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INDEX – GJRMI - Volume 7, Issue 5, May 2018 INDIGENOUS MEDICINE Ayurveda – Rasa Shastra – Short Communication STANDARDIZATION OF SAPTAMRITA LOHA VATI - A MODIFIED COMPOUND HERBOMINERAL PREPARATION OF AYURVEDA Hiremath Virupaxayya, Kulkarni Reena*, Koppala Narayan Sunil Kumar, Patil Amrit
COVER PAGE PHOTOGRAPHY: DR. HARI VENKATESH K R, PLANT ID – INFLORESCENCE OF BIXA ORELLANA L.* OF THE FAMILY BIXACEAE PLACE – KOPPA, CHIKKAMAGALUR DISTRICT, KARNATAKA, I NDIA *BOTANICAL NAME VALIDATED FROM www.theplantlist.org AS ON 30/05/2018
68–74
Global J Res. Med. Plants & Indigen. Med. | Volume 7, Issue 5 | May 2018 | 68–74 ISSN 2277-4289│ www.gjrmi.com │International, Peer reviewed, Open access, Monthly online Journal
Short Communication STANDARDIZATION OF SAPTAMRITA LOHA VATI - A MODIFIED COMPOUND HERBOMINERAL PREPARATION OF AYURVEDA Hiremath Virupaxayya1, Kulkarni Reena2*, Koppala Narayan Sunil Kumar3, Patil Amrit4 1
Consultant, Sammruddhi Clinic, 2nd main, Vinayaka Nagar, Davanagere, PIN-577006, Karnataka, India. Professor and Head, Department of Kaumarabhritya, SDM Institute of Ayurveda and Hospital, Anchepalya, Kumbalgodu Post, Bengaluru, PIN- 560074, Karnataka, India. 3 Researh Officer and HOD (Pharmacognosy), Siddha central Research Institute, (Central council for Research in Siddha, Ministry of AYUSH, Govt. of India), Anna Hospital, Arumbakkam, Chennai, India, PIN- 600106 4 Consultant, Sanjeevani children’s clinic, Nandre, Miraj taluk, Sangli Dt., PIN-416423, Maharashtra, India *Corresponding Author: E-mail: drreenakulkarni@gmail.com; Mob-+91-9480478639 2
Received: 10/04/2018; Revised: 21/05/2018; Accepted: 28/05/2018
ABSTRACT Saptamrita loha is a compound herbomineral formulation widely used in Ayurvedic clinical practice with multi fold benefits, specifically to improve eye sight and haemoglobin by its properties namely chakshusya (good for eye sight) and raktavardhaka (haematinic). The available standardization works on saptamrita loha have highlighted about the bhasma (calcined ash) preparation by puta paka (incineration) method and about shodhana (Purification) aspect. This study highlights physico-chemical characterization, HPTLC profiling of saptamrita loha which can be applied for authentication of this herbo-mineral formulation. Four plant medicines namely, Haritaki (Terminalia chebula Retz.), Bibitaki (Terminalia bellirica (Gaertn.) Roxb.), Amalaki (Phyllanthus emblica L.) Yastimadhu (Glycyrrhiza glabra L.), were authenticated botanically. Loha bhasma, Madhu (Honey) and Ghrita (Ghee) were purchased from authenticated pharmacy. Tablets were prepared by combining all these drugs and subjected for detailed physico-chemical and HPTLC analysis. Set of standardization parameters were derived for the compounded tablet containing four herbs and one mineral by physico-chemical characterisation. The tests proposed would serve as diagnostic parameters to identify this herbo-mineral formulation. HPTLC fingerprint profile which can serve as a fingerprint for the identification of the formulation has been developed. The proposed method of making tablet from herbo mineral drug will aid in uniform dosage form, better palatability and easy acceptability in children. Standards for the herbo-mineral formulation have been developed for the quality check of the formulation. KEYWORDS: HPTLC densitogram, herbo-mineral preparation, physico-chemical characterization, saptamrita loha, standardization
Cite this article: Hiremath Virupaxayya, Kulkarni Reena*, Koppala Narayan Sunil Kumar, Patil Amrit (2018), STANDARDIZATION OF SAPTAMRITA LOHA VATI - A MODIFIED COMPOUND HERBOMINERAL PREPARATION OF AYURVEDA, Global J Res. Med. Plants & Indigen. Med., Volume 7 (5): 68–74
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Global J Res. Med. Plants & Indigen. Med. | Volume 7, Issue 5 | May 2018 | 68–74
