The European Journal of Hospital Pharmacy Science Volume 12 2006 Issue 5 P.

P. 91-95 2006 The European Association of Hospital Pharmacists. All rights reserved 1781-7595 20 www.ejhp.org

Stability and compatibility of the ready-to-use solution of paracetamol admixed with phloroglucinol for intravenous infusion
Nicolas K Kambia, PharmD, PhD, Michel Luyckx, PharmD, Thierry Dine, PharmD, Thrse Dupin-Spriet, PharmD, Bernard Gressier, PharmD, Claude Brunet PharmD ABSTRACT The chemical stabilities of paracetamol and phloroglucinol, when mixed together for intravenous infusion, were studied. We performed this study to establish whether an admixture containing the ready-to-use solution for injection of paracetamol (Perfalgan 10 mg/mL), and phloroglucinol (Spasfon ampoules of 40 mg/4 mL), was stable and compatible for a useful period (30 min) or more (48 hours). According to the recommendations of the manufacturer, Perfalgan must be administered alone and should not be mixed with other drugs. However, combinations of the new ready-to-use solution of paracetamol and adjuvant drug solutions are often used in clinical practice while little information is available on the chemical stability. So, the chemical stability and compatibility of both paracetamol and phloroglucinol, when mixed together for intravenous infusion, were studied by a stability-indicating HPLC assay for up to 48 hours at room temperature without any protection against light. The admixtures were also monitored for precipitation, colour change and pH. Over the 48-hour period, the mean concentration of paracetamol at all samples studied was within 98.40% +/- 2.90% to 102.30% +/- 1.20% of the initial concentration. The mean concentration of phloroglucinol at each assay time was 98.10% +/2.10% to 102.30% +/- 0.90% of the initial concentration. More than 98% of the initial paracetamol and phloroglucinol concentrations remained at each assay interval. There was no visible evidence of precipitation, gas formation, or colour change. No appreciable change in pH occurred during the study period. In conclusion, paracetamol and phloroglucinol, prepared by adding phloroglucinol to a glass bottle of paracetamol ready-to-use solution, were stable and compatible for up to 48 hours at room temperature without any protection against light. KEYWORDS Paracetamol, phloroglucinol, drug stability, compatibility, high-performance liquid chromatography INTRODUCTION Paracetamol is a peripherally acting analgesic commonly used in multimodal post-operative pain management. It is a popular analgesic and antipyretic drug that is used for the relief of fever, headaches and other minor aches and pains [1]. It is commonly administered in tablet, in injection form, liquid suspension, or suppository form. A new, ready-to-use solution for injection of paracetamol (Perfalgan 10 mg/mL) without previous reconstitution has been developed. It is a diluted paracetamol solution stable enough to be marketed as a ready-to-use container for intravenous infusion. The presentation offers a saving of time and a safety of use in comparison with the old marketed reference formulation for injection propacetamol (Pro-Dafalgan 1 g). The easy use of Contact for correspondence: Dr Nicolas K Kambia Perfalgan is very favoured by nurses who are very interested Pharmacology, Pharmacokinetic and Clinical Pharmacy Department in its consequences on the reduction of their workload and Faculty of Pharmaceutical and Biological Sciences are aware of the better safety of this unit-dose [2]. Perfalgan 3 rue du Professeur Laguesse, BP 83 is available in both adult and paediatric formulations and is F-59006 Lille Cedex, France indicated for the short-term treatment of moderate pain, Tel: +33 3 20 96 40 40 especially following surgery, and for the short-term treatment Fax: +33 3 20 96 97 52 of fever, when administration by intravenous route is clinically nicolas.kambia@libertysurf.fr justified by an urgent need to treat pain or hyperthermia and/or when other routes of administration are not possible.
Received: 10 March 2006; Revised manuscript received: 9 October 2006; Phloroglucinol is a non atropinic antispasmodic agent which Accepted: 12 October 2006

