research article self-microemulsifying drug delivery...

Hindawi Publishing CorporationJournal of PharmaceuticsVolume 2013 Article ID 328769 6 pageshttpdxdoiorg1011552013328769

Research ArticleSelf-Microemulsifying Drug Delivery System Formulation andStudy Intestinal Permeability of Ibuprofen in Rats

Bharat Bhushan Subudhi1 and Surjyanarayan Mandal2

1 Department of Pharmaceutical Analysis School of Pharmaceutical Sciences SOA University Khandagiri SquareBhubaneswar Orissa 751030 India

2Mercury Pharmaceutical Limited Gujarat India

Correspondence should be addressed to Bharat Bhushan Subudhi bharatbhusansgmailcom

Received 20 December 2012 Accepted 23 May 2013

Academic Editor Hadi Valizadeh

Copyright copy 2013 B B Subudhi and S MandalThis is an open access article distributed under the Creative Commons AttributionLicense which permits unrestricted use distribution and reproduction in any medium provided the original work is properlycited

The study was aimed at developing a self-microemulsifying drug delivery system (SMEDDS) of Ibuprofen for investigating itsintestinal transport behavior using the single-pass intestinal perfusion (SPIP) method in ratMethods Ibuprofen loaded SMEDDS(ISMEDDS) was developed and was characterized The permeability behavior of Ibuprofen over three different concentrations(20 30 and 40 120583gmL) was studied in each isolated region of rat intestine by SPIP method at a flow rate of 02mLmin Thehuman intestinal permeability was predicted using the Lawrence compartment absorption and transit (CAT) model since effectivepermeability coefficients (119875eff) values for rat are highly correlated with those of human and comparative intestinal permeability ofIbuprofenwas carried outwith plain drug suspension (PDS) andmarketed formulation (MF)ResultsThedeveloped ISMEDDSwasstable emulsified upon mild agitation with 444 nm plusmn 213 and 9886 plusmn 121 as globule size and drug content respectively Higher119875eff in colon with no significant 119875eff difference in jejunum duodenum and ileum was observed The estimated human absorptionof Ibuprofen for the SMEDDS was higher than that for PDS and MF (119875 lt 001) Conclusion Developed ISMEDDS would possiblybe advantageous in terms of minimized side effect increased bioavailability and hence the patient compliance

1 Introduction

In recent years much attention has been focused on usingemulsion drug delivery system for the purpose of improvingthe solubility and oral absorption of poorly water-solubledrugs [1ndash3] SMEDDSs are defined as isotropic mixturesof natural or synthetic oils solid or liquid surfactants oralternatively one or more hydrophilic solvents and cosol-ventssurfactants that have a unique ability of formingfine oil-in-water (ow) microemulsions upon mild agitationfollowed by dilution in aqueous media such as GI fluids[4] SMEDDSs spread readily in the GI tract and thedigestive motility of the stomach and the intestine providethe agitation necessary for self-emulsification SMEDDS isoptically clear and thermodynamically stable system with adroplet size less than 100 nm This system has been shown toimprove absorption of drugs due to small droplet size and

promotes intestinal lymphatic transport due to its specificcomponents [5] Ibuprofen (2-(4-(2-methylpropyl)phenyl)propanoic acid) is a nonsteroidal anti-inflammatory drug(NSAID) used for relief of symptoms of arthritis and feverIbuprofen is a poor water-soluble drug and absorption inrats was shown to occur mainly from the intestine and to alesser though significant extent from the stomach [6] SinceIntestinal permeability is necessary for oral administrationso in this study SMEDDS was developed to improve thesolubility and oral absorption of Ibuprofen Labrafil M1944CS Tween 80 and Transcutol P were used as oil phasesurfactant and cosurfactant respectively The intestinal per-meability of Ibuprofen was determined using the single-passintestinal perfusion (SPIP) in rats since this method providesunique advantages of experimental control (eg compoundconcentration and intestinal perfusion rate) ability to studyregional differences an intact intestinal blood supply and

2 Journal of Pharmaceutics

a functional intestinal barrier [7] Intestinal permeabilitystudy of Ibuprofen through ISMEDDS using SPIPwas a novelway to study the permeation behavior of Ibuprofen since itcontrols the experimental variables and also provides clear-cut idea of the permeation pattern in different regions ofintestine

