Online : ISSN 2349-669X J.Pharm.Chem CODEN:JPCOCM Journal ...jpc.stfindia.com/images/current_issue/JPC 13_1.pdf · Development and Validation of Reverse Phase Hplc Method for Estimation

J.Pharm.Chem CODEN: JPCOCM

Journal of Pharmacy and Chemistry(An International Research Journal of Pharmaceutical and Chemical Sciences)

Indexed in Chemical Abstract and Index Copernicus (IC Value 5.28)

www.stfindia.comwww.jpc.stfindia.com

Editor-in-chiefProf. K.N. JAYAVEERA

Jawaharlal Nehru Technological University Anantapur,Anantapur, Andhra Pradesh -515001.

Associate EditorDr. K.V. Madhusudhan

Executive EditorDr. K. Balaji

Editorial Board

Dr. B.M. Vrushabendra Swamy Dr. A. Venkateshwar Reddy Dr. G. S. Kumar Dr. G. Madhu Dr. S. Subramanyam Dr. K. Yogananda Reddy

Dr. K. Bhaskar Reddy Dr. E. Sreedevi Dr. K.C. Chaluvaraju

Editorial Advisory Board

Prof. Nagarapu Lingaiah India Prof. G. Krishna Mohan India

Prof. T.R. Rao India Prof. M.L.N.Rao India

Prof. R.Nageshwara Rao India Prof. S. Srihari India

Prof. K.V.S.R.G. Prasad India Prof. Y. Rajendra Prasad India

Prof. K. Kannan India Prof. Yeoh Peng Nam IMU, Malaysia

Prof. D.R. Krishna U.S.A Prof. K.C.Naidu India

Prof. Jonathan R Dimmock Canada Prof. Ananth. P. Haridas India

Prof. Helton Max M. Santos Portugese Prof. Damaris Silveira Brazil

Prof. Mustafa Iraz Turkey Prof. Abdul Naser B Singab Egypt

Prof. Ali Asgarh hemmati Iran Prof. N. Devanna India

Prof. K.R.S. Sambasiva Rao India Prof. R. Shyam Sunder India

Dr. Nitin Mahukar India Prof. Arun Goyal India

Prof. Sarangapani India Prof. Sunil K. Khare India

Prof. Y. Narasimha Reddy India Dr. S. Narasimha Murthy U.S.A

Dr. Girish Gowda Saudi Arabia Dr. K. Suresh Babu India

Online : ISSN 2349-669XPrint : ISSN 0973-9874

January - March 2019 2 Journal ofPharmacyandChemistry •Vol.13 • Issue.1

ISSN 0973 – 9874 J.Pharm.Chem CODEN: JPCOCM

Journal of Pharmacy and Chemistry(An International Research Journal of Pharmaceutical and Chemical Sciences)

Volume 13 • Issue 1 • January – March 2019

F

VIEWS

The views and opinions expressed in this journal are those of the contributors; Science-Tech Foundation does not necessarily concur with the same. All correspondence should be addressed to the Editor-In-Chief (Hon.), Journal of Pharmacy and Chemistry (Science-Tech Foundation), Plot No 22, Vidyut Nagar, Anantapur - 515 001, Andhra Pradesh, India. • e-mail:[email protected]. Send your queries at www.jpc.stfindia.com, www.stfindia.com

CONTENTS

DevelopmentAndValidationOfReversePhaseHplcMethodForEstimationOfGatifloxacin InBulkDrugAndMicroparticle Loaded InSituGel .................................................................... 3

MADHURI PUJARI, SWAPNA B, GEETHA M AND VIJAYA G JOSHI

FormulationandEvaluationofPregabalinExtendedReleaseTablet ................................................................ 10

M. VAMSHI KRISHNA, DR.P.SATYA NARAYANA RAO, DR.K.YOGANANDA REDDY, A.NIRANJAN, K.PRAGATHI AND N.AKHILA

ColourPropertiesOfCatechuHeartwoodExtract .................................................................................................. 17

SWAPNIL Y. JADHAO AND JAYARAJ RAO

AReviewOnPathogenesisOfObesity .................................................................................................................... 27

DR. C. GIRISH AND CH. JL PRASANNA

Instruction to Authors

Online : ISSN 2349-669XPrint : ISSN 0973-9874


Development and Validation of Reverse Phase Hplc Method for Estimation of Gatifloxacin in Bulk Drug and Microparticle Loaded

In Situ Gel

Madhuri Pujari1, Swapna B2, Geetha M3*, Vijaya G Joshi4

1, 3, 4 Department of Pharmaceutics, Government College of Pharmacy, Bengaluru-560027, Karnataka, India2, Department of Pharmacology, Government College of Pharmacy, Bengaluru-560027, Karnataka, India

ABSTRACT

A reverse phase HPLC method was developed for the estimation of gatifloxacin in bulk drug and microparticle loaded in-situ gel formulation. The optimized conditions of chromatographic system include column Agilent ZORBAX SB-C18 (5µm, 4.5×250mm), isocratic pump mode with UV detection at 285nm, mixture of water:acetonitrile (75:25)v/v as mobile phase and a flow rate of 1ml/min. The system suitability tests were done before starting the validation as per ICH guidelines and parameters were found within the acceptance criteria. The method exhibited retention time of 4.6 min and linearity between the concentration range of 5 and 35µg/ml with a correlation coefficient of 0.999 which indicated best fit line. LOD and LOQ values were found to be 0.618 and 1.874µg/ml respectively. The calculated %RSD values for intra-day, inter-day, method precision and repeatability were found to be within the limits of ˂2%, which indicated that the method is precise and rugged. The % recovery ranged between 76.7 to 119.4 % showed that the method is accurate. Robustness was indicated by negligible variations in assay values with deliberate variations in flow rate, mobile phase ratio and detection wavelength. All the above mentioned findings indicated that the proposed method was proved to be specific, simple, fast, sensitive, precise and robust. Hence, the method was successfully used for the estimation of gatifloxacin in micro-particle loaded in-situ gel preparation.Keywords: Reverse phase, HPLC, Gatifloxacin, Microparticle loaded in situ gel

*Correspondance : E-mail : [email protected]

Introduction Fluoroquinolones are one of the promising groups of anti-bacterial agents widely used in the treatment of ocular infections [1]. Fourth generation fluoroquinolones have been developed with an intention to reduce the problem of emerging bacterial resistance and to generate therapeutic moieties with a broad spectrum of activity [2]. In addition, these new generation fluoroquinolones show superior ocular penetration as compared to previous generation fluoroquinolones [3]. Gatifloxacin (GTX) is a member of fourth generation fluoroquinolone antibiotic family. It contains a substitution of a methoxyl group at position 8 of the quinolone ring, which allows for simultaneous inhibition of both DNA gyrase and topoisomerase IV in gram-positive bacteria. This structural modification is responsible for increased potency of GTX against gram-positive bacteria while maintaining the broad spectrum of gram-negative activity observed with older quinolones. It is commercially available in the form of an eye drops and ointment. It is reported that the ocular use of gatifloxacin is safe and effective against a broad spectrum of bacteria, including intracellular bacteria and anaerobes [4,5].

GTX has a molecular weight of 375.39 Da, logP value of 2.6 and solubility is pH dependent [6]. Chemically, 1-cyclopropyl-6-fluoro-8-methoxy-7-(3-methylpiperazin-1-yl)-4-oxo-1,4-dihdroquinoline-3-carboxylic acid [7].

Fig.1: Structure of Gatifloxacin Based on the literature survey, various analytical methods have been reported for the estimation of GTX in bulk, pharmaceutical dosage forms and biological fluids [8] which include, UV spectrophotometric method for GTX [9] and in combination [10-15], HPLC method for

N

NH

CH3

OCH3

F

N

O O

OH


GTX [16-18] and also in combination with other drugs [19], spectroflurimetric method [20], micellar liquid chromatography [21], stability indicating HPLC method [22]. However, no specific HPLC method has been reported for the estimation of GTX in microparticle loaded in situ gel. Hence an attempt was made to develop a simple, rapid, sensitive and precise HPLC method for the estimation of GTX in bulk drug and microparticle loaded in situ gel.MethodsReagents Gatifloxacin was obtained as gift sample from Micro labs Pvt Ltd Bengaluru, India. HPLC grade acetonitrile was purchased from Fischer scientific chemicals. HPLC grade water was prepared using Milli-Q water purification system. Orthophosphoric acid and triethylamine used were of AR grade and were purchased from Merck specialities Pvt Ltd and Thomas Baker chemicals Pvt Ltd, Mumbai respectively. Class A glassware was used throughout the experiment.Instrumentation The method development was carried out using Shimadzu LC (Grace Smart RPc18 model) which consisted of a column Agilent ZORBAX SB-C18 (5µm, 4.5×250mm), isocratic pump mode.Chromatographic conditions Flow rate of 1 mL/minute, UV detector wavelength at 285nm and the injection volume 20µl. The mobile phase selected was water and acetonitrile in the ratio of 75:25. All solutions were degassed by ultra-sonication. Mobile phase was filtered through 0.45µ nylon filter.Preparation of standard stock solution Accurately 10mg of GTX was weighed, transferred in to a clean and dry 10mL volumetric flask and dissolved in sufficient amount of mobile phase. The final volume was made up to the mark using mobile phase to get concentration of 1000µg/mL of GTF (SS I). Further 1ml of SS-I was taken in 10ml volumetric flask and the volume was made up using mobile phase (SS II- 100 µg/mL).

Mobile phase A mixture of water and acetonitrile in the ratio of 75:25 v/v was used in the preparation of mobile phase, to this 0.35ml of tri-ethylamine was added and the pH was adjusted to pH-3.5 using dilute ortho-phosphoric acid. The above prepared mixture was used as mobile phase and was filtered through 0.45µ nylon membrane filter and degassed by 30min sonication.Preparation of sample stock solution The microparticle loaded in situ gel formulation equivalent to 3mg of GTX was dispersed in 3 mL mobile phase, sonicated for 10 min to extract the drug, filtered and the concentration was found to be 1000µg/mL. Further the 0.2 mL of the above solution was diluted to 10 mL having concentration of 20 µg/mL. The solution was filtered using 0.45µm membrane filter and degassed by sonication. The

sample was injected into the chromatograph in triplicate and the chromatogram was recorded. The developed method was validated for various parameters such as specificity, linearity, sensitivity, precision, accuracy and robustness.Specificity It was evaluated by injecting analytical placebo containing all ingredients of the formulation except the analyte which is prepared according to sample preparation procedure. To identify the interference by these excipients, a mixture of the inactive ingredients (placebo) was spiked with GTX and chromatogram was recorded. Linearity The linearity of the method was determined by multipoint calibration method consisting of seven different concentrations of analyte and the data was statistically analyzed by performing regression analysis. The standard dilutions of GTX were prepared by pipetting aliquots of 0.5mL, 1mL, 1.5mL, 2mL, 2.5mL, 3mL, and 3.5mL of standard stock solution-II in to 10mL volumetric flask and diluted to the volume with mobile phase to get a concentration between 5 to 35µg/mL. About 20µL of standard dilutions of GTX were injected in to the chromatograph. Retention time and peak area obtained were recorded and standard curve was plotted.Sensitivity LOD and LOQ of GTX were determined based on standard deviation of the response and slope. The LOD and LOQ were calculated as: LOD = 3.3σ/S and LOQ=10σ/S; where σ is the standard deviation of the lowest standard concentration and S is the slope of the curve. Precision Precision between a series of measurements expresses the closeness of agreement obtained under prescribed conditions of the same homogeneous sample from multiple sampling of an analytical method. The repeatability, intra-day precision, inter-day precision of the analytical method were assessed by analyzing the dilution containing 20 µg/ml of GTX (n=6). Peak area of all six replicates was determined and %RSD was calculated.Accuracy The accuracy of the proposed method was tested by determining the percentage recovery of the analyte in microparticles loaded situ gel formulation by assaying the known amounts of analyte added. It was carried out at three different levels LQC, MQC and HQC by spiking the formulation with standard drug substance at 80%, 100%, and 120% respectively. All the measurements were carried out in triplicate and the recovery was expressed in terms of analyte found in spiked sample.Robustness Robustness of the current method was studied by estimating the effect of deliberate changes such as variation


in mobile phase composition (water: acetonitrile, 73:27 and 77:23 v/v), flow rate (0.9-1.1mL/min) and detection wavelength (284-287nm) on analyte determination. The process of method development started with optimizing various chromatographic conditions in terms of solubility of GTX in different solvent systems, suitability of the method, cost, ease of preparation and applicability of the method to different purposes. Specifically, the mobile phase composition was optimized by trial and error basis using different solvent mixture systems with varying proportions, among which a mixture of water and acetonitrile in the ratio of 75:25 v/v was selected as suitable system with the

addition of triethylamine and the pH adjusted to 3.5 using orthophosphoric acid. Well resolved peak was obtained with the addition of triethylamine and orthophosphoric acid. Further, suitability of the method was tested by analyzing system suitability parameters viz theoretical plates, HETP and tailing factor; all these parameters found within the limits (table 1). The retention time of the drug was found to be 4.6 min (fig.2)Specificity is described as the ability of a method to discriminate the analyte from all interfering substances. From the fig.3 and 4, it is clear that the excipients used in the formulation did not interfere with the GTX peak

