDIC Conference Series. 9; 2009: 1-8.

26. Stainer RY, Ingraham JL and Wheelis ML. General Microbiology, 5th edition. The McMillan Press Ltd, London. 1986.

27. Sandhya VR and Biradar SD. Preliminary phytochemical screening and antimicrobial activity of Lagenaria siceraria (mol). Indian journal plant science. 2(1); 2012: 126-130.

28. Shi ZHK. Brassicaceae (Cruciferae). Flora of China. 8; 2001: 1–193.

29. D’agaro E. Utilization of water cress (Nasturtium officinale ) in noble crayfish (astacus astacus) feeding. Bull. Fr Pêche Piscic. 380-381; 2006: 1255-1260.

30. Pradhan S, Manivannan S and Tamang JP. Proximate, mineral composition and antioxidant properties of some wild leafy vegetable. Journal of Scientific and Industrial Research. 74; 2015: 155-159.

31. Yulan R and Bingren X. Determination of total ash and acid insoluble ash of Chinese herbal medicine Prunellae spica by Near Infrared spectroscopy. The Pharmaceutical Society of Japan. 129 (7); 2009: 881-889.

32. Harinarayan SC, Pathak AK and Mukul T. Standardization of some herbal anti-diabetic drugs in polyherbal formulation. Pharmacognosy Research. 3(1) ; 2011:49–56.

33. Gerard LP and Celia PM. Antibacterial activity of extracts of twelve common medicinal plants from the Philippines. Journal of Medicinal Plant Research. 5(16); 2011: 3975-3981.

34. Billore KV, et al. Database on Medicinal Plants used in Ayurveda; Vol.7. Central Council for Research in Ayurveda and Siddha, New Delhi. 2005. pp. 52.

35. McCance RA. McCance and Widdowson’s: The Composition of Foods, 6th summary edition. RSC, Cambridge. 2002.

36. Cowan M. Plant products as antimicrobial agents. Clinical Microbiology Review. 12; 1999: 564.

Received on 22.07.2015 Accepted on 09.09.2015

Asian J. Res. Pharm. Sci. 5(3): July-Sept.; Page 168-174

DOI: 10.5958/2231-5659.2015.00025.9

Bacterial species	*Lepidium* *sativum* Inhibition zone (mm)		*Nastrutium* *officinale* Inhibition zone¹ (mm)		Azithromycin Inhibition zone (mm)¹
Bacterial species	ETH^a	METH^b	ETH	METH	Azithromycin Inhibition zone (mm)¹
P. aeruginosa(ATCC27853)	R³	R	R	25 ± 0.001	40 ± 0.1
E. coli (ATCC 35218)	R	R	R	R	32 ± 0.001
S. aureus(ATCC 25923)	R	R	R	R	40 ±0.0577
S. pyogenes (ATCC 19615)	R	R	R	R	20 ±0.1527
S.pneumoniae (ATCC6303)	R	R	R	R	30 ±0.57