Pushpa Jain* et al. /International Journal Of Pharmacy & Technology Available Online through www.ijptonline.com ISSN: 0975-766X CODEN: IJPTFI Research Article DAHALIA FLOWER SAP A NATURAL RESOURCE AS INDICATOR IN ACIDIMETRY AND ALKALIMETRY Poonam Gupta, Pushpa Jain*, Pramod Kumar Jain** *Professor, Botany, Mata Jija Bai Govt. Girls Post Graduate College, Indore **Professor, Chemistry, Govt. Holkar Science College, Indore. Email: saral_1@yahoo.com Received on 09-11-2012 Accepted on 12-12-2012 Abstract Dahlia pinnata is a species of the genus dahlia, belonging to the family Asteraceae. The present study reported the use of dahlia pinnata flower extract as an acid base indicator in different types of acid base titrations. The equivalence point obtained by the flower extract coincided with the equivalence point obtained by standard indicators. The results obtained by the flower extract matched with the results obtained by mixed indicator in case of strong acid and weak base, weak acid and strong base, strong acid and strong base, weak acid and weak base titration. This natural indicator was found to be a very useful, economic, simple and accurate for the said titration. Keywords: Dahlia pinnata, acid-base indicator, natural indicator. Introduction Flowers are symbolic; they are often regarded as symbol of love or as a token of well wishes. Even animal kingdom is not untouched by the magic of flowers. Many herbivores feed on them while few birds and insects suck nectar from these flowers and even provide final product like honey to the world. Flowers also attract the insects for pollination. Thus flowers are miracle of the nature, by the God, for the world. ‘Titrimetric analysis’ refers to quantitative chemical analysis carried out by determining the volume of a solution of accurately known concentration which is required to react quantitatively with a measured volume of a solution of the substance to be determined. The process of adding the standard solution until the reaction is just complete is termed as titration. Acidimetry and alkalimetry include the titration of free bases with a standard acid and the titration of free acids with a standard base respectively. The reactions involve the combination of hydrogen and hydroxyl ions to form water. In Europe and America, prior to the discovery of Insulin in 1923, diabetics as well IJPT | Jan-2013 | Vol. 4 | Issue No.4 | 5038-5045 Page 5038 Pushpa Jain* et al. /International Journal Of Pharmacy & Technology as consumptives were often given a The discovery of Insulin in 1923, diabetics as well as consumptives were often given a substance called Atlantic Starch or diabetic sugar, derived form inulin, a naturally occurring from of fruit sugar, extracted from dahlia tubers. Inulin is still used in clinical tests for kidney functionality (Williams 1895). The extract was preserved in tightly closed container and stored away from direct sunlight (Vishwas et al 2008). Anthocyanidins (anthocyanins) are notable plant pigments, which are responsible for red-violet blue colour seen in plant flowers. (Harborne J.B. 1998). The intention behind this study is simply to bring in market the use of plant pigments and to increase the wealth of traditional medicinal system of India which is mostly plant based and to help farmers regarding cultivation collection of plants as well as to industry regarding preparation of natural indicators. (Pathade K.S. 2009). The dahlia was declared the national flower of Mexico in 1963. (Harvey, Marian 1987). The appearance of red color is due to presence of flavonoids, phenols and anthocyanins (P.S. Vankar 2008). Red-fleshed peaches are rich in anthocyanins (Bolivar A. Cevallous -Casals, 2006). The anthocyanins, anthocyanidins with sugar group (s), are mostly 3-glucosides of the anthocyanidins. The anthocyanins are subdivided into the sugar free anthocyanidin aglycones and the anthocyanin glycosides. As of 2003 more than 400 anthocyanins had been reported (Kong JM, 2003). Flavonoids have also been found to inhibit a wide range of enzymes involved in oxidation system such as 5lipoxygenase, cyclooxygenase, monooxygenase, or xanthine oxidase (J.Tinoi 2006). As flavonoids, anthocyanins are present in flowers of Dahlia pinnata and are pH sensitive therefore the flower extract could be utilized as an indicator for different types of acid base