Pharm Res (2010) 27:1318–1324

DOI 10.1007/s11095-010-0114-x

RESEARCH PAPER

Isolation and Characterization of Cetirizine Degradation
Product: Mechanism of Cetirizine Oxidation
Tatyana Dyakonov & April Muir & Hassen Nasri & Dana Toops & Aqeel Fatmi

Received: 20 May 2009 / Accepted: 24 November 2009 / Published online: 31 March 2010
# Springer Science+Business Media, LLC 2010

ABSTRACT INTRODUCTION
Purpose The goal of the study was to isolate and analyze a
cetirizine degradation product, formed within a PEG-containing Cetirizine hydrochloride (cetirizine), the active component
formulation and to elucidate the mechanism of oxidation of of ZYRTEC® tablets and syrup, is an orally active and
cetirizine. selective H1-receptor antagonist used for the treatment of
Methods Cetirizine, formulated in PEG-containing matrix, allergy symptoms. Soft gelatin capsules (Softgels) offer the
was subjected to forced degradation conditions in the pH possibility of delivering a liquid in a solid oral dosage form.
range from 3 to 10, and the product was analyzed by HPLC A softgel can therefore contain the active ingredient in
and LC-MS/MS. Additionally, pure cetirizine was subjected to solution, suspension or emulsion, which may inherently
selective oxidization by hydrogen peroxide and sodium lead to better absorption of the active ingredient compared
percarbonate. The reaction mixture was purified, and the to the delivery in a tablet or powder. Many problems asso-
isolated material was analyzed by 1H NMR. ciated with tableting, including poor compaction and lack of
Results Oxidation process was investigated in order to model content or weight uniformity, can be eliminated if drug is
the degradation of cetirizine in PEG-containing formulation. Site incorporated into this dosage form. Softgel dosage forms are
of oxidation is proposed based on correlation of the results of perceived by customers as easier to swallow, help to mask the
forced degradation with ionization scheme of cetirizine. The taste of the medicine and provide faster onset of action (1). In
finding was verified by spiking of cetirizine degradation sample order to develop this highly water soluble and sensitive
with cetirizine N-oxide reference standard. compound in soft gelatin dosage form, cetirizine was
Conclusions Degradation of cetirizine in polyethylene glycol formulated in PEG-containing matrix. However, cetirizine
arose from the reaction between the drug and the reactive formulated in PEG-containing solution for Softgels was
peroxide intermediates such as peroxyl radicals formed found to degrade during manufacture and storage.
through oxidation of PEG. Selective oxidation of cetirizine Our preliminary results, supported by the literature
and isolation/characterization of the oxidation product allowed reports (2), showed that cetirizine does not degrade under
the identification of the oxidation product as cetirizine N-oxide. acidic or basic conditions. It is known that cetirizine is
The mechanism of oxidation is proposed. susceptible to oxidation under chemical (hydrogen peroxide),
biochemical (enzymatic) and electrochemical conditions (3).
KEY WORDS cetirizine . oxidation . PEG The metabolism of levocetirizine (R enantiomer of cetirizine)
was studied (3) and found to proceed through the metabolic
pathways: hydroxylation, O-dealkylation, N-oxidation and N-
dealkylation (Fig. 1).
T. Dyakonov (*) : A. Muir : H. Nasri : D. Toops : A. Fatmi Excipients containing polyoxyethylene chains, for example
Banner Pharmacaps, polyethylene glycol (PEG) 400 and polysorbate, are known to
4125 Premier Drive,
High Point, North Carolina 27265, USA contain some residual peroxides, which accumulate during
e-mail: tadyakonov@banpharm.com storage and could potentially affect the stability of oxidation-