INTRODUCTION Ayurveda, the Indian system of medicine is the first recorded medical science widely practiced in India since ancient times (Reena, K., Abhimanyu, K., & Kumar, K. N. S. 2013). Considering the significance of traditional practices in global health care, WHO has been encouraging and promoting traditional practices since few decades. Hence the standardization of raw drugs, processing, finished products, verification of claims, mechanism of actions, products that are free of microbial contamination etc, have become major issues, which are to be taken in to consideration in order to increase the global acceptability of herbomineral drugs and prove their respective clinical efficacy. It is very tedious to standardize Ayurvedic herbo-mineral formulations due to a number of factors. The non availability of reference standard also hurdles the study. WHO has evolved guidelines for the validation of plant based drugs for developing countries like India (Kimmatkar, N et al., 2003). These can be used as baseline standards to work on further drug development and standardization. Saptamrita loha (SL) is one of the herbomineral preparations widely used in shula (pain in abdomen), timira (disease of eye), amlapitta (dyspepsia) etc. (Ambikadatta Shastri,. 2004). Vati (tablet) is a widely accepted dosage form in the present scenario due to its palatability, easy administration, shelf life etc. Keeping this in view Saptamrita loha was subjected for analytical and physiochemical study. Available market samples of Saptamrita loha vati do not contain ghee and honey in their formulation. SL is usually administered either with plain water. Most of the time when given alone there could be chances of gastric irritation with the drug due to the iron content in the drug. Keeping this in mind it was planned to prepare Saptamrita loha vati as described in the original reference including ghee and honey in its contents and evaluate analytically. The available standardization works on Saptamrita loha have highlighted about the bhasma (calcination process) preparation by puta paka (special method of incineration)
method and about shodhana (purification) aspect (Jadhao Ujwal Ashokrao., Ingole Rajesh Kundlikrao., 2014). This study highlights physico-chemical characterization, TLC and HPTLC densitogram profiling of saptamrita loha vati which can be applied for authentication of this herbo-mineral formulation. MATERIALS AND METHODS The individual ingredients of Saptamrita loha were procured from the pharmacy of SDM college of Ayurveda and hospital Hassan and authenticated in pharmacognosy laboratory of SDM College of Ayurveda Hassan. SL was prepared from 320 gram powders of each of Amalaki (Phyllanthus emblica L.), Haritaki (Terminalia chebula Retz.), Vibhitaki (Terminalia bellirica (Gaertn.) Roxb.,) Yashtimadhu (Glycyrrhiza glabra L.) and Loha bhasma along with 250 ml of honey (Agmark, SDM brand), and 250 ml Ghee (Nandini milk federation, Government of Karnataka undertaking). (Sample code: 591/15032301). METHOD OF PREPARATION Equal quantity of powder of above mentioned ingredient except honey and ghee were taken in a khalva yantra (porcelain mortar and pestle) mixed thoroughly and triturated with honey and ghee to obtain fine paste consistency. Paste was put in to a tablet cutting machine to obtain uniform sized drug heap which was later rolled in to pills of weight 500 mg after drying as recorded on a digital weighing scale. Instrumentation and techniques Physico-chemical determination of total ash, acid insoluble ash and water soluble ash, loss on drying at 110° C and water soluble extractive were done as per API (AYUSH study group 2008) standards. Disintegration time of the tablets was assessed as per (IP Indian Pharmacopoeia study group 1996) HPTLC studies were done as per standard procedure (Stahl I. 1969, Sethi PD. 1996 and WHO study group, 2002). The standardization studies were carried out at SDM Centre for Research in Ayurveda and Allied Sciences, Kuthpady,
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Global J Res. Med. Plants & Indigen. Med. | Volume 7, Issue 5 | May 2018 | 68–74
Udupi, Karnataka state, India as per standard procedures (Sample code: 591/15032301). To develop HPTLC fingerprinting, One gram of powdered samples of SL was dissolved in 10 ml ethanol and kept for cold percolation for 24h and filtered. 3, 6 and 9 Âľl of the above samples were applied on a pre-coated silica gel F254 on aluminum plates to a band width of 7 mm using CAMAG (Muttenz, Switzerland) Linomat 5 TLC applicator. The plate was developed in Toluene: ethyl acetate; acetic acid: water (3:2:0.8:0.2) in CAMAG twin-trough chamber. The developed plates were visualized under short and long UV (UV 254nm, 366nm) and then derivatised with vanillin sulphuric acid reagent, prior to derivatisation in CAMAG Photo documentation unit (Stahl I. 1969, Sethi PD. 1996 and WHO study group, 2002). The plate was scanned under UV 254 and 366 nm using CAMAG Scanner 4. HPTLC the Rf values and densitometric data were documented using win CATS Planar Chromatography Manager version 1.4.6 (CAMAG Muttenz, Switzerland).