Table 1: System Suitability parameters

System suitability parameters

Gatifloxacin (concentration 20µg/mL)

Acceptance criteria

Theoretical plates 7052 ˃2000

HETP 154710 -

Tailing factor 1.230 ˂2

Fig.2: HPLC chromatogram of Gatifloxacin

Fig.3: Chromatogram of placebo


Fig.4: Chromatogram of GTX loaded microparticle loaded in-situ gel formulation

Table 2: Statistical data of calibration curve of GTX

Parameters Gatifloxacin

Linearity 5-35μg/ml

Regression equation y = 421897x + 34394

Correlation coefficient (R2) 0.9991

Limit of detection (LOD) (μg/ml) 0.619

Limit of quantification (LOQ) (μg/ml) 1.874


indicating specificity of the method. Linearity is important for the confirmation of method’s sensitivity for the analysis of analyte’s concentration with a defined range. The concentration range established was within 5-35 μg/mL (fig.5) and the linear regression model gave the R2 value of 0.999 which indicated good linear relationship between analyte concentration and the response, peak area. The developed HPLC method was highly sensitive to detect and determine the GTX content as the LOD and LOQ was found to be 0.619 and 1.874 μg/ml. The precision measurements were expressed in % RSD. The % RSD of repeatability (table

Table 3: Repeatability (n=6)

Parameters Gatifloxacin Concentration 20 μg/ml

Mean ± standard error 1084682 ± 1033.38

Standard deviation (SD) 2531.27

Relative standard deviation (%) 0.42

Table 4: Intra-day precision

GTX concentration 20 μg/mlTime(h) Area Retention time

1 1086440.17 4.49

2 1086419.67 4.47

3 1086041.83 4.48

4 1086051 4.45

5 1086011.5 4.41

6 1086151.17 4.44

Average 1086185.89 4.46

SD 194.87 0.029

% RSD 1.79 0.65

Table 5: Inter-day precision

GTX Concentration 20 μg/mlDay Area Retention time

Day 1 1078101.5 4.42Day 2 1070511 4.48Day 3 1088039 4.36Average 1078883.83 4.42SD 8790.15 0.058% RSD 0.81 1.30

3) and intermediate precision (inter-day, intra- day) (table 4 and 5) were found to be <2% which indicated the precision of the method as it complies with acceptable limits. The % recovery was determined by standard addition method and ranged between 76.7 to 119.4 % (table 6) showed that the method is accurate. The assay values of 91.50 to 104.06 % (table 7) with deliberate changes in set parameters of the method describe the robustness of the method. Therefore, the method was successfully adopted for the estimation of GTX in micro-particle loaded in-situ gel preparation


4. Schultz C. Ophthalmol Eye Dis 2012;4:65–70.5. Cervantes LJ, Mah FS. Clin Ophthalmol 2011;5:495.6. Pawar P, Katara R, Mishra S, Majumdar DK. Expert

Opin Drug Del 2013;10(5):691-711.7. Al-Dgither S, Alvi SN, Hammami MM. J Pharm

biomed anal 2006;41(1):251-5.8. Teng R, Tensfeldt TG, Liston TE, Foulds GJ.

Chromatogr B 1996;675:53-59.9. Sridhar S, Rayala K, Muvvala S. Asian J Pharm Clin

Res 2015;8(1):242-6.10. HRN Salgado and CLCG Oliveira. Die Pharmazie Int J

Pharm Sci 2005;60:263-64.11. Sayed RA, Hassan WS, El – Mammli MY, Shalaby AA.

Orient J Chem 2012;28: 639-50.12. El-Didamony AM, Abo-Elsoad MO, J Saudi Chem Soc

2013. http://dx.doi.org/10.1016/j.jscs.2013.10.00313. Sversut RA, Alcaˆ ntara IC, Rosa AM, Baroni ACM,

Rodrigues PO, Singh AK, Amaral MS, Kassab NM. Arab J Chem 2017;10:604–10.

14. Patel P, Patel D, Desai S, Meshram D. Int J Pharm Sci Invent 2014;3( 4).

15. Abdel-Razeq SA, Fouad MM, Darwish MK, Zaazaa

ConclusionThe RP-HPLC method for analysis of GTX in microparticle loaded in situ gel was successfully developed and validated in terms of validation parameters as per ICH guidelines. Hence, it can be concluded that the proposed method can be used for routine analysis of GTX in pharmaceutical dosage forms.

AcknowledgementThe authors are thankful to Microlabs Pvt limited, Bangalore for providing gatifloxacin as gift sample. The authors express deepest sense of gratitude to American Association of Government College of Pharmacy (AAGCPA) for providing monetary fund to carry out the research work.

References 1. Blondeay JM. Surv Opthalmol 2004;49(2):S73-8.2. Donnenfeld E, Perry HD, Chruscicki DA, Bitterman

A, Cohn S, Solomon R. Curr Med Res Opin 2004;20(11):1753-8.

3. Duxfield L, Sultana R, Wang R, Englebretsen V, Deo S, Rupenthal ID, Al-Kassas R. Drug Dev Ind Pharm 2016;42(1):1-11.

Table 6: Results of accuracy (Recovery Studies) (N=3)

Sl.

No

Percentage level of stan-

dard

Peak area SD %RSD Sample con-centration

(µg/mL)

Std concen-tration

µg/mL

Amount of standard (µg/mL)

Recovery (%)

Spike Found

1 80% 1316948.96 6429.173 0.4881 10 6 16 15.338 76.7%

2 100% 1706552.0 2581.640 0.1512 10 10 20 19.876 99.3%

3 120% 20514417 4704.249 0.2293 10 14 24 23.892 119.4%

Table 7: Results of robustness

Change in Assay (%)

Flow rate

0.9 100.50

1.1 91.81

Wavelength

287 107.26

283 91.50

Mobile phase ratio(water:acetonitrile)

(73:27) 104.06

(77:23) 96.43


HE, Nasr ZA. Indo Am J Pharm Res 2015:5(09).3048-5716. Rao AL, Ravi Kumar BNV, Girija Sankar G. J Pharm

Res Health care 2011;3(3):72-76.17. Salgado HR , Lopes CC. J AOAC Int 2006;89(3):642-

5.18. Bera AK , De AK, Pal B. Int J Pharm Scu Res

2014;5(9):3741-45.19. Kashyap R , Srinivasa U. Int J Drug Dev Res

2014;6(1):99-112.

20. Venugopal K, Movva S, Saha RN. Ind J Pharm Sci 2006;68(6):726-30.

21. El Gammal R, Hammoudal MEA, El-Wasseef DR, El-Ashry1 SM. J Chromatogr Sci 2018;56(4):367–74.

22. Grace1 AC, Prabha1 T, Jagadeeswaran M, Srinivasan K, Sivakumar T. J Pharm Sci Res 2019;11(5):1814-20.


Formulation and Evaluation of Pregabalin Extended Release Tablets

M. VAMSHI KRISHNA*, DR.P.SATYA NARAYANA RAO, DR.K.YOGANANDA REDDY, A.NIRANJAN, K.PRAGATHI N.AKHILA.

Simson Life Sciences, Shed No. 6A, TYPE III, TSIIC, Prashanthi Nagar,

Kukatpally, Hyderabad, Telangana, INDIA, Pin: 500072.

ABSTRACT

The current paper was an attempt to design a extended release dosage form using various grades of Hypromellose and Povidone as extended release polymers.

Laboratory scale batches of nine tablet formulations were prepared by direct compression micromeritics properties of the Powder was evaluated prior to compression. Tablets were characterized as crushing strength, friability, weight variation, thickness, drug content and evaluatedforin-vitroreleasepatternfor24hoursin0.06NHClat37±0.5°C.Thein-vitrorelease mechanism was evaluated by kineticmodeling.

The results obtained revealed that Hypromellose 100000, Hypromellose 15000 at a concentration of26%informulation(F6)wasabletoExtendedthedrugreleasefor24handfollowedtheHiguchi pattern quasi-Fickian diffusion. With that, combined effect of Hypromellose 100000, Hypromellose 15000 as Dissolution data were compared with innovator for similarity factor (f2), andexhibitedanacceptablevalueof≥50.Threeproductionvalidationscalebatchesweredesigned based on lab scale best batch and charged for stability testing, parameters were within the limitof acceptance. There was no chemical interaction found between the drug and excipients during compatibility studies conducted at 40°C/75%RH and 25°C/60%RH for1month.

Hence, combination of Hypromellose 100000 and Hypromellose 15000 was compatible with PregabalinAPIandthesetwopolymersatasuitableconcentrationwaseffectivelyusedtoachieve desired Dissolutionprofile.

Keywords: Pregabalin, Micromeritics properties, Extended release tablet and Dissolution.

*Corresponding address: [email protected]

INTRODUCTION

TheInternationalAssociationfortheStudyofPaindefinesneuropathicpain(NP)aspain“initiated or caused by a primary lesion or dysfunction in the nervous system.” The characteristic abnormality in NP is a lesion or dysfunction of the sensory pathway, with associated hyper excitability of the denervatedzone.

The pain can be of high intensity and long duration and can have a profound effect on the mood, personality, and social relationships of a person. Long duration of pain causes concomitant depression, fatigue, anxiety, sleep disturbances, and decreased overall physical functioning. Peripheral neuropathy has various etiologies, which also differ depending on the location of the nerve damage. A single mechanism can rarely be claimed for the development of signs and symptoms of NP.

Peripheral neuropathy is very common in type 2 diabetes mellitus (T2DM), with diabetic peripheral neuropathy

(DPN) affecting up to half of the patients suffering from diabetes. One study, conducted in the rural part of India, reported peripheral neuropathy (PN) as the most common complication of diabetes mellitus, affecting around 60% of patients. Apart from pain other important clinical manifestation of DPN is insensitivity, which increases the risk of burns, injuries, and foot ulcerations. Poor glycemic control, duration of diabetes, hyperlipidemia (particularly hypertriglyceridemia), elevated albumin excretion rate, and obesity have been implicated as risk factors for the development of DPN.

Thus, an attempt has been made to formulate the Extended-release tablets of Pregabalin tablets to treat neuropathic pain.

MATERIALS AND METHODS

Materials

Pregabalin API was procured from (Divi’s Laboratories Limited) Microcrystalline cellulose pH - 102(MCC PH-102) Hypromellose 100000, Hypromellose 15000, Povidone


K-30, Magnesium stearate All Excipients and solvents used were of pharmaceutical grade.

Method for preparation of tablets

Pregabalin, MCC PH-102, Hypromellose 100000, Hypromellose 15000, Povidone K-30 were passed through #30 mesh and collected separately in a polyethylene bag. Direct compression technique was applied for the batch preparation of Pregabalin tablets.All the materials were Mixed to Octagonal Blender and mixed for 20 min at optimized speed. Further lubricated with, Magnesium stearate (sifted through #60 mesh) for 5 min in Octagonal Blender. Finally, the lubricated blend was compressed to formulate tablets using a tablet compression machine with 22 mm × 10.80 mm almond-shaped punches. The compositions of various formulations designed in the present study are given in Table1.

