Long term stability study of cisplatin in polyolefin infusion bags in dose banding conditions using eco-friendly hydrophilic interaction chromatography and colorimetry methods

Abstract

Purpose

The aim of this study was to develop and validate a green and sustainable hydrophilic interaction chromatography (HILIC) stability indicating assay for measuring the concentration of cisplatin and to evaluate its long-term stability at standardized rounded doses in polyolefin (POF) infusion bags at 25°C ± 3°C, 60% ± 5% relative humidity and protected from daylight. Physical stability, including colorimetry was periodically performed.

Methods

The HILIC with diode array detector method was fully validated and responds perfectly to degradation tests. Diluted cisplatin infusion solutions were aseptically prepared by further dilution of cisplatin stock solution with NaCl 0.9% in POF infusion bags (508 mL) at banded doses of 30 mg, 200 mg, and 400 mg. The POF bags were stored at 25°C in the dark. Physical and chemical stabilities were evaluated respectively through visual inspection, turbidity, particulate contamination, colorimetry measurements and through chromatographic assays, pH and osmolality monitoring.

Results

. The long-term stability of cisplatin was confirmed for at least 25 weeks at 25°C and in the dark. In addition, it was shown that a potential exposure of the cisplatin infusion bags in the natural light, the time to fall to 95% (90%) of the initial concentration would be estimated at eight (sixteen) minutes.

Conclusions

A simple, accurate, and eco-friendly stability-indicating HILIC method was developed to determine cisplatin concentrations in dose-banding conditions associated for the first time with color variation investigation. This study supports a centralized production of cis-platine in accordance with the studied conditions.

Keywords

Green sustainable eco-frienly HILIC long-term stability colorimetry infusion bags cisplatin

Introduction

Cisplatin is a platinum-containing antineoplastic agent approved for the treatment of various solid tumors.¹ Dose banding is a method of dose individualization in which all patients with similar characteristics are allocated to the same dose.^2–4 This allows the anticancer drug to be prepared in advance in a secure production environment.⁵ Prior to administration, cisplatin Mylan 1 mg/mL must be diluted with sodium chloride-containing vehicle solutions. In solutions with low chloride concentrations, cisplatin degrades rapidly. According to the Summary of Product Characteristics (SmPC), chloride concentrations in diluted cisplatin mylan infusion solutions should not fall below 0.45%.⁶ If diluted with 0.9% sodium chloride infusion solutions, physicochemical stability is ensured for 24 h at room temperature, when protected from light. At refrigerated temperatures, precipitation may occur.⁶ The physicochemical stability of cisplatin brand products diluted with 0.9% sodium chloride solution is established for up to 28 days when stored protected from light.^7–9 Cisplatin concentrations in infusion solutions exposed to light decline more rapidly.¹⁰ To the best of our knowledge no studies have focused on the cis platin stability at the dose-banding conditions of 30 mg, 200 mg and 400 mg in POF bags beyond 30 days. For monitoring the evolution of cisplatin concentration over time, green and sustainable analytical chemistry is pivotal in minimizing the environmental footprint ensuring safer and more efficient analytical methods. Reversed-phase HPLC (RP-HPLC) is the most commonly applied HPLC mode,¹¹ and is known for their high solvent consumption.¹² Thus, the high amounts of harmful organic solvents used in HPLC mobile phases represent the biggest impact regarding sustainability. Acetonitrile is the most commonly used solvent in RP-HPLC and is classified as ‘problematic’according to the CHEM21 classification to establish safety, health and environment criteria of solvents.¹³ In recent years, the application of ethanol in HPLC has grown significantly in popularity due to the need for eco-friendly analytical methods. Indeed ethanol is easy produced from renewable sources via fermentation and its biodegradation occurs in few days.¹⁴ An HILIC column contains a silica stationary phase modified with ammonium sulfonic acid ligands which makes it an excellent tool to enhance the retention and separation of hydrophilic, polar, and ionic analytes that would be poorly retained in traditional reversed-phase HPLC separations.¹⁵ In HILIC chromatography, best results are obtained with high concentrations of organic solvent.¹⁵ In this work, a green and sustainable HILIC stability indicating assay was developed to evaluate the stability of cisplatin in POF infusion bags over a long period of time up to 220 days at 25°C ± 3°C and 60% ± 5% relative humidity (RH) in the dark. The parameters concerning the physicochemical stability such as turbidity, particulate contamination, osmolality, and pH measurements were also analysed. As well, in this work, and for the first time, the color variations of cisplatin in infusion bags during the long-term stability study were analyzed by a colorimetric analysis.