2 Materials and Methods

21 Materials Ibuprofen was obtained as gift sample fromAbbott India Ltd Goa India Capmul MCM Labrafac CCCremophor EL Cremophor RH 40 Labrafil M 1944CS andTranscutol P were kindly provided by Gattefosse FranceIsopropyl Myristate (IPM) Tween 60 PEG 600 PEG 400ethanol and glycerol were purchased from National Chemi-cals (Baroda India) Acetonitrile (HPLC grade) ammoniumacetate methanol (HPLC grade) and sodium citrate werepurchased from Suvidhinath Laboratories (Baroda India)All other chemicals were reagent grade

22 Animals Male albino rats (250 plusmn 20 g) were used for thecomparative in vivo studies The animals were maintained attemperature (25 plusmn 2∘C) and humidity (60 plusmn 5) and weresupplied with food and water ad libitum All experimentsconducted on animals were approved by the Animal EthicalCommittee and animal ethical committee registration no984a06CPCSEA

23 Solubility Studies Solubility of Ibuprofen in variousoils (Oleic acid Isopropyl Myristate Soya bean oil LabrafilM 1944CS and Capmul CMC) surfactants (Labrafac CCTween 60 Cremophor EL and Cremophor RH 40) andcosurfactants (PEG 400 PEG 600 glycerol and ethanol) wasdetermined A total of 5mL of each of the selected vehicle wasadded to each cap vial with excess amount of Ibuprofen Aftersealing the mixture was heated at 40 plusmn 2∘C in a water bathto facilitate the solubilization and then mixed using a vortexmixerMixtures were then shakenwith orbital shaker at roomtemperature for 48 hrs Then each vial was centrifuged at4000 rpm for 5min and insoluble aswell as soluble Ibuprofenwas quantified by UV-VIS Spectrophotometer [8]

24 Pseudoternary Phase Diagram Study Pseudoternaryphase diagrams were constructed to obtain the appropriatecomponents and their concentration ranges that result inlarge existence area of microemulsion For convenience thephase diagrams were constructed by drawing ldquowater dilutionlinesrdquo representing increasing water content and decreasingsurfactant-cosurfactant levels If turbidity appeared followedby a phase separation the samples were considered to bebiphasic If clear and transparent mixtures were visualizedafter stirring the samples were considered monophasic Thesamplesweremarked as points in the phase diagramas shownin Figure 1 The area covered by these points was consideredto be the microemulsion existence region After performingthe solubility study and phase diagram study componentswere selected for microemulsion formulation The effect ofIbuprofen in the phase diagrams was also investigated [9]

25 Preparation of SMEDDS of Ibuprofen ISMEDDS wasprepared by emulsification method [10] Ibuprofen was firstdissolved in the premeasured volume of oil by stirring on amagnetic stirrer Amixture of the surfactant and cosurfactantat a fixed ratio (vv) was added to the above resulting mixturein order to develop SMEDDS Emulsification time was alsodetermined The optimized ISMEDDS was also subjected foraccelerated stability study

26 Optical Clarity and Emulsification Time Optical clarityof ISMEDDS was determined by the refractive index of thesystem and transmittance Refractive index was measuredby an Abbe refractometer (Bausch and Lomb Optical Com-pany Rochester NY USA) by placing one drop of solutionon the slide Transparency of microemulsion formulationwas determined by measuring percentage transmittance at650 nm with purified water taken as blank through UV-VISSpectrophotometer (UV-1601-220X Shimadzu) [11] 1mL ofthe formulation was put into 100mL of distilled water andthe dissolution time was measured as emulsification time

27 Globule Size and Zeta Potential Measurement Globulesize and zeta potential of ISMEDDS were carried out bydynamic light scattering through Zetasizer 300 ZS-NanoMalvern Instruments Corp UK [12]

28 Single-Pass Intestinal Perfusion (SPIP) of the Rat SegmentsBefore the SPIP experiment sufficient Krebs-Ringerrsquos buffersolution was added to adjust the formulations to a designedconcentration The intestinal permeability experiments ofISMEDDS with different concentrations of Ibuprofen wereinvestigated and compared to bothPDS andMAof equivalentdrug concentration Preliminary studies are necessary beforecommencing the SPIP studies to ensure that the loss of drugfrom perfusion is due to absorption and not by other lossesthat is the binding of the intestinal wall or degradation[13] Approximately 10 cm from the rat intestine was clippedintestinal sacs were turned over and put into 50mL Krebs-Ringerrsquos buffer solution in which Ibuprofen (20 120583gmL) wasincubated at 37 plusmn 2∘C for 3 hrs Chromatographic separa-tion was achieved isocratically on a C18 column (InertsilC18 5m 150mm times 46mm) utilizing a mobile phase ofacetonitrilephosphate bufferwater (60 30 10 vv pH 70)at a flow rate of 1mLmin with UV detection at 264 nmSamples of 40 120583L were withdrawn at 3 hrs interval dilutedwith mobile phase and assayed by HPLC SPIP studieswere performed according to previously described methods[14ndash16] After an overnight fast male albino rats weighingapproximately 250 plusmn 20 gm were anesthetized with 2mgkgim injection of ketamine Body temperature wasmaintainedusing a surgical lamp kept under the ratrsquos cage The abdomenwas opened with a midline incision an intestinal segmentof approximately 10 cm was chosen and separated Intestinalsegment was then rinsed with isotonic saline (37∘C) untilthe outlet solution was clear Using an infusion pump theintestinal segmentswere perfused at a flow rate of 02mLminfor 30min with Krebs-Ringerrsquos buffer solution Then thepump was connected to the reservoir containing Ibuprofen

