International Research Journal of Environment Sciences________________________________ ISSN 2319–1414Vol. 1(5), 58-61, December (2012) Int. Res. J. Environment Sci. International Science Congress Association 58 Chemical Estimation of Air Pollutants and Its Impact on the Flavonoid Content of Adhatoda vasica, ocimum sanctum; and Aloe VeraSharma Manju and PandeyAnish Chandra Department of P.G. Studies and research in Chemistry, Government SLP P.G. College (Jiwaji University), Gwalior, INDIAAvailable online at: www.isca.in Received 22nd October 2012, revised 30 October 2012, accepted 8 November 2012 Abstract The present investigation was carried out to establish a correlation environmental pollution especially the SO, Nox, RSPM, SPM and O3 pollution with plant chemistry. In this present study chemically estimate SO, No, RSPM, SPMand O3 at SLP govt. PG College, Maharaj bada and Deendayal Nagar. Samples of three medicinal plantsAdhatoda vasica, ocimum sanctumand aloe Vera were analyzed for their phytochemical composition, flavonoid contents the result revealed the presence of bioactive constituents Flavonoid. The result of this study show that the Flavonoid content in Adhatoda vasica and aloe veraplant decreases with increases pollutant and ocimum sanctum show positive correlation with pollutants. Keywords: ozone(0), sulphur dioxide(SO), oxides of nitrogen (No), respirable suspended particulate matter(RSPM), suspended particulate matter ( SPM), flavonoid content, adhatoda vasica, ocimum sanctum and Aloe Vera.Introduction Environmental pollution and its impact on plant have well recognized. The role of air pollutants causing injury to plants either by direct toxic effect or modifying the host physiology. The environment defined as to include water, air and land, the interrelationship which exists among and between water, air and land, and human being, plant, other living creatures, micro-organisms. Environment is continuously polluted due to increasing the concentration of NO, SO, SPM, and O. Pollution means the presence of undesirable substances in any segment of environment, primary due to human activity discharging by products, waste product or harmful secondary products, which are harmful to man, vegetation or other organism. Ozone in the lower atmosphere is a highly reactive secondary pollutant photo chemically formed in the presence of primary pollutants like NO, SO, Suspended particulate matter (SPMandPM), HC and CO etc, which are (NO, SO, O and SPM) the major constituents of automobile exhaust. Atmosphere plays a significant role in global processes supporting life on earth. It serves as an efficient heat reservoir. Troposphere is a zone closest to earth. It is an important zone for living world. Stratosphere is next zone to the troposphere. SO, NO, SPM, and Ozone are present in the troposphere and stratosphere. While ozone in the presence of stratosphere protects the entire biosphere from the lethal ultraviolet radiation by absorption it in the troposphere it acts as a toxic green house gas by an absorbing terrestrial radiation5 and contribute to the global warming. Highly reactive Ozone binds to plasma membrane, alters metabolism and inhibited stomatal photosynthesis. Ozone reacts with O and produces reactive oxygen species, including hydrogen peroxide (H), Super oxide (O), Singlet oxygen, and the hydroxyl radical. These denature proteins and nuclic acid, and cause lipid per oxidation, which break down lipids in membrane. Material and Method Plant material: Samples of Adhatoda vasica, Ocimum Sanctumand Aloe Vera were collected from SLP PG college morar, Deen Dayal Nagar and Maharaj bada sampling sites. Test for flavonoids: A portion of crude powder was heated with 10 ml of ethyl acetate over a steam bath for 30 min. The mixture was filtered and 4 ml of the