ISSN (Online) 2249-6084 (Print) 2250-1029 International Journal of Pharmaceutical and Phytopharmacological Research (eIJPPR) [Impact Factor – 0.7826] Research Article Journal Homepage: www.eijppr.com HERBAL INDICATORS: AN ALTERNATIVE TO SYNTHETIC INDICATORS Kavitha G*, Chinju Sara George, Ancy Alex, Rakhi Raju, Rini Biju, Deepa T Vasudevan NAZARETH COLLEGE OF PHARMACY, DEPARTMENT OF PHARMACEUTICAL CHEMISTRY, OTHERA, THIRUVALLA * Corresponding author: kavithaamritha@gmail.com 1 ABSTRACT The pH indicators are used to show sharp color changes at intervals of pH.Natural pigments in plants are highly colored substances and may show color change with variation of pH. An attempt has been made to investigate the indicator activity of water extract of flower and fruit pigments and to replace synthetic indicators, as they cause chemical pollution and are of high cost. The water extract of Caesalpinia sappan, Caesalpiniaceae and Methanolic extract of Dahlia pinnata, Asteraceae showed sharp and intense color change in acidic and basic medium. These herbal indicators are evaluated by using strong acid- strong base, strong acid- weak base, weak acid- strong base and weak acid – weak base titration. In all these titrations, the extract was found to be very useful and accurate for indicating the equivalence point as the synthetic indicators. The isolation of pure compounds possessing indicator properties helps to know the mechanism by which they show indicator properties and new theories of indicators could be established. KEYWORDS-Synthetic indicators, Caesalpinia sappan, Dahlia pinnata, Herbal indicators 2 1. INTRODUCTION A pH indicator is something that can tell you whether a substance is an acid or a base. We often notice the color changes that we see in the nature all around us. Many flowers, fruits and vegetables contain chemical substances that changes color in the solutions of different pH. These substances are known as natural pH indicators. All pH indicators change color depending on whether they donate or accept protons. Therefore, pH indicators are themselves acids or bases1. Dahlia pinnata (Asteraceae) is a yearly flowering plant and easily available in India and elsewhere in high quantity and present in almost every garden to enhance their beauty2. Caesalpinia sappan (Caesalpiniaceae) whose heartwood is widely being used in drinking water for its anti-thirst, blood purifying, ant diabetic and several other properties is nowadays being used as a coloring agent for wine, meat, fabric etc..As such, Sappan wood was a major trade good during the 17th century when it was exported from South-east Asian nations aboard Red seal ships to Japan3. Presently available acid-base indicators like phenolphthalein and methyl orange are synthetic indicators which produce chemical hazards, availability problems and high cost4,5. The present study is to investigate the indicator activity of Caesalpinia sappan and Dahlia pinnata and to establish analytical applications. 2. MATERIALS AND METHODS 2.1 Plant materials Fresh flowers of Dahlia pinnata (Asteraceae) and Heart wood of Caesalpinia sappan (Caesalpiniaceae) were collected from Central Travancore. 2.2 Reagents Analytical grade reagents like hydrochloric acid (HCl), Sodium hydroxide (NaOH), Acetic Acid (CH3COOH), Ammonia (NH3) and phenolphthalein were used. The reagents and volumetric solutions were prepared as per Indian Pharmacopoeia (IP 1996)6. 3.3 Methodology The petals of the flower –Dahlia pinnata were cleaned by using distilled water and cut into small pieces .Flower petals of 10g were shaken for an hour in 100 ml methanol7. The extract was filtered to obtain a clear liquid. Heartwood of 10g sappan wood was boiled with 100ml water for 3 minutes and filtered. The extracts were preserved in a tightly closed glass container and stored away from direct sunlight8. The original pH of the indicators, indicator range9 and color of the indicators prepared in acid (1M HCl) and base (1M NaOH) were tested. The chemical test for Anthocyanins10 was performed for the prepared indicators. The analytical applications of prepared indicators were confirmed by neutralization titrations11, 12. 