Journal of Pharmaceutics 3

10

20

30

40

50

60

70

80

9010

20

30

40

50

60

70

80

90

10 20 30 40 50 60 70 80 90SurCosurWater

Oil

(a)

10

20

30

40

50

60

70

80

9010

20

30

40

50

60

70

80

90

10 20 30 40 50 60 70 80 90SurCosurWater

Oil

(b)

Figure 1 Shaded area of pseudoternary Phase diagram ((a) 3 1 and (b) 35 1 ratio of Sur to Cosur) representing the microemulsion area

Table 1 The net water flux (NWF) in each intestinal segment (119899 = 4)

Time (min) Duodenum Jejunum Ileum Colon60 minus0584 plusmn 0121 minus0144 plusmn 0175 minus0102 plusmn 0056 minus0129 plusmn 0018

80 minus0555 plusmn 0212 minus0117 plusmn 0128 minus0238 plusmn 0078 minus0197 plusmn 0056



loaded SMEDDS with the phenol red (marker) When thebuffer solution was completely pushed out time was set as119905 = 0 Each perfusion experiment lasted for 3 hrs and theperfusate was quantitatively collected in 60 90 120 135 150165 and 180min Samples were stored in a refrigerator atminus20∘C until analysis by HPLC

29 HPLC Analysis of Samples All samples were dilutedto 5mL with the mobile phase and measured by HPLC(Shimadzu Japan) system consisting of membrane degasserbinary solvent delivery system a pump a rheodyne injec-tor equipped with a 20120583L sample loop and UV detector(Shimadzu Japan) [17] Aliquots of 20 120583L were injected ontothe HPLC system which was used to analyze Ibuprofen at264 nm Chromatographic separations were achieved usingan Hypersil ODS C18 column (250 times 46 id times 5 120583m) Themobile phase used for the sample analysis was acetoni-trilephosphate bufferwater (60 30 10 vv pH 70) with theflow rate of 10mLmin Water absorption or secretion (flux)was measured by gravimetric method [18]The net water flux(NWF) per cm of each segment was calculated using thefollowing equation and results were tabulated in Table 1

NWF (120583Lmincm) = [(1 minusCPRoutCPRin) times119876in119871] times 1000

(1)

where CPRin and CPRout are inlet and outlet concentrationsof phenol red respectively 119876in was the flow rate of the

perfusion solution entering the intestinal segment and 119871wasthe length of the intestinal segment (cm)

NWF value will be negative if intestinal lumen absorbswater while NWF is positive if intestinal lumen secreteswater however under normal condition intestinal lumenabsorbs water except when the high osmotic solution isadministered [19] The effective permeability (119875eff) was cal-culated based on the inlet and outlet concentrations ofIbuprofen (119862in and 119862out resp) and result was shown inFigure 2 Steady state was considered to be achieved when theconcentrations of phenol red and Ibuprofen were constantThe permeability values are calculated after steady state wasachieved using the following equation [20]

119875eff =[119876 ln (119862in119862out)

1015840

]

2120587119903119871 (2)

where 119876 is the flow rate in mLmin 119903 is radius of theintestine (018 cm in rat and 175 cm in man) 119871 is the lengthof the perfused intestinal segment (cm) and (119862in119862out)

1015840 is theconcentration ratio corrected for fluid flux Using ANOVAthe statistical significances of the differences among groupmeans were assessed with the least significant difference(LSD) test and a value of119875 lt 005was considered statisticallysignificance

3 Results

Labrafil M 1944CS was selected as the oil phase for for-mulation development because it provided higher solubility

4 Journal of PharmaceuticsIn

tens

ity (

)

Size distribution by intensity16141210

86420

01 1 10 100 1000 10000

Record 3 drug

Size (dmiddotnm)