Materials and methods

Materials

The High-Performance Liquid Chromatography (HPLC) apparatus consisted of an Agilent 1260 Infinity I system coupled with a diode area detector (DAD). The Nucleodur HILIC 2 mm×150 mm column was furnished by Macherey Nagel (Paris, France). pH, osmolality chromaticity data and particulate size measurements were carried out using respectively a MA235Advanced pH - Meter (Mettler Toledo, Béthune, France), a freezing point osmometer Loeser TYP15 (Löser Messtechnik, Berlin, Germany), a UV – visible spectrophotometer (Cary 60 UV-Vis spectrophotometer, Agilent) and a LS 13 320 XR Laser diffraction particle size analyser (Beckman Coulter). Polyolefine (POF) infusion bags containing a nominal mean volume of 508 mL NaCl 5% vehicle solutions were from Freeflex, Fresenius Kabi (Germany).

Chemicals

The pharmaceutical specialities of cisplatin for injection (1 mg/mL) and NaCl 0.9% used for pharmaceutical formulation were obtained from Mylan^® (France). Cisplatin analytical standard reference (Sigma Aldrich) was used for the preparation of the validation and the calibration standard solutions. Potassium trichloroamine platinate II (TriCAP) and Potassium tetrachloroplatinate II (TetraCP) were obtained from sigma Aldric. Standard solutions of mono-hydrated cis-platine (MHC) was obtained following reference.¹⁶ Phosphoric acid, ACN and ethanol analytical grade were purchased from Sigma Aldrich (Saint Quentin Fallavier, France). All the other chemical products were of analytical grade.

Preparation and storage of cisplatine infusion bags

The diluted cisplatin infusion solutions were prepared using the cisplatin commercial solution from Mylan at room temperature inside the Qube sterility isolator from Bioquel (Thiais, France) (This Qube is an aseptic processing workstation with an integrated Hydrogen Peroxide Vapor (HPV) decontamination system). The three cisplatin standard doses (30 mg, 200 mg, 400 mg) were prepared in triplicate in POF infusion bags containing the nominal mean volume of 508 mL NaCl 0.9% vehicle solutions measured per unit. The corresponding cisplatin standard concentrations in the infusion bags were thus equal to 0.0590 mg/mL, 0.3970 mg/mL, and 0.7874 mg/mL.

Study design for the stability of the cisplatin solutions in unopened infusion bags at 25°C ± 3°C and 60% ± 5% relative humidity

The corresponding infusion bags containing cisplatin were stored at 25°C ± 3°C and 60% ± 5% RH) in the dark. On the following designated time points (just after preparation, i.e., Day 0 (D₀)), D₈, D₃₂, D₅₇, D₉₉, D₁₄₃, D₁₆₅, D₂₂₀), three units (n=3) of each standard dose were submitted to chemical and physical analyses.

Analyses performed on the cisplatin solutions

Visual inspection, colorimetry mesurements

Each sample was emptied into glass tubes, and a visual exam was carried out in a double-blind randomized study. To confirm the visual examination, by using a UV-visible spectrophotometer, the colorimetry i.e., chromaticity, and lightness of the cisplatin solutions were determined. In this study, the CIE La*b* color space (L*, a*, b*) was used to represent the color changes.^17–19 In this system, chromaticity was presented as a two-dimensional diagram (a* and b* axes) describing the entire color system independently of lightness L* defined as the visual sensation of luminosity of a surface. L* values varied from 0 (no luminosity) from 100 (maximum luminosity). The values of each color parameter are expressed as the mean of three different samples. Before analyzing the samples, a blank sample was prepared with reference deionized water to obtain 100% of transmittance. Samples were transferred into the quartz cell previously cleaned twice with purified water and twice with the sample, then subjected to the color analysis. Transmittance spectra were obtained using the Agilent Cary 60 UV-Vis spectrophotometer comprises a double beam, Czerny-Turner monochromator, 190–1100 nm wavelength range, 1.5 nm fixed spectral bandwidth, full spectrum Xenon pulse lamp single source, dual silicon diode detectors, quartz overcoated optics, scan rates up to 24,000 nm/min, 80 data points/sec maximum measurement rate, nonmeasurement phase stepping wavelength drive, room light immunity, central control by PC with Microsoft^® Windows^® operating system. Color analysis of spectra were performed using the cary WinUV color application.

