---
title: "One-month physicochemical stability of idarubicin diluted in polyolefin (Freeflex) infusion bags"
id: "plos-one-21-one-month-physicochemical-stability-study-of-idarubicin-prepared-in-polyolefin"
canonical_url: "https://medichelpline.com/clinical-feed/plos-one-21-one-month-physicochemical-stability-study-of-idarubicin-prepared-in-polyolefin"
content_type: "clinical_feed_article"
specialty: "Oncology"
source_name: "PLOS ONE (Medicine)"
source_url: "https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0356906"
published_at: "2026-08-28T14:00:00.000Z"
evidence_level: "Journal Feed"
license: "CC-BY-NC-4.0 / Informational Use"
---
# One-month physicochemical stability of idarubicin diluted in polyolefin (Freeflex) infusion bags
## Provenance & Clinical Metadata
- **Canonical URL:** https://medichelpline.com/clinical-feed/plos-one-21-one-month-physicochemical-stability-study-of-idarubicin-prepared-in-polyolefin
- **Specialty:** [Oncology](https://medichelpline.com/clinical-feed/oncology.md)
- **Primary Source:** PLOS ONE (Medicine)
- **Source URL:** [Original Journal Publication](https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0356906)
- **Published At:** 2026-08-28T14:00:00.000Z
- **Evidence Rating:** Journal Feed
## Executive GIST (TL;DR)
- Study evaluated the **physicochemical stability** of idarubicin diluted in polyolefin-type infusion bags (Freeflex + ®) for intravenous use in hematology. - Preparations used Zavedos® idarubicin diluted in 100 mL bags of **0.9% sodium chloride (NaCl)** or **5% dextrose (Dex)** at two target concentrations: **0.04 mg/mL** and **0.2 mg/mL**. - Eight storage conditions were tested: combinations of diluent (NaCl or Dex), concentration (0.04 or 0.2 mg/mL) and temperature (5 ± 3 °C refrigerated or 22 ± 3 °C room temperature); three replicate bags per condition for a total of **24 bags**. - Samples were protected from UV in photoprotective secondary packaging and prepared aseptically in an ISO 5 isolator. - Chemical stability was assessed using a validated **stability-indicating HPLC-UV/VIS diode array detector** method; assays performed on days D0, D1, D2, D7, D15, D22 and D30. - Physical stability checks (visible and sub-visible particles, pH, osmolality) were performed on days D0, D15 and D30. - Over 30 days, measured idarubicin concentrations remained above **95%** of initial values; no degradation products were detected by the assay. - No visible color change or precipitation observed; sub-visible particle counts stayed below recommended limits; pH and osmolality values did not change from baseline. - Study supports the stability of idarubicin diluted between 0.04–0.2 mg/mL in 0.9% NaCl or 5% Dex in Freeflex + ® bags, stored at either refrigerated or room temperature, for up to 30 days under the tested conditions. - The work addresses gaps in prior literature by testing clinically relevant concentrations, polyolefin infusion bags rather than polypropylene tubes, and including contemporary physicochemical stability endpoints.
## Clinical Analysis & Structured Key Points
One-month physicochemical stability study of idarubicin prepared in polyolefin-type bags for the treatment of hematological diseases | PLOS One Browse Subject Areas ? Click through the PLOS taxonomy to find articles in your field. For more information about PLOS Subject Areas, click here . Article Authors Metrics Comments Media Coverage Peer Review Reader Comments Figures Figures Abstract Idarubicine is a mainstay of the treatment strategy for hematological cancer. The literature lacks stability data that comply with current preparation practices and recommendations for conducting stability studies. The aim of the present work is to establish the stability of idarubicin preparations diluted in polyolefin-type bags (Freeflex + ®) for intravenous administration in hematological clinical settings. Preparations were produced by diluting Zavedos® idarubicin in Freeflex + ® bags containing 0.9% sodium chloride (NaCl) or 5% dextrose (Dex) at concentrations of 0.04 mg/mL and 0.2 mg/mL. These preparations were stored at 5 ± 3 °C or 22 ± 3 °C. Three bags were prepared to evaluate each condition, for a total of 24 bags. To assess chemical stability, a stability-indicating high pressure liquid chromatography-ultraviolet/visible diode array detector assay was used. Samples were assayed on days D0, D1, D2, D7, D15, D22 and D30. A physical stability study was carried out on days D0, D15 and D30, looking for visible and sub-visible particles, measuring pH and osmolality. During the study, sample concentrations remained