International Journal of Current Research and Review
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IJCRR - 6(9), May, 2014

Pages: 18-29

Date of Publication: 12-May-2014


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VALIDATION OF NEWLY FORMULATED LAPORTEA ARISHTA BY USING DIFFERENT ANALYTICAL METHODS

Author: Deepa Deepa Deepa Deepa Deepa Deepa P., Seena

Category: General Sciences

Abstract:PurposPurposPurposPurposPurposPurpose: Traditional medicines have nurtured the knowledge of natural remedies against diseases since ages. Growing awareness about harmful side effects of modern medicine has led to interest in Ayurveda. Method: Laporteainterrupta (L) Chew of family Urticaceae having common name Hawaii woodnettle is an herb having many traditional uses. physicochemical analysis of crude drug, Laporteainterrupta-leaves were performed,Laporteainterrupta leaves extracted in soxhlet apparatus using ethyl acetate, petroleum ether, chloroform, methanol and water for identification of constituents by qualitative phytochemical analysis (tests for protein, carbohydrates, phenols and tannins, flavanoids, saponins, glycosides, steroids, terpenoids and alkaloids) and quantitative phytochemical analysis (total phenol content and total flavanoid content). The collected leaves of Laporteainterrupta was used to prepare arishta. After preparing the arishta the organoleptic characteristics (color, odor, taste and appearance), the physicochemical analysis ( pH, acid value, alcohol content, total solid content, viscosity and refractive index), quantitative analysis (alcohol content by spectrophotometry by dichromate method, total reducing sugar, total phenol content and total flavanoid content), anti-oxidant activity determination ( Ferric Thiocyanate method, Thiobarbituric acid method and Total Antioxidant Activity by FRAP method) were performed. LCMS was performed for determining various constituents (Qualitative analysis) and standardization of formulation was done by using UV and HPLC. The above formulated LAPORTEA ARISHTA was validated by using different analytical LAPORTEA ARISHTA was validated by using different analytical LAPORTEA ARISHTA was validated by using different analytical LAPORTEA ARISHTA was validated by using different analytical LAPORTEA ARISHTA was validated by using different analytical LAPORTEA ARISHTA was validated by using different analytical LAPORTEA ARISHTA was validated by using different analytical LAPORTEA ARISHTA was validated by using different analytical LAPORTEA ARISHTA was validated by using different analytical LAPORTEA ARISHTA was validated by using different analytical LAPORTEA ARISHTA was validated by using different analytical LAPORTEA ARISHTA was validated by using different analytical LAPORTEA ARISHTA was validated by using different analytical LAPORTEA ARISHTA was validated by using different analytical LAPORTEA ARISHTA was validated by using different analytical LAPORTEA ARISHTA was validated by using different analytical LAPORTEA ARISHTA was validated by using different analytical LAPORTEA ARISHTA was validated by using different analytical LAPORTEA ARISHTA was validated by using different analytical LAPORTEA ARISHTA was validated by using different analytical LAPORTEA ARISHTA was validated by using different analytical LAPORTEA ARISHTA was validated by using different analytical LAPORTEA ARISHTA was validated by using different analytical LAPORTEA ARISHTA was validated by using different analytical LAPORTEA ARISHTA was validated by using different analytical LAPORTEA ARISHTA was validated by using different analytical LAPORTEA ARISHTA was validated by using different analytical LAPORTEA ARISHTA was validated by using different analytical LAPORTEA ARISHTA was validated by using different analytical LAPORTEA ARISHTA was validated by using different analytical LAPORTEA ARISHTA was validated by using different analytical LAPORTEA ARISHTA was validated by using different analytical LAPORTEA ARISHTA was validated by using different analytical LAPORTEA ARISHTA was validated by using different analytical LAPORTEA ARISHTA was validated by using different analytical LAPORTEA ARISHTA was validated by using different analytical LAPORTEA ARISHTA was validated by using different analytical LAPORTEA ARISHTA was validated by using different analytical LAPORTEA ARISHTA was validated by using different analytical LAPORTEA ARISHTA was validated by using different analytical LAPORTEA ARISHTA was validated by using different analytical LAPORTEA ARISHTA was validated by using different analytical LAPORTEA ARISHTA was validated by using different analytical LAPORTEA ARISHTA was validated by using different analytical LAPORTEA ARISHTA was validated by using different analytical LAPORTEA ARISHTA was validated by using different analytical LAPORTEA ARISHTA was validated by using different analytical LAPORTEA ARISHTA was validated by using different analytical LAPORTEA ARISHTA was validated by using different analytical LAPORTEA ARISHTA was validated by using different analytical LAPORTEA ARISHTA was validated by using different analytical LAPORTEA ARISHTA was validated by using different analytical LAPORTEA ARISHTA was validated by using different analytical LAPORTEA ARISHTA was validated by using different analytical LAPORTEA ARISHTA was validated by using different analytical LAPORTEA ARISHTA was validated by using different analytical LAPORTEA ARISHTA was validated by using different analytical LAPORTEA ARISHTA was validated by using different analytical LAPORTEA ARISHTA was validated by using different analytical LAPORTEA ARISHTA was validated by using different analytical LAPORTEA ARISHTA was validated by using different analytical equipmentequipmentequipmentequipmentequipmentequipmentequipmentequipmentequipment's as per ICH guidelines.guidelines.guidelines.guidelines.guidelines.guidelines.guidelines.guidelines.guidelines.guidelines.guidelines. Results: Results: All results are coming between standard ranges. results are coming between standard ranges. results are coming between standard ranges. results are coming between standard ranges. results are coming between standard ranges. results are coming between standard ranges. results are coming between standard ranges. results are coming between standard ranges. Conclusion: Conclusion: Conclusion: Laporteainterrupta (L) Chew of family Urticaceae found to have number of active constituents from LCMS reports, which may can take as a proof for its traditional uses. Laporteainterruptaleaves have been successfully formulated and determined its anti-oxidant activity and validated using HPLC.

