Eight species Convunvulus prostrates, Portulaca quadrifida, Taraxacum officinale Albizia lebbeck, Olea ferruginea, Salvadora oleoides, Suaeda fruticosa and Vitex negundo were analyzed for macro and micro minerals in three phenological stages i-e Pre-reproductive, reproductive and post reproductive stages which showed that Ca, Al, P, N, S, Na, K, Mg were macro and Fe, Si, Cu and Cl were micro-nutrients. Nutritional analysis showed that moisture, ash contents, crude protein, crude fiber, crude fat and carbohydrate contents are non significant at three phenological stages of herbs and woody species.

Determination of range animal productivity depends upon the amount and nutritive quality of vegetation available to grazing animal. Age and their physiological function like gestation, growth maintenance, fattening, location and determination of the nutritional demands of livestock. Plant material is divided into fibrous and non fibrous contents. Due to overstocking most rangelands of Pakistan may have sufficient forage but of low palatability.¹ Proteins are fundamental components of all living cells because it is building unit of enzymes, hormones, and antibodies which are necessary for the proper functioning of an organism. For growth and repair of tissue proteins are essential in the diet of animals. All the nitrogenous compounds present in forage feed is reliable source of overall nutritional status referred as crude protein. They are directly related to digestibility, calcium, vitamins and phosphorus contents.^2,3 Macronutrients are important constituents of body fluids as electrolytes in order to protect and maintain the structural components of the body organs and tissues. In growth, reproduction, health and proper functioning of the animal's body minerals play a vital role. About 30 million herds of livestock support by rangelands, which play an important role in Pakistan annual export income.⁴ Visible symptom of Al toxicity is Inhibition growth of root and shoot. The earliest symptoms appear on roots. Shoots with Al observed are less affected for Mn toxicity.⁵ Root with a consequence of Al-induce the elongation of root. Roots are usually become strong and brittle and tips of root and lateral roots become thick and turn to brown.⁶ Al does not affect the seed germination, but impair the growth of new roots and establishment of seedling.⁷ The common responses of shoots to Al are: ultrastructural and cellular changes in leaves, increased resistance in rates of diffusion, stomatal aperture reduction, chlorosis and necrosis of leaves, total decrease in size and number of leaf and shoot biomass decrease.⁸ Sulphur is an important element, which take active part in protein forming nutrients. Sulphur deficiency can also result in Nitrogen deficiency. Due to S deficiency cereals and forage grasses, yellowing of newly emerging leaves occur. S deficiency also leads to cupping and purpling of leaves.⁹ Chlorine is an important element frequently accumulating in undesirable quantities, particularly in semiarid regions so its absence to the seas along with other salts causes a problem. However, in agricultural areas, chlorine is a useful element to crops because of less supply from natural sources. The nutritional disease due to chlorine deficiency yellowing of the leaves chlorosis and finally death necrosis of leaf tissue occur. Growth was exceedingly restricted due to chlorine deficiency and plants fails to set fruit.¹⁰ Plants typically absorb bio available silicon in the form of silicate known as monosilicic or ortho silicic acid. Silicon in plants can stimulate plant photosynthesis, nutrient uptake, decrease susceptibility to disease and insect damage, alleviate water and various mineral stresses and also decrease the toxic effects of aluminium. “Silicon is taken up by plants as silic acid through the root system and moves upwards in the transpiration stream and then move to sites of strong evapo- transpiration where it transformed into insoluble polymers”.¹¹

Chemical analysis of some forage plants

Eight palatable plants species were collected from the research area. For mineral and proximate analysis these plants were dried, powdered and stored in plastic bags.

Mineral composition

Plant samples were dried at 70°C in air tight oven for 48 hour following method of AOAC.¹² For mineral composition of Ca, Mg, Fe, Mn, Zn, Cu, etc the powdered plant materials were analyzed by using Spectrometer of Atomic Absorption.^13–15

Statistical analysis

Statistically t-test was applied for chemical contents comparison of herbaceous and woody plant species.

Proximate analysis

Determination of the moisture

Equipment and glassware: Electric balance, Electric oven, Petri dish and desiccators

Procedure: In a clean weighted Petridish about 2gram of respective eight plants samples were taken respectively (W1). The Petri dishes were partially covered with lid, placed in electric oven at temperature of 105°C for 4-6hours, and was then transferred these petri dishes to desiccators for 30 minutes to cool down; after that, weighted again (W2) of these Petri dishes. The following formula for calculating percent moisture contents was as follow.¹⁶

% Moisture = X/wt of sample ×100

Where
X=W₂ - W₁= Weight of the sample (after heating)

W₂=Wt of empty Petri dish+after heating of sample

W₁=Empty Petri dish Wt

Ash Contents: At 550°C- 600°C for 8 hrs in the muffle furnace one to two grams of plant sample was ignited and ash contents of samples were determined by following method AOAC.¹² Ash contents percentage were calculated by following formula:

%Ash Content=Wt of ashWt of fresh Sample×100

Nitrogen/Crude Protein

Determination of proteins by “Macrojeldahl distillation method”

Reagents: 32% NaOH, Conc. H₂SO₄, 4% Boric Acid, K2SO4, CuSO4 and 0.1 N standard HCl solution.