The detector N N wavelength was set at 230 nm.07. 5 µm.0 mL/min. The flow rate was 1. PEG 400 was obtained from BASF. as Reagents and Chemicals described by Hamburger et al. and Cetirizine was oxidized by aqueous hydrogen peroxide. N N OH OH O LC-MS/UV and LC-MS/MS/UV Method Cl Cl LC-MS/UV and LC-MS/MS-UV data were obtained (c) (d) using an Agilent 1200 series HPLC system. In order to identify cetirizine degrada.6 mm (Waters Corporation). caused by PEG-containing formulations. Spotting was done on precoated silica gel aluminum Cetirizine hydrochloride was supplied by Matrix Laboratories plate 60 F254. and sodium percarbonate were of the ACS grade and were purchased from JT Baker. 5 µm. was pumped at the flow rate of 1. calibrated HPLC system equipped with a binary pump and UV detector was used. It was reported (5. HTS PAL autosampler and MDS Sciex/ Fig. cetirizine hydrochloride (1 g) was exposed to 30% aqueous solution of hydrogen peroxide for 5 h at 70– 80°C. ammonia (JT Baker) and acetonitrile (EMD and Burdick & O Jackson) were of the HPLC grade. Column temperature was Cl Cl maintained at 25°C.13. magnesium sulfate. The resulting degradation Method 1 product was isolated and analyzed. (5) with some modifications.5. 1 Metabolic products of levocetirizine oxidation produced by (a) Applied Biosystems QTRAP 4000 mass spectrometer with Hydroxylation (b) N-oxidation (c) O-dealkylation (d) N-dealkylation (3). The mobile phase consisted of 0. and the injection volume was N N O O O O 25μL. then the reaction mixture was extracted with ethyl MATERIALS AND METHODS acetate.0 mL/min.6) that Corporation).Mechanism of Cetirizine Degradation 1319 HO chromatograms were acquired using Chemstation LC 3D O Rev A 10. Organic phase was separated and concentrated. The HPLC column was Xterra-RP C18. Cetirizine N-oxide was obtained from LGC GmbH (Lot 380. Since the cetirizine soft gelatin capsule matrix contains PEG. auto-oxidation can progress through formation of hydro. Potassium phosphate monobasic monohydrate. and the injection volume was 10 μL.4. we selectively oxidized cetirizine. 250× OH OH 4. investigate. Trifluoroacetic acid and NMR solvent were purchased from Sigma-Aldrich. both at pH 2.02 software (Agilent Technologies). methanol (EMD). . The solvent system consisted of ethyl Limited (Lot CTP 0171106). The crude material was purified by preparatory TLC. we believe that this may be the main cause for Synthesis and Purification of Cetirizine Degradation cetirizine oxidation.1% trifluoroacetic acid and peroxides and peroxyl radicals in polyoxyethylene linkages acetonitrile in the ratio of 75:25 (v/v)–0. this reaction. Column temperature was maintained at 25°C.1% trifluoroacetic containing substances such as polysorbate surfactants and acid and acetonitrile in the ratio of 25:75 (v/v). OH HPLC/UV Chromatographic Method Cl m/z 201 An Agilent 1200 series. Hence.5). equipped with UV detector. the aim of this work was to identify a degradation product. Analyst 1. hydrogen per- N N oxide. A binary mixture of potassium phos- (a) (b) phate buffer: acetonitrile 70:30 (v/v) (solvent A) and potassium phosphate buffer: acetonitrile 35:65 (v/v) (solvent B).03). PEG. Product tion products. The analytical column used was Xterra-RP C18.2 software. Ethyl acetate (Fisher Scientific).6 mm (Waters sensitive pharmaceuticals (4. For establish the mechanism of the degradation. Phosphoric acid (NF FCC) O O was also purchased from JT Baker. 250×4. 2 Estimated structure of degradation product (m/z 405/407). The Fig.

. ate. dried over magnesium sulfate. acetate-methanol-ammonia (80:15:5.1320 Dyakonov et al. v/v/v) mixture. The kinetics of oxidation was monitored by HPLC/ 1 Fig. concentrated and analyzed by HPLC/UV Cetirizine was selectively oxidized with sodium percarbon- and 1H NMR. 3 H NMR spectra of cetirizine (a) and degradation product (b). Method 2 Scraped spots were extracted with ethyl acetate.