Figure 2 Globule size of Ibuprofen loaded SMEDDS indicating thenanosizing of the formulation

than other oils and oral compatibility Tween 80 as surfactantand Transcutol P as cosurfactant were used on the basisof their drug solubilising capacity and might influence thetight junctions of the epithelial cells [21] From the resultsof the pseudoternary phase diagram 3 1 ratio of Tween 80and Transcutol P was selected formicroemulsion preparationsince maximum microemulsion existing zone was observedas shown in Figure 1

The optimal formulation of ISMEDDS was 4 LabrafilM 1944CS as oil 215 Tween 80 as surfactant and 75Transcutol P as cosurfactant with globule size 444 nm plusmn 213(PdI-0266) as shown in Figure 2 PdI value was below 04suggesting the monodisperse property of the formulation

Zeta potential and drug content of the formulation wereminus919mV plusmn 12 indicating the stability of the formulationand 9886 plusmn 121 respectively Developed SMEDDS wasspontaneously forming microemulsion upon mild agitationin simulated gastric fluid at room temperature transmit-tance and refractive index of the resulted formulation werefound to be 979 plusmn 131 and 1337 plusmn 013 indicating thetransparency as well as the nanosizing of the formulationMoreover developed formulation was stable for 6 months atambient temperature

31 Validation of HPLC Assay The regression equation forthe concentration of Ibuprofen (ngmL) vs response in theperfusion fluid ranging from 50 ngmL to 500 ngmLwas119875 =9132119862ndash473 (1198772 = 09998) The mean recovery of Ibuprofenwas 1008 plusmn 113The intraday and interday RSDs were lessthan 2

32 Water Absorption or Secretion of Intestinal Lumens TheNWF for all intestinal segments was of negative value aslisted in Table 1 which suggested that the water was mainlyabsorbed in the intestine tract The NWF for the duodenumwas significantly higher than that for the jejunum ileum andcolon (119875 lt 005) andNWF for the jejunum ileum and colonwas not significantly varied (119875 gt 005)

33 SPIP Studies The amount of Ibuprofen was 9932 plusmn123after incubated in intestinal sacwith theKrebs-Ringerrsquosbuffer for 3 hrs which was not significantly decreased which

002040608

11214

0 50 100 150 200Time (min)

IbuprofenPhenol red

C outC

in

Figure 3 Concentration ratio of 119862out and 119862in with time forphenol red and Ibuprofen indicating the steady state permeabilityin intestine

suggested that neither Ibuprofen markedly bound to theintestinal wall nor the potential metabolites were formedThe intestinal permeability of Ibuprofen was studied as afunction of concentration in each segment of the intestineand 30 120583gmL concentrationwas chosen because of its limitedaqueous solubility Steady state is confirmed by plotting theconcentration of Ibuprofen and phenol red versus time asshown in Figure 3

Steady state which was assessed by the constant con-centrations of both phenol red and Ibuprofen was reachedabout 50min after the beginning of the perfusion Thepermeability values were calculated only after steady statewas achieved in the experiments The permeability values ofSMEDDS for each intestinal segment and concentrations arelisted in Table 2 119875eff in the jejunum at 10 120583gmL (0388 plusmn0021 cms) was significantly higher than 119875eff at 20120583gmL(0229 plusmn 0037 cms 119875 lt 001) There was no statisticaldifference in 119875eff at three investigated concentrations inother segments (including duodenum ileum and colon)Comparisons across each segment revealed that 119875eff at eachconcentration in colon was higher than all other segments(119875 lt 001) Moreover the permeability values in duodenumjejunum and ileumwere not statistically different (119875 gt 005)

Permeability values of Ibuprofen in ISMEDDS PDS andMF for each segment at 10120583gmLwere shown in Figure 4119875effof ISMEDDS was significantly higher than PDS and MF ineach segment (119875 lt 001) Except for the duodenum (119875 lt005) 119875eff of microemulsion drug delivery system and MFin the jejunum ileum and colon did not all reach statisticaldifference (119875 gt 005)

4 Discussion

Characterization data and microemulsifying study in sim-ulated gastric fluid revealed that the developed ISMEDDSswas of nanosize stable and was forming microemulsionspontaneously 119875eff in the colon at each concentration wassignificantly higher than those in other segments of ratintestine as shown in Table 2 There are various reportsshowing the regional difference in the expression and activityof P-glycoprotein throughout the gastric intestinal tract Theorder of the expression and activity of P-glycoprotein is