Keywords: Arishta, Physicochemical analysis, organoleptic characteristics

Full Text:

INTRODUCTION
Herbal medicine (or "Herbalism") is the study and use of medicinal properties of plants. They have the ability to synthesize a wide variety of chemical compounds that are used to perform important biological functions, and to defend against attack from predators such as insects, fungi and herbivorous mammals. Ayurveda - The Natural Way of Treating Disease Ayurveda is a form of alternative medicine that uses different parts of herbs, plants and parts of animals to cure diseases and ailments. Arishta are medicinal preparations made by soaking the drugs in coarse powder form or in the form of decoction (Kashaya), in a solution of sugar or jaggery, as the case may be, for a specified period of time, during which it undergoes a process of fermentation generating alcohol, thus facilitating the extraction of the active principles contained in the drugs. Laportea interrupta (L) Chew of family Urticaceae having common name Hawaii woodnettle is an herb having many traditional uses like Allergies, Alopecia, Amenorrhea, Arthritis, Asthma, Bedwetting/incontinence, Female Hormones, Fibromyalgia, Libido, Longevity/tonics, Menorrhagia, Nutrition, Osteoporosis, PMS, Prostate, Rheumatoid arthritis. Flavanoids have been reported to exert wide range of biological activities like anti-inflammatory, antibacterial, antiviral, anti-allergic, cytotoxic antitumor, treatment of neurodegenerative diseases, vasodilatory action. Standardization is the development and implementation of concepts, doctrines, procedures and designs to achieve and maintain the required levels of compatibility, interchangeability or commonality in the operational, procedural, material, technical and administrative fields to attain interoperability. Validation is documenting that a process meets its pre-determined specifications and quality attributes the objective of validation of an analytical procedure is to demonstrate that it is suitable for its intended purpose. Typical validation characteristics which should be considered are accuracy, precision, repeatability, intermediate precision, specificity, detection limit, quantisation limit, linearity, range.

MATERIALS AND METHOD
Collection of plant material: The leaves of Laportea interrupta were collected and washed well then dried under shade and coarsely powdered. Physicochemical analysis of crude drug: Total ash value, Acid insoluble ash, Water insoluble ash, Sulphated ash, Moisture content. Extraction of plant material: The plant material was dried and coarsely powdered and then extracted in Soxhlet Apparatus using 250ml of ethyl acetate (70-80°c), 250ml of Petroleum Ether (60-80°c), 250ml of Chloroform (50.5-61.5°c ), 250ml of methanol (60-70°c) and 250ml of Distilled water (0.25%) by continuous hot percolation according to their polarity. (Harborne, J.B., 2007). Qualitative phytochemical analysis: The extracts were subjected to qualitative tests for identification of phytochemical constituents such as proteins ( Million’s test and Ninhydrin test), carbohydrates ( Fehling’s test, Benedict’s test, Molisch’s test and Iodine test), phenols and tannins (Ferric Chloride test and Lead Acetate test), flavanoids ( Shinoda test and Alkaline Reagent test), saponins, glycosides (Liebermann’s test, Salkowski’s test and Keller-Kilani), steroids, terpenoids, alkaloids ( Mayer’s test and Wagner’s test) present in it. (RNS Yadev et al. 2011) . Quantitative phytochemical analysis Total Phenol Content: The amount of phenol in the ethyl acetate extract was determined by Folinciocalteu reagent method with some modification and then absorbance were measured at 765nm. Total Flavanoid Content: The amount of phenol in the ethyl acetate extract was determined by Aluminium chloride colorimetric method with some modification and then absorbance were measured at 420nm.