Composition of Tablet formulation (mg)

Table 1

Ingredients F1 F2 F3 F4 F5 F6 F7

Intragranular Portion

Pregabalin 330mg 330mg 330mg 330mg 330mg 330mg 330mg

MCC PH 102 400mg 400mg 400mg 400mg 400mg 400mg 400mg

HPMC100000 100mg 125mg 150mg 175mg 200mg 210mg 215mg

HPMC15000 150mg 125mg 100mg 75mg 50mg 40mg 35mg

Povidone-K30 60mg 60mg 60mg 60mg 60mg 60mg 60mg

Extragranular portion

Magnesium

stearate10mg 10mg 10mg 10mg 10mg 10mg 10mg

Total weight 1050mg 1050mg 1050mg 1050mg 1050mg 1050mg 1050mg

Micromeritics properties of prepared granules

Prior to compression Blend was evaluated for their characteristic parameters such as Bulkdensity (BD) and tapped density (TD) were determined by Cylinder method, and Carr’s index (CI) was calculated using the followingequation

CI = (TD-BD)/TD × 100

Hausner’s ratio (HR) was calculated by the following equation

HR = TD/BD

Physical characterization of Extended release tablets

Thephysicalpropertiessuchascrushingstrength,friability,weightvariation,thickness,andassay of compressed tablet for each formulation were determined. Tablet crushing strength was determined for 10 tablets using digital tablet hardness tester (Electro lab) and the data reported is the mean of three individual determinations. Friability test was performed to assess the effect of frictionandshocks,whichmayoftencausethetablettochip,cap,orbreak.Preweighed,randomly selected twenty tablets were placed in a friability tester (Electro lab) and operated for 4 min at 25 rpm. Compressed tablets should not lose more than 1% of their weight. A weight variation test was performed according to United States

Pharmacopeia (USP) 43 NF38 on 20 tablets by taking samples from a batch after production of every 100 tablets and randomly from a total batch of 300 tablets using an electronic balance (Contech Instruments CA224, India).The thickness esof tablets was measured by Vernier calipers (Ocean). The drug content in terms of assay of each batch was determined in triplicate. Foreach batch, 20 tablets were weighed and crushed to fine pow derusing mortar and pestle. Anaccurately weighed 1050mg of the powder was taken and suitably dissolved in water and analyzed by High performance liquid chromatography (HPLC) after making appropriate dilutions. The procedure was carried out on Shimadzu LC-2030C (Ecosil) C18; 250 mm × 4.60 mm with flow rate of 1.0 mL/min at ambient temperature.


In-vitro dissolution studies

Release rate of all designed formulations were studied up to 24hrs.The procedure was determined using USP dissolution testing apparatus II (paddle method). The dissolution test was performed using 900 mL of 0.06N HCl at 37 ± 0.5°C and 50 rpm. A sample of 10 mL of the solution was withdrawn from the dissolution apparatus at 1hr intervals with the replacement of fresh dissolution medium for 24 hrs. The samples were passed through a 0.45-μm membrane filter and diluted to a suitable concentration with phosphate buffer.

Comparison of dissolution profile

The similarity factor (f2) given by scale up and post approval Changes guidelines for a modified release dosage form was used as a basis to compare dissolution profile. The dissolution profiles are considered to be similar when f2 is between 50-100 and dissimilarity factor (f1) lies between 0 to 15. The dissolution profiles of product were compared to Innovator using f2 and f1 whichare calculated from the followingformulae,

Where, n is the number of dissolution sample times and Rt and Tt are the individual or mean percent dissolved at each time point, t, for the innovator and test dissolution profiles respectively.

Statistical analysis

Analysis of variance followed by Tukey's test was used for

statistical comparison of the data. Significance level was fixed at P < 0.05

RESULTS AND DISCUSSION

Micromeritics properties of granules

Result shows that all the formulations produced optimal flow properties calculated in terms of compressibility. Table 2depicts the micromeritics properties of the designed formulations. In addition to that, the TD and BD for all formulation granules ranged between 0.48-0.50 and 0.37- 0.41, respectively, whereas HR was obtained as 1.32-1.35.

Micromeritics’ properties of prepared granules

Physical characterization of Extended release tablets

The physical properties of the designed tablets are presented in Table 3. These properties were studied by determining crushing strength, friability, weight variation, drug content, and thickness of the prepared tablets. Crushing strength of the prepared tablets ranged from 12 to 15 newton. It was observed that among all the formulations containing this could be due to the higher binding capacity of Hypromellose100000, Hypromellose15000. All the tablet formulations showed acceptable pharmacotechnical properties and complied with the USP specifications for weight variation, drug content, hardness, and friability. All the formulations showed uniform thickness. In a weight variation test, the pharmacopeial limit for the percentage deviation for tablets ofmore than 1050 mg is ± 5%. The average percentage deviation of all tablet formulations was found to be with in the above limit, and hence all formulations passed the test for uniformity of weight as per official requirements. The average weigh to feach formulation table tranged from1030 to1060 mg. Satisfactory uniformity in drug content was found among different batches of the tablets, and the percentage of drug content was more than 98%

Table 2

Formulation code Bulk density Tapped density Hausner’s ratio Carr’s index

F1 0.38g/ml 0.50g/ml 1.31% 24.48%

F2 0.41g/ml 0.54g/ml 1.31% 24.52%

F3 0.39g/ml 0.48g/ml 1.23% 24.50%

F4 0.37g/ml 0.49g/ml 1.32% 24.53%

F5 0.39g/ml 0.50g/ml 1.28% 23.48%

F6 0.37g/ml 0.49g/ml 1.32% 25.48%

F7 0.41g/ml 0.54g/ml 1.31% 24.52%


Table 3

Physical characterization of the designed formulation:

Formulation codeAverage

weight of (mg)Thickness(mm) Crushing strength Friability (%)

F1 1055 6.20 14.5-19.0 kp 0.12%

F2 1050 6.20 14.5-19.0 kp 0.10%

F3 1060 6.70 23.0-28.0 kp 0.12%

F4 1045 7.00 12.0-15.0 kp 0.11%

F5 1050 7.00 12.0-15.0 kp 0.10%

F6 1055 7.00 12.0-15.0 kp 0.12%

F7 1050 7.00 12.0-15.0 kp 0.12%

Dissolution Data (RLD vs Test):

Time RLD F5 F4 F6 F7

1 17 18 25 12 10

2 26 28 36 22 15

4 39 43 55 28 25

6 49 55 70 35 30

8 58 65 80 50 45

10 65 73 85 58 55

12 71 80 90 68 66

16 81 91 96 78 78

24 96 101 100 92 91

Similarity factor (f2) 59 40 57 49

In-vitro dissolution studies

Microcrystalline cellulose PH -102, Hypromellose 100000, Hypromellose 15000, Povidone K-30 were used to formulate Pregabalin ER tablets and those formulations were subjected to in- vitro drug dissolution studies. The dissolution studies were performed in 0.06N HCl the result showed that approximately 17% of the drug was released within 60 min for all formulations and more than 80% of the drug was found to release at the end of 24hrs.

Comparison of dissolution profile

The comparative results of the selected formulation batch F5 was compared with the theoretical dissolution profile (Innovator) using the similarity factor f2 test and dissimilarity factor f1 test to as sure the best batch.The results of the similarity tests showed that formulation F5 containing 200 mg of HPMC 100000 and 50mg of HPMC 15000 in intragranular and Magnesium stearate in extragranular had an f2 value > 50, i.e., indicating the closest fit to the dissolution profile of innovator


Drug-Excipient compatibility studies

The compatibility studies were carried out in order to study the possible interactions between activepharmaceuticalingredientandinactiveingredients.TostudythecompatibilityofPregabalin with inactive ingredients, physical mixtures were prepared in the defined

ratio as tabulated below and taken in a sealed glass vial and evaluated for chemical stability by storing the compatibility blends in stability at 40°C/75%RH and 25°C/60 %for 1 month. Various excipients were selected depending upon the reference product composition to formulate Tablet formulation. The results are tabulatedbelow

Drug-Excipient compatibility Initial Results

RELATED SUBSTANCES

S.NO Name Of TheExcipient

BinaryMixture Code Unknown-1 Unknown-2 Unknown-3 Lactum

ImpurityTotal

Impurity

1 Pregabalin API PRE-DEC-01 0.05 0.02 ND BQL 0.08

2

Pregabalin API+

Microcrystalline cel-lulose 102

PRE-DEC-02 0.09 0.05 ND BQL 0.14

3

Pregabalin API+

Povidone k30 PRE -DEC-03 0.04 ND 0.04 BQL 0.07

4

Pregabalin API+

Hypromellose 100000 PRE -DEC-04 0.04 0.03 ND BQL 0.08

5

Pregabalin API+

Hypromellose 15000 PRE -DEC-05 0.05 0.03 ND BQL 0.08

6

Pregabalin API+

Magnesium stearate PRE -DEC-05 0.09 0.04 ND BQL 0.13

Pregabalin Tablets of Dissolution profile ( RLD vs TEST)

100

80

60

40

20

0 0 2 4 6 8 10 12

Time in hrs

14 16 18 20 22 24

RLD F5 F4 F6 F7


Drug-Excipient compatibility (40°C/75% 1M) Results

RELATED SUBSTANCES (40°C/75% 1M)

S.NOName Of The

Excipient

Binary

Mixture CodeUnknown-1 Unknown-2 Unknown-3

Lactum

Impurity

Total

Impu-rity


2

Pregabalin API

+ Microcrystalline

cellulose PH 102

PRE-DEC-02 0.20 0.07 ND BQL 0.23

3

Pregabalin API

+ Povidone k30PRE -DEC-03 0.14 ND ND ND 0.14

4

Pregabalin API

+ Hypromellose 100000

PRE -DEC-04 0.14 0.05 ND BQL 0.20

5

Pregabalin API

+ Hypromellose 15000

PRE -DEC-05 0.15 0.04 ND BQL 0.21

6

Pregabalin API

+ Magnesium stearate

PRE -DEC-05 0.15 0.07 ND BQL 0.22


RELATED SUBSTANCES (25°C/60% 1M)

S.NOName Of The

ExcipientBinary

Mixture CodeUnknown-1 Unknown-2 Unknown-3

LactumImpurity

TotalImpurity


2

Pregabalin API+

Microcrystalline cellulose PH

102

PRE-DEC-02 0.35 0.13 ND BQL 0.49

3

Pregabalin API+

Povidone k30PRE -DEC-03 0.19 ND ND ND 0.19

4

Pregabalin API+

Hypromellose 100000

PRE -DEC-04 0.21 0.05 ND BQL 0.27

5

Pregabalin API+

Hypromellose 15000

PRE -DEC-05 0.22 0.07 ND BQL 0.30

6

Pregabalin API+

Magnesium stea-rate

PRE -DEC-05 0.22 0.08 ND BQL 0.30

Conclusion :

From the present investigation, it was concluded that the formulation (F5) having concentrations of HPMC 100000 (19%) and HPMC 15000 (4.8%) polymers and drug release from Extended- release tablet was comparable with Innovator product with having similarity factor more than 50% and with desirable drug release pattern.

REFERENCES1. H. Salem. E-Journal of Chemistry 2009, 6(2):332-340.2. L. Arnold, L. Pauer, E. Whalen, J. Barrett, T. Leon. J Pain 2008,

9(4):48.3. A Ghosh; KS Gupta. Asian J Pharm Res Health Care. 2010, 2(3),

222-227.4. SK Singh; V Sharma; K Pathak. Int J Pharm Pharm Sci. 2012,

4,168-174.5. MA Kumar; PK Lakshmi; J Balasubramanium. Int J Pharm Tech

Res. 2011, 3(2),968-979.6. SR Vachhani; JJ Patel; D Patel; ST Prajapati; CN Patel. J Chem

Pharm Res. 2010, 2(2),57-64.

7. DN Kapoor; OP Katare; S Dhawan. Int J Pharm. 2012, 426(1),132-143.

8. RS Gujral; SM Haque; S Kumar. Afr J Pharm Pharmacol. 2009, 3(6),327-334.

9. M.S. Reza, M. Abdul Quadir, S.S. Haider. J Pharm Pharm Sci 2003, 6:282-291.

10. Kastler A, Aubry S, Piccand V, Hadjidekov G, Tiberghien F, Kastler B. Radiofrequency neurolysis versus local nerve infiltration in 42 patients with refractory chronic inguinal neuralgia. Pain Physician.2012;15(3):237–244.