titrations. Hence the flavonoids were extracted. Material and Methods Identification and collection of that Dahlia pinnata Dahlia pinnata was identified from the flora of Botany Department of the Holkar Science College, Indore. Flowers were collected during spring season. Dahlia pinnata was collected for the purpose of study of natural indicator. Fresh petals were collected in the month of February and March because it is the blooming season of these plants. Natural indicator can be isolated from the Dahlia pinnata flower. Several plant pigments like Anthocyanin, Flavonoids occurs in petals of these flowers which acts like natural indicator. IJPT | Jan-2013 | Vol. 4 | Issue No.4 | 5038-5045 Page 5039 Pushpa Jain* et al. /International Journal Of Pharmacy & Technology Method Flowers were cleaned by distilled water, petals of these flowers were kept in strong sunlight until they get completely withered. The dried petals were grinded into fine powder with a mechanical blender. Dried powder of petals were soaked in 40 ml methanol for 48 hours and then triturated in mortal and pestal and the resulting solution was filtered through muslin cloth. The resulted methanolic extract was further used as natural indicator for acidimetry and alkalimetry. The extract was preserved in light closed container and stored away from direct sunlight. Following the similar process aqueous extract of dried pulvarized petals was prepared, filtered and used as natural aqueous indicator. Analytical grade HCl, NaOH, CH3COOH, NH4OH,phenolphthlein,methylorange,and phenol red were made available by the Department of Chemistry, Government Holkar Science College, Indore. Reagents and volumetric solutions were prepared as per standard. The experimental work was carried out by using the same set of glasswares for all types of titration. As the same aliquots were used for both titrations the standard indicator, floral extract and the reagents were not calibrated. The equinormal titrations were performed using 10 ml of titrant with five drops of natural indicator. A set of four experiments each for all the types of acid base titrations were carried out. The t-value and standard deviation for each type of acid base titrations were calculated from results obtained. The anthocyanins which are water soluble pigments, generally occur in the aqueous cell sap, and are responsible for the large variety of colours in flowers such as red, violet, blue. Anthocyanin pigments are amphoteric, their acid salts are usually red and their metallic salts are usually blue and in neutral solution it is violet. In addition to anthocyanins, the colour of flower depends on the prescence of co-pigments such as flavones, flavonols etc. and to metal chelation particularly with iron and aluminium. Geissman et al., (1952) have applied the term flavonoids to embrace all compounds whose structure is based on flavone. Thus the anthocyanins are group of flavonoid compounds. Singh et al., (2010) used Nerium Oleander (Apocynaceae), Tecoma Stans (Bignonaceae), Calatropis gigantea (Asclepiadaceae), Albizia lebbeck (Mimosaceae) and Cassia fistula (Caesalpiniaceae) flowers as acid base indicator. The titrant v/s titrand used were NaOH/HCl, HCl/NH4OH, CH3COOH/NaOH and CH3COOH/NH4OH. Ramling et al., (2010) used flower extract of Bombax malabaricum (Malvaceae) as indicator in acid base titration. The equivalence point obtained by the flower extract coincided with the equivalence point obtained by IJPT | Jan-2013 | Vol. 4 | Issue No.4 | 5038-5045 Page 5040 Pushpa Jain* et al. /International Journal Of Pharmacy & Technology standard indicators. In case of weak acid and weak base titration, the results obtained by the flower extract matched with the results obtained by mixed indicator. WEI You - haun et al., (2004) studied the juices of Green tea’s and Ku Ding Teas changing colour in the solution of acid and alkali. The research workers compared the result of the Tea with phenolphthalein and methyl orange in acid base titration as indicator. The result concluded that the juice can be used as a substitute for acid base indicator similarly FANG Rong – Mei (2001) studied the juice of radish red pigment as an indicator for acid alkali titration. Wadkar et al., (2008) used methanolic extract of the leaves of careya arborea as an acid base indicator in acid base titrations. Promising