above 95% of the initial concentration, and no degradation products were detected. No color change or precipitation were observed with the naked eye. Sub-visible particles count remained below recommended limits. The pH and osmolality values remained unchanged from D0. This study demonstrates the stability of idarubicin, diluted to concentrations between 0.04 and 0.2 mg/mL in solutions of 0.9% NaCl and 5% Dex, stored at 5 ± 3 °C or 22 ± 3 °C for 30 days. Citation: Bouguéon G, Jouvien A, Bernadou J-M, Venet A, Berroneau A, Mora P, et al. (2026) One-month physicochemical stability study of idarubicin prepared in polyolefin-type bags for the treatment of hematological diseases. PLoS One 21(8): e0356906. https://doi.org/10.1371/journal.pone.0356906 Editor: Christophe Curti, Assistance Publique - Hôpitaux de Marseille, FRANCE Received: February 17, 2026; Accepted: August 10, 2026; Published: August 28, 2026 Copyright: © 2026 Bouguéon et al. This is an open access article distributed under the terms of the Creative Commons Attribution License , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Data Availability: All relevant data are within the paper and its Supporting Information files. Funding: The author(s) received no specific funding for this work. Competing interests: No authors have competing interests. Introduction Idarubicin, a semi-synthetic anthracycline ( S1 Fig ), remains a mainstay of the treatment strategy for newly-diagnosed acute myeloblastic leukemia in children and adults, or for patients with relapsed or refractory disease, ahead of other anthracyclines. Idarubicin is also used in the treatment of recurrent acute lymphoblastic leukemia in children and adults [ 1 – 3 ]. For solid tumors, idarubicin is used in gastroenterology in transarterial chemoembolization (TACE) therapies mixed with contrast medium emulsions (e.g., Lipiodol®) for the treatment of intermediate-stage liver carcinoma [ 4 ]. According to the current guidelines, idarubicin is used in the treatment of acute leukemia in children and adults at dosages ranging from 5 to 12 mg/m 2 administered daily intravenously in regimens lasting 3–5 days. For administration to patients, idarubicin must be diluted in bags or syringes with 0.9% sodium chloride (NaCl) or 5% Dextrose (Dex) [ 1 ]. Given that solvent bag dilution is commonly used in the treatment of hematological diseases, this study will focus on this preparation method, syringe dilution being used in a chemoembolization context. In centers specialized in hematology and providing care for a large number of patients, the number of idarubicin preparations needed can be high, requiring advance preparation for organizational and logistical reasons. Hospital pharmacists responsible for producing sterile cytotoxic preparations, in accordance with applicable good preparation practices, must therefore have reliable data on the stability of these preparations after dilution in solvents, beyond the 24 hours recommended by pharmaceutical firms [ 1 , 5 – 7 ]. Regarding the dilution of idarubicin in infusion bags, the only study available was conducted by Beijnen et al. in 1985 [ 8 ]. In this article, the stability of idarubicin diluted to a concentration of 0.1 mg/mL in 0.3% and 0.9% NaCl, 3.3% and 5% Dex and Ringer Lactate was assessed. The study was conducted at room temperature and samples were protected from light. Beijnen et al . concluded that idarubicin was stable for a period of 28 days. However, this work has limitations and lacks specific details regarding the criteria currently recommended for stability study procedures [ 6 , 9 , 10 ]. Indeed, this study was not conducted under routine clinical conditions of use, idarubicin having been diluted in polypropylene tubes and not in bags of diluents for infusion. Furthermore, fundamental criteria on physicochemical stability (determination of visible, sub-visible particles, osmolality, and pH monitoring) are missing or lack details. Moreover, idarubicin concentrations used in routine practice frequently deviate from the target value of 0.1 mg/mL studied or may be stored in the refrigerator by accident. Extrapolating data from stability studies to conditions other than those studied (e.g., concentrations, solvent, storage conditions) involves risks and engage the responsibility of the pharmacists who produce these preparations [ 6 , 9 , 10 ]. For these reasons, we propose to study the stability of idarubicin preparations under conditions of use in line with current practices: dilution in polyolefin-type bags (Freeflex + ®) in 0.9% NaCl and 5% Dex, at concentrations