Preparation of formulation: An earthen pot sufficiently large and strong is chosen. The proportion of the different ingredients are 32 seers of distilled water, boiled and added 1.25 seers dried plant leaf powder jaggery 12.5 seers, honey 6.25 seers and sugar 6.25 seers, mixed well to form a uniform solution and pour into it . Pot is buried in a pit made in the soil for 30 days. Physicocemical analysis of formulation: Determination of PH of formulation, Determination of acid value, Determination of alcohol content, Total solid content, Determination of viscosity of formulation, Determination of refractive index. Quantitative analysis of formulation Alcohol content by spectrophotometry (Dichromate Method): Alcohol was estimated by caputi et al (1968) and measured the absorbance at 620nm. Total reducing sugar: Total phenol content: The amount of phenol in the formulation was determined by Folin-ciocalteu reagent method with some modification and then absorbance were measured at 765nm and the results were determined from the standard curve and expressed in gallic acid equivalents (mg/g of extracted compound).. Total flavanoid content: The amount of phenol in the formulation was determined by Aluminium chloride colorimetric method with some modification and then absorbance were measured at 420nm and the results were determined from the standard curve and expressed in Quercetin equivalents (mg/g of extracted compound) Anti-oxidant activity determination of formulation: Ferric Thiocyanate (FTC) method, Thiobarbituric Acid (TBA) method, Total Antioxidant activity by FRAP method. Qualitative analysis of formulation by LCMS: Specifications of LCMS are LC Column: REVERSE PHASE C-18, Pump: SPD 10 AVP, Mobile Phase: 2% OPA IN WATER: METHANOL (50:50), Ionization Mode: ELECTRONIC SPRAY IONIZATION, Mode: BOTH POSITIVE, Injection Volume: 10 µL, Flow Rate: 2 ML/MIN, Column Temperature: 250°C, Column: PHENOMENEX RP 18, Column Dimension: 25CM 2.5 MM,LC Detection: 254NM,m/z range: 50- 1000,Soft Ware: CLASS V P INTEGRATED AND Library: METWIN 2.0. Standardisation of formulation using UV: Specifications of UV are Model: ELICO SL 164, Instrument: UV-Visible Double Beam Spectrophotometer, Wavelength Range: 190-999.9 nm, Accuracy: +/- 0.5nm, Repeatability: +/- 0.2nm, Resolution: 0.1nm, Bandwidth: 0.5-6.0 nm (Variable at an Interval of 0.1nm), Photometric range: -2.5 TO +2.5 Abs, Accuracy: +/- 0.005 Abs at 0.1Abs from 200-850nm, Repeatability: +/- 0.002 Abs at 1.0 Abs from 200-850nm, Stray light: less than 0.05% at 220-340nm,Base line correction: Automatic Base Line Correction, Scan speed: Slow, Medium and Fast, Data interval: Depend Upon Wavelength, Sample holder: 5 Position, Automatic positioning far 10mm and Sample Cuvette and Fixed position for Reference Cuvette, Source: Tungsten- Halogen lamp (310-999.9nm) and Deuterium lamb with Quartz window (190-340nm) and Detectors: Photomultiplier Tube (PMT).Validation experiments were performed to demonstrate linearity, precision, accuracy, robustness, ruggedness, LOD and LOQ as per ICH guidelines. Standardisation of formulation using HPLC: Specification of HPLC is Instrument: Schimadzu, LC Column: Reverse Phase C-18 Phenomenex C18, Pump: SPD 10 AVP, Mobilephase: Acetonitrile: Methanol (40:60), Injection volume: 10 µl, Flow Rate: 2.0 ml/min, Column Temperature: 25°C, Column: Phenomenex RP 18, Column Dimension: 5cm 1.5 cm,LC detection: 254nm and Soft Ware: Class V P Integrated.Validation experiments were performed to demonstrate system suitability, linearity, precision, accuracy study and robustness as per ICH guidelines.