11. Jensen TS, Baron R. Translation of symptoms and signs into mechanisms in neuropathic Pain. 2003;102(1–2):1–8.

12. M.M. Talukdar, P. Rommbaut, R. Kinget. Int J Pharm 1996, 129:233-241..

13. A. Sharma, S. Sharma, K.K. Jha. The Pharma Research 2009, 1:15-22.

14 Smith HS, Argoff CE. Pharmacological treatment of diabetic neuropathic pain. Drugs. 2011;71(5):557–589

15 Tesfaye S, Selvarajah D. Advances in the epidemiology, pathogenesis and management of diabetic peripheral neuropathy. Diabetes Metab Res Rev. 2012;28 Suppl1:S8–S14

Drug-Excipient compatibility (25°C/60% 1M) Results


Colour Properties of Catechu Heartwood Extract

SWAPNIL Y. JADHAO1, JAYARAJ RAO, K2*. Dairy Technology Section, ICAR-National Dairy Research Institute (Southern Regional Station), Adugodi, Bangalore – 560 030, India

Jayaraj Rao, K., Principal Scientist, Dairy Technology Section, ICAR-National Dairy Research Institute

(Southern Regional Station), Adugodi, Bangalore – 560 030, India

ABSTRACT

Acacia catechu (Khair) is well known for its heartwood extract called as katha, commonly used in betel leaf preparation (paan) by people in Indian subcontinent. The extract gives dark red to reddish brown colour and is known to possess several medicinal properties because of its phytoconstituents. It is also being used in leather and textile industry as colouring agent. However, it has not yet been tried in food industry as colouring agent though it is a good source of natural colour and can be used to replace or supplement artificial colours. In this project, colour was extracted from catechu heartwood by heat extraction method using water as the medium. The extract was heat concentrated to get a Catechu colour concentrate (CCC).Efforts were made to enhance the colour intensity by different additives, but with little success. Processing treatments such as thermisation, boiling and sterilisation had no adverse effect on colour intensity of CCC.

Keywords: Acacia catechu, colour concentrate, heartwood extract, colour properties, additives

INTRODUCTION

* Correspondance : E-mail: [email protected]

Colour is actually a result of sensations perceived and collated by eye and brain by means of a mechanism overhauled in evolutionary chain. In fact the colourful world perceived by humans is not as colourful to many living beings. It is based on the human perception and acceptance that colour systems what we use today have been evolved. Probably, it is aquatic living beings that first possessed colour sensations which served evolutionary purpose, but later terrestrial plants dominated production of colours in myriad hues. What purpose plants serve by producing colours is not yet clear, though many reasons have been attributed like hiding from enemy, reproduction purpose, defence against enemy etc. Whatever be the reason, no doubt humans have tremendously benefitted from these colours. Quantification of colours has evolved slowly. Newton in 16th Century laid foundation for basic physical mechanism of colours. Till 18th Century, the colour was measured by qualitative terms, but later need was felt to measure it in quantifiable terms. Munsell colour system developed in 1929 was a great step in this direction. It is based on the primary colours red, blue and green. Though it is a systematic approach in measurement of colours, there was a scope for further refinement of the system. This led to the development of L, a and b system in 1976 which saw the colour in a three dimensional space. This system attained revolutionary proportion by introduction of computers in measurement and scanning of colours. Now it is possible to accurately quantify any colour combination and reproduce it without flaws. If the developments in measurement of colours took years to arrive at the latest system of measurement, development of suitable colouring

material per se has been pursued consistently by humans with constant look out for better and better colour sources or dye sources. This pursuit is continuing even today with industry still in search of better and better sources for colour extraction. In fact, many colour sources remain unexplored even till today, including plant and animal sources. In recent years, this assumed great importance in view of health benefits of natural colours and health risks posed by artificial and synthetic colours. Scientists have been exploring new sources of colours from plant kingdom like leaves, flowers, roots etc. One of these pursuits has led us to Catechu plant. Catechu is a plant belonging to Mimosaceae family whose heartwood finds several applications in industries. It yields a red to dark brown dye in water which is used in textile and leather industries. The heartwood of the plant reportedly has several phytochemicals including catechins and tannins, which are good antioxidants. Catechu extract has not yet been tried in any food products, but the dried catechu heartwood extract is popularly used in paan not only for developing red colour in mouth but also for good digestibility attribute of paan. Catechu heartwood is well known in India as its extract in the form of dry powder is used in ‘paan’ widely eaten in India and subcontinent after a meal. Since eating of the ‘paan’ along with the catechu powder has been in vogue for centuries, catechu powder is considered as safe for consumption. The catechu powder is known to paint mouth and tongue with red colour when chewed along with betel leaves. Though it is known that catechu has colouring property, it was not used for colouring purpose for unknown reasons. In this study, we used catechu


heartwood as a source for colour and studied its some basic extraction and properties so that its use in suitable foods including dairy foods may be explored.

A few natural colours of seed, vegetable and fruit origin are being used in dairy products, for example annatto seeds, turmeric, mango, berries etc., but catechu has not been tried so far in dairy products

MATERIALS AND METHODS

Catechu (Acacia catechu)

Catechu (Acacia catechu) heartwood was procured from Buldhana district of Maharashtra, India. Wood of reddish brown colour was selected and cut from the plant and brought to the laboratory. It was further cut into ~ 4cm pieces (Fig. 1) and used for colour extraction.

METHODOLOGY

Extraction of catechu heartwood

Two extraction methods were tried: hot extraction and cold extraction. About 20 g of catechu heartwood was taken in a vessel and boiled in 100 ml water for 5 min. Then the wood pieces were filtered out and boiled further in fresh lot of water second time. In this manner, the wood pieces were extracted five times till all the colour got extracted from the pieces. The extent of colour extraction was measured by OD of the pooled solution at 500 nm. Similarly, known amount of catechu heartwood pieces were soaked in water for about 24 h and extraction of colour into the medium observed. Alcohol was also tried as a medium of extraction. The OD measured at 500 nm indicated the efficiency of the extraction method.

Preparation of CCC

About 100 g of catechu heartwood pieces (4-5 cm) were washed under tap water to remove adhering dust. They were then boiled in 500 ml water for about 20 min. The catechu pieces were removed from the solution by filtration and again boiled in a fresh 500 ml lot of water. In such a manner, 6-7 extractions were collected. All the extracts obtained were pooled in a container and heat concentrated to a final volume of about 25 ml. This was called catechu colour concentrate (CCC) which contained about 10% TS.

Effect of various additives on catechu colour

Various additives are known to enhance the colour intensity of dyes. With this assumption, different additives were tried to enhance the colour intensity of catechu extract.

The catechu extract was prepared as follows: Two per cent solution of colour concentrate of catechu was used. The 10 ml of the extract was taken in test tubes and added with a little quantity of the following commonly used additives and any change in the colour of the extract was noted (betel leaf extract, calcium hydroxide, calcium chloride and turmeric).

The present study being the first study on catechu colour, the objective here was to find out qualitatively whether any substance darkened or enhanced the colour of catechu extract. Any changes were recorded by digital imaging.

Stability of extracted catechu colour under various conditions

Ten ml of catechu extract was taken in test tubes and kept in an incubator at 37°C for 10 days. The various colour parameters RGB and L were measured by SAP (Scanner – Adobe Photoshop) method at the end of 10 days. These values were compared with the zero day values to know the effect storage at 37oC on colour intensity. Some tubes were left open exposed to atmospheric air and some tubes were filled to the brim and closed tightly with rubber stopper. The any effect of dissolved oxygen was neglected. Similarly some tubes closed with rubber stoppers were stored in dark. The R,G, B and L colour parameters at the end of 10 days storage at 37ºC were measured by SAP method.

Effect of heat treatment on catechu colour

The catechu extract was taken in test tubes and subjected to thermization, boiling and sterilisation treatments. Any change in color was evaluated by measuring RGB and L by SAP method.

Effect of different processing treatments on catechu colour

Milk was added with catechu colour and subjected to thermization, boiling and sterilisation treatments. Colour parameters were measured by SAP method.

Scanner-Adobe Photoshop (SAP) method

The colour of samples was measured by the method described by Vyawahare and Rao (2011). All the samples were tempered to about 30oC before measurement. The samples were prepared as follows: Lassi – Samples of lassi were directly poured into scratch-less petri plate (~70 mm diameter) which has been thoroughly cleaned and dried earlier. Yoghurt and shrikhand– With the help of a spatula, yoghurt was scooped into petri plate and the surface evened out. Any air gaps were removed by spatula and by gentle tapping the plate. Flavoured milk – it was directly poured into petri plate without leaving any air bubbles. The petri dish containing the sample was placed on the bed of the scanner and was covered by scanner top. Scanning was performed and the image was saved as JPEG file. Scanning of the sample was performed under the following scanner parameters: resolution: 75 dpi, sharpness: medium; image quality: medium; background of the sample: white; thickness of sample: 1 cm. Colour analysis software namely Adobe Photoshop Version CS3 running under the Microsoft Windows XP environment was used to extract and analyse colour information from the scanned image. The scanned image in JPEG format was opened in Adobe Photoshop


(Fig. 2) and the following parameters were measured in RGB mode: Lightness (L), red (R) , green (G) and blue (B) (Fig. 2).

RESULTS AND DISCUSSION

Studies on extraction of catechu heartwood colour and its properties

Catechu plant has a hard interior wood which possesses a reddish brown colour. Most of this colour is water soluble, so can be extracted in water medium. However, there are reports of extraction of many natural colours from plant source using different media other than water like ethanol, methanol etc. Moreover, the extent of colour extraction depends on factors like temperature, pH etc. Hence, water and ethanol were tried as medium of extraction and colour quality studied.

Effect of type of extraction on catechu colour

Catechu heartwood was extracted in water as well as other medium like alcohol and methanol. Colour extraction was found to be maximum by hot extraction and very less in cold extraction as indicated by OD values presented in Table 1. Further, colour extraction was maximum in aquous medium than in alcohol medium. These observations could have stemmed from two facts: firstly the catechu compounds are mostly water soluble and secondly viscosity of water less high temperatures and thereby surface tension. These facilitated easy extraction of catechu colour compounds. Catechu colour was found to be water soluble, slightly soluble in alcohol and insoluble in oils. Hence, all further trials were conducted using hot extraction.

Effect of various additives on catechu colour

The colour quality of natural colours depends on the composition of the medium of extraction. Presence of additives might alter their colour quality as well as intensity. Hence, effect of various additives was studied.

Effect of betel leaf extract and calcium hydroxide on catechu colour

It is common practice to use catechu powder in paan which is a betel leaf preparation. It is believed that catechu powder together with betel leaf and lime combinedly develops a red colour in mouth when chewed. Since, paan contains betel leaf, lime and catechu powder combinedly and gives bright red on tongue, lime and betel leaf extracts were added to catechu colour extract and seen for any red colour development. But surprisingly use of betel leaf extract resulted in light yellow colour, which slowly faded off to no colour on storage for 4 days at ambient temperature (Fig. 3A). So far, colour development on chewing of pan has not been explained though mention has been made in some sources (Lakshmi et al., 2011; Patel, 2011). The extract added with only lime first turned dull, light brown and on storage for 4 days at ambient temperature got discoloured. However, settling of colour particles was observed at the

bottom (Fig. 3A). Even on thorough mixing the colour was not restored. Betel leaf extract (betel leaves brought from market were washed in running tap water, cut into pieces and ground in pestle and mortar with minimum amount of water, filtered through multi-layered muslin cloth and the extract collected) alone was added to catechu heartwood extract and seen for any change in colour. It was observed that there was no improvement in colour by addition of betel leaf extract, rather the colour became light and dull (Fig. 3B). Similar were the observations when calcium hydroxide alone was used along with catechu extract (Fig. 3C). Based on these observations, whether colour development in the mouth as a result of chewing of paan has anything to do with any enzyme action can be explored.

Effect of calcium chloride on catechu colour

On addition of calcium chloride, there was no improvement in the colour of the extract (Fig. 3D).

Effect of acidification on catechu colour

On acidification with HCl, the extract turned from dark reddish brown to light yellowish brown (Fig. 3E).