RESULTS AND DISCUSSION Standardization of SL performed was as per AYUSH testing protocol for Vati. SL pills were pinkish brown and nearly round in shape with sweetish odour and taste. The average weight of SL was 500 mg. Uniformity in the size of the tablet was ensured through use of tablet cutting machine to maintain the consistent drug dosage. The slight variation in the size and shape of the rolled pills could be attributed to manual pill rolling. Never the less it did not affect the size by weight. The tablet disintegration time (70 min) indicating proper dissemination of the active ingredients for targeted action. This is slightly higher than the acceptable norms for coated tablets as per British Pharmacopoeia (BP) (Pharmacopoeia, C. 2013). The drug did not show any variations in its physical characteristics and consistency for a period of 8 months indicating acceptable shelf life (1year as per standard) (Parashurama shastri, 2013). Physico-chemical standards for herbomineral formulation Saptamrita loha are presented in (Table 1, figure 1).
Table 1: Physico-chemical characters of Saptamrita loha vati Parameters Total Ash Acid Insoluble Ash Water soluble ash Disintegration time (min)
Results n=3 %w/w 14.214 7.654 1.896 70
Figure 1: Saptamrita loha vati
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Rf values of the spots and their colour by HPTLC photo documentation of ethanol extracts of Saptamrita loha were developed (Figure 2). Study revealed 5 spots under short UV (0.04 D green, 0.07 green, 0.40 D green, 0.72 L green, and 0.78 green); 9 spots under long UV (0.07 F violet, 0.16 F L violet, 0.42 F violet, 0.54 F blue, 0.66 F blue, 0.72 F blue, 0.76 F violet, 0.78 F L violet, 0.87 F green), 3 spots under white light (0.07 brown, 0.14 L brown, 0.42 brown) and 12 spots under white light after derivatisation (0.04 grey, 0.07 grey, 0.14 L violet, 0.22 L violet, 0.42 brown, 0.45 L violet, 0.42 brown, 0.45 L violet, 0.54 L violet, 0.64 L violet, 0.69, 0.76 pink) (Table 2). Densitometric scan SL (Figure 3) at 254 nm revealed 7 peaks corresponding to different compounds in the ethanol extract. At 366 nm there were 7 peaks again of which 4 major peaks were at Rf 0.03 (12.80%), 0.07 (15.09%), 0.47 (42.37%) and 0.84 (22.17%). At 540 nm, 3major peaks were seen at Rf 0.04 (19.93%), 0.07 (37.51%) and 0.47(42.57%). HPTLC is an important tool in standardisation and quality control of polyherbal formulations. As there is more than one
ingredient, qualitative HPTLC fingerprinting can be used for development of quality standards for poly herbal formulations (Reena, K., Abhimanyu, K., & Kumar, K. N. S. 2013; Kumar, K. N. S. et al., 2016). The major phyto constituents of Haritaki, Vibhitaki and Amalaki are Gallic acid, rutin and quercetin. The observed Rf values of the same corresponds to that of active phytoconstituents as per the standards of previous study 0.11 to 0.16 (standard Rf 0.09–0.17) for gallic acid (Sai Prasad, A. J. V, Ratna Manikyam B., Trimurtulu. G., Reddy, K. N., Naidu, M. L., 2014), 0.34 and peak area 15143. (Garg, S., Mishra, A., & Gupta, R., 2013) and rutin and quercetin (0.13 and 0.93) (Ashok, K., Lakshman, K., Jayaveera, K. N., Tripathi, S. M., & Satish, K. V., 2010). There were three peaks of Saptamritaloha in this range at both 254 and 366 nm. There was one peak band 0.42 at 366 nm and 540 nm and after derivatisation which corresponds to standard glycyrrhizin ammonical hydrate band at Rf 0.42. (Syed, Y.H., Khan, M., Bhuvaneshwari, J., Sayyed, Mateen., 2014).