 

CONCLUSION
Laportea interrupta (L) Chew of family Urticaceae found to have number of active constituents from LCMS reports, which may can take as a proof for its traditional uses. Laportea interrupta leaves have been successfully formulated and determined its anti-oxidant activity and validated using UV and HPL

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A Study by Jabbar Desai et al. entitled "Prevalence of Obstructive Airway Disease in Patients with Ischemic Heart Disease and Hypertension" is awarded Best Article for Vol 12 issue 17
A Study by Juna Byun et al. entitled "Study on Difference in Coronavirus-19 Related Anxiety between Face-to-face and Non-face-to-face Classes among University Students in South Korea" is awarded Best Article for Vol 12 issue 16
A Study by Sudha Ramachandra & Vinay Chavan entitled "Enhanced-Hybrid-Age Layered Population Structure (E-Hybrid-ALPS): A Genetic Algorithm with Adaptive Crossover for Molecular Docking Studies of Drug Discovery Process" is awarded Best article for Vol 12 issue 15
A Study by Varsha M. Shindhe et al. entitled "A Study on Effect of Smokeless Tobacco on Pulmonary Function Tests in Class IV Workers of USM-KLE (Universiti Sains Malaysia-Karnataka Lingayat Education Society) International Medical Programme, Belagavi" is awarded Best article of Vol 12 issue 14, July 2020
A study by Amruta Choudhary et al. entitled "Family Planning Knowledge, Attitude and Practice Among Women of Reproductive Age from Rural Area of Central India" is awarded Best Article for special issue "Modern Therapeutics Applications"
A study by Raunak Das entitled "Study of Cardiovascular Dysfunctions in Interstitial Lung Diseas epatients by Correlating the Levels of Serum NT PRO BNP and Microalbuminuria (Biomarkers of Cardiovascular Dysfunction) with Echocardiographic, Bronchoscopic and HighResolution Computed Tomography Findings of These ILD Patients" is awarded Best Article of Vol 12 issue 13 
A Study by Kannamani Ramasamy et al. entitled "COVID-19 Situation at Chennai City – Forecasting for the Better Pandemic Management" is awarded best article for  Vol 12 issue 12
A Study by Muhammet Lutfi SELCUK and Fatma entitled "Distinction of Gray and White Matter for Some Histological Staining Methods in New Zealand Rabbit's Brain" is awarded best article for  Vol 12 issue 11
A Study by Anamul Haq et al. entitled "Etiology of Abnormal Uterine Bleeding in Adolescents – Emphasis Upon Polycystic Ovarian Syndrome" is awarded best article for  Vol 12 issue 10
A Study by entitled "Estimation of Reference Interval of Serum Progesterone During Three Trimesters of Normal Pregnancy in a Tertiary Care Hospital of Kolkata" is awarded best article for  Vol 12 issue 09
A Study by Ilona Gracie De Souza & Pavan Kumar G. entitled "Effect of Releasing Myofascial Chain in Patients with Patellofemoral Pain Syndrome - A Randomized Clinical Trial" is awarded best article for  Vol 12 issue 08
A Study by Virendra Atam et. al. entitled "Clinical Profile and Short - Term Mortality Predictors in Acute Stroke with Emphasis on Stress Hyperglycemia and THRIVE Score : An Observational Study" is awarded best article for  Vol 12 issue 07
A Study by K. Krupashree et. al. entitled "Protective Effects of Picrorhizakurroa Against Fumonisin B1 Induced Hepatotoxicity in Mice" is awarded best article for issue Vol 10 issue 20
A study by Mithun K.P. et al "Larvicidal Activity of Crude Solanum Nigrum Leaf and Berries Extract Against Dengue Vector-Aedesaegypti" is awarded Best Article for Vol 10 issue 14 of IJCRR
A study by Asha Menon "Women in Child Care and Early Education: Truly Nontraditional Work" is awarded Best Article for Vol 10 issue 13
A study by Deep J. M. "Prevalence of Molar-Incisor Hypomineralization in 7-13 Years Old Children of Biratnagar, Nepal: A Cross Sectional Study" is awarded Best Article for Vol 10 issue 11 of IJCRR
A review by Chitra et al to analyse relation between Obesity and Type 2 diabetes is awarded 'Best Article' for Vol 10 issue 10 by IJCRR. 
A study by Karanpreet et al "Pregnancy Induced Hypertension: A Study on Its Multisystem Involvement" is given Best Paper Award for Vol 10 issue 09

List of Awardees

A Study by Ese Anibor et al. "Evaluation of Temporomandibular Joint Disorders Among Delta State University Students in Abraka, Nigeria" from Vol 13 issue 16 received Emerging Researcher Award


A Study by Alkhansa Mahmoud et al. entitled "mRNA Expression of Somatostatin Receptors (1-5) in MCF7 and MDA-MB231 Breast Cancer Cells" from Vol 13 issue 06 received Emerging Researcher Award


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International Journal of Current Research and Review (IJCRR) provides platform for researchers to publish and discuss their original research and review work. IJCRR can not be held responsible for views, opinions and written statements of researchers published in this journal

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