between 0.04 mg/mL and 0.2 mg/mL, stored at room temperature or refrigerated, and protected from UV. Materials and methods Materials Idarubicin 10 mg/10 mL (Zavedos®, Pfizer, New York City, NY, US) (batches CA7906 exp: 2022 03 and DX0725 exp: 2023 03) was used to prepare idarubicin bags of 100 mL 0.9% sodium chloride (NaCl) and 100 mL 5% dextrose (Dex). For solution dilution, Freeflex + ® bags were purchased from Fresenius Kabi (Bad Homburg vor der Höhe, Germany). The infusion bags were made of polyolefin (primary internal film: polypropylene; secondary external film: polypropylene/polyethylene admixture). The following materials were used for the analytical experiments: idarubicin hydrochloride powder (Sigma Aldrich (Merck KGaA, Darmstadt, Germany)(batch 0000424922 exp: 2024 11), distilled water provided in-site with a Milli-Q® de-ionization system (PURELAB® Flex 1, ELGA CA, High Wycombe, UK), acetonitrile (ACN) HPLC-S gradient grade (Biosolve, Deuze, France), potassium dihydrogen phosphate (KH 2 PO 4 ) (Prolab, Sion, Switzerland), 30% hydrogen peroxide (H 2 O 2 ) (Cooper, Melun, France), AnalaR Normapur 37% hydrochloric acid (HCl) and 30% sodium hydroxide (NaOH) (VWR Chemicals Avantor, Radnor, PA, US). Choice of concentrations for the stability study To guide the choices of concentrations for the stability study, a preliminary analysis of injectable anticancer drug production data from University Hospital of Bordeaux was conducted. The data were extracted from Chimio® production management software (Computer Engineering, Paris, France) and were centered on idarubicin production in bags between 2016 and 2023 intended for adult and pediatric oncohematology care units. University Hospital of Bordeaux makes approximately 80,000 injectable preparations per year. Over the study period, 2,172 idarubicin bags were made as part of acute lymphoid leukemia, acute myeloid leukemia and lymphoma treatment regimens ( S8 Fig ). Preparations intended for adults represented over 95% of production. As the dose banding approach was not applied for idarubicin dose standardization, 23 different concentrations between 0.02 and 0.28 mg/mL were produced, of which close to 87% were centered between 0.09 and 0.15 mg/mL. The limits of the stability study were defined between 0.04 mg/mL and 0.2 mg/mL, with concentrations beyond these limits concerning fewer than 10 preparations over the 8 years of analysis. Idarubicin bag preparation Idarubicin bags were prepared from the brand-name drug Zavedos® idarubicin, 10 mg/10 mL and diluted in 100 mL polyolefin-type infusion bags (Freeflex + ®). Three variation factors were studied and crossed: the diluent (0.9% NaCl or 5% Dex), the final concentration (0.04 mg/mL or 0.2 mg/mL) and the storage temperature (room temperature 22 ± 3 °C or refrigerated conditions 5 ± 3 °C). In this way, eight conditions were studied. For each condition, the bags were prepared in triplicate, for a total of 24 bags. All the preparations were produced under aseptic conditions in an ISO 5 (International Organization for Standardization) isolator at negative pressure (JCE Biotechnology, Hauterive, France). The preparations were then stored protected from UV in a photoprotective secondary packaging (200300UVS+ (SLB, Genas, France)). Collective and individual protective measures against cytotoxic risks All staff involved in sample preparation and quality control during the stability study have received training and are authorized to handle cytotoxic medicines. The preparation of cytotoxic drugs was performed in an ISO 5 negative-pressure isolator located within an ISO 7 controlled-atmosphere area. Operators wore appropriate protective clothing for work in these areas: reusable fabric scrubs and gowns, single-use caps, and single-use non-sterile gloves. During the control steps, sample preparation was performed in areas with an unclassified atmosphere. Appropriate collective protective measures were taken: handling under a microbiological safety cabinet class 2, use of sterile drapes. Technicians wore appropriate protective clothing: non-sterile gloves, a single-use lab coat, and a single-use mask. Stability-indicating high pressure liquid chromatography-ultraviolet/visible diode array detector (HPLC-UV/VIS DAD) assay method The idarubicin assay was conducted with a high-performance liquid chromatographic method adapted from the method described by Kaushik et al [ 11 ]. The apparatus used was a Thermo Scientific TM UltiMate TM 3000 HPLC-UV/VIS system, coupled with a photodiode array detector (DAD, diode array detector) (Thermo