Mixed indicator: Dissolve 0.016g of methyl red and 0.03g of bromocresol green in 100 ml of alcohol.

Apparatus: Kjeldahl flask, apparatus of digestion and distillation, burette etc

Digestion of Plant: Determination of all nutrients involved wet digestion of plant samples. One gm plant sample add in concentrated selenium sulphuric acid and hydrogen peroxide was added to each digestion tube for digestion. On heating blocks these digestion tubes with sample was heated. In order to remove the color digestion was continued at 350°C. Then these prepared solutions were diluted with distilled water and stored in tubes. These solutions were used for the analysis of nitrogen /crude protein, crude fiber etc by using following methods.

Procedure: By Macro kjeldahl method Protein (% Nx6.25) was determined. One gram of dry ground plants samples were taken in digestion flask repectively. Digestion mixture (Cu SO4, K2SO4 and ferrous sulphate in the ratio of 5, 94 and 1 respectively) then added 25 ml of conc Sulphuric acid to the flask and digested in digestion flask (kjeldatherm) for 6 hours. Then the flask was removed, cooled and then transferred to 250 ml flask. Distilled water was added in order to make the volume level to 50 ml of the above solution. Strong alkali 10 ml was added to make it alkaline and then added 50ml of 4% Boric Acid solution. Then transferred it to the distillation flask and mixed 3-5 drops of indicator. Then 50 ml water and 60 ml of 32% NaOH solution were added to it. After distillation, for titration it was then collected in flask. Add 0.1 N HCl in burette to the content of the flask. Noted the reading and the percentage of protein was determined using the following formula.¹⁵

(N%)=(V1−V2)×14.01×0.5×100(sample in mg)

 V₁= Reading of sample after titration

V₂= Reading of blank after titration

14.01= Nitrogen Atomic weight (N)

Contents of crude protein (%) were calculated for all the plant samples by multiplying the nitrogen content of the sample by 6.25

Protein (%) = Percent of Nitrogen × 6.25.

Crude Fiber

Fat determination (ether extract)

Chemicals, Equipment and glassware: H.T (Tecator), Petroleum ether B.P (40-60°C), Soxhlet extraction apparatus, Extraction thimbles, water bath, heating mantle.

Procedure: For the extraction of crude Fat Soxhlet apparatus was used¹⁷ 2gram of each plant sample was packed in filter paper (cellulose extraction thimble) and placed in apparatus of extraction chamber. A clean and dried pre weighted round bottom flask of 250ml filled with Petroleum ether and connected to the extraction tube containing thimble. The apparatus of soxhlet run for 5-6hours. The solvent extract in the round bottom flask was evaporated by using water bath and then reweighted (W2). Percentage of fats was then calculated by the following formula.¹⁵

% Crude fibers=XWt of Sample×100

 X = W2 - W1= Wt of the fats

W1 = Empty flask Wt

W2 = Empty flask Wt + sample Wt after solvent evaporation.

Determination of crude fiber

Glassware and Equipment: Muffle furnace, apparatus of crude fiber extraction (Fiber Tec System M. Tecator), Suction pump and oven.

Reagents: Sulphuric Acid (H₂SO₄) 0.255N, Sodium Hydroxide (NaOH) 0.313 N, Asbestos, Ethyl Alcohol and Petroleum Ether.

Procedure: 2gms of these residue materials remaining from crude fat were transferred to digestion flask along with 0.5g asbestos and about 200 ml boiling 0.255 N, H₂SO₄ was added. For 30minutes the flask was connected to the condenser and boiled. These contents were filtered through linen cloth in fluted funnel. Residues were washed to remove the acids and then transferred again to digestion flask and boiled with 0.313N of NaOH. Addition of NaOH was continued till the volume reached to 200ml. For 30minutes the flask was then connected to the reflux condenser and boiled. This hot residue was then filtered separately through Gooch crucible prepared with asbestos mat. Residue was then thoroughly washed with boiling water followed by Ethyl Alcohol (15ml). The residue was transferred to crucible and dried at 110°C in hot air oven (W₁). These crucibles were then transferred to the muffle furnace, ignited till it converted into white grey powder (W₂). Crude fibers were then calculated by following formula.¹⁵

% Crude fibers=W2−W1Wt of sample×100

Carbohydrates contents: Carbohydrates contents were calculated by subtracting the sum of the weights of proteins, fat, crude fibers, ash, and moisture contents from 100.