4.31 7. Varian spectrometer.0.05 15.88 11. as described in the Method 1.50 150 31.94 38. spiked with cetirizine N-oxide (c).59 200 40.5. Triplicate aliquots of each solution were transferred into Fig.Mechanism of Cetirizine Degradation 1321 Response Cetirizine 700 650 600 Cet N-oxide 550 500 450 Cetirizine Cet N-oxide 400 350 300 250 (a) 31.2334.5. cetirizine N-oxide standard (b). Samples were withdrawn peri- 1 H NMR odically and analyzed by HPLC for cetirizine and related impurities.5 and 10.0. UV. 6. 4. 7. 5 The effect of pH on the oxidation of Cetirizine in PEG solution separate scintillation vials and purged with nitrogen. 2 days 6 days 16 days 28 days 48 days 5.5.81 41. % 50 pH–Stability Profile 40 30 A cetirizine solution was prepared by dissolving 8 g of the 20 drug in 8.72 Peg Esters Peg Esters 50 (c) 2. 80°C. 70 60 Conversion. The 10 0 solution was stirred for 1 h at room temperature then 2 4 6 8 10 transferred into 10 separate vials. The reaction mixture was purified by preparatory samples were then incubated for 48 days at 30.28 6. All solutions were purged with nitrogen each The NMR measurements were carried out on a 400 MHZ time the vials were opened.30 for methanol.00 (b) 100 16. and stressed cetirizine solution. 6.0 using NaOH.93 11. 5.97 23. the under forced degradation conditions.29 0 5 10 15 20 25 30 35 40 Retention time Fig. The pH of the solution pH in each vial was adjusted individually to 3. The NMR spectra were measured in methanol-d4 and D2O. Aliquots of the solutions were also stored at room temperature as controls.6 g of DI water and 260 g of PEG 400. 4 Chromatograms of cetirizine (a). .0. 60 and TLC.29 35.0. Chemical shifts are reported on δ scale (ppm) by assigning the residual solvent peak to 3. 7.74 33.0.

8 Cetirizine zwitterionic form HX (predominant specie between pH 3 to 8). several oxidative methods for deshielding influence of the N-oxide function. The PEG-containing formulation. aromatic and aliphatic fragment of molecular ion. Fig. The number of protons in the aromatic region of the 1 H NMR of the degradation product was similar to that of Selective Oxidation of Cetirizine and Identification cetirizine. Termination 2RO2 inactive products RO2 + R inactive products preparing the N-oxide of tertiary amines were considered. Results value of m/z 405 for piperazine containing aliphatic showed that in the presence of sodium percarbonate.89) in retention time of oxidative peak of interest (Rt =24 min by PEG-containing formulation showed the monochlorinated HPLC) formed during forced degradation. water) and different catalysts (sodium tungstrate. 6 Auto-oxidation process. The N-oxide effect can be observed by the difference in the chemical shifts of the proximal In order to model the degradation process of cetirizine in a protons adjacent to the N-oxide carbons (Fig. molybdenum trioxide) (7). As a first attempt.1322 Dyakonov et al. cetirizine was selectively oxidized with m/z 201. 3).. 7 Proposed mechanism for the oxidation of cetirizine to form N-oxide. 3). generated - HO HO H O H - O O . All methods used hydrogen peroxide in different media (acetic acid. cluster of peaks at m/z 405/407 (M+16) and a base peak at Additionally. The retention time of the The preliminary LC/MS and LC/MS/MS results from the compound isolated from TLC plates corresponded to the study of the stressed solution of cetirizine (MW 388. The 1H NMR spectrum of isolated cetirizine the aliphatic portion of cetirizine. indicating formation of N-oxide. suggesting that oxidation occurs in 80°C. 2. The The kinetics of oxidation were monitored by HPLC. fragment can be attributed to the monohydroxylated conversion of cetirizine approached 50% during 2 h at product or N-oxide. . cetirizine RESULTS hydrochloride was successfully oxidized with 30% aqueous hydrogen peroxide. leading to a possible structure as shown in Fig. rather than of Oxidation Product monohydroxylation. Initiation X- H3X++ H2X+ HX RH + light or catalist R +H A B Propagation C + O2 N N R ROO O O OH ROO + RH ROOH + R Cl ROOH ROO +H Fig. Reaction mixture was purified by Preliminary LC/MS Results preparative TLC as described in the literature (2) and analyzed by HPLC/UV. N N N N + N N O O O O O O + H 2O OH OH OH Cl Cl Cl Fig. The intense ion at m/z 201 was assumed to be sodium percarbonate with higher selectivity and yield of formed due to the fission of >C-N< bond between oxidation product than with conventional hydrogen peroxide. There was no evidence of a oxidation product was compared to NMR of cetirizine m/z fragment of 217 (oxidation on the aromatic fragment) hydrochloride (Fig.