3 3. RESULTS AND DISCUSSION The color of natural indicators prepared, Ph, color changes in acid and base, indicator range and chemical test are indicated in table no 1. The color changes obtained in neutralization titrations with different Titrants and titrates of 0.1 M are given in table no-2.The titre values in comparison with synthetic indicator phenolphthalein are given in table no-3 For all types of titrations, equivalence point obtained by the aqueous extract of Caesalpinia sappan either exactly coincided or very close with equivalence point obtained by standard indicator phenolphthalein .These represents the usefulness of the extract as an indicator in acidbase titrations. The use of Dahlia pinnata in strong acid- strong base and weak acid – strong base titrations was found to be more significant over standard indicator as it gives sharp color change at equivalence point. It is observed that the indicators act reversibly and give sharp color change in both directions. The results obtained prove that the routinely used indicators can be replaced successfully by plant extracts as they are simple, accurate TABLE NO – 1 INDICATOR RANGE AND COLOR CHANGE OF HERBAL INDICATORS PARAMETERS DAHLIA PINNATA 1.pH 2.color of the indicator 3.Color in acid 4.Color in base 5.Indicator range 6.Test for Anthocyanins 5.55 Violet CAESALPINIA SAPPAN 6.2 Red Pink Green 3.5-7 Positive Yellow Pink 5.3-7.4 Negative TABLE NO – 2 COLOR CHANGES IN NEUTRALIZATION TITRATIONS INDICATOR Dahlia pinnata Dahlia pinnata Dahlia pinnata TITRANT HCl CH3COOH HCl COLOR Pink Pink Pink TITRATE NaOH NaOH NH3 Dahlia pinnata CH3COOH NH3 Caesalpinia sappan Caesalpinia sappan Caesalpinia sappan Caesalpinia sappan HCl CH3COOH NH3 NH3 No visible color change Yellow Yellow Pink Pink 4 NaOH NaOH HCl CH3COOH COLOR Green Green No visible color change No visible color change Pink Pink Yellow Yellow TABLE NO – 3 COMPARISON WITH SYNTHETIC INDICATOR CHEMICALS USED TITRANT TITRATE HCl CH3COOH HCl CH3COOH NaOH NaOH NH3 NH3 VOLUMES OF TITRATES REQUIRED FOR EQUIVALENCE POINT WITH TITRANT(25 ML) WITH INDICATOR STANDARD DAHLIA SAPPAN INDICATOR INDICATORS WOOD INDICATOR 25.1±0.2 24.4±0.43 24.2±0.37 24.5±0.35 24.8±0.28 24.3±0.40 24.8±1.02 24.6±0.44 25.0±0.40 25.6±0.52 4. CONCLUSION In this study herbal indicators are prepared from Caesalpinia sappan and Dahlia pinnata by simple procedures and the analytical applications were recorded. The presence of coloring pigments anthocyanin in Dahlia pinnata and brazilin in Caesalpinia sappan may be responsible for the indicator action. The isolation13 of pure compounds possessing indicator properties helps to know the mechanism by which they show indicator properties and new theories of indicators could be established. REFERENCES 1. Ingrid J, Natural pH indicators: What are they and how do they work, Chapter 8: Acids and Base, Nelsons science 10, 2001. 2. Orient Longmann, Indian medicinal plants, 3rd, 5th edition, 2003, 1,396. 3. http://wonderherbs.org/index.html 4. NTP, Toxicology and Carcinogen studies of phenolphthalein,F344/n Rats and B365 Mice, Research Triangle Park,NC: national toxicology program, 1996,354. 5. Hazardous substances database, National library of medicine http://toxnet.nlm.nihgov/cgibin/sis/hmtlgen?HSDB, 2001. IndianPharmacopoeia,1996,volume II,A,211 6. Luis, ER, Ronald EW, Extraction, Isolation and Purification of Anthocyanins, Current protocol in food analytical chemistry 2001, (F1.1.1-F1.1.11) 7. Bakhshayeshi MA et al, “The effects of Light, Storage temperature, pH, Variety on Stability of Anthocyanin pigments in four Malus Varieties”, Pak.J Biol. Sci, 2006, 9(3): 428 – 433. 5 8. Beckett HH and Stenlake. Text book of Practical chemistry, 4th edition, 2005, 96. 9. Harborne JB, Phytochemical methods, 3rd edition, 1998, 72-74. 10. Mendhan J, et al Vogel’s text book of quantitative chemical analysis 6th edition, 2002, 374- 375. 11. Vishwas CB, “Herbal indicator as a substituent to synthetic indicator” ijgp, 2008, (2): 162 – 163. 12. Lioe HN, Adawiyah DR, Anggraeni R.,”Isolation and characterization of the major natural component of Brazil wood (Caesalpinia sappan L.)” International Food Research Journal, 2012, 19(2):537-542 6