Table 2: Rf values ethanolic extract of Saptamrita loha vati At 254 nm 0.04 (D Green) 0.07 (Green) 0.40 (D Green) -
At 366 nm 0.07 (F Violet) 0.16 (F L Violet) 0.42 (F Violet)
At 540 nm 0.07(Brown) 0.14(L brown) 0.42(Brown)
After derivatisation 0.04 (Grey) 0.07 (Grey) 0.14 (L Violet) 0.22 (L Violet) -
0.72 (L Green) 0.78 (Green) -
0.54 (F Blue) 0.66 (F Blue) 0.72 (F Blue) 0.76 (F Violet) 0.78 (F L Violet) 0.87 (F Green)
-
0.42 (Brown) 0.45 (L Violet) 0.54 (L Violet) 0.64 (L Violet) 0.69 (L Violet) 0.76 (Pink) 0.83 (L Violet) 0.87 (L Violet)
L-Light, D-Dark, F-Fluorescence
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Figure 2:
HPTLC photo documentation of ethanol extract of Saptamrita loha vati
Under long UV
|Under short UV
Under white light
Under white light post derivatisation
TRACK 1- Saptamrita loha vati – 3 µl; TRACK 2 - Saptamrita loha vati – 6 µl; TRACK 3 - Saptamrita loha vati – 9 µl
Figure 3: HPTLC photo documentation of ethanol extract of Saptamrita loha vati densitogram of ethanol extract of Saptamrita loha vati
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CONCLUSION Standardization is very important aspect of every pharmaceutical preparation, despite the advent of modern technology in standardization of compound formulations, only a few Ayurvedic herbomineral medicines are standardized so far. The well-known compound formulation Saptamrita loha in the form of vati will be better accepted in terms of palatability and standardized dosage form. Addition of honey and ghee in the tablet preparation enhances palatability and hence better acceptability. It also did not affect the shelf life of the tablet. The physicochemical characterization would serve as preliminary test for the standardization of the formulation. SL showed peaks coinciding with reference standards of the individual ingredients. The unique Rf values, densitometric scan and densitogram obtained at different wavelengths pre- and post-derivatisation can be used as fingerprint to identify the herbo-mineral
preparation Saptamrita loha. The current investigation can be used as standardization test for other compound formulation for haematinic action. Further, detailed macro and microscopic examination of the raw drug as whole and powder forms in comparison with the formulation would add to the standardization tests of the saptamrita loha. ACKNOWLEDGEMENTS Authors are highly grateful to our revered President, Dr. D. Veerendra Heggade and Dr. B. Yashoverma, Secretary, SDM Educational Society for the encouragement. Authors highly regard the constant support of Dr. Prasanna N Rao, Principal, Dr Shailaja U, HOD Kaumarabhritya, Dr. Girish K J, Research coordinator SDM College of Ayurveda, Hassan. Authors thank Dr. B. Ravishankar, Director, SDM Centre for Research in Ayurveda and Allied Sciences for providing the facilities and Mr. Puneeth for help in carrying out the studies.
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Parashurama shastri (editor). (2013), Sharngadhara samhita of Sharngadhara with Deepika commentary of Adamalla and Gudartha deepika of Kashiram vaidya, Chaukhambha Krishnadas academy, Varanasi: 1:52 pp13. Pharmacopoeia, C. (2013). Commission. British Pharmacopoeia. Reena, K., Abhimanyu, K., & Kumar, K. N. S. (2013). Formulation and standardization of Medhya Rasayana–A novel Ayurvedic compound nootropic drug. Pharmacognosy Journal, 5(2), 72–76. Sai Prasad, A. J. V, Ratna Manikyam B., Trimurtulu. G., Reddy, K. N., Naidu, M. L. (2014) Analytical Standardization of Ayurvedic FormulationAqueous extracts of Hedichium spicatum Ham.Ex Smith, Sassurea lappa C.B.Clarke, Emblica officinalis Gaertn and Curcuma
Source of Support: NIL
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