RP-HPLC Method Development and Validation of Gallic …

1 2013 Kamal Kardani et al. This is an open access article distributed under the terms of the Creative Commons Attribution License -NonCommercial-ShareAlike Unported License ( ). Journal of Applied Pharmaceutical Science Vol. 3 (05), pp. 037-042, May, 2013 Available online at DOI: ISSN 2231-3354 RP-HPLC Method Development and Validation of Gallic acid in Polyherbal Tablet Formulation Kamal Kardani1, Nilesh Gurav2*, Bhavna Solanki2, Prateek Patel2, Bhavna Patel1 1 Post Graduate Department of Pharmaceutical Sciences, Sardar Patel University, Vallabh Vidyanagar, Gujarat- India.

2 2 Vasu Research Centre, A Division of Vasu HealthCare Pvt Ltd. Vadodara, Gujarat- India. ARTICLE INFO ABSTRACT Article history: Received on: 04/04/2013 Revised on: 30/04/2013 Accepted on: 14/05/2013 Available online: 30/05/2013 A new simple, accurate, precise, sensitive and validated RP-HPLC Method was developed for the estimation of Gallic acid in bulk and pharmaceutical tablet dosage form. The chromatographic conditions used for the separation was Phenomenex Luna C18 (2) ( x 250mm, 5 ), rheodyne manual injector with capacity of 20 L and mobile phase comprised of Water: Acetonitrile (80: 20%v/v) and pH is maintained at using O-phosphoric acid (OPA).

3 The flow rate was with detection at 272nm. The retention time was found to be The linearity was found to be in the range of g/mL for Gallic acid with correlation coefficient of The proposed Method is accurate with - % recovery and precise (%RSD of repeatability, intra-day and inter-day variations were , , ). The Limit of Detection (LOD) and Limit of Quantification (LOQ) were found to be g/mL and g/mL respectively. The amount of Gallic acid in Polyherbal tablet was found to be Key words: Gallic acid, RP-HPLC , Validation , Polyherbal formulation.

4 INTRODUCTION Gallic acid is a Poly phenolic compound having an antioxidant property. Chemically it is a 3, 4, 5-trihydroxybenzoic acid [Figure-1]. Gallic acid and its derivatives are a group of naturally occurring polyphenol antioxidants which have recently been shown to have potential health effects. Gallic acid and its derivatives have antioxidant activities, and neuroprotective effects with free radical scavenging effects (Zhongbing et al, 2006 & Kubo et al 2010). OHOHOHOHO Fig. 1: Structure of Gallic acid.

5 Herbs that contains Gallic acid as a major constituent are citrus fruits like Phyllanthus emblica (Amalaki), Terminalia bellirica (Bibhitaki), Terminalia chebula (Haritaki), etc. which is useful in common cold and fever, diuretic, laxative, liver tonic, . * Corresponding Author Nilesh Gurav, Research & Development Department, Vasu Research Centre, A Division of Vasu Healthcare Pvt. Ltd., Vadodara, Gujarat- India. Phone: +91-9979184312, 0265-2657701, 02 refrigerant, stomachic, restorative, alterative, antipyretic, anti-inflammatory (Kroes et al, 1992), hair tonic, to prevent peptic ulcer and dyspepsia, and as a digestive.

6 Preclinical studies have shown that amla possesses antipyretic, analgesic (Perianayagam et al, 2004), antitussive, antiatherogenic, adaptogenic (Sai Ram et al, 2003), cardioprotective, gastroprotective, antianemia, antihypercholesterolemia, cytoprotective (Gulati et al, 1995), wound healing, antidiarrheal, antiatherosclerotic, hepatoprotective (Jose et al, 2001), Antimutagenic, anticarcinogenic (Calixto et al, 1998), nephroprotective, memoryenhancing activity and neuroprotective properties.

7 In addition, experimental studies have shown that amla and some of its phytochemicals such as Gallic acid, ellagic acid, pyrogallol, some norsesquiterpenoids, corilagin, geraniin, elaeocarpusin, and prodelphinidins B1 and B2 (Yadav, 1987) also possess antineoplastic effects. Amalaki is also reported to possess radiomodulatory (Ghoshal et al, 1996), chemomodulatory, chemopreventive effects, free radical scavenging (Jose & Kuttan, 1995), antioxidant (Miller, 1996), anti-inflammatory, antimutagenic and immunomodulatory activities and properties that are efficacious in the treatment and prevention of cancer.

8 Polyherbal formulation contains more than one herb or a mixture of different herbal powders or extracts, so being difficult to identify a particular component. 038 Kardani et al. / Journal of Applied Pharmaceutical Science 3 (05); 2013: 037-042 This Polyherbal Trichup tablet is used as essential nutrition to hair roots, stimulates repair of hair follicles thus enhancing natural hair growth, helps control hair fall, helps prevent premature greying and falling of hair, which contains Amla and Triphala as a major constituent of Gallic acid, so an attempt is being made to develop a specific, accurate, precise and simple Method for estimation of particular compound from Polyherbal tablet formulation.

9 Validation of developed Method was carried out as per ICH Q2R1 guideline to maintain the reproducibility and consistency of the Method . MATERIALS & methods Apparatus The chromatography was performed on a Shimadzu LC-20AT, SPD-20A hplc instrument equipped with UV detector UV-20A with wavelength Range 190-700nm and Spinchrom LC Solution software. Phenomenex Luna C18(2) ( x 250mm, 5 particle size) was used as stationary phase. Rheodyne manual injector with 20 L capacity loop was used. Shimadzu Unibloc AUX220 with Capacity of 10mg -220g as an analytical balance.

10 Toshcon pH meter and Sonicator Toshcon SW-2 were used in the study. Reagents and Materials The reference standard of Gallic acid was purchased from Hi-media Laboratories Pvt. Ltd. Mumbai. hplc grade Water, Methanol, Acetonitrile and OPA made of Merck specialities Pvt. Ltd. Mumbai were used in study. Trichup tablet (An Ayurvedic Proprietary Formulation) is manufactured and provided by Vasu Healthcare Pvt. Ltd., Vadodara- Gujarat., which contains extracts of Eclipta alba (Bhringraj), Glycyrrhiza glabra (Yashtimadhu), Emblica officinalis (Amalaki), Centella asiatica (Mandukparni), Hibiscus rosa-sinensis (Japa), Tinospora cordifolia (Guduchi), Tribulus terrestris (Gokshur), and Powder of Triphala Churna and Shukti Bhasma.