Table 2 Comparison of effective permeation coefficients (119875eff times 10minus4) of different intestinal segments with three concentrations Results are

given as mean plusmn SD (119899 = 4)

Conc of Ibuprofen (120583gmL) 119875eff in duodenum (cms) 119875eff in jejunum (cms) 119875eff in ileum (cms) 119875eff in colon (cms)10 0421 plusmn 0032 0458 plusmn 0044 0475 plusmn 0052 0671 plusmn 0067

20 0388 plusmn 0035 0434 plusmn 0046 0439 plusmn 0067 0598 plusmn 0062


00102030405060708

Duodenum Jejunum Ileum Colon

P efftimes10minus4

(cm

s)

Figure 4 Comparison of 119875eff of the SMEDDS PDS and MF indifferent intestinal segments (119899 = 4) at concentration of theIbuprofen as 10120583gmL

ileum gt duodenum and jejunum gt proximal and distal colonAs a multidrug resistance-reversing agent the intestinalabsorption of Ibuprofen was limited due to the presence ofthe P-glycoprotein throughout the intestinal tract [22] Thelower expression activity of P-glycoprotein was conducedto be increasing the permeability of Ibuprofen in colon ascompared to other sections (Table 2) It has been shownthat there is a selectivity of permeation depending of thegut sections Moreover small intestine has more cationicselectivity whereas the colon has more anionic selectivity[23] Another possible mechanism for higher 119875eff valuesin colon was due to the higher crypt surface area in thecolonic mucosa compared with the small intestine whichcould account for its high paracellular permeability Theextent of drug absorption in human can be predicted fromrat intestinal permeability experiments Human intestinalpermeability values may be estimated using the Lawrencecompartmental absorption and transit (CAT) model [24]

Fa(fraction of dose absorbed)=1minus [1+054 119875eff(man)]minus7

119875eff(man) = 36 times 119875eff(rat) + 003 times 10minus4

(3)

The estimated fractions of Ibuprofen absorbed forISMEDDS MF and PDS were 855 plusmn 19 593 plusmn 31 and812 plusmn 22 respectively It suggested that the estimated oralabsorption in human for microemulsion drug delivery sys-tems would be higher than that for PDS and MF (119875 lt 001)SMEED is known to improve the oral absorption of lipophilicdrugs The main mechanism reported includes increasingmembrane fluidity opening tight junction inhibiting P-glycoprotein andor CYP450 by surfactants and stimulatinglipoproteinchylomicron production by lipid [25]

5 Conclusions

Self-microemulsifying drug delivery system of Ibuprofen wasamonodisperse stable system capable of formingmicroemul-sion systems in simulated gastric fluid upon mild agitationwhich will provide sustained drug release in intestinal part assay was above 95 indicating the systemrsquos suitabilitywith drug Ibuprofen loaded SMEDDS and its dilutions werestable The 119875eff in jejunum at 10 120583gmL was significantlyhigher than that at 20120583gmL (119875 lt 001) Compared tothe jejunum duodenum and ileum the higher 119875eff in colonwas observed at the same concentration The estimatedabsorption of Ibuprofen in human for the microemulsiondrug delivery system was higher than that for PDS and MF(119875 lt 001)

References

[1] N H Shah M T Carvajal C I Patel M H Infeld and A WMalick ldquoSelf-emulsifying drug delivery systems (SEDDS) withpolyglycolyzed glycerides for improving in vitro dissolution andoral absorption of lipophilic drugsrdquo International Journal ofPharmaceutics vol 106 no 1 pp 15ndash23 1994

[2] W Wu Y Wang and L Que ldquoEnhanced bioavailability ofsilymarin by self-microemulsifying drug delivery systemrdquoEuro-pean Journal of Pharmaceutics and Biopharmaceutics vol 63no 3 pp 288ndash294 2006

[3] T R Kommuru B Gurley M A Khan and I K ReddyldquoSelf-emulsifying drug delivery systems (SEDDS) of coenzymeQ10 formulation development and bioavailability assessmentrdquoInternational Journal of Pharmaceutics vol 212 no 2 pp 233ndash246 2001

[4] J-L Tang J Sun and Z-G He ldquoSelf-emulsifying drug deliverysystems strategy for improving oral delivery of poorly solubledrugsrdquo Current Drug Therapy vol 2 no 1 pp 85ndash93 2007

[5] W N Charman and V J Stella ldquoTransport of lipophilicmolecules by the intestinal lymphatic systemrdquo Advanced DrugDelivery Reviews vol 7 no 1 pp 1ndash14 1991