Effect of alkalisation on catechu colour

On addition of NaOH, the extract turned darker indicating at alkaline pH the colour becomes dark (Fig. 3F).

Effect of turmeric on catechu colour

On addition of turmeric, there was no change in the colour, rather it turned slightly turbid (Fig. 3G).

Effect of turmeric and calcium hydroxide on catechu colour

On addition of turmeric along with lime, the extract turned reddish yellow, however there was settling of colour particles (Fig. 3H).

Stability of extracted catechu colour under various conditions

The stability of the extracted colour is very important if it has to be utilised in dairy and food products. Hence, colour stability was studied. Two per cent solution of concentrated catechu colour was used for the stability tests.

At 37°C storage

It may be observed from the table that there were changes in R,G,B and L values of the extract during storage of 10 days at ambient temperature. The L value slightly decreased indicating that the extract became slightly darker. The recreated colours using R, G and B values shown in Table 2 also indicated the same.

Exposure to atmospheric air

In presence of atmospheric air and by storage at room temperature for 10 days, colour became dark, but in the absence of atmospheric air, the colour further darkened as indicated by lower L values (Table 3 a and b).


Exposure to light

It was observed that the catechu colour became darker in presence of light than in absence of light as indicated by L values (Table 4 a and b). This indicates that catechu colour can be stored even under light without impacting its colour intensity.

Effect of heat treatment on catechu colour

In order to determine the stability of catechu colour to different heat treatments, the colour was extracted from catechu heartwood by hot extraction method and subjected to thermization (60°C), boiling and sterilisation (~121°C) treatments. Initially, the extracted colour showed brown colour (Fig. 4a). It became slightly darker after thermisation treatment (60oC). After boiling treatment, the solution became further dark (Fig. 4 b and c) and darkest after sterilisation treatment (Fig. 4d). The R,G,B and L values are presented in Table 4. The R,G,B and L values of fresh extract were 194.09, 138.67, 68.37 and 147.46, respectively which significantly changed because of heat treatments (Fig. 5).

The fact that the solutions became darker is indicated by L values which decreased to 142.06, 121 and 105.94 respectively by thermization, boiling and sterilisation treatments (Table 5). From these results, it may be inferred that the intensity of catechu colour increased by heat treatment, which seems to be a unique property for a natural pigment. Many of the natural pigments like betanins, carotenes, chlorophyll, anthocyanin’s etc. are heat sensitive (Rahman, 2007) and lose their colour on heating, but catechu seems to be highly heat stable. It was reported that tea epicatechins are remarkably stable to heat in acidic environment: at pH 5, only about 15% is degraded after seven hours in boiling water (therefore, adding lemon juice to brewed tea does not cause any reduction in their content) (Anonymous, 2015 a).

Effect of different processing treatments on catechu colour

The catechu extract was added to milk @ 1% and subjected to thermization, boiling and sterilisation temperatures. Immediately after the addition of the extract, the milk looked brown (Table 6) whose L value was 200.63. On thermization, there was no change in L value (201.44), but on sterilisation, there was slight increase in L value (204.67) indicating milk became light brown. By boiling of milk also, there was no effect on both samples but slightly luminosity of coloured sample increased from 200 to 204. This shows that processing has little effect on the heat stability of the colour. The decrease in colour may be attributed to effect of milk proteins.

CONCLUSION

Hot extraction was found to be more suitable for colour extraction from the catechu heartwood yielding dark brown colour. Several additives were tried to enhance the brown colour intensity of the extract, but none of them was found favourable. It was found that catechu colour on heat treatment like thermization, pasteurisation and sterilisation showed darkening trend as indicated by lower lightness values measured by SAP method. This showed a good heat stability of catechu colour to heat treatments. Acidification or alkalisation of medium showed a significant effect on the colour. At low pH, the extract a showed yellowish colour, whereas under alkaline conditions, it displayed brown colour. At about neutral pH, the catechu extract showed a desirable darker brown colour. A catechu colour concentrate (CCC) was prepared by extraction of catechu heart wood 6-7 times in water and heat concentrating the pooled extract to a minimum volume to obtain CCC which can be utilised to colour the products like flavoured milk.

Declarations of conflict of interest

The authors report no declarations of conflict of interest.

Authors’ contributions

Swapnil Jadhao performed the experiments, analyzed the data and coordinated the data collection. K. Jayaraj Rao guided Swapnil Jadhao in conducting experiments and supported in writing and proof reading of the research paper.

REFERENCES

Anonymous 2015a. Catechins: definition, structure, green tea, black tea, cocoa. www.tuscany-diet.net/2014/02/.../catechins-definition-structure-green-tea...

Anonymous 2015b. http://www.phytotech.in/www3/html/conv.

Galli, A., L. Franzetti and D. Briguglio. 1985. Antimicrobial properties in vitro of essential oils and extract of spices used for food. Ind. Aliment. 24: 463–466.

Lakshmi, T., R.V. Geetha and Anitha Roy. 2011. In vitro evaluation of anti bacterial activity of Acacia catechu willd heartwood extract. Int. J. Pharma Biosci. 2(1): 235-240.

Patel, N.K. 2011. Natural dye based sindoor. Life Sci. Leaflets 11: 355-361.

Rahman, S.M. 2007. Handbook of food preservation, 2nd Edition, CRC Press, Taylor & Francis Group, Boca Raton, USA., p.600.

Ramadan, F.M., R.T. El-Zanfaly, F.A. El-Wakeil and A.M. Allian. 1972. On the antibacterial effects of some essential oils I. Use of agar diffusion method. Chem. Mikrobiol. Technol. Lebensm. 2: 51–5.

Tassou, C.C., G.J.E. Nychas and P.N. Skandamis. 2004. Herbs and spices and antimicrobials. In: “Handbook of herbs and spices” (Peter, K.V., ed.), Woodhead Publishing Ltd., Cambridge, England, p. 35-42.

Wilkins, K.M. and R.G. Board. 1989. Natural antimicrobial systems. In: Mechanisms of action of food preservation procedures (Gould, G.W., ed.), Elsevier, London, pp. 285–362.


Fig 1. Acacia catechu heartwood pieces

Fig. 2. Measurement of colour parameters by SAP method (a) JPEG format of scanned image in

Adobe-Photoshop window (b) Measurement of colour parameters under RGB mode

(a)

(b)


i ii i ii 0 day 4th day

i-Catechu extract; ii- Catechu extract +betel extract+ calcium hydroxide


i-Catechu extract; ii-Catechu extract + betel extract


i-Catechu extract; ii-Catechu extract + calcium hydroxide


i-Catechu extract; ii-Catechu extract + calcium chloride


i-Catechu extract; ii-Catechu extract + HCl


i-Catechu extract; ii-Catechu extract + NaOH


i-Catechu extract; ii-Catechu extract + turmeric


i-Catechu extract ; ii-Catechu extract + turmeric+ calcium hydroxide

Fig 3. Effect of various additives on catechu colour

A B

C D

E F

G H


(a) Fresh extract (b) Thermised extract (c) Boiled extract (d) Sterilised extract

Fig 4. Colour of catechu extract as affected by various heat treatments

Fig 5. Instrumental colour parameters of catechu extract as influenced by various heat

treatments

194.09 194182.08

154.68138.67 132.9

109.2993.68

68.3750.25

17.1839.83

147.46 142.06121

105.94

0

50

100

150

200

250

Cold Thermization Boiling Sterilization

R G B L


Table 1: Optical density (measured at 500 nm) of catechu extracts

Method of extraction Medium used OD Hot Potable water 1.56 Cold Potable water 0.72 Cold Alcohol 0.53 Cold Methanol 0.21

Table 2: Effect of storage at 37oC on colour of catechu extract

Days R G B L Recreated colour*

0 198.83 149.04 97.06 158.13

10 202.23 144.87 51.01 151.71

Table 3: Effect of exposure to air on the colour of catechu extract

(a) Exposed to atmospheric air


0 198.83 149.04 97.06 158.13

10 208.79 140.39 35 149.32

(b) Air tight closure


0 198.83 149.04 97.06 158.13

10 192.35 109.25 9.8 123.26

* The colour in column was recreated by R, G and B values of respective row using shading option on toolbar of MS Word software


Table 4: Effect of exposure to light on the colour of catechu extract

(a) Presence of light


0 198.83 149.04 97.06 158.13

10 195.62 123.84 34.41 135.49

(b) Absence of light


0 198.83 149.04 97.06 158.13

10 201.14 130.27 48.6 142.47

Table 5: Instrumental colour parameters of catechu extract as influenced by various heat treatments

Treatment R G B L Recreated colour*

Cold (20° C) 194.09 138.67 68.37 147.46 Thermization 194 132.9 50.25 142.06 Boiling 182.08 109.29 17.18 121 Sterilization 154.68 93.68 39.83 105.94



Table 6: Effect of various processing treatments on catechu colour

(a)

Product At 20° C Recreated colour* R G B L

Plain milk (Control) 254.52 254.63 243.64 252.43 Milk added with catechu colour

223.74 194.51 171.17 200.63

(b)

Product Thermization ( 60° C ) Recreated colour* R G B L


225.69 194.91 171.10 201.44

(c)

Product Boiling Recreated colour* R G B L


225.96 199.23 176.50 204.67

(d)

Product Sterilisation (121° C) Recreated colour* R G B L


226.69 199.48 173.09 204.66



A Review on Pathogenesis of Obesity

DR. C. GIRISH1, CH. JL PRASANNA2.

1, 2 S.V.U. College of Pharmaceutical Sciences, Sri Venkateshwara University, Tirupati - 517502. A.P, India.

Running Title: Antianaphylactic Activity of Ocimum sanctum

ABSTRACT

The lifestyle of people made them for taking fast food rather than healthy food. But the people are not aware that fast food leads to the disease like obesity, diabetes, dyslipidemia etc. Obesity is the widespread disease; approximately 23% of adult are obese and cannot be controlled within time. The etiology of obesity is involving a complex interaction among genetics, hormones and the environment and the main factor that causes obesity is imbalance between intake of energy and the energy expenditure. According to WHO around 1.7 billion adults were overweight and 400 million people were obese. Obesity develops because of energy required to body adipose tissue that secrets triglycerol during excess food intake and releases free fatty acid FFA thus the excess fat store in the adipose tissues in the body and the mechanism of body action fails to maintain the balance of body fat that is body mass index. Hence, there is the essential need to target the obesity. In this review we focused on the various causes, pathophysiology and management for obesity.

Keywords: Obesity, Pathogenesis and Management,

INTRODUCTION:

Correspondance : E.Mail: [email protected]

Overweight and obesity are defined as abnormal or excessive fat accumulation that presents a risk to health. A crude population measure of obesity is the body mass index (BMI), a person’s weight (in kilograms) divided by the square of his or her height (in metres). A person with a BMI of 30 or more is generally considered obese. A person with a BMI equal to or more than 25 is considered overweight.

Overweight and obesity are major risk factors for a number of chronic diseases, including diabetes, cardiovascular diseases and cancer. Once considered a problem only in high income countries, overweight and obesity are now dramatically on the rise in low- and middle-income countries, particularly in urban settings.

Obesity is a medical condition in which excess body fat has accumulated to an extent that it may have a negative effect on health. [1] People are generally considered obese when their body mass index (BMI), a measurement obtained by dividing a person’s weight by the square of the person’s height, is over 30 kg/m2; the range 25–30 kg/m2 is defined as overweight. Some East Asian countries use lower values.[2] Obesity increases the likelihood of various diseases and conditions, particularly cardiovascular diseases, type 2 diabetes, obstructive sleep apnoea, certain types of cancer, osteoarthritis, and depression.[3][4]

Obesity is most commonly caused by a combination of excessive food intake, lack of physical activity, and genetic susceptibility. [5] A few cases are caused primarily by genes, endocrine disorders, medications, or mental disorder.[6]The

view that obese people eat little yet gain weight due to a slow metabolism is not medically supported.[7]On average, obese people have greater energy expenditure than their normal counterparts dueto the energy required to maintain an increased body mass.[8]

Obesity is mostly preventable through a combination of social changes and personal choices. Changes to diet and exercising are the main treatments. Diet quality can be improved by reducing the consumption of energy-dense foods, such as those high in fat or sugars, and by increasing the intake of dietary fibre.

