Shamshad et al. Journal of Analytical Science and Technology 2014, 5:22 http://www.jast-journal.com/content/5/1/22 RESEARCH Open Access Spectroscopic characterization of in vitro interactions of cetirizine and NSAIDS Hina Shamshad1*, M Saeed Arayne1 and Najma Sultana2 Abstract Background: Cetirizine (anti-allergy agent) and non-steroidal anti-inflammatory drugs (NSAIDs; anti-inflammatory agents) are co-administered drugs. Cetirizine, a P-glycopretein substrate may be affected by the use of NSAIDs. In the present work, quantification of cetirizine in the presence of commonly co-administered NSAIDs such as diclofenac sodium, ibuprofen, flurbiprofen, tiaprofenac acid, meloxicam and mefenamic acid were studied using the RP-HPLC technique. Methods: A high-throughput HPLC method for the analysis of cetirizine was developed, validated in the presence of NSAIDs and was further used to study the interactions of cetirizine in the presence of NSAIDs at four different pH levels. Purospher Star, C18 column (5 μm, 25 cm × 0.46 cm) with a mobile phase of methanol/water (90:10 v/v, pH adjusted to 3.5) at a flow rate of 1.0 mL/min and a wavelength of 240 nm was used. Results: The synthesis of cetirizine in the presence of NSAIDs was carried out, and complexes were characterized using infrared (IR) and nuclear magnetic resonance (NMR) techniques. The method was found to be applicable in serum and was found useful for therapeutic purposes. The differences in availabilities of cetirizine with NSAIDs at three pH levels clearly indicated the interactions of afore mentioned drugs. Conclusions: The findings suggested further studies on the co-administration of these two drugs simultaneously in order to know the pharmacokinetic and pharmacodynamic profiles of the drugs. It is highly recommended to have a suitable time lapse between the oral uses of these drugs. Keywords: Cetirizine; Drug interaction; HPLC method; NSAIDs Background the treatment of respiratory disorder (Philip and Philip Second- and third-generation antihistamines are P- 2010). Nimesulide and cetirizine combination was also glycoprotein substrates. Drugs that affect the P-glyco- prepared which was found to possess anti-leukotriene, protein may lead to drug-drug interactions (Scott and antihistamine, anti-allergy, and anti-inflammatory actions Kelly 2003; Jarkko et al. 2006; Renwick 1999). Cetirizine (Singh and Jain 2003). Drug interactions of cetirizine have being a P-glycoprotein substrate may interact with com- also been reported in literature (Ihsan et al. 2005; Arayne monly co-administered drugs such as NSAIDs that may et al. 2010a; Sultana et al. 2010). affect the P-glycoprotein expression (Lee et al. 2007), as Simultaneous HPLC method for the determination of celecoxib and diclofenac, were found to inhibit the func- paracetamol, phenylpropanolamine hydrochloride, and tion of P-glycoprotein (Wageh et al. 2004). It has been cetirizine was reported; however, the mobile phase con- established that synergistic effect was exhibited by the sisted methanol and disodium hydrogen phosphate di- combined use of cetirizine with nimesulide (Rewari and hydrate buffer which could be corrosive to the column Gupta 1999). Moreover, cetirizine was found to possess (Suryan et al. 2011). Another simultaneous method of analgesic activity in mice (Priya et al. 2013). Cetirizine paracetamol, acetyl salicylic acid, mefenamic acid, and and mefenamic acid combination was also developed for cetirizine was achieved using disodium hydrogen phos- phate buffer and acetonitrile combination which could * Correspondence: [email protected] once again be corrosive and expensive altogether (Freddy 1 Research Institute of Pharmaceutical Sciences, Department of and Dharmendra 2010). Separation method of ibuprofen, Pharmaceutical Chemistry, Faculty of Pharmacy, University of Karachi, Karachi 75270, Pakistan phenylephrine HCl, and cetirizine from soft gels was Full list of author information is available at the end of the article © 2014 Shamshad et al.; licensee Springer. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Shamshad et al. Journal of Analytical Science and Technology 2014, 5:22 Page 2 of 8 http://www.jast-journal.com/content/5/1/22 reported using the mobile phase of acetonitrile and hexa- spectrometer (Madison, WI, USA) using TMS as an in- nosulphonate (Cosmes and Rodriguez 2012). Simultaneous ternal standard. determination of cetirizine and aceclofenac was achieved by using mobile phase of acetonitrile and heptane sul- Method development phonic acid (Padmavathi and Niranjan 2011). Nimesulide, Optimized conditions for separation cetirizine, and pseudoephedrine hydrochloride separation The chromatographic analysis was performed at laboratory was done by using acetonitrile/phosphate buffer/triethyl temperature (25°C) with isocratic elution. The optimized amine as the mobile phase (Jain et al. 2012). Our group conditions for the separation of the eluents were achieved by also reported methods of simultaneous determination of using the mobile phase of methanol/water 90:10 monitored − cetirizine with anti-diabetic drug (Arayne et al. 2010b) and by UV detection 240 nm at a flow rate of 1 ml min 1 with with H2 receptor antagonists (Sultana et al. 2010). pH 3.5 adjusted by orthophosphoric acid. The samples from It is clearly evident from the above literature that most these solutions were injected in to the system six times. of the simultaneous methods used acetonitrile as solvent which is quite expensive. In others, buffers were also Reference standard solutions used which is column corrosive. In order to quantify the Stock reference standard solutions of all drugs were pre- drugs and understand the degree of interactions, a fast, pared by dissolving appropriate amounts of each drug in a − least expensive, efficient HPLC method was required, mobile phase to yield concentrations of 100 μgml1. For which could be used for therapeutic purposes and could calibration curve studies, serial dilutions in the concentra- − also be employed for routine analysis work. tion range of 50 to 3.12 μgml 1 were injected in triplicate. So, in the present work, cetirizine method was developed and validated in the presence of NSAIDs such as diclofenac Pharmaceutical dosage form samples sodium, ibuprofen, flurbiprofen, meloxicam, and mefe- Commercially available pharmaceutical formulations namic acid using the RP-HPLC method. The method was of the respective brands were evaluated by groups of further used to study the interactions of the cetirizine in 20 tablets for each drug and dissolved in mobile phase thepresenceofNSAIDsatthreepHlevels,i.e.,4,7.4,and according to the labeled claim. The samples from this 9. Cetirizine complexes were also synthesized with interact- solution were injected into the system in triplicate. ing drugs. The synthesized complexes were characterized by their physical parameters as well as by the techniques of Standard drug plasma solutions infrared (IR) and nuclear magnetic resonance (NMR). The supernatant obtained by centrifuging blood samples was deprotonated by acetonitrile, spiked daily with wor- Methods king solutions, and chromatographed. Materials Raw materials used were of pharmaceutical purity and for- Method development protocol mulations as cetirizine (Zyrtec), mefenamic acid (Ponstan), Certain parameters such as mobile phase composition and diclofenac sodium (Voren), flurbiprofen (Froben), meloxi- pH, flow rate, diluents of solutions, and wavelength of ana- cam (Melfex), tiaprofenic acid (Surgam), and ibuprofen lytes were altered in order to achieve symmetrical and (Brofen) were purchased from local markets. Analytical well-resolved peaks at a reasonable retention time. grade reagents were used during the whole experimental procedures. Methanol of HPLC grade (TEDIA®, USA) and Method validation other reagents included were hydrochloric acid, sodium All validation steps were carried out according to the ICH hydroxide, sodium chloride, potassium dihydrogen ortho- guidelines such as system suitability, selectivity, specificity, phosphate, disodium hydrogen orthophosphate, ammonium linearity (concentration-detector response relationship), chloride, 10% NH3 solution, phosphoric acid 85% (Merck, accuracy, precision, and sensitivity, i.e., detection and Darmstadt, Germany) were utilized. quantification limit. Equipment System suitability Shimadzu HPLC system (Kyoto, Japan) equipped with LC- System suitability was assessed by examining the six rep- 10 AT VP pump, with a 20-μl loop, Purospher® Star, RP-18 licates of the drugs at a specific concentration for repeat- endcapped (5 μm) column and SPD-10 A VP UV-vis de- ability, peaks symmetry (symmetry factor), theoretical tector was utilized. IR studies were carried out using a Shi- plates, resolution, and capacity factors. madzu Model Fourier transform infrared (FTIR) Prestige- 21 spectrophotometer. Spectral treatment was performed Specificity using Shimadzu IR solution 1.2 software. The 1H-NMR The drugs were spiked with pharmaceutical formulations spectra were recorded on a Bruker AMX 500 MHz containing different excipients. Shamshad et al. Journal of Analytical Science and Technology 2014, 5:22 Page 3 of 8 http://www.jast-journal.com/content/5/1/22 Figure 1 Representative chromatograms of (a) cetirizine and (b) meloxicam. Linearity