100 – (Protein +fats +crude fiber +ash + moisture contents) = % Carbohydrate

Elemental analysis

The present data showed that at three phonological stages Convunvulus prostratesL. (0.91ppm), Portulaca quadrifidaL.,(0.19ppm), Taraxacum officinale (0.40ppm) while woody plants like Albizia lebbeck L. (0.46ppm), Olea ferruginea Royle (0.45ppm), Salvadora oleoides(1.15ppm), Suaeda fruticosa Forssk. (0.56ppm) and Vitex negundo L. was 0.31ppm. Convunvulus prostrates L. (0.05 ppm), Portulaca quadrifidaL.,(0.90 ppm), Taraxacum officinale (0.84 ppm) while woody plants like Albizia lebbeck L. (0.11ppm), Olea ferruginea Royle (0.45ppm), Salvadora oleoides(0.04 ppm), Suaeda fruticosa Forssk. (0.63ppm) and Vitex negundo L. was 0.18ppm (Table 1 and 2). Portulaca quadrifolia L.,(0.30ppm), Taraxacum officinale (0.21ppm) while woody plants like Suaeda fruticosa (0.40ppm) and Vitex negundo L. was 0.34ppm. The average P contents in herbs and woody plants 0.01ppm (Convunvulus prostrates), 0.05ppm (Portulaca quadrifida), 0.20ppm(Taraxacum officinale)while woody plants like Albizia lebbeck (0.05ppm), Olea ferruginea (0.01ppm), Salvadora oleoides(0.02ppm), Suaeda fruticosa (0.03ppm) and Vitex negundo was 0.03ppm. The average Nitrogen content 2.21ppm (Convunvulus prostrates), 5.19ppm (Portulaca quadrifida), 3.93ppm(Taraxacum officinale)while woody plants like Albizia lebbeck (3.82ppm), Olea ferruginea (3.72ppm), Salvadora oleoides(5.95ppm), Suaeda fruticosa(4.11ppm) and Vitex negundo was 2.67ppm (Table 1 and 2). Mg content 0.09ppm (Convunvulus prostrates), 0.34ppm (Portulaca quadrifida), 0.18ppm(Taraxacum officinale)while woody plants like Albizia lebbeck (0.20ppm), Olea ferruginea (0.18ppm), Salvadora oleoides(0.23ppm), Suaeda fruticosa(0.60ppm) and Vitex negundo was 0.13ppm. Mg content 0.09ppm (Convunvulus prostrates), 0.34ppm (Portulaca quadrifida), 0.18ppm(Taraxacum officinale)while woody plants like Albizia lebbeck (0.20ppm), Olea ferruginea (0.18ppm), Salvadora oleoides(0.23ppm), Suaeda fruticosa(0.60ppm) and Vitex negundo was 0.13ppm. Al content in tested plants 0.1ppm (Convunvulus prostrates), 0.09ppm (Portulaca quadrifida), 0.19ppm(Taraxacum officinale)while woody plants like Albizia lebbeck (0.06ppm), Olea ferruginea (0.10ppm), Salvadora oleoides(0.09ppm), Suaeda fruticosa(0.07ppm) and Vitex negundo was 0.17ppm.Sulphur content in tested species were 0.05ppm (Convunvulus prostrates), 0.06ppm (Portulaca quadrifida), 0.38ppm(Taraxacum officinale)while woody plants like Albizia lebbeck (0.06ppm), Olea ferruginea (0.1ppm), Salvadora oleoides(0.44ppm), Suaeda fruticosa(1.14ppm) while in Vitex negundo 0.07ppm. Chlorine is present in 0.07ppm (Convunvulus prostrates), 0.1ppm (Portulaca quadrifida), 0.35ppm(Taraxacum officinale)while woody plants like Albizia lebbeck (0.1ppm), Olea ferruginea (0.04ppm), Salvadora oleoides(0.86ppm), Suaeda fruticosa(0.21ppm) while in Vitex negundo chlorine was absent. Si contents in tested plants 0.15% (Convunvulus prostrates), 0.20% (Portulaca quadrifida), 0.59%(Taraxacum officinale)while woody plants like Albizia lebbeck (0.18%), Olea ferruginea (0.26%), Salvadora oleoides(0.18ppm), Suaeda fruticosa(0.14ppm) while in Vitex negundo was 0.54ppm. Recent results showed that 0.01% (Convunvulus prostrates) at pre reproductive stage, Portulaca quadrifida and(Taraxacum officinale have no copperwhile woody plants like Albizia lebbeck (0.1ppm), Olea ferruginea (0.13ppm), Salvadora oleoides(0.05ppm), Suaeda fruticosa(0.06ppm) while in Vitex negundo 0.12ppm. Bahadur et al.¹⁸ evaluate the elemental analysis of some fodder plant species like Amaranthes viridus, Chenopodium album, Medicago denticulata, Setaria viridus and Sonchus arvrnsis. Elemental composition of aerial parts was determined by using Atomic Absorption Spectrophotometer (AAS). A total of 16 elements; Na, Mg, Rb, Al, Si, P, S, K, Ca, Fe, Ti, Ni, Cl, Mn, Cu and Zn were observed. Tahira et al.¹⁹ evaluate the elemental values from aerial part of five medicinal plants Convolvulus arvensis L., Rumex dentatus L., Physalis divaricata D. Don, Achyranthes asperaL. and Chenopodium ambrosiodes L., of District Swabi Khyber Pakhtoon Khwa, Pakistan. Elements were determined by Atomic Absorption Spectrophotometer (AAS); a total 10 elements five micro and five macro elements like Na, Al, Fe, Mn, Zn and K, P, Mg, Ca and S were measured. Ghani et al.²⁰ evaluate mineral contents of four medicinal plants like Achryanthus aspera, Solanum nigrum, Peganum hermlaand Mentha longifolia of Khushab Valley,Pakistan. For elemental analysis AbsorptionSpectrometric method was used for essential elements such as Cu, Na, Fe, Cd, Mn, Ni, Pb and Crwere present in medicinal plants(Figures 1-8).