[6] S G Kapsi and J W Ayres ldquoProcessing factors in developmentof solid solution formation of Ibuprofen for enhancement ofdrug dissolution and bioavailabilityrdquo International Journal ofPharmaceutics vol 229 pp 193ndash203 1999

[7] L Salphati K Childers L Pan K Tsutsui and L TakahashildquoEvaluation of a single-pass intestinal-perfusion method in ratfor the prediction of absorption in manrdquo Journal of Pharmacyand Pharmacology vol 53 no 7 pp 1007ndash1013 2001

[8] N Kaewnopparat S Kaewnopparat A Jangwang DManeenaun and T Chuchome ldquoIncreased solubilitydissolution and physicochemical studies of curcumin-pol-yvinylpyrrolidone K-30 solid dispersionsrdquo World Academy ofScience Engineering and Technology vol 55 pp 229ndash234 2009


[9] R Aboofazeli and M J Lawrence ldquoInvestigations into the for-mation and characterization of phospholipid microemulsionsII Pseudo-ternary phase diagrams of systems containing water-lecithin-isopropyl myristate and alcohol influence of purity oflecithinrdquo International Journal of Pharmaceutics vol 106 no 1pp 51ndash61 1994

[10] B M Patel S Mandal and K S Rajesh ldquoFormulation andkinetic modeling of curcumin loaded intranasal mucoadhesivemicroemulsionrdquo Journal of Pharmacy and Bioallied Sciencesvol 4 no 5 pp 81ndash83 2012

[11] A A Date and M S Nagarsenker ldquoDesign and evaluation ofmicroemulsions for improved parenteral delivery of propofolrdquoAAPS PharmSciTech vol 9 no 1 pp 138ndash145 2008

[12] D Patel and K K Sawant ldquoOral bioavailability enhancementof acyclovir by self-microemulsifying drug delivery systems(SMEDDS)rdquo Drug Development and Industrial Pharmacy vol33 no 12 pp 1318ndash1326 2007

[13] T J Cook and S S Shenoy ldquoIntestinal permeability of chlor-pyrifos using the single-pass intestinal perfusion method in theratrdquo Toxicology vol 184 no 2-3 pp 125ndash133 2003

[14] P Zakeri-Milani H Valizadeh H Tajerzadeh et al ldquoPredictinghuman intestinal permeability using single-pass intestinal per-fusion to ratrdquo Journal of Pharmacy and Pharmaceutical Sciencesvol 10 no 3 pp 368ndash379 2007

[15] J You Q-P Li Y-W Yu and F-D Cui ldquoAbsorption of zedoaryoil in rat intestine using in situ single pass perfusion modelrdquoActa Pharmacologica Sinica vol 39 no 10 pp 849ndash853 2004

[16] M Grassi and G Cadelli ldquoTheoretical considerations on thein vivo intestinal permeability determination by means of thesingle pass and recirculating techniquesrdquo International Journalof Pharmaceutics vol 229 no 1-2 pp 95ndash105 2001

[17] J C Tsao andT S Savage ldquoHigh-performance liquid chromato-graphic determination of ibuprofen in bulk drug and tabletsrdquoDrug Development and Industrial Pharmacy vol 11 no 5 pp1123ndash1131 1985

[18] S C Sutton M T Rinaldi and K E Vukovinsky ldquoComparisonof the gravimetric phenol red and 14C-PEG-3350 methodsto determine water absorption in the rat single-pass intestinalperfusion modelrdquo AAPS PharmSci vol 3 no 3 p E25 2001

[19] JMDeSesso andC F Jacobson ldquoAnatomical and physiologicalparameters affecting gastrointestinal absorption in humans andratsrdquo Food and Chemical Toxicology vol 39 no 3 pp 209ndash2282001

[20] U Fagerholm M Johansson and H Lennernas ldquoComparisonbetween permeability coefficients in rat and human jejunumrdquoPharmaceutical Research vol 13 no 9 pp 1336ndash1342 1996

[21] S Mandal S S Mandal and K S Rajesh ldquoDevelopmentcharacterization and evaluation of microemulsion gel for trans-dermal delivery of furosemiderdquo Journal of Pharmacy Researchvol 4 no 7 pp 3133ndash3135 2011

[22] T Ikegawa F Ushigome N Koyabu et al ldquoInhibition of P-glycoprotein by orange juice components polymethoxyflavonesin adriamycin-resistant human myelogenous leukemia(K562ADM) cellsrdquo Cancer Letters vol 160 no 1 pp 21ndash282000