Medications can be used, along with a suitable diet, to reduce appetite or decrease fat absorption.[9]If diet, exercise, and medication are not effective, a gastric balloon or surgery may be performed to reduce stomach volume or length of the intestines, leading to feeling full earlier or a reduced ability to absorb nutrients from food.[10][11]

Obesity is a leading preventable cause of death worldwide, with increasing rates in adults and children. [12] In 2015, 600 million adults (12%) and 100 million children were obese in 195 countries. Obesity is more common in women than men. Authorities view it as one of the most serious public health problems of the 21st century. [13]Obesity is stigmatized in much of the modern world (particularly in the Western world), though it was seen as a symbol of wealth and fertility at other times in history and still is in some parts of the world. [14] In 2013, several medical societies, including the American Medical Association and the American Heart


Association, classified obesity as a disease.[15][16][17]

EPIDEMOLOGY:

In earlier historical periods obesity was rare, and achievable only by small elite, although already recognised as a problem for health. But as prosperity increased in the early modern period, it affected increasingly larger groups of the population. [18]

In 1997 the WHO formally recognized obesity as a global epidemic. [19]As of 2008 the WHO estimates that at least 500 million adults (greater than 10%) are obese, with higher rates among women than men.[20] The percentage of adults affected in the United States as of 2015–2016 is about 39.6% overall (37.9% of males and 41.1% of females). [21]

The rate of obesity also increases with age at least up to 50 or 60 years old[22] and severe obesity in the United States, Australia, and Canada is increasing faster than the overall rate of obesity.[23][24][25] The OECD has projected an increase in obesity rates until at least 2030, especially in the United States, Mexico and England with rates reaching 47%, 39% and 35% respectively.[26]

Once considered a problem only of high-income countries, obesity rates are raising worldwide and affecting both the developed and developing world. [27] These increases have been felt most dramatically in urban settings. The only remaining region of the world where obesity is not common is sub-Saharan Africa.

PREVALENCE OF OBESITY:

A number of factors influence body fat, including age, sex race and physical activity. At birth, the human body contains about 12% fat, a content that is higher than in any other mammal except the whole. During the new born period, the body fat content rises rapidly to reach a peak of about 25% at 6 months of age and then declines to 15% to 18% body fat and girls 20% to 25%. The amount of body fat increases in both sexes after puberty and during adult life rises to between 30% and 40% of body weight. Between ages 20 and 50, the fat content in men approximately doubles and that in women increases by about 50%. Total body weight, however, rises by only 10% to 15%. The increased fat is accounted for in part by an increase in body weight and in part by a reduction in lean body mass.[28] The composition of the body is also influenced by the level of physical activity. During physical training, body fat usually decreases the lean tissues increases. After training ends, however this process is reversed. The shifts between body fat and lean tissue can occur without any changes in body weight, but if the regular activity is maintained throughout adult life, the increase in body fat that usually occurs when body weight is stable may be prevented. Racial differences in body weight may exist but are often difficult to separate racial from environmental factors.[29]

Socio-economic conditions, however, clearly play an important role in the development of obesity. Excess body

weight is 7 to 12 times more frequent in women from lower social classes than in women from upper social classes. Among men, social class has a much smaller relationship to being overweight. The patterns of body weight in men and women are changing.

CLASSIFICATION:

BMI (kg/m2) Classification BMI (kg/m2)underweight <18.5

normal weight 18.5-25

overweight 25-30

class I obesity 30-35

class II obesity 35-40

class III obesity >40

A “super obese” male with a BMI of 53 kg/m2: weight 182 kg (400 lb), height 185 cm (6 ft 1 in). He presents with stretch marks and enlarged breasts

Obesity is a medical condition in which excess body fat has accumulated to the extent that it may have an adverse effect on health.[30]It is defined by body mass index (BMI) and further evaluated in terms of fat distribution via thewaist-hip ratio and total cardiovascular risk factors.[31][32]BMI is closely related to both percentage body fat and total body fat.[33] In children, a healthy weight varies with age and sex. Obesity in children and adolescents is defined not as an absolute number but in relation to a historical normal group, such that obesity is a BMI greater than the 95thpercentile.[34]The reference data on which these percentiles were based date from 1963 to 1994, and thus have not been affected by the recent increases in weight [35].BMI is defined as the subject’s weight divided by the square of their height and is calculated as follows.

BMI = mass(kg)/height(m2)

= mass(lb)/height(in)2×703

Where m and h are the subject’s weight and height respectively.

BMI is usually expressed in kilograms of weight per metre squared of height. To convert from pounds per inch squared multiply by 703 (kg/m2)/(lb/sq in).[36] The most commonly used definitions, established by the world health organisation(WHO) in 1997 and published in 2000, provide the values listed in the table.[37][38]Some modifications to the WHO definitions have been made by particular organizations.[39]The surgical literature breaks down class II and III obesity into further categories whose exact values are still disputed.[40]

• Any BMI ≥ 35 or 40 kg/m2 is severe obesity.

• A BMI of ≥ 35 kg/m2 and experiencing obesity-related health conditions or ≥40–44.9 kg/m2 is morbid obesity.

• A BMI of ≥ 45 or 50 kg/m2 is super obesity.


As Asian populations develop negative health consequences at a lower BMI than Caucasians, some nations have redefined obesity; Japan has defined obesity as any BMI greater than 25 kg/m2 while China uses a BMI of greater than 28 kg/m2.

OBESITY AND INSULIN RESISTANCE:

Obesity-associated insulin resistance is a major risk factor for type 2 diabetes and cardiovascular disease. In the past decade, a large number of endocrine, inflammatory, neural, and cell-intrinsic pathways have been shown to be deregulated in obesity. Although it is possible that one of these factors are independent, and it is likely that their dynamic interplay underlies the pathophysiology of insulin resistance. Understanding the biology of these systems will inform the search for interventions that specifically prevent or treat insulin resistance and its associated pathologies.

ETIOLOGY OF OBESITY:

To maintain a healthy weight, your energy IN and OUT doesn’t have to balance exactly every day. It’s the balance over time that helps you maintain a healthy weight.

• The same amount of energy IN and OUT over time =


weight stays

• More energy IN than energy OUT over time = weight gain.

• More energy OUT than energy IN overtime = weight loss.

Overweight and obesity happen over time when you take in more calories than you use.

At an individual level, a combination of excessive food energy intake and a lack physical activity is thought to explain most cases of obesity. [41] A limited number of cases are due primarily to genetics, medical reasons, or psychiatric illness. In contrast, increasing rates of obesity at a societal level are felt to be due to an easily accessible and palatable diet,[42] increased reliance on cars, and mechanized manufacturing.[43][44]

A 2006 review identified ten other possible contributors to the recent increase of obesity:

1. Insufficient sleep

2. Endocrine disruptors (environmental pollutants that interfere with lipid metabolism)

3. Decreased variability in ambient temperature

4. Decreased rates of smoking, because smoking suppresses appetite

5. Increased use of medications that can cause weight gain (e.g., atypical anti psychotics)

6. Proportional increases in ethnic and age groups that tend to be heavier

7. Pregnancy at a later age (which may cause susceptibility to obesity in children)

8. Epigenetic risk factors passed on generationally

9. Natural selection for higher BMI and

10. Assortative mating leading to increased concentration of obesity risk factors (this would increase the number of obese people by increasing population variance in weight).[45]

According to the Endocrine Society, there is “growing evidence suggesting that obesity is a disorder of the energy homeostasis system, rather than simply arising from the passive accumulation of excess weight”.[46]

PATHOPHYSIOLOGY:

There are many possible pathophysiological mechanisms involved in the development and maintenance of obesity. [47]This field of research had been almost approached until the leptin gene was discovered in 1994 by J. M. Friedman’s laboratory. [48] Whileleptin and ghrelin are produced peripherally, they control appetite through their actions on the central nervous system. In particular, they and other appetite-related hormones act on the

hypothalamus, a region of the brain central to the regulation of food intake and energy expenditure. There are several circuits within the hypothalamus that contribute to its role in integrating appetite, the melanocortin pathway being the best understood. The circuit begins with an area of the hypothalamus, the arcuate nucleus, that has outputs to the lateral hypothalamus (LH) and ventromedial hypothalamus (VMH), the brain’s feeding and satiety centres, respectively.[49]The arcuate nucleus contains two distinct groups of neurons.The first group coexpresses neuropeptide y (NPY) and agouti-related peptide (AgRP) and has stimulatory inputs to the LH and inhibitory inputs to the VMH. The second group expresses pro-opiomelanocortin (POMC) and cocaine and amphetamine-regulated transcript (CART) and has stimulatory inputs to the VMH and inhibitory inputs to the LH. Consequently, NPY/AgRP neurons stimulate feeding and inhibit satiety, while POMC/CART neurons stimulate satiety and inhibit feeding. Both groups of arcuate nucleus neurons are regulated in part by leptin. Leptin inhibits the NPY/AgRP group while stimulating the POMC/CART group. Thus a deficiency in leptin signalling, either via leptin deficiency or leptin resistance, leads to overfeeding or may account for some genetic and acquired forms of obesity.

MANAGEMENT:

The main treatment for obesity consists of weight loss via dieting and physical exercise. [50][51] Dieting, as part of a lifestyle change, produces sustained weight loss, despite slow weight regains over time. [52][53][54] Intensive behavioural interventions combining both dietary changes and exercise are recommended. [55]

Several diets are effective. In the short-term low carbohydrate diets appear better than low fat diets for weight loss.[56] In the long term; however, all types of low-carbohydrate and low-fat diets appear equally beneficial.[57] A 2014 review found thatthe heart disease and diabetes risks associated with different diets appear to be similar. [58] Promotion of the Mediterranean diets among the obese may lower the risk of heart disease. Decreased intake of sweet drinks is also related to weight-loss.Success rates of long-term weight loss maintenance with lifestyle changes are low, ranging from 2–20%.[59] Dietary and lifestyle changes are effective in limiting excessive weight gain in pregnancy and improve outcomes for both the mother and the child.[60] Intensive behavioural counselling is recommended in those who are both obese and have other risk factors for heart disease.[61]

Five medications have evidence for long-term use orlistat, lorcaserin, liragutide, phentermine-topiramate, and naltrexone-bupropion.[62] They result in weight loss after one year ranged from 3.0 to 6.7 kg (6.6-14.8 lbs) over placebo. Orlistat, liraglutide, and naltrexone–bupropion are available in both the United States and Europe, whereas lorcaserin and phentermine–topiramate are available only in the United States.[63] European regulatory authorities rejected the latter two drugs in part because of associations of heart valve problems with lorcaserin and more general heart and blood


vessel problems with phentermine–topiramate. Orlistat use is associated with high rates of gastrointestinal side effects [64] and concerns have been raised about negative effects on the kidneys.[65] There is no information on how these drugs affect longer-term complications of obesity such as cardiovascular disease or death.

The most effective treatment for obesity is bariatric surgery. The types of procedures include laparoscopic adjustable gastric banding, Roux-en-y gastric bypass, vertical-sleeve gastrectomy, and biliopancreatic diversion. Surgery for severe obesity is associated with long-term weight loss, improvement in obesity-related conditions,[66] and decreased overall mortality. One study found a weight loss of between 14% and 25% (depending on the type of procedure performed) at 10 years, and a 29% reduction in all-cause mortality when compared to standard weight loss measures.[67] Complications occur in about 17% of cases and reoperation is needed in 7% of cases. Due to its cost and risks, researchers are searching for other effective yet less invasive treatments including devices that occupy space in the stomach.[68] For adults who have not responded to behavioural treatments with or without medication, the US guidelines on obesity recommend informing them about bariatric surgery.

CLINICAL PROTOCOLS :

Much of the western world has created clinical practice guidelines in an attempt to address rising rates of obesity. Australia, Canada, the European Union, and the United States have all published statements since 2004.[69]

In a clinical practice guideline by the American college of physicians, the following five recommendations are made.[70]

1. People with a BMI of over 30 should be counselled on diet, exercise and other relevant behavioural interventions and set a realistic goal for weight loss.