5 and 6. chain tion of PEG. respectively. 8. The aliquots of the product were result.5.7 and 1. 60 and 80°C). N-oxide formation increased cetirizine oxidation under stressed conditions was evaluated at pH>7 and higher N-oxide formation was observed in our previous work (12). where R in our case hindered piperazine nitrogen. 4. formed through oxida- predominantly consists of the reactions of initiation.5 and 6. addition of BHA reduced the rate of oxidation of cetirizine. Mechanism of Cetirizine Oxidation The polyoxyethylene-based excipients contain some resid.5 could be explained by the protection effect of explained by N-oxidation of piperazine nitrogen by protons on tertiary amine (9). The effect of anti-oxidant (BHA) on the rate of at pH 4. by HPLC. As a atures (30. A well-established auto. Cetirizine degradation product was synthe- propagation and termination.2) and chain. Only butylated hydroxytoluene (BHT) and hydroxyl anisole 0.27). 8). ethoxy (Δδ=0.Mechanism of Cetirizine Degradation 1323 on the nitrogen bearing the ethoxyacetic acid chain. 7. one could obtain another minimum in the oxidation withdrawn at different times and analyzed by HPLC/UV. The previous studies (8) reported three N-oxide (Fig. 3 to 8 (Fig. The peroxide formation in PEG-containing formulations Cetirizine proved to be significantly more stable at pH can be inhibited by including an anti-oxidant. located on the nitrogen bearing the ethoxyacetic acid causes a down-field shift of the piperazine (Δδ=0. Based on carboxylic acid. The results obtained at 80°C are presented in Fig.5.5 and 6. The autoxidation starts with decompo. The mechanism of oxidation was proposed. Cetirizine. Chromatograms of two standards—cetirizine (a) nearest to the aromatic rings and pKa 2. peroxides formation. 6.5 during 48 days exposure at 80°C. The ionization scheme of cetirizine of stressed solution of cetirizine spiked with cetirizine is shown in Fig. N.52. formulated in PEG. 5. CONCLUSION ual peroxides and could potentially affect the stability of oxidation-sensitive pharmaceuticals. for cetirizine and cetirizine-oxidation products. happen because the neutral HOO˙ as an oxidizer would was subjected to the forced degradation at different temper. not have the Coloumb attraction as a driving force. The minimum of the oxidation rate at these results.9—to the and cetirizine N-oxide (b)—are shown in Fig. zwitterionic form HX is predominant between pH polyoxyethylene chains.5–7. 4c) showed retention times 19 and 25 min macro-pKa values for cetirizine (1.oxidation The formation of N-oxide progresses according to was found to be the major pathway for cetirizine Fig. We assume that between these values  of pH the  Effect of pH on Oxidation of Cetirizine in PEG protonation of superoxide  radical O2 around pH∼6 þ  formulation (11) O 2 þ H ! HOO may result in the elimination of the charged O 2 species and therefore decrease the The formation of cetirizine N-oxide in PEG was investigated oxidation rate of the protonated tertiary amine. The qualitative analyses is –(OCH2CH)O–. producing cetirizine N-oxide with the oxygen atom transformation. 2. According to the literature hydroperoxides formed from oxidative decomposition of (8. . the lowest of which was attributed to the nitrogen tively. rate at pH below 6. respec.5) protons. such as 4. sized by the selective oxidation of the sterically less oxidation process is shown in Fig. The presence of The results of degradation of cetirizine in PEG-containing hydroperoxides and peroxyl radical formed in cetirizine formulation indicated that cetirizine is susceptible to soft gelatin capsule matrix could be the main reason for oxidation.10). This can in a pH range from 3 to 10.34–5. Degradation of cetirizine in polyethylene cetirizine oxidation. LC/MS/MS along with 1H NMR were used sition of alkyl polyoxyethylene chain (RH) and subsequent to identify the oxidation product as cetirizine N-oxide. The data indicated that the between pH 4. The results suggested that the degradation of cetirizine in polyethylene glycol arose from the reaction between the DISCUSSION drug and reactive peroxide intermediates.92 and 8. glycol arose from the reaction between the drug and The radical-driven process of cetirizine auto-oxidation reactive peroxide intermediates. This site of oxidation in the molecule was proposed This result agreed with the LC/MS data and was based on the correlation of results of degradation in pH verified by spiking the cetirizine degradation sample range from 3 to 10 with ionization scheme of cetirizine with cetirizine N-oxide standard. formed through oxidation of PEG.6% of cetirizine N-oxide was formed (BHA). the cetirizine oxidation process could be pH of 6. The chromatogram and 1H NMR data.

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