[23] T Y Ma D Hollander R A Erickson H Truong H Nguyenand P Krugliak ldquoMechanism of colonic permeation of inulin Israt colon more permeable than small intestinerdquo Gastroenterol-ogy vol 108 no 1 pp 12ndash20 1995

[24] L X Yu and G L Amidon ldquoA compartmental absorption andtransitmodel for estimating oral drug absorptionrdquo InternationalJournal of Pharmaceutics vol 186 no 2 pp 119ndash125 1999

[25] X Sha G Yan Y Wu J Li and X Fang ldquoEffect of self-microemulsifying drug delivery systems containing Labrasol ontight junctions inCaco-2 cellsrdquoEuropean Journal of Pharmaceu-tical Sciences vol 24 no 5 pp 477ndash486 2005

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Pharmaceutics


MEDIATORSINFLAMMATION

of


a functional intestinal barrier [7] Intestinal permeabilitystudy of Ibuprofen through ISMEDDS using SPIPwas a novelway to study the permeation behavior of Ibuprofen since itcontrols the experimental variables and also provides clear-cut idea of the permeation pattern in different regions ofintestine

2 Materials and Methods

21 Materials Ibuprofen was obtained as gift sample fromAbbott India Ltd Goa India Capmul MCM Labrafac CCCremophor EL Cremophor RH 40 Labrafil M 1944CS andTranscutol P were kindly provided by Gattefosse FranceIsopropyl Myristate (IPM) Tween 60 PEG 600 PEG 400ethanol and glycerol were purchased from National Chemi-cals (Baroda India) Acetonitrile (HPLC grade) ammoniumacetate methanol (HPLC grade) and sodium citrate werepurchased from Suvidhinath Laboratories (Baroda India)All other chemicals were reagent grade

22 Animals Male albino rats (250 plusmn 20 g) were used for thecomparative in vivo studies The animals were maintained attemperature (25 plusmn 2∘C) and humidity (60 plusmn 5) and weresupplied with food and water ad libitum All experimentsconducted on animals were approved by the Animal EthicalCommittee and animal ethical committee registration no984a06CPCSEA

23 Solubility Studies Solubility of Ibuprofen in variousoils (Oleic acid Isopropyl Myristate Soya bean oil LabrafilM 1944CS and Capmul CMC) surfactants (Labrafac CCTween 60 Cremophor EL and Cremophor RH 40) andcosurfactants (PEG 400 PEG 600 glycerol and ethanol) wasdetermined A total of 5mL of each of the selected vehicle wasadded to each cap vial with excess amount of Ibuprofen Aftersealing the mixture was heated at 40 plusmn 2∘C in a water bathto facilitate the solubilization and then mixed using a vortexmixerMixtures were then shakenwith orbital shaker at roomtemperature for 48 hrs Then each vial was centrifuged at4000 rpm for 5min and insoluble aswell as soluble Ibuprofenwas quantified by UV-VIS Spectrophotometer [8]

24 Pseudoternary Phase Diagram Study Pseudoternaryphase diagrams were constructed to obtain the appropriatecomponents and their concentration ranges that result inlarge existence area of microemulsion For convenience thephase diagrams were constructed by drawing ldquowater dilutionlinesrdquo representing increasing water content and decreasingsurfactant-cosurfactant levels If turbidity appeared followedby a phase separation the samples were considered to bebiphasic If clear and transparent mixtures were visualizedafter stirring the samples were considered monophasic Thesamplesweremarked as points in the phase diagramas shownin Figure 1 The area covered by these points was consideredto be the microemulsion existence region After performingthe solubility study and phase diagram study componentswere selected for microemulsion formulation The effect ofIbuprofen in the phase diagrams was also investigated [9]

25 Preparation of SMEDDS of Ibuprofen ISMEDDS wasprepared by emulsification method [10] Ibuprofen was firstdissolved in the premeasured volume of oil by stirring on amagnetic stirrer Amixture of the surfactant and cosurfactantat a fixed ratio (vv) was added to the above resulting mixturein order to develop SMEDDS Emulsification time was alsodetermined The optimized ISMEDDS was also subjected foraccelerated stability study

26 Optical Clarity and Emulsification Time Optical clarityof ISMEDDS was determined by the refractive index of thesystem and transmittance Refractive index was measuredby an Abbe refractometer (Bausch and Lomb Optical Com-pany Rochester NY USA) by placing one drop of solutionon the slide Transparency of microemulsion formulationwas determined by measuring percentage transmittance at650 nm with purified water taken as blank through UV-VISSpectrophotometer (UV-1601-220X Shimadzu) [11] 1mL ofthe formulation was put into 100mL of distilled water andthe dissolution time was measured as emulsification time