Turbidity and particulate contamination measurements

Turbidimetry has been used in numerous stability studies, and Y-site compatibility studies to evaluate the subvisual aspect of physical stability.^20,21 In our study, we have used turbidimetry at the wavelength 350 nm, 410 nm and 550 nm suggested by the European Guidelines for the practical stability studies.²² Transmittance measurement method from the Cary 60 UV-Vis spectrometer was used to measure the absorbance of each sample with the use of a 50 mm light pathlength rectagular cell. Purified water filtrated using 0.2 μm membrane filter is used as blank sample. The absorbance of more than 0.010 AU was considered as an evidence of turbidity, providing a quantitative determination of incompatibility.²² An absorbance reading less than 0.010 AU was considered to be a noise level. However, turbidimetry must be considered as a global measurement of particles suspended in a liquid and does not bring information on the size of the particulates. Therefore the laser diffraction particle size analysis was carried out using the LS 13 320 XR (Beckman Coulter) with its enhanced PIDS technology and an extended measurement range (10 nm - 3500 μm). Particulate contamination was tested at the beginning and the end of the stability study.²³ For content of more than 100 mL the preparation complies with the test if, in each unit tested, the number of particles present per unit that are 10 μm or larger in size does not exceed 25 per milliliter and the number of particles that are 25 μm or larger in size does not exceed 3 per milliliter.²⁴

pH and osmolality measurements

pH measurements were carried out for each unit of studied cisplatin solutions, and the osmolality was determined on 20 μL samples. pH measurements were considered acceptable if they did not vary by more than one pH unit from the initial value.^23,24 Ideally, injectable products should be formulated as isotonic solutions (osmolality of about 300 mOsm/kg) and no more than 600 mOs/kg must be used to prevent pain.²⁵

Development of a new HILIC stability-indicating assay method

HILIC experimental conditions

In all the chromatographic experiments carried out, the mobile phase was a mixture of phase (A) (water + 0.05% H₃PO₄), and (B) (Ethanol). For only one experiment (in triplicate), ACN was used instead of ethanol. The detection wavelength, the mobile phase, its flowrate, and column temperature were optimized to obtain cisplatin and its degradation products (DPs) without interferences with appropriate resolution values (Rs >1.5).

Forced degradation studies of the cisplatin solutions

To exclude potential interferences of degradation products with cisplatin quantification, and to respect the recommended degradation range^22,24 which varied between 20–30%, cisplatin solutions were subjected to the following forced degradation conditions. All experiments were performed using a 4mL glass vial with a PTFE cap (Thermoscientific, USA).

For the basic stress 500 μL of the cisplatin solutions were introduced in a amber glass vial, and 500 μL of 1M NaOH was added to adjust the pH to 10. The basic pH stressed cisplatin solutions were stocked at room temperature for 3 h before analysis.

For the acid stress 500 μL of the cisplatin solutions were introduced in a amber glass vial, and 500 μL of 1M HCL was added to adjust the pH to 1. The acid pH stressed cisplatin solutions were stocked at room temperature for 2 h before analysis.

For the study of the photodegradation kinetic of cisplatin, a series of three units (n=3) of a cis platin POF infusion bags were stocked at 25°C over a period of 20 days under the normal laboratory light. On the following designated time points (just after preparation, i.e., Day (D₀)), D₁, D₂, D₃, D₁₀, D₁₅, D_20, these units were submitted to cisplatin quantification by our optimized HILIC method.