results were obtained when it was tested against standard synthetic indicators. Titration showed sharp colour change at the equivalence point. The indicator was useful in all type of acid – base titration except weak acid and weak base titration. Okonkwo et al., (2010) used Hibiscus Sabdariffa petals extract for acid – base titration. The results were compared with those of standard end point indicators as phenolphthalein and methyl red. Prominent absorption in the 500 – 550 nm wavelength region of the UV/Visible spectrum of methanolic extract confirmed the presence of anthocyanidins. Sidana Jaspreet et al., (2011) used methanolic extract of Antirrhinum majus (Scrophulariaceae) and Dianthus plumaris (Caryophyllaceae) which gave sharp and intense colour change as compared to phenolphthlein and methyl orange. Phytochemical investigation and chemical test of both show prescense of anthocyanins and were the main reason for the activity of indicator. ZHANG et al., (2006) studied the condition to extract violet cabbage pigment and found that the best extraction was by using distilled water as extraction reagent with the ratio of liquid to solid being 5:1 (ml/g) under the condition of pH 2 with 2 hour of extraction at 50OC. The yield was 17.1%. Lu Rongrong et al., (2008) studied purple cabbage pigment as indicator for acid – base titration. The pigment accurately indicated the end point with sensitive colour change. Agrawal et al., (2011) investigated the indicator activity of methanolic fruit extract of Punica granatum (Punicaceae). The results were compared with synthetic indicators in different types of acid – base titration. The extract was found to be very useful and accurate for indicating the neutralization point. IJPT | Jan-2013 | Vol. 4 | Issue No.4 | 5038-5045 Page 5041 Pushpa Jain* et al. /International Journal Of Pharmacy & Technology Jadhav et al., (2009) used Ixora Chinensis flower extract as acid – base indicator. The result obtained by extract was similar to the result obtained by synthetic indicator. In case of weak acid and weak base titration the results obtained matched with the result obtained, matched with the results obtained by mixed indicator. Results and Discussion The flower was screened for its use as an indicator in acid base titration and the results were compared with the results obtained by standard indicator phenolphthelin,Methylorange,phenolred. The results of the screening for strong acid-strong base (H2SO4 & KOH),strong acid- weak base (H2SO4 & NH4OH), weak acid-strong base (CH3COOH & KOH),weak acid –weak base (CH3COOH & H) are listed in table A & B. Table-A: Colour Change during Titration. Indicator colour changes and PH range Titrant Titrand Standard H2SO4 KOH Colourless to pink (PH) H2SO4 NH4OH Pink to yellow (MO) CH3COOH KOH Colourless to pink (PH) CH3COOH NH4OH Yellow to red (PR) Methanolic floral extract of Dahlia pinnata orange to wine red Pink to red Aqueous floral extract of Dahlia pinnata orange to wine red Pink to red Pink to yellow Pink to yellow orange to yellow orange to yellow Keyword: PH = Phenolphthlein, MO = Methyl orange, PR = Phenolred. In the present study aqueous extract of dahlia pinnata was taken as a natural indicator for acid base titration. The strength of acids and bases (HCL ,H2SO4, CH3COOH,NaOH,KOH, NH4OH) taken were 0.1N,0.5N and 1.0N.Four different types of titration performed were strong acid/strong base, strong acid/weak base, weak acid/strong base, and weak acid/weak base. The titration v/s titrand were KOH v/s H2SO4, NH4OH v/s H2SO4, KOH v/s CH3COOH, NH4OH v/s CH3COOH. The colour changed from orange to wine red for aqueous and methanolic floral extract of dahlia pinnata. The standard deviation ranged from + 0.05 to + 2.9 in dahlia pinnata, the results obtained showed that the routinely used IJPT | Jan-2013 | Vol. 4 | Issue No.4 | 5038-5045 Page 5042 Pushpa Jain* et al. /International Journal Of Pharmacy & Technology indicator can be replaced successfully by floral extract. The table-B represents t-value of four titrations + standard deviation. Table-B: Titration with Methanolic and Aqueous Floral Extract of Dahlia Pinnata as Natural Indicator. Titration Titrant V/s Titrand KOH v/s H2SO4 NH4OH v/s H2SO4 KOH v/sCH3COOH NH4OH v/s CH3COOH Strength in normality Methanolic floral extract of Dahlia t value + S.D* pinnata as