27 Globule Size and Zeta Potential Measurement Globulesize and zeta potential of ISMEDDS were carried out bydynamic light scattering through Zetasizer 300 ZS-NanoMalvern Instruments Corp UK [12]

28 Single-Pass Intestinal Perfusion (SPIP) of the Rat SegmentsBefore the SPIP experiment sufficient Krebs-Ringerrsquos buffersolution was added to adjust the formulations to a designedconcentration The intestinal permeability experiments ofISMEDDS with different concentrations of Ibuprofen wereinvestigated and compared to bothPDS andMAof equivalentdrug concentration Preliminary studies are necessary beforecommencing the SPIP studies to ensure that the loss of drugfrom perfusion is due to absorption and not by other lossesthat is the binding of the intestinal wall or degradation[13] Approximately 10 cm from the rat intestine was clippedintestinal sacs were turned over and put into 50mL Krebs-Ringerrsquos buffer solution in which Ibuprofen (20 120583gmL) wasincubated at 37 plusmn 2∘C for 3 hrs Chromatographic separa-tion was achieved isocratically on a C18 column (InertsilC18 5m 150mm times 46mm) utilizing a mobile phase ofacetonitrilephosphate bufferwater (60 30 10 vv pH 70)at a flow rate of 1mLmin with UV detection at 264 nmSamples of 40 120583L were withdrawn at 3 hrs interval dilutedwith mobile phase and assayed by HPLC SPIP studieswere performed according to previously described methods[14ndash16] After an overnight fast male albino rats weighingapproximately 250 plusmn 20 gm were anesthetized with 2mgkgim injection of ketamine Body temperature wasmaintainedusing a surgical lamp kept under the ratrsquos cage The abdomenwas opened with a midline incision an intestinal segmentof approximately 10 cm was chosen and separated Intestinalsegment was then rinsed with isotonic saline (37∘C) untilthe outlet solution was clear Using an infusion pump theintestinal segmentswere perfused at a flow rate of 02mLminfor 30min with Krebs-Ringerrsquos buffer solution Then thepump was connected to the reservoir containing Ibuprofen


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(1)




119875eff =[119876 ln (119862in119862out)

1015840

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2120587119903119871 (2)



3 Results



tens

ity (

)


86420

01 1 10 100 1000 10000

Record 3 drug

Size (dmiddotnm)








002040608

11214

0 50 100 150 200Time (min)

IbuprofenPhenol red

C outC

in





4 Discussion








00102030405060708


P efftimes10minus4

(cm

s)





(3)


5 Conclusions


References































Volume 2014

ToxinsJournal of

VaccinesJournal of















AddictionJournal of






Autoimmune Diseases




Journal of


Pharmaceutics



of


10

20

30

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80

9010

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80

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10 20 30 40 50 60 70 80 90SurCosurWater

Oil

(a)

10

20

30

40

50

60

70

80

9010

20

30

40

50

60

70

80

90

10 20 30 40 50 60 70 80 90SurCosurWater

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(b)










(1)




119875eff =[119876 ln (119862in119862out)

1015840

]

2120587119903119871 (2)



3 Results



tens

ity (

)


86420

01 1 10 100 1000 10000

Record 3 drug

Size (dmiddotnm)








002040608

11214

0 50 100 150 200Time (min)

IbuprofenPhenol red

C outC

in





4 Discussion








00102030405060708


P efftimes10minus4

(cm

s)





(3)


5 Conclusions


References































Volume 2014

ToxinsJournal of

VaccinesJournal of















AddictionJournal of






Autoimmune Diseases




Journal of


Pharmaceutics



of


tens

ity (

)


86420

01 1 10 100 1000 10000

Record 3 drug

Size (dmiddotnm)








002040608

11214

0 50 100 150 200Time (min)

IbuprofenPhenol red

C outC

in





4 Discussion








00102030405060708


P efftimes10minus4

(cm

s)





(3)


5 Conclusions


References































Volume 2014

ToxinsJournal of

VaccinesJournal of















AddictionJournal of






Autoimmune Diseases




Journal of


Pharmaceutics



of







00102030405060708


P efftimes10minus4

(cm

s)





(3)


5 Conclusions


References































Volume 2014

ToxinsJournal of

VaccinesJournal of















AddictionJournal of






Autoimmune Diseases




Journal of


Pharmaceutics



of























Volume 2014

ToxinsJournal of

VaccinesJournal of















AddictionJournal of






Autoimmune Diseases




Journal of


Pharmaceutics



of

research article self-microemulsifying drug delivery...

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