Fischer, MA, US). Separation was conducted on an XDB-C18 column (250 mm x 4.6 mm; particle size 5 µm) (Agilent, Santa Clara, CA, US) preceded by a 0.5 µm frit (Thermo Fischer Scientific, Waltham, MA, US). The mobile phase was a 60:40 mixture composed of a 10 mM pH 2.8 KH 2 PO 4 phosphate buffer and acetonitrile. The analyses were conducted under isocratic elution conditions (1 mL/min) at a temperature of 25 °C. Samples awaiting analysis were stored in the autosampler at a temperature of 4 °C. The volume of sample injected was 5 µL. Data were acquired at 254 nm. Data acquisition and peak purity match were performed using Thermo Scientific TM Chromeleon TM software Version 7.2.7 (Thermo Fischer Scientific, Waltham, MA, US). HPLC-UV/VIS DAD assay method validation The samples used to validate the assay method were prepared each day, extemporaneously from idarubicin hydrochloride powder (purity 100%, water content 2.1%) diluted in distilled water provided in-site with a Milli-Q® de-ionization system (PURELAB® Flex 1, ELGA CA, High Wycombe, UK). The validation of the assay method was conducted in accordance with international conference on harmonization (ICH) Q2 (R2) guidelines [ 12 ]. The linearity of the method was assessed by producing a serie of five-point concentrations (0.010, 0.025, 0.050, 0.100 and 0.250 mg/mL) from independent test samples. This serie was repeated on 3 different days. The linear regression analysis was conducted using Chromeleon TM software. The method was considered linear if all the correlation coefficients (R 2 ) calculated were greater than 0.99. The accuracy (expressed as %) and the precision (expressed as a coefficient of variation CV%) were assessed through the intraday and interday repeatability, using 3 quality controls prepared at concentrations of 0.02, 0.15 and 0.2 mg/mL. The intraday repeatability (precision and accuracy) was assessed using the three quality control points, each prepared and analyzed 6 times. The interday repeatability (precision and accuracy) was determined by repeating the same assay on 3 different days. The accuracy was considered acceptable if the values obtained were between 98 and 102%. The precision was considered acceptable if the CV values obtained were not greater than 2%. The detection limit was determined from the background noise and was considered to correspond to the concentration at which the signal to noise ratio was 3:1. The quantitation limit was determined based on a signal to noise ratio of 10:1. Forced degradation study In order to be able to detect any degradation products, the ability of the assay method to be stability-indicating was assessed according to ICH Q1A(R2) and SFPC/ GERPAC guidelines [ 9 , 10 ]. To this end, one-mL samples prepared from idarubicin hydrochloride powder were exposed to different forced degradation conditions. For the heating stress conditions, one mL of idarubicin was heated for 4 hours at 80 °C in a Jouan water bath (Thermo Electron, ThermoFischer Scientific, Nantes, France) and analyzed undiluted. For oxidative stress conditions, one mL of idarubicin was mixed with one mL of 7.5% (w/w) hydrogen peroxide (H 2 O 2 ) for 4 hours (sample diluted 1:2). For acidic stress conditions, one mL of idarubicin was mixed with one mL of a hydrochloric acid (HCl) solution 0.1 M and heated to 80 °C in a water bath for 5 min; before analysis, the samples were neutralized with one mL of sodium hydroxide solution (NaOH) 0.1 M (sample diluted to one-third). For alkaline stress conditions, one mL of idarubicin was mixed with one mL of a sodium hydroxide (NaOH) solution 1 M and heated to 80 °C in a water bath for 1 min; before analysis, the samples were neutralized with one mL of hydrochloric acid solution (HCl) 1 M (sample diluted to one-third). For photolytic stress conditions, four mL samples of idarubicin were exposed to a fluorescent lamp emitting 29,000 lux at 15 cm and at room temperature of 22 ± 3 °C for 2 hours (Sylvania Sylfast SSE T5, Osram Sylvania, Munich, Germany) and analyzed undiluted. All the chromatograms obtained were compared to the reference chromatograms obtained with the assay method described above, using samples not subjected to stress conditions. The degradation parameters were optimized to ensure effective separation down to the baseline, or to minimize overlap between the peaks of the degradation products and the idarubicin peak. All tests were performed in triplicate. Stability study design and acceptance criteria The chemical stability of the idarubicin preparations over time, under the 8 study conditions, was determined