Nutrinional analysis

Moisture content showed that herbs and woody plants have 53.46% (Convunvulus prostrates), 37.83% (Portulaca quadrifida), 73.63%(Taraxacum officinale)while woody plants like Albizia lebbeck (58%), Olea ferruginea (57.2%), Salvadora oleoides(61.6%), Suaeda fruticosa (42.63%) and in Vitex negundo was 47.2%. ash contents of herbs and woody plants were 10.16% (Convunvulus prostrates), 8.2% (Portulaca quadrifida), 7.5%(Taraxacum officinale)while woody plants like Albizia lebbeck (10.4%), Olea ferruginea (9.3%), Salvadora oleoides(9.56%), Suaeda fruticosa(8.26%) while in Vitex negundo 08%. crude proteins in herbs and woody species were 6.9% (Convunvulus prostrates), 6.4% (Portulaca quadrifida), 10.26%(Taraxacum officinale)while woody plants like Albizia lebbeck (8.23%), Olea ferruginea (8.5%), Salvadora oleoides(9.6%), Suaeda fruticosa(10.86%) while in Vitex negundo 11.16%. crude fiber of herbs and woody species were 4.06% (Convunvulus prostrates), 12.16% (Portulaca quadrifida), 11.2%(Taraxacum officinale)while woody plants like Albizia lebbeck (12.4%), Olea ferruginea (6.93%), Salvadora oleoides(8.13%), Suaeda fruticosa(11%) while in Vitex negundo high crude fiber 21.13%. high fat contents were present in woody plants in which Olea ferruginea with 20.1%. Fat content of herbs showed that 5.13% (Convunvulus prostrates), 0.83% (Portulaca quadrifida), 10.16%(Taraxacum officinale) while woody species like Albizia lebbeck (8.13%), Salvadora oleoides (10.7%), Suaeda fruticosa (12.03%) and Vitex negundo with 12.46%. showed that 79% (Convunvulus prostrates), 73.23% (Portulaca quadrifida), 71.03%(Taraxacum officinale)while woody plants like Albizia lebbeck (68.96%), Olea ferruginea (75.2%), Salvadora oleoides(72.63%), Suaeda fruticosa(69.86%) and Vitex negundo 59.83% (Table 3 and 4). Bahadur et al.¹⁸ evaluate the nutritional analysis of some fodder plant species like Amaranthes viridus, Chenopodium album, Medicago denticulata, Setaria viridus and Sonchus arvrnsis. Proximate composition of crude fibers, proteins, fats and oils, ash, moisture and carbohydrates contents of aerial parts was determined by using Atomic Absorption Spectrophotometer (AAS). Tahira et al.¹⁹ evaluate the nutritional values from aerial part of five medicinal plants Convolvulus arvensis L., Rumex dentatus L., Physalis divaricata D. Don, Achyranthes asperaL. and Chenopodium ambrosiodes L., of District Swabi Khyber Pakhtoon Khwa, Pakistan. Nutritional analysis like total ash, crude protein, crude fiber, nitrogen free extracts, acid detergent fiber, neutral detergent fiber, hemi-cellulose, carbohydrate and moisture contents of wild medicinal plant species were determined and showed significant results. Ghani et al.²⁰ evaluate nutritional contents of four medicinal plants like Achryanthus aspera, Solanum nigrum, Peganum hermlaand Mentha longifolia of Khushab Valley,Pakistan. Proximate analysis showed that in Mentha longifolia protein (7.491%), ash (22.79%) was highestand in Peganum hermla, fats (12.595%) carbohydrate (75.23%) and in Achryanthus aspera moisture (6.82%) was present.