2. If the goals are not achieved, pharmacotherapy can be offered. The person needs to be informed of the possibility of side effects and the unavailability of long term safety and efficacy data

3. Drug therapy may consist of sibutramine, orlistat, phentermine, diethylpropion, fluoxetine, and bupropion. For more severe cases of obesity, stronger drugs such as amphetamine and methamphetamine may be used on a selective basis. The evidence is not sufficient to recommend sertraline, topiramate or zonisamide.

4. In people with a BMI over 40 who fail to achieve their weight loss goals (with or without medication) and who develop obesity-related complications, referral for bariatric surgery may be indicated. The person needs to be aware of the potential complications.

5. Those requiring bariatric surgery should be referred to high volume referral centres, as the evidence suggests that surgeons who frequently perform these procedures have fewer complications.

A clinical practice guideline by the US Preventive Services Task Force (USPSTF) concluded that the evidence is insufficient to recommend for or against routine behavioural counselling to promote a healthy diet in unselected people in primary care settings, but that intensive behavioural dietary counselling is recommended in those with hyperlipidemia and other known risk factors for cardiovascular and diet-related chronic disease. Intensive counselling can be delivered by primary care clinicians or by referral to other specialists, such as nutritionists or dieticians.

Canada developed and published evidence-based practice guidelines in 2006.The guidelines attempt to address the prevention and management of obesity at both the individual and population levels in both children and adults. The European Union published clinical practice guidelines in 2008 in an effort to address the rising rates of obesity in Europe. Australia came out with practice guidelines in 2004.

REFERENCES:1. “Obesity and overweight Fact sheet N°311”. WHO. January 2015.

Retrieved 2 February 2016.

2. Kanazawa M, Yoshiike N, Osaka T, Numba Y, Zimmet P, Inoue S. “Criteria and Classification of Obesity in Japan and Asia-Oceania”. Nutrition and Fitness: Obesity, the Metabolic Syndrome, Cardiovascular Disease, and Cancer. World Review of Nutrition and Dietetics. 2005; 94. pp. 1–12.

3. Haslam DW, James WP. “Obesity”. Lancet (Review). October 2005; 366 (9492): 1197–209.

4. Luppino FS, de Wit LM, Bouvy PF, Stijnen T, Cuijpers P, Penninx BW, Zitman FG. “Overweight, obesity, and depression: a systematic review and meta-analysis of longitudinal studies”. Archives of General Psychiatry. March 2010; 67 (3): 220–9. doi:10.1001/archgenpsychiatry.2010.2.

5. Yazdi FT, Clee SM, Meyre D. “Obesity genetics in mouse and human: back and forth, and back again”. PeerJ. 2015; 3: e856.

6. Bleich S, Cutler D, Murray C, Adams A. “Why is the developed world obese?”. Annual Review of Public Health (Research Support). 2008; 29: 273–95.

7. Oxford Handbook of Medical Sciences (2nd ed.). Oxford: OUP Oxford. 2011. p. 180.

8. Kushner R. Treatment of the Obese Patient (Contemporary Endocrinology). Totowa, NJ: Humana Press. 2007; p. 158. Retrieved 5 April 2009.

9. Yanovski SZ, Yanovski JA. “Long-term drug treatment for obesity: a systematic and clinical review”. JAMA (Review). January 2014; 311 (1): 74–86.

10. Colquitt JL, Pickett K, Loveman E, Frampton GK. “Surgery for weight loss in adults”. The Cochrane Database of Systematic Reviews (Meta-analysis, Review). August 2014; 8 (8): CD003641.

11. Imaz I, Martínez-Cervell C, García-Alvarez EE, Sendra-Gutiérrez JM, González-Enríquez J. “Safety and effectiveness of the intragastric balloon for obesity. A meta-analysis”. Obesity Surgery. July 2008; 18 (7): 841–6.

12. Encyclopedia of Mental Health (2 ed.). Academic Press. 2015. p. 158.

13. Dibaise JK, Foxx-Orenstein AE (July 2013). “Role of the gastroenterologist in managing obesity”. Expert Review of Gastroenterology &Hepatology (Review). July 2013;7 (5): 439–51.

14. Woodhouse R. “Obesity in Art – A Brief Overview”. Obesity in art:


a brief overview. Frontiers of Hormone Research. 2008; 36. pp. 271–86.

15. Jensen, MD; Ryan, DH; Apovian, CM; Ard, JD; Comuzzie, AG; Donato, KA; Hu, FB; Hubbard, VS; Jakicic, JM; Kushner, RF; Loria, CM; Millen, BE; Nonas, CA; Pi-Sunyer, FX; Stevens, J; Stevens, VJ; Wadden, TA; Wolfe, BM; Yanovski, SZ; Jordan, HS; Kendall, KA; Lux, LJ; Mentor-Marcel, R; Morgan, LC; Trisolini, MG; Wnek, J; Anderson, JL; Halperin, JL; Albert, NM; Bozkurt, B; Brindis, RG; Curtis, LH; DeMets, D; Hochman, JS; Kovacs, RJ; Ohman, EM; Pressler, SJ; Sellke, FW; Shen, WK; Smith SC, Jr; Tomaselli, GF; American College of Cardiology/American Heart Association Task Force on Practice, Guidelines.; Obesity, Society. “2013 AHA/ACC/TOS guideline for the management of overweight and obesity in adults: a report of the American College of Cardiology/American Heart Association Task Force on Practice Guidelines and The Obesity Society”. Circulation. 24th June 2014; 129 (25 Suppl 2): S102–38.

16. Pollack A. “A.M.A. Recognizes Obesity as a Disease”. New York Times. Archived from the original on 24 June 2013.

17. Weinstock, Matthew. “The Facts About Obesity”. H&HN. American Hospital Association. 21stjune 2013; Retrieved 24 June 2013.

18. Haslam D. “Obesity: a medical history”. Obesity Reviews (Review). march 2007; 8 Suppl 1: 31–6.

19. Caballero B. “The global epidemic of obesity: an overview”. Epidemiologic Reviews. 2007; 29: 1–5.

20. “Obesity and overweight”. World Health Organization. Retrieved 8 April 2009.

21. Hales CM, Carroll MD, Fryar CD, Ogden CL. “Prevalence of Obesity Among Adults and Youth: United States, 2015–2016”. October 2018; NCHS Data Brief (288): 1–8.

22. Seidell 2005 p.5

23. Sturm R.”Increases in morbid obesity in the USA: 2000–2005”. public Health. July 2007; 121 (7): 492–6.

24. Howard NJ, Taylor AW, Gill TK, Chittleborough CR. “Severe obesity: Investigating the socio-demographics within the extremes of body mass index”. Obesity Research & Clinical Practice. March 2002; 2 (1): I–II.

25. Tjepkema M (6 July 2005). “Measured Obesity–Adult obesity in Canada: Measured height and weight”. Nutrition: Findings from the Canadian Community Health Survey. Ottawa, Ontario: Statistics Canada.

26. “Obesity Update 2017”(PDF). Organisation for Economic Co-operation and Development. Retrieved 6 October 2018.

27. Tsigos C, Hainer V, Basdevant A, Finer N, Fried M, Mathus-Vliegen E, Micic D, Maislos M, Roman G, Schutz Y, Toplak H, Zahorska-Markiewicz B.”Management of obesity in adults: European clinical practice guidelines”(PDF). Obesity Facts. April 2008; 1 (2): 106–16. Archived fromthe original(PDF)on 26 April 2012.

28. Kern, PA Ranganathan, S; Li; C; wood, L; Ranganathan G (2001).

29. Gray DS, Fujioka K. “Use of relative weight and body mass index for the determination of adiposity”. J ClinEpidemiol. 1991;44 (6): 545-50.

30. Sturm R.”Increases in morbid obesity in the USA: 2000–2005”. Public Health. July 2007; 121 (7): 492–6.

31. Lau DC, Douketis JD, Morrison KM, HramiakIM, Sharma AM, Ur E. “2006 Canadian clinical practice guidelines on the management and prevention of obesity in adults and children [summary]”. CMAJ (Practice Guideline, Review). April 2007; 176 (8): S1–13.

32. Drewnowski A, Specter SE. “Poverty and obesity: the role of energy density and energy costs”. The American Journal of Clinical Nutrition (Review). January 2004; 79 (1): 6–16.

33. Nestle M, Jacobson MF. “Halting the obesity epidemic: a public health policy approach”. Public Health Reports (Research Support). 2000; 115 (1): 12–24.

34. James WP. “The fundamental drivers of the obesity epidemic”. Obesity Reviews (Review). march 2008; 9 Suppl 1 (Suppl 1): 6–13.

35. Keith SW, Redden DT, Katzmarzyk PT, Boggiano MM, Hanlon EC, Benca RM, Ruden D, Pietrobelli A, Barger JL, Fontaine KR, Wang C, Aronne LJ, Wright SM, Baskin M, Dhurandhar NV, Lijoi MC, Grilo CM, DeLuca M, Westfall AO, Allison DB. “Putative contributors to the secular increase in obesity: exploring the roads less traveled”.

International Journal of Obesity (Review). November 2006; 30 (11): 1585–94.

36. Schwartz, MW; Seeley, RJ; Zeltser, LM; Drewnowski, A; Ravussin, E; Redman, LM; Leibel, RL. “Obesity Pathogenesis: An Endocrine Society Scientific Statement”. Endocrine Reviews (Professional society guidelines). 1 August 2017; 38 (4): 267–296.

37. WHO 2000 p.9

38. Nikcevic AV,Kuczmierczyk AR, Bruch M. Formulation and Treatment in Clinical Health Psychology. Routledge. 2009.

39. Caballero B. “The global epidemic of obesity: an overview”. Epidemiologic Reviews.2007; 29: 1–5.

40. “Obesity and overweight”. World Health Organization. Retrieved 8 April 2009.

41. Hales CM, Carroll MD, Fryar CD, Ogden CL. “Prevalence of Obesity Among Adults and Youth: United States, 2015–2016”. October 2017; NCHS Data Brief (288): 1–8.

42. Seidell 2005 p.5

43. Sturm R. “Increases in morbid obesity in the USA: 2000–2005”. Public Health. July 2007; 121 (7): 492–6.

44. Howard NJ, Taylor AW, Gill TK, Chittleborough CR. “Severe obesity: Investigating the socio-demographics within the extremes of body mass index”. Obesity Research & Clinical Practice. March 2008; 2 (1): I–II.

45. Tjepkema M. “Measured Obesity–Adult obesity in Canada: Measured height and weight”. Nutrition: Findings from the Canadian Community Health Survey. Ottawa, Ontario: Statistics Canada. 6th July 2005.

46. “Obesity Update 2017”(PDF). Organisation for Economic Co-operation and Development. Retrieved 6 October 2018.

47. Flier JS. “Obesity wars: molecular progress confronts an expanding epidemic”. Cell (Review). January 2004; 116 (2): 337–50.

48. Zhang Y, Proenca R, Maffei M, Barone M, Leopold L, Friedman JM. “Positional cloning of the mouse obese gene and its human homologue”. Nature (Research Support). December 1994; 372 (6505): 425–32.

49. Boulpaep, Emile L.; Boron, Walter F. Medical physiology: A cellular and molecular approach. Philadelphia: Saunders. 2007; p. 1227.

50. US Department of Health and Human Services. “2015–2020 Dietary Guidelines for Americans - health.gov”. health.gov. Skyhorse Publishing Inc. 2017; Retrieved 30 September 2019.

51. Arnett, Donna K.; Blumenthal, Roger S.; Albert, Michelle A.; Buroker, Andrew B.; Goldberger, Zachary D.; Hahn, Ellen J.; Himmelfarb, Cheryl D.; Khera, Amit; Lloyd-Jones, Donald; McEvoy, J. William; Michos, Erin D.; Miedema, Michael D.; Muñoz, Daniel; Smith, Sidney C.; Virani, Salim S.; Williams, Kim A.; Yeboah, Joseph; Ziaeian, Boback. “2019 ACC/AHA Guideline on the Primary Prevention of Cardiovascular Disease”. Circulation. 17 March 2019; 140 (11): e596–e646.

52. Strychar I. “Diet in the management of weight loss”. CMAJ (Review). January 2006; 174 (1): 56–63.

53. Shick SM, Wing RR, Klem ML, McGuire MT, Hill JO, Seagle H. “Persons successful at long-term weight loss and maintenance continue to consume a low-energy, low-fat diet”. Journal of the American Dietetic Association. April 1998; 98 (4): 408–13.