by assessing the idarubicin concentration on 7 study days: D0, D1, D2, D7, D15, D22 and D30. Each day, three bags of each condition were analyzed, and the mean concentration and standard deviation calculated. The preparations were considered stable if the individual concentrations measured at the different time points during the study were within the 90–110% interval of the initial concentration (D0) [ 9 ]. pH determination The pH was determined using a Seven Compact TM pH-meter equipped with an Inlab® Micro-Pro-ISM 51344163 probe (Mettler Toledo, Colombus, Ohio, US). Three-point calibration were conducted with the pH 4.01, 7.00 and 9.21 buffer solutions available from Mettler Toledo. The assays were conducted on 2 mL samples in triplicate. Throughout the study, it was verified that the pH variation did not exceed one unit. Osmolality determination The osmolality was measured using a Type 15 cryoscopic osmometer (Löser Messtechnik, Berlin, Germany). Calibration was conducted at 0, 300 and 900 mOsm/kg using water for injections (point 0 mOsm/kg, PROAMP, Aguettant, Lyon, France) and reference solutions available from Löser Messtechnik. The measurements were conducted on 100 µL samples in triplicate. Sub-visible particles count The sub-visible particles assay was performed in accordance with the European Pharmacopoeia (Test 2.9.19 Particulate contamination: sub-visible particles, test 1.B) and was conducted using an HIAC 9703 + particle counter coupled with an HRLD 150 sensor (Beckman Coulter, Brea, California, US). The sample volume used was 25 mL. For each sample, the apparatus calculated a mean of 4 successive measurements. Samples were compliant if the number of the average number of particles present in each bag tested does not exceed 6,000 particles of ≥10 μm or 600 particles of ≥25 μm in accordance with the European Pharmacopoeia [ 13 ]. Organoleptic inspection The idarubicin bags were observed visually on a STETDMLED12 visual inspection table (STERIGEN, Upton, Canada), alternately for 5 seconds in front of a white panel followed by 5 seconds in front of a black panel to detect visible particles or a change of color. The comparison was made with a preparation produced extemporaneously on the day of observation [ 9 , 13 ] Results Assay method validation The chromatograms obtained showed a well-defined and symmetrical peak with a mean retention time of 3.4 min and baseline only exhibiting a low level of background noise ( Fig 1 ). The calibration curves produced on 3 different days, were linear with a correlation coefficient (R 2 ) of 0.999. Both the intraday and interday repeatability have produced satisfactory results ( Table 1 ). The intraday and interday accuracies ranged between 98.09 and 101.73% and between 99.45 and 101.24% respectively. Similarly, the CV% values ranged between 0.04 and 0.37% and between 0.41 and 1.47% respectively. The limit of detection (LOD) was assessed at 6.01 x 10 −5 mg/mL and the limit of quantification (LOQ) at 20.03 x 10 −5 mg/mL. The HPLC-UV/VIS DAD assay method complies with ICH Q2 (R2) requirements [ 12 ]. Download: PNG larger image TIFF original image Table 1. Intra and interday validation of the chromatographic method. https://doi.org/10.1371/journal.pone.0356906.t001 Download: PNG larger image TIFF original image Fig 1. Idarubicin 0.04 mg/mL reference chromatogram from the chromatographic stability indicating method developed to detect degradation products (total analysis time: 22 min). Abbreviation: IDA: idarubicin. https://doi.org/10.1371/journal.pone.0356906.g001 Stability-indicating method Regarding the forced degradation study of idarubicin, different degradation levels were obtained according to the exposure conditions ( Table 2 ). The chromatograms were compared to the reference chromatogram illustrated by Fig 1 . Download: PNG larger image TIFF original image Table 2. Forced degradation study results. https://doi.org/10.1371/journal.pone.0356906.t002 Under heating conditions, the degradation of idarubicin was calculated at 38.2% and the chromatogram obtained detected degradation products at the retention times of 3.98, 4.48 and 7.32 min annotated as DEG H1, DEG H2 and DEG H3 respectively ( S2 Fig ). Under oxidative conditions, the degradation of idarubicin obtained was 22,7% % and one degradation product was detected at 2.47 min annotated as DEG OX1 ( S3 Fig ). The sensitivity of idarubicin to acidic stress conditions showed degradation levels of 34.3%. However, under these conditions, few changes in the chromatogram compare
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