Figure 1 Portulaca olaraceae.

Figure 2 Albezia lebbek.

Figure 3 Taraxicum officinale.

Figure 4 Vitex nugundu

Figure 5 Olea ferruginea

Figure 6 Convulus prostrate.

Figure 7 Suaeda fruticosa

Figure 8 Salvadora oleoides

None.

The author declares no conflict of interest.

1. Hussain F, Durrani MJ. Seasonal availability, palatability and animal preferences of forage plants in Harboi arid range land. Pak J Bot. 2009;41(2):539–554.
2. Ganskopp D, Bohnert D. Mineral concentration dynamics among 7 northern Great Basin grasses. J Range Management. 2003;56:174–184.
3. Anonymous. Economic Survey. Government of Pakistan, Finance Division, Islamabad, Pakistan; 2006.
4. Chang YC, Yamamoto Y, Matsumoto H. Accumulation of aluminium in the cell wall pectin in cultured tobacco (Nicotiana tabacum L.) cells treated with a combination of aluminium and iron. Plant Cell Environ. 1999;22:1009–1017.
5. Mossor–Pietraszewska T, Kwit M, £êgiewicz M. The influence of aluminium ions on activity changes of some dehydrogenases and aminotransferases in yellow lupine. Biol Bull Poznañ. 1997;34:47–48.
6. Nosko P, Brassard P, Krame JR, et al. The effect of aluminum on seed germination and early seedling establishment, growth and respiration of white spruce (Picea glauca). Can J Bot. 1988;66(11):2305–2310.
7. Thornton FC, Schaedle M, Raynal DL. Effect of aluminum on the growth of sugar maple in solution culture. Can J For Res. 1986;16(5):892–896.
8. Sahota TS. Importance of Sulphur in crop production. Ontario Farmer. 2005;38(42):25.
9. Perry R Stout, Johnson CM, Broyer TC. Chlorine in Plant Nutrition experiments with plants in nutrient solutions establish chlorine as a micronutrient essential to plant growth. California agriculture; 1956. 10 p.
10. Smith A, Nutrifert. Silicon’s key role in plant growth. Australian Grainp; 2011. 35 p.
11. AOAC. Official Methods of Analysis. 15th ed. Washington, DC, USA: Association of Official Analytical Chemists; 1990;5:3–10.
12. Anonymous. Manual for Feed Analytical Laboratory. Islamabad, Pakistan: PARC; 1982.
13. Anonymous. Nutrient requirements of domestic animals. No5. Nutrient requirements of sheep. Washington DC, USA: Nat Acad Sci Nat Res Council; 1985.
14. Galyean M. Techniques and Procedures in Animal Nutrition Research. Department Animal & Range Conditions: New Mexico State University; 1985.
15. AOAC (Association of Official Analytical Chemists). Official methods of analysis. Gaithersburg, MD, Washington, USA; 2000.
16. AOAC. Official Methods of Analysis. 11th ed. Washington, DC, USA: Association of Official Analytical Chemists; 1984.
17. Zarnowski R, Suzuki Y. Expedient Soxhlet extraction of resorcinolic lipids from wheat grains. Jour Food Comp & Anal. 2004;17(5):649–663.
18. Bahadur A, Chaudhry Z, Jan G, et al. Nutritional and elemental analyses of some selected fodder species used in traditional medicine. African Journal of Pharmacy and Pharmacology. 2011;5(8):1157–1161.
19. Tahira M, Bahadur A, Shah Z, et al. Elemental and Nutritional analysis and Ethnomedicinal study of selected wild plants species of district Swabi. Journal of Pharmacy Research. 2012;5(9):4910–4913.
20. Ghani A, Ali Z, Islam T, et al. Nutrient evaluation and elemental analysis of four selected medicinal plants of soon valley Khushab. Pak J Pharm Sci. 2014;27(3):597–600.