filtrate was shaken with 1 ml of dilute ammonia solution and observed a yellow coloration. Estimation of Pollutants: In this study Sulphur dioxide, Nitrogen dioxide, Suspended particulate matter, Respirable suspended particulate matter and surface ozone concentration along with air temperature, relative humidity, wind speed, wind direction and traffic density has been monitored in Gwalior at three sites. SO and NOx estimation has been done spectrophotometrically by Systronic 108 UV visible sppectrophotometer. West-Gaeke and Jacob- Hocheiser(1958)method has been used for the determination of SO and NOx respectively.GF/A whatman’s filter paper No. 1 used to collect Suspended particulate matter. Ozone collected by bubbling through borosilicate glass impingers containing 1% KI solution (pH 7.0). Ozone estimation was done colorimetrically by the method of Byers Saltzman8 with the modification suggested by Boyd et al.,. Ozone concentrations have been expressed as ppbv. It’s determined colorimetrically at 352nm. Extraction of flavonoids: Accurate 10 g of raw plant material was extracted with 25 ml of 95% methanol under 200 rpm shaking for 24 hour. After filtration, the filtrate was adjusted to International Research Journal of Environment Sciences______________________________________________ ISSN 2319–1414 Vol. 1(5), 58-61, December (2012)Int. Res. J. Environment Sci. International Science Congress Association 59 25 ml with 80% methanol and stored in an amber bottle at 4C. The flavonoids constituents extracted are quantified spectrophotometric method. Quantification of total flavonoids content in plants by Spectroscopic method: Total flavonoids content was determined as per the method described by Chang et.al. (2002)10. each plant extracts (0.5 ml of 1:10g/l) of each plant in methanol was separated mixed with 1.5 ml of methanol,0.1 ml of 10% aluminium chloride,0.1 M potassium acetate and 2.8 ml of distilled water. After keeping the mixture at the room temperature for 30 minutes, the absorbance of the reaction mixture was measured at 415nm with a double beam UV/Visible Spectrophotometer. The calibration curve was prepared by using quercetin and rutin as a standard solution. Results and Discussion The average data of ambient air quality with lower and upper limits are given in table 1 and results are shown figure 4.a to 4.c. In this present study the lower and upper limits of air pollutants at different locations were observed for RSPM (10.2g/m825g/m), SPM (26g/m- 956g/m), SO (1.9g/m-33 g/m), NO (3.2g/m- 46g/m) and (0 ppbv – 22 ppbv). The level of RSPM was maximum (825 g/m) at Maharaj bada location during August, which is more than four times of CPCB level (200 g/m) prescribed for residential area. The lowest level of RSPM was recorded at SLP in August (10.2g/m). Similarly, highest (956 g/m) level of SPM was recorded at Deen Dayal Nagar during August and lower for SLP 26 g/m3 in August month. The NO2 level at Maharaj bada was highest (46 g/m) in April month, while the lowest (3.2g/m) level was recorded in August at SLP College. The level of SO was found to be lower than NO at all study locations through out the year. SO was highest (33g/m) for Maharaj bada during November while lowest (1.9g/m) at SLP during August Month. Ozone level was highest (22g/m) in April at Maharaj bada, while lowest (0g/m) for SLP college in January and August months. From the average data of ambient air quality concluded that Maharaj bada sampling site is more polluted due to SO, NOand O at all the seasons and SLP college site is less polluted. Deen Dayal Nagar is highly polluted due to total Suspended particulate matter. Table-1 Ambient air pollutants at different location during different representative months from 2008-2011 Sampling sites January April August November Range Mean Range Mean