54. Tate DF, Jeffery RW, Sherwood NE, Wing RR. “Long-term weight losses associated with prescription of higher physical activity goals. Are higher levels of physical activity protective against weight regain?”. The American Journal of Clinical Nutrition (Randomized Controlled Trial). April 2007; 85 (4): 954–9.

55. US Preventive Services Task Force. “Behavioral Weight Loss Interventions to Prevent Obesity-Related Morbidity and Mortality in Adults US Preventive Services Task Force Recommendation Statement”. JAMA. 2018; 320 (11): 1163–1171.

56. Services, Swedish Agency for Health Technology Assessment and Assessment of Social Services (SBU) (1987). “Dietary treatment of obesity”. Annals of the New York Academy of SciencesRetrieved 17 June 2016. 499 (1): 250–263.

57. Johnston BC, Kanters S, Bandayrel K, Wu P, Naji F, Siemieniuk RA, Ball GD, Busse JW, Thorlund K, Guyatt G, Jansen JP, Mills


EJ. Comparison of weight loss among named diet programs in overweight and obese adults: a meta-analysis. JAMA. September 2014 312 (9): 923–33.

58. Naude CE, Schoonees A, Senekal M, Young T, Garner P, Volmink J. “Low carbohydrate versus isoenergetic balanced diets for reducing weight and cardiovascular risk: a systematic review and meta-analysis”. PLOS ONE (Research Support). 2014; 9 (7): e100652. Bibcode:2014PLoSO...9j0652N.

59. Wing RR, Phelan S. “Long-term weight loss maintenance”. The American Journal of Clinical Nutrition (Review). July 2005; 82 (1 Suppl): 222S–225S.

60. Thangaratinam S, Rogozinska E, Jolly K, Glinkowski S, Roseboom T, Tomlinson JW, Kunz R, Mol BW, Coomarasamy A, Khan KS. “Effects of interventions in pregnancy on maternal weight and obstetric outcomes: meta-analysis of randomised evidence”. BMJ (Meta-analysis). May 2012; 344: e2088.

61. LeFevre ML. “Behavioralcounseling to promote a healthful diet and physical activity for cardiovascular disease prevention in adults with cardiovascular risk factors: U.S. Preventive Services Task Force Recommendation Statement”. Annals of Internal Medicine. October 2014; 161 (8): 587–93.

62. Heymsfield SB, Wadden TA. “Mechanisms, Pathophysiology, and Management of Obesity”. The New England Journal of Medicine. JANUARY 2017; 376 (3): 254–266.

63. Wolfe SM. “When EMA and FDA decisions conflict: differences in patients or in regulation?” August 2013; BMJ. 347: f5140.

64. Rucker D, Padwal R, Li SK, Curioni C, Lau DC. “Long term pharmacotherapy for obesity and overweight: updated meta-analysis”. BMJ (Meta-analysis). December 2007; 335 (7631): 1194–9.

65. Wood, Shelley. “Diet Drug Orlistat Linked to Kidney, Pancreas Injuries”. Medscape. Medscape News. Retrieved 26 April 2011.

66. Chang SH, Stoll CR, Song J, Varela JE, Eagon CJ, Colditz GA. The effectiveness and risks of bariatric surgery: an updated systematic review and meta-analysis, 2003–2012. JAMA Surgery (Meta-analysis, Review). March 2014; 149 (3): 275–87.

67. Sjöström L, Narbro K, Sjöström CD, Karason K, Larsson B, Wedel H, Lystig T, Sullivan M, Bouchard C, Carlsson B, Bengtsson C, Dahlgren S, Gummesson A, Jacobson P, Karlsson J, Lindroos AK, Lönroth H, Näslund I, Olbers T, Stenlöf K, Torgerson J, Agren G, Carlsson LM. Effects of bariatric surgery on mortality in Swedish obese subjects. The New England Journal of Medicine (Research Support). August 2007; 357 (8): 741–52.

68. Weintraub, Karesn. “New allies in war on weight”. The Boston Globe. The Boston Globe. Retrieved 30 June 2014.

69. Imaz I, Martinez-Cervell C, Garcia-Alvarez EE, sendra-Gutiterrez JM, Gonzalez-Enrsquez. Safety and effectiveness of the intragastricballon for obesity. ObesSurg, July 2008; 18(7):841-6.

70. Snow V, Barry P, Fitterman N, Qaseem A, Weissk. pharmacological and surgical management of obesity in primary care: A clinical practice guidelines from the American college of physicians. Annals of Internal medicine, 2005;142(7):521-31


April - June 2019 31 Journal ofPharmacyandChemistry •Vol.13 • Issue.2January - March 2018 23 Journal ofPharmacyandChemistry •Vol.12 • Issue.1January - March 2012 31 Journal of Pharmacy and Chemistry • Vol.6 • Issue.1

Science – Tech Foundation

Journal of Pharmacy and Chemistry

(An International Research Journal of Pharmaceutical and Chemical Sciences)(Indexing in CAS)

Plot No. 22, Vidyut Nagar, Anantapur – 515 001(A.P), INDIA

MEMBERSHIP FORMDear Editor,

I/We wish to be Annual Member / Life Member of Journal of Pharmacy and Chemistry (AnInternational Research Journal of Pharmaceutical and Chemical Sciences) and agree to abide by your rules andregulations

1. Name in full: _________________________________________________ Male Female

(In block letters)

2. Nationality: _______________________________ 3. Date of birth:___________________________

3. Corresponding Address: _____________________________________________________________

____________________________________________________________________________________

____________________________________________________________________________________

5. Institutional / Business Address: _____________________________________________________

6. Phone : (O): ______________ (R): _______________ E-mail :______________________________

7. Academic Qualifications : ___________________________________________________________

8. Profession and Experience : __________________________________________________________

9. Present Position / Post : _____________________________________________________________

10. Pharmaceutical / Chemical interest: _____________________________________________________

Membership Fees LifeIndividual Rs. 8000/- US $ 800

Institutional Rs. 20,000/- US $ 2000

AnnualRs. 2000/- US $ 150

Rs. 2500/- US $ 200

Rs. / $ ________________________________ remitted by Bank / Cash

DD No. ________________ Date _____________ Banker’s Name___________________________

DD’s are drawn in favour of Science-Tech Foundation, Payable at Anantapur.

Place: ____________________

Date: _____________________

(Signature of the applicant)

Note: (i) All authors should be member of this Journal, (ii) Saperate Photostat copy may be filled and send.


April - June 2019 32 Journal ofPharmacyandChemistry •Vol.13 • Issue.2January - March 2018 24 Journal ofPharmacyandChemistry •Vol.12 • Issue.1

InstRUCtIon to AUtHoRsGENERAL REQUIREMENTS: Journal of Pharmacy and Chemistry (ISSN 0973-9874) is a quarterly Journal, Indexing in CAS(Coden:JPCOCM) which publishes original research work that contributes significantly to further the scientific knowledge in Pharmaceutical Sciences (Pharmaceutical Technology, Pharmaceutics, Biopharmaceutics, Pharmacokinetics, Pharmaceutical Chemistry, Computational Chemistry and Molecular Drug Design, Pharmacognosy and Phytochemistry, Pharmacology, Pharmaceutical analysis, Pharmacy Practice, Clinical and Hospital Pharmacy, Cell Biology, Genomics and Proteomics, Pharmacogenomics, Stem Cell research, vaccines & Cera, Bioinformatics and Biotechnology of Pharmaceutical Interest) and in Chemical Sciences (Inorganic, Soil, Forensic, analytical, Nano, environmental, Polymer, Physical, agricultural, Medicinal, Biochemistry, organic, Computational, Food, Pesticides etc). Manuscripts are accepted for consideration by Journal of Pharmacy and Chemistry on the condition that they represent original material, have not been published previously, are not being considered for publication elsewhere, and have been approved by each author. review articles, research papers, short communication and letters to the editor may be submitted for publication.

SUBMISSION OF MANUSCRIPTS: Typewritten manuscripts prepared using MS word should be submitted in triplicate and rw-CD to Prof. Dr. K.N Jayaveera, editor-in-Chief of Journal of Pharmacy and Chemistry , Plot No 22, vidyut Nagar, Ananthapur- 515 001, Andhra Pradesh, India. e-mail: [email protected]

all components of the manuscript must appear within a single electronic file: references, figure legends and tables must appear in the body of the manuscript.

TYPING INSTRUCTION: The following detailed instructions are necessary to allow direct reproduction of the manuscript for rapid publishing. If instructions are not followed, the manuscript will be returned for retyping. The following typefaces, in 12 points size, are preferred: Times Roman.

GENERAL FORMAT: The typing area must be exactly 6 5/8” (168 mm) wide by 9 7/8” (250 mm) long. Justify margins left and right (block format). The entire typing area of each page must be filled, leaving no wasted space. Text should be double-spaced, special care should be taken to insure that symbols, superscripts and subscripts are legible and do not overlap onto lines above or below. Make sure text lines are equidistant.

TITLE: On the first page of the manuscript, start title 1” (25 mm) down from top text margin. Type title in all capital letters, centred on the width of the typing area and single-spaced if more than one line is required. The title should be brief, descriptive and have all words spelled out. Double-space, then type the author(s) name(s), single-spaced if more than one line is required. Double-space, than type author(s) address(es), single-spaced, capitalizing first letters of main words. Quadruple-space before abstract.

ABSTRACT: Centre, type and underline abstract heading, capitalizing the first letter. a double-space should separate the heading from the abstract text. Indent abstract text approximately 1/2” (13 mm) from both left and right margins. The abstract should be intelligible to the reader without reference to the body of the paper and be suitable for reproduction by abstracting services. Introduction to the text (without a heading) should being four spaces below the abstract using full margins.

KEY WORDS: Three or more key words must be provided by authors for indexing of their article. Key words will be listed directly below the abstract. abbreviated forms of chemical compounds are not acceptable. Spell out entirely, using the official nomenclature. Example: L-dihydroxyphenylalanine (L-DoPa)

MAJOR HEADINGS: Papers must include the major headings: Introduction, Methods, results, Discussion, acknowledgments and references. Capitalize first letter, underline, and centre headings on width of typing area.

TABLES/FIGURES: Incorporate tables and/or figures (B & w) with their legends into the main body of text.

REFERENCES: referneces should be referred to a number [1] in the text and be listed according to this numbering at the end of the paper. only papers and books that have been published or in press may be cited; unpublished manuscripts or manuscripts submitted to a journal but which have not been accepted may not be cited.

The references shoulld comprise the following information and in the given order and with given punctuation as given in the example below: Author name (s), Initials (s), Publication Title, Page Number, year of Publication.

Standard Journal Article: [1] Bhattacharyya D, Pandit S, Mukherjee r, Das N, Sur TK.

Indian J Physiol Pharmacol 2003; 47:435.[2] Skottova N, Krecman V. Physiol Res 1998; 47:1.

Book:[1] Ghosh MN. Fundamentals of experimental Pharmacology,

2nd ed. Calcutta Scientific Book Agency, 1984:154.

Proofs will be sent to the corresponding author. These should be returned as quickly as possible.

The facts and view in the article will be of the authors and they will be totally responsible for authenticity, validity and originality etc. the authors should give an undertaking while submitting the paper that the manuscripts submitted to journal have not been published and have not been simultaneously submitted or published elsewhere and manuscripts are their original work.

www.jpc.stfindia.comwww.stfindia.com

ThisQuarterly Journal isPrinted,Published,OwnedandEditedbyDr.K.N. Jayaveera,Published fromScience-TechFoundation,PlotNo.22,DoorNo. 12/3/925,VidyutNagar,Anantapur - 515001.Send your queries at www.stfindia.comandPrintedatViplaComputerServices (Designers&MultiColourOffsetPrinters) # 2-3-756/2,Golnaka,Amberpet,Hyderabad - 500013.

Documents

Online : ISSN 2349-669X J.Pharm.Chem CODEN:JPCOCM Journal ...jpc.stfindia.com/images/current_issue/JPC 13_1.pdf · Development and Validation of Reverse Phase Hplc Method for Estimation