Analytical method validation

The method linearity was studied following the instructions from reference,²⁶ and the validation of the approach was conducted in accordance with the International Council for Harmonisation of technical requirements for Pharmaceuticals for Human Use.²⁷ Calibration and validation standard cisplatin solutions were prepared by dissolving cisplatin analytical standard reference from Sigma Aldrich (10 mg) in 10 mL graduated flasks with NaCl 0.9% to obtain 1 mg/mL primary stock solutions. Calibration solutions were prepared by diluting stock solutions with NaCl 5% to obtain calibration standards at concentrations of 0.05 mg/mL, 0.10 mg/mL, 0.25 mg/mL, 0.45 mg/mL, 0.65 mg/mL, 0.85 mg/mL and 1 mg/mL. This range between 0.05 mg/mL and 1 mg/ml has been validated to quantify the molecule of interest in the infusion bags corresponding to the dose banding and provides acceptable precision and accuracy at the extreme values. All samples were examined in triplicate on three different days. The standard curve of cisplatin was plotted and analyzed using the linear least-squares regression. The method is considered to have an optimal linearity when the coefficient of determination (r²) is not less than 0.999. The limits of detection (LOD) and quantification (LOQ) can be calculated as LOD = 3.3σ/S and LOQ = 10σ/S where σ is the standard deviation of y-intercepts of the linear regression and S is the slope of the standard calibration curve. The precision of the analytical method refers to the consistency, and reliability of the results obtained from repeated measurements under specified conditions evaluated in terms of repeatability (intra-day), and reproducibility (inter-day). For this, concentrations of validation standard cisplatin solutions (0.06 mg/mL; 0.2 mg/mL, 0.4 mg/mL; 0.5 mg/mL; 0.8 mg/mL), prepared by diluting analytical reference standard stock solutions with NaCl 0.9%, were injected using the developed HILIC method. The intra-day repeatability was evaluated by analyzing three replicates of validation standard cisplatin solutions in one consecutive experiment, while the inter-day reproducibility was performed by repeating over three consecutive days. The precision of the method was expressed as a relative standard deviation (%RSD), calculated from the mean concentration of cisplatin obtained from the experiment, which should be less than 2%. As per test procedure Q2(R2),²⁷ the recovery study for the assay of drug product shall be conducted from 80% to 120% for two concentrations of the validation standard (0.06 mg/mL and 0.80 mg/mL). The accuracy solution at 80%, 100% and 120% can be prepared, analysed, and percent recovery shall be calculated. The method is considered to be accurate when the percentage recovery is in the range of 98–102% with a RSD value less than 2%. To verify the robustness of the HILIC method, retention time changes were analyzed after doing minor variations in those chromatographic conditions affecting more the HILIC method parameters, such as, the % of phase A (water + 0.05% H₃PO₄). Chemical stability was defined as not less than 95% of the initial concentration of cisplatin.

Results

Search of the optimal chromatographic conditions of the cisplatin HILIC method: highlighting the factors of instability of the cisplatin solutions

The UV absorbance spectrum of cisplatin exhibited a maximum absorption at 210 nm (Supplementary data file, Figure S1). The UV detection wavelength was thus fixed at 210 nm. The composition of the mobile phase was A (water + 0.05% H₃PO₄) (10%(v/v)) + B (Ethanol) (90%(v/v)) at a flowrate of 0.3 mL/min with a column temperature equal to 20°C. The corresponding chromatogram of a stress-free solution was given in Figure 1. The chromatogram obtained when ethanol was replaced by ACN was also given in Figure 1. The chromatograms obtained after the acid, basic, and photodegradation stress were presented in Supplementary data file, Figure S2 for the eco friendly (water + 0.05% H₃PO₄) (10%(v/v)) + B (Ethanol) (90%(v/v)) mobile phase. The cisplatin peak appeared distinct on the chromatogram and was separated from the peak of degradation products (DPs) (Rs >1.5). The determination of the mass balance and peak purities were evaluated with the Agilent software processing method and presented in supplementary data file, Table S1. The nature of the DPs were corroborated by injection in the chromatographic system of standard solution of TriCAP, TetraCP and MHC. As well, concerning the photodegradation stress, Figure S2 showed a strong decrease of the cisplatin peak area between Day 0 and Day 20. The experimental data at 25°C corresponding to the LnC values versus time (t) dependencies could be fitted to a linear plot LnC= -κt+