Range Mean Range Mean SO (µg/m 3 ) a.Maharaj bada b.Deendayal Nager c.SLP College 4.5-20.41 4 – 16.6 2.1-11.7 12.50 9.297 5.471 5 – 32.7 4 – 17 3– 14.78 12.12 10.357 7.9245 4.5– 25.25 4.0 -17 1.9– 14.25 11.462 10.12 7.009 5 - 33 5 - 25 3- 12.12 12.578 10.918 6.2445 NOx(µg/m 3 ) a.Maharaj bada b.Deendayal Nager c.SLP College 5.9– 35.7 8.3– 35.7 4.0– 18.46 17.4236 18.163 9.078 7.1-46 7.9 -36.6 5 - 20.34 19.050 20.983 11.6345 6.8-32.7 7.1 -37 3.2-20.1 18.211 18.875 10.518 7.1 -39 9.5-35.7 5.2-18.79 18.574 19.025 11.5035 (ppbv) a.Maharaj bada b.Deendayal Nager c.SLP College 1.03-8.02 0.64 -8.0 0 -2.32 7.84 6.77 1.39 4.01 -22 2.96 -18 0.32 -5.95 19.86 16.63 5.41 0 -6.95 0 -6.28 0 -1.6 6.21 4.99 0.84 1.67 -10 1.02-12.3 0.84-4.91 9.3 9.576 3.07 RSPM (µg/m 3 ) a.Maharaj bada b.Deendayal Nager c.SLP College 48 -664 62 -602 52 - 380 233.86 265.64 207.2875 59 -669 73 -605 43-395.72 237.643 293.714 206.1617 21 -825 12 -695 10.2-296.3 224.38 279.52 134.02 53 -597 42 -624 24-276.3 186.689 270.172 161.776 SPM (µg/m 3 ) a.Maharaj bada b.Deendayal Nager c.SLP College 127 -754 123 -782 96 - 402 352.65 371.71 288.6333 126 -736 130 -750 114-408.23 341.671 411.142 292.5233 63 -824 43 -956 26-280.3 294.72 376.67 177.4192 120 -809 98 -680 89.3-378.2 306.47 394.716 230.766 International Research Journal of Environment Sciences______________________________________________ ISSN 2319–1414 Vol. 1(5), 58-61, December (2012)Int. Res. J. Environment Sci. International Science Congress Association 60 Table-2 Total flavonoid contents of Aloe vera, ocimum sanctum and Adhatoda vasica at three sampling sites during different seasons No. Plant Seasons Sampling sites MRB DDN SLP 2010 2010 2010 MRB DDN SLP 2011 2011 2011 1 Aloe vera (mg) January April August November 2.02 2.34 3.49 0.74 1.01 1.67 3.01 2.49 4.94 1.34 1.48 2.08 1.96 1.96 2.69 0.48 0.98 1.42 2.63 2.01 3.16 1.26 1.32 1.94 2 ocimum sanctum µg/gJanuary April August November 262.15 255.3 148.4 210.3 186.4 99.63 85.3 56.2 29.3 282.3 269.3 135.0 283.62 263.70 182.9 235.4 255.31 101.46 119.3 156.34 55.32 262.8 226.2 120.01 3 Adhatoda vasica (mg/g) January April August November 38.49 36.58 46.01 20.14 24.23 28.39 46.32 40.32 62.33 30.05 25.58 38.54 32.41 22.04 40.58 20.01 22.49 26.31 40.04 28.60 60.66 28.49 20.02 34.32 Figure-1 Total RSPM/SPM/SO/NO at MRB sampling site in different months Figure-2 Total RSPM/SPM/SO/NO at DDN Sampling site in different months Figure-3 Variation of RSPM/SPM/SO/NO at SLP College in different months Total flavonoids content of Adhatoda vasica, Ocimum sanctumand Aloevera in differen representative months has been shown in table 2. Total flavonoids contents from Adhatoda vasica, Ocimum sanctum and Aloevera have been determine by aluminium chloride colorimetric method described by change et al. Quercetin was used to perform the calibration curve (standard solution) in methanol for adhatoda vasica. The absorbance of the reaction mixture was measured at 415nm. The total flavonoid contain for adhatoda vasica was expressed as quercetin equivalent using the standard curve equation and result were expressed as mg/g. The flavonoid contain were found between 20.01 to 62.33mg/g and it was found maximum 62.33mg/g at SLP in August month in 2010. For ocimum sanctum rutin was used to perform the calibration curve in methanol. The absorbance of the reaction mixture was measured at 415 nm. Total flavonoid contain for ocimum  \n \r   !  "#  !  "#  !  "$  !  $  % \r  && !  "$ && !  $ && !  "# && !  "#   \r  "'# !  "$ "'# !  $ "'# !  "# "'# !  "# International Research Journal of Environment Sciences______________________________________________ ISSN 2319–1414 Vol. 1(5), 58-61, December (2012)Int. Res. J. Environment Sci. International Science Congress Association 61 sanctum was expressed as rutin equivalent and result were expressed as µg/g. The flavonoid content was found between 29.3 to 283.62µg/g and was found maximum 283.62 at Maharaj bada sampling sites. For Aloevera plant rutin was used to perform the calibration curve in methanol. The absorbance of reaction mixture was measured at 415nm, and result was expressed as rutin equivalent in mg. The total flavonoid contents were found between 4.94 to 0.48mg and it was found maximum at SLP College. Conclusion From the result it conclude that the vehicular pollution resulted in significant increase in the total flavonoid content in ocimum sanctum plant (table-19) .for ocimum sanctum plant, Maharaj bada sampling site recorded highest value of total flavonoid during April season and lowest value of during all seasons at SLP college. These findings are in agreement with the finding of A. Qayoom Mir et al11 who has reported increase in total flavonoid and phenolics in catharanthus roseus L. and ocimum sanctum L. as bio markers of urban auto pollution. This finding can also be attributed to the finding of Nikolova and Ivancheva12, who have reported increase total flavonoids in Artemisia vulgaris L; and veronicachamaedrys L; in relation to air pollution stress.But opposite results were found for Aloevera and adhatoda vasica plant. In both plant the total flavonoid content decreases with increasing pollution level (i.e. NO, SO2, SPM, RSPM and Ozone). In both plants, SLP sampling sites recorded the highest value of total flavonoid content during all seasons, which decreased significantly along the site attaining minimum value at Maharaj bada sampling sites and total flavonoid content continuously to decreases from 2010 to 2011. These findings are in contrary to the observation of A. Qayoom Mir et al11. In the present investigation it was observed that the total chlorophyll content in adhatoda vasica and Aloe vera were found decreases in more polluted habitats and in ocimum sanctum total flavonoids increases in polluted habitats. References 1.Samal A.C. and Santra S.C., Air quality of kalyani township (Nadia, West Bengal) and its impact on surrounding vegetation, Indian J. Environ Helth,44(1), 71-76 (2002)2.Saxena N.C., Sing Gurdeep and Ghosh Rekha, Environmental management in mining areas scientific publishers (India) Jodphur (2002)3.Pandey A.C., Das R.R. and Murty B.P., Ozone pollution in Gwalior, Research Link,12, 33-37 (2004) 4.Botkin D. and Keller E., Environmental science: Earth is a living planet, John Wiley and sons, Von Haffmen press (1995)5.Jackson A.R.W. and Jackson J.M., Environmental Science: The natural Environment and human impact, Logan G. Singapore Publishers Pvt. Ltd. (1996)6.West W. Philip and Gaeke G.C., Reference method for the determination of SO in the atmosphere (Pararosaniline method), Analytical Chemistry, 28, 1816-1819 (1956)7.Jacob M.B. and Hochheiser S., Continuous sampling and ultra micro determination of nitrogen dioxide in air, Anal, Chem.,30, 426-428 (1958)8.Byers D.H. and Saltman B.E., Determination of O in air by neutral and alkaline iodide procedure, Adv. Che. Ser.,21, 93-101 (1969)9.Boyed A.W., Willis C. and Cry R., New determination of stochiometry of the odometric method for Ozone analysis at PH 7.0, Analyt, che.,110, 670-672 (1970)10.Chang C., Yang M., Wen H. and Chern J., Estimation of total flavonoid content in propolis by two complemetry colorimetric methods, J. food Drug Analysis10, 178-182 (2002)11.Qayoom Mir A., Yazdani T., Ahmad S. and Yinus M., Total flavonoid and phenolics in catharanthus roseus L. and Ocimum sanctum L. asa Biomarkers of urban Auto pollution, Caspian J.Env. Sci.,7(1), 9-16 (2009)12.Nikolova M.T. and Ivancheva S.V., Quantitative flavonoid variation of Artemisia vulgaris L. and veronica chamaedrys L. in relation to altitude and polluted environment, Act.Biol. Szeged., 49(3-4), 29-32 (2005)