natural indicator Aqueous floral extract of Dahlia t value + S.D* pinnata as natural indicator 1.0 PH v/s MFE 2.1213 + 0.17 PH v/s AFE 0.4472+0.27 0.1 PH v/sMFE 1.3422 +0.09 PH v/s AFE 2.6805+0.09 0.5 PH v/s MFE 1.3422 +0.18 PH v/s AFE 1.9424+0.16 1.0 MO v/s MFE 0.8965 +0.09 MO v/s AFE 1.2659+0.12 0.1 MO v/s MFE 2.1276 +0.05 MO v/s AFE 2.2405+0.09 0.5 MO v/s MFE 1.4349 +0.36 MO v/s AFE 0.8965+0.09 1.0 PH v/s MFE 0.8965+0.09 PH v/s AFE 0.4475+0.09 0.1 PH v/s MFE 2.1276+0.05 PH v/s AFE 2.1537+0.09 0.5 PH v/s MFE 2.1276 +0.05 PH v/s AFE 2.2375+0.09 1.0 PR v/s MFE 2.0018 +0.20 PR v/s AFE 1.4556 +0.16 0.1 PR v/s MFE 0.04475+0.09 PR v/s AFE 0.7223+0.05 0.5 PR v/s MFE 2.1276+0.05 PR v/s AFE 0.6334+0.12 *All values are t value + S.D. for n = 4 Keyword: H2SO4: Sulphuric acid, CH3COOH: Acetic acid, KOH: Pottasium hydroxide, NH4OH: Ammonium hydroxide, PH: Phenolphthlein, MO: Methyl orange, PR: Phenolred, MFE: Methanolic floral extract, AFE: Aqueous floral extract. Conclusion The standard deviation calculated for synthetic indicator and aqueous and methanolic floral extract of natural indicator shows very less variation in the results. Thus statistically also the use of natural indicator in acid base titration is proved. As the aqueous and methanolic extract provide similar results hence aqueous and methanolic floral extract can be used with cent per cent reliability and accuracy for acid base titration. Thus the use of natural indicator in acid base titration is more beneficial because of its economy, easy to prepare, simplicity, easy availability, pollution free inert and accurate results. IJPT | Jan-2013 | Vol. 4 | Issue No.4 | 5038-5045 Page 5043 Pushpa Jain* et al. /International Journal Of Pharmacy & Technology Acknowledgement Creation is quite a difficult task in this world, it becomes more hideous without God’s inspiration, elder’s blessing, younger’s love and cooperation of friends. The completion of any interdisciplinary project depends upon the cooperation, coordination and combined efforts of several sources of knowledge, skill, labour and time and hence the precious guidance and spiritual help of a guide cannot be fulfilled just by a mere word, thanks. Dr. Rooplekha Vyas Head, Department of chemistry, Govt. Holkar Science College, Indore often went out of the way to see if I had completed this work with case. She encouraged me on several occassions and so my heart feels sincere gratitude to her. I am thankful to Principal Dr. R.K. Tugnawat of Govt. Holkar Science College, Indore for this scholarly encouragement of my academic activities. He has extended all the necessary help needed for my work in his capacity being the principal of college. References 1. Agrawal S., Raj N.R., Chouhan K., Raj C.N., Jain S. and Balasubramanian A. (2011). Isolation of herbal acid – base indicator from the seeds of Punica granatum. 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IJPT | Jan-2013 | Vol. 4 | Issue No.4 | 5038-5045 Page 5044 Pushpa Jain* et al. /International Journal Of Pharmacy & Technology 7. Ramling Patrakar, Deshpande A., Walsangikar S., Niranjane K. and Gadgil A. (2010). Use of Bombax Malabaricum Flower Extract as a Natural Indicator in Acid Base Titration. Der Pharmacia Lettre, 2(4): 520-524. 8. Sidana Jaspreet, Arora K., Nain P. and Deswal G. (2011). Herbal Indicators as an upcoming trend in Titremetric analysis. International Research Journal of Pharmacy 2(4), 177-179. 9. Singh Sudarshan, Bothara S.B., Singh Sangeeta, Patel R. and Mahobia N. (2010). Preliminary Pharmaceutical characterization of flowers as natural indicator: Acid base Indicator. The Pharma Research Journal, 4: 83-90. 10. Wadkar K.A., Mugdum C.S., and Kondawan M.S. (2008). Use of Careya arborea Roxb. Leaf extract as an indicator in Acid Base titrations. Research J. Pharm. and Tech. J. (4). Oct-Dec. 2008. 535-536. 11. WEI You-huan, WEI Liang-xing, WANG Li-ping, LUN Zhen-biao, QIN xiong-jun, LIANG Su-heng (2004). A study on Using Tea Juice as substitute for acid – base indicator: Journal of Nanning Junior Teachers’ College, 021. 12. ZNANG Yu-min, GUO Yan-hua, and CHEN Yi-feng (2006). Study on the best condition of the extraction of violet cabbage pigment. Journal of Central China Normal University. 02-017. Corresponding Author: Pushpa Jain*, Email: saral_1@yahoo.com IJPT | Jan-2013 | Vol. 4 | Issue No.4 | 5038-5045 Page 5045