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Stability and compatibility of paracetamol injection admixed with phloroglucinol Nicolas K Kambia et al The European Journal of Hospital Pharmacy Science

is used by oral route, rectal route, intramuscular or intravenous injection for its spasmolytic properties against digestive or urinary tract. Phloroglucinol (Spasfon) is primarily used to treat the pains in the digestive functional disorders, in the renal colics (pains related to urinary conditions) and in certain pains in gynaecology. The drug was extensively used in the past for augmentation of labour [3]. One of its ethers, the trimethylphloroglucinol, is also used. As a chemical compound, phloroglucinol is used as an analytical reagent and as a decalcifier of bone specimens for microscopic examination. Paracetamol and phloroglucinol have different mechanisms and sites of action. Certain clinical situations require a combined therapy of drugs to improve the quality of analgesia. Currently, there are no commercially-available pre-packed or ready-to-use mixtures. So, combinations of analgesics and adjuvant drugs are often used in clinical practice while little information is available on their chemical stability. According to the recommendations of the manufacturer, Perfalgan must be administered alone and should not be mixed with other medicinal products [4]. Unfortunately, this is not always respected in certain clinical situations. Thus, the aim of this study was to investigate the physical compatibility and chemical stability of the ready-to-use solution for injection of paracetamol (Perfalgan 10 mg/mL) admixed with phloroglucinol (Spasfon ampoules of 40 mg/4mL) stored at room temperature (2025C) without any protection against light for 48 hours. The mixture paracetamolphloroglucinol was prepared by adding 2 ampoules of phloroglucinol to a glass bottle of paracetamol ready-to-use solution. Immediately after preparation and at different time intervals, samples were visually inspected, tested for pH, and the concentration of each component in the binary mixture was determined by a stability-indicating liquid chromatographic method. METHODS Chemicals and drugs HPLC-grade acetonitrile was obtained from Prolabo (Paris, France). The water used to prepare aqueous buffers and dilutions was de-ionized and purified by Milli-Q Academic water purification (Millipore, Saint Quentin en Yvelines, France). The drugs studied were the commercial products suitable for clinical use. They were provided in injection forms. Paracetamol was the new, ready-to-use solution for injection not requiring reconstitution, Perfalgan 10 mg/mL. It was generously donated by Denain Hospital Pharmacy (Denain, France). Phloroglucinol was the current clinical for-

mulation, Spasfon intravenous injection and was generously donated by Denain Hospital Pharmacy (Denain, France) in ampoules of 40 mg diluted in 4 mL of injectable solvent. Ceftazidime, Fortum was used as both paracetamol and phloroglucinol internal standard and was obtained from GlaxoSmithKline Laboratory (Marly-le-Roi, France). All the others reagents used were analytical grade. Chromatographic apparatus and conditions A high performance liquid chromatographic (HPLC) method and pH-meter were used to conduct the analyses. Chromatographic analyses were performed using a HP 1090 HPLC system (Hewlett Packard, Orsay, France) equipped with a variable volume injector, an automatic sampling system and a Hewlett Packard 79994 linear photodiode array UV detector operating at suitable wavelengths. The output from the detector was connected to a Hewlett Packard 9000 model 300 integrator to control data acquisition and integration. Retention times and peaks areas were determined by a computer connected to a Hewlett Packard Thinjet terminal printer. Drugs analyses were performed on a 5 m Hypersil ODS C18 column (150 mm x 4.6 mm) operating at room temperature. pH determination The pH-meter, used to measure the pH of the mixture during storage was a model HI 8520 N microprocessor equipped with a Micro pH electrode HI 1083 (Hanna Instruments, Lingolstein, France) and calibrated with standard buffer solutions at pH 4.00, 7.00 and 10.00. Stability study Paracetamol and phloroglucinol were analysed using the modified HPLC methods described elsewhere [5, 6]. Paracetamol concentrations were determined by a stabilityindicating HPLC assay. The mobile phase was a degassed and filtered (0.45 m, Millipore) acetonitrile-buffer (0.05 % orthophosphoric acid solution) mixture (10/90, v/v). The HPLC pump was set at a flow rate of 0.8 mL/min, volumes of 20 L were injected and the detector was set at 254 nm. Phloroglucinol analyses were carried out at 210 nm using the same mobile phase previously described. A total of 20 l of each sample was injected into the analytical column. Calibration curves Paracetamol and phloroglucinol calibrations curves were constructed at concentration ranges of 25100 g/mL and 540 g/mL respectively. Standard stock solutions (1 mg/mL) of both paracetamol and phloroglucinol were prepared in water. A series of dilutions was made to prepare

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Stability and compatibility of paracetamol injection admixed with phloroglucinol Nicolas K Kambia et al The European Journal of Hospital Pharmacy Science

the suitable standard solutions of desired concentrations before injection into the column. The precision of the assays were validated by establishing the within-day and the between-day coefficients of variation. Admixture components and storage conditions The stability of phloroglucinol in ready-to-use solution for injection of paracetamol at room temperature was studied for use as an intravenous admixture. The admixture was prepared in triplicate and kept at room temperature without any protection against light for 48 hours. Samples were removed immediately and at various time points. The physical compatibility was evaluated on the basis of the colour, clarity and pH. The pH of the commercial formulation of paracetamol ready-to-use solution and phloroglucinol was measured, and the pH of the mixture was assessed at each time over the study period. The stability (the concentration of each component in the binary mixture) was determined by a stability-indicating HPLC after suitable dilution in the mobile phase. RESULTS AND DISCUSSION Chromatography Drug concentrations were determined by using a stabilityindicating HPLC assay. All assays were performed isocrati-cally at ambient temperature. The compounds were resolved with a satisfactory baseline separation under developed Figure 1: Chromatograms of paracetamol solution under two pH conditions: acidic (E), alkaline (F) and after heating both solutions (E, F) at 60C for 30 min

Figure 3: Plot showing the evolution of the remaining amount of both paracetamol and phloroglucinol in mixtures

-Paracetamol 5min 1h 10min 20 rnin 30min

-Phloroglucinol' 24 h Time (min, h) 43 h

conditions. After various treatments (forced degradation), the secondary compound peaks of both paracetamol and phloroglucinol were adequately separated. No degradation product interfered or was eluted with the same retention time of the intact parent drugs peaks (Figures 1 and 2). Paracetamol calibration curve was constructed at a concentration range of 25100 g/mL. A good linear response was

Figure 2: Chromatograms of phloroglucinol solution under two pH conditions: acidic (G), alkaline (H) and after heating both solutions (G, H) at 60C for 30 min

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Stability and compatibility of paracetamol injection admixed with phloroglucinol Nicolas K Kambia et al The European Journal of Hospital Pharmacy Science

found with a correlation coefficient better than 0.9998. The within-day coefficients of variation for replicate analysis (n = 4) of three different concentrations (25, 50 and 100 g/mL) averaged 1.82%, 1.30% and 0.85%, respectively. The between-day coefficient of variation (n = 4) was estimated on the calibration solution of 50 g/mL. It was less than 2.3%. The retention times for paracetamol and the internal standard were 4.40 min and 5.80 min, respectively. In the same way, a good linear response was found for phloroglucinol assays. The linear regression analysis of the peak area of the drug concentration yielded a correlation coefficient (r) better than 0.9996 (concentrations ranging from 5 to 40 g/mL). The within-day coefficients of variation (n = 4) of two different concentrations (5, 40 g/mL) averaged 1.30% and 1.10%, respectively. The between-day coefficient of variation (n = 4) estimated on the calibration solution of 40 g/mL was less than 1.94%. The retention times for phloroglucinol and the internal standard were 2.90 min and 6.00 min, respectively. Stability of paracetamol The analysis of each sample was performed by HPLC after a suitable dilution in the mobile phase in order to fit the calibration curve. At time zero, the initial concentration of paracetamol was designated as 100% and all subsequent measured concentrations were expressed as percentages of the initial concentration (Table 1). Where applicable, the stability studies followed the European guidelines on stability of existing active substances
Table 1: Results from the determination of paracetamol phloroglucinol concentrations in the mixture after storage at room temperature Storage time (min., h) Paracetamol: 1g/100 mL Paracetamol conc. (%) +/- SD a (n = 3) 100.00 +/- 0.00 98.40 +/- 2.90 99.90 +/- 0.80 101.30 +/- 1.80 101.80 +/- 1.50 100.10 +/- 1.30 102.30 +/- 1.20 99.80 +/- 1.70 98.90 +/- 2.40 Phloroglucinol: 80 mg/8 mL Phloroglucinol conc.
(%) +/- SD

and related finished products [7]. The stability was defined as a concentration within 90105% of the initial one. Therefore a decrease from mean initial concentration (T0) of more than 10% was considered to represent a significant loss of drug [8]. Evolution of these concentrations was assessed throughout the observation period (up to 48 hours). As shown in Figure 3, the chemical stability of paracetamol when mixed with phloroglucinol for intravenous infusion, the variations in drug concentration never exceeded 10% of the initial concentration over the study period. There was no substantial difference between paracetamol concentrations at zero time and at any subsequent time intervals. No additional peak corresponding to degradation products was observed on chromatograms. Stability of phloroglucinol The initial concentration (T0) of phloroglucinol and all subsequent measured concentrations were assessed by HPLC method. Evolution of these concentrations was measured throughout the study period. Data were expressed in percentage of the initial drug concentration (Table 1). The drug concentration never fell below 98% of the initial concentration over the 48 hours study period. Therefore, phloroglucinol and paracetamol, prepared by adding 2 ampoules of phloroglucinol to a glass bottle of paracetamol ready-to-use solution, were stable for 48 hours when stored at room temperature without any protection against light. For all samples, there was no visible evidence of precipitation, gas formation, or colour change throughout the observation period. Average pH measurements are given in Table 2. For all aliquots, the pH decreased slightly over 48 hours and the changes in pH were 0.40 pH unit without loss of the stability of the mixture. Forced degradation of both paracetamol phloroglucinol solutions were conducted to confirm separation of the parent drugs from their secondary products formed during
Table 2: Mean pH measurements of the mixture paracetamol phloroglucinol after storage at room temperature pH data (%) +/- SD (n = 3) Ready-to-use solution of paracetamol Mixture paracetamol -phloroglucinol 5.40 +/- 1.20 T0 T30 min T1 h T24 h T48 h 4.98 +/- 1.60 5.05 +/- 1.30 4.70 +/- 0.85 4.63 +/- 1.15 4.58 +/- 1.40

(n = 3) 100.00 +/- 0.00 101.00 +/- 1.10 99.80 +/- 1.80 102.30 +/- 0.90 98.10 +/- 2.10 99.70 +/- 1.50 100.80 +/- 0.60 100.90 +/- 2.50 101.80 +/- 1.90

T0
5 min. 10 min. 20 min. 30 min. 1 h. 4 h. 24 h. 48 h.
a Initial conce b Initial conce

ntration (100%) before storing: g/100 mL +/- 0.024 1.09 ntration (100%) before 5 mg/8 mL +/- 0.122 storing: 78.9

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Stability and compatibility of paracetamol injection admixed with phloroglucinol Nicolas K Kambia et al The European Journal of Hospital Pharmacy Science

the assay. Results obtained show that both compounds are completely degraded in alkaline medium. Therefore, it will be necessary to avoid any mixture with a basic compound. Moreover, heat is without effect on the stability of the two molecules (Figures 1 and 2). Within these limits, phloroglucinol and paracetamol, prepared by adding 2 ampoules of phloroglucinol to a glass bottle of paracetamol ready-to-use solution may be prepared and REFERENCES
1. Holmer Pettersson P, Owall A, Jakobsson J. Early bioavailability of paracetamol after oral or intravenous administration. Acta Anaesthesiol Scand 2004; 48: 867-70. 6. Schmitt E, Vainchtock A, Nicoloyannis N, Locher F. Injectable acetaminophen now ready to-use : easier, more accurate and lowering costs. Pharm Hosp 2001; 36: 9-18 7. Tabassum S, Afridi B, Aman Z. Phloroglucinol for acceleration of labour: double blind, randomized controlled trial. J Pak Med Assoc 2005; 55: 270-3. 8. Bristol-Myers Squibb Pharmaceuticals Ltd, Perfalgan, document last updated on December 2005. 9. Arayne M.S, Sultana N, Siddiqui F.A. In vitro availability of trimethylphlorogluci-

stored in advance at room temperature without any protection against light before use in clinical units. CONCLUSION Admixtures of the ready-to-use solution for injection of paracetamol with phloroglucinol at clinically relevant concentrations were physically compatible and chemically stable for up to 48 hours at room temperature without any protection against light.

nol and its degradation product from dosage formulations by RP-HPLC. Pak J Pharm Sci 2005; 18: 7-12 10.Rao R.N, Narasaraju A. Rapid separation and determination of process-related substances of paracetamol using reversed-phase HPLC with photodiode array as a detector. Anal Sci 2006; 22: 287-92 11. Guideline on stability testing: stability testing of existing active substances and related finished products, CPMP/QWP/122/02, European Agency for the Evaluation of Medicinal Products (2003). 8. Gury C, Dolizy I, Aymard N, Sellali Y, Rochet S. Stability of citalopram hydrochloride in PVC bags for I.V. solutions and compatibility in the presence of dipotassium clorazepate. Eur J Hosp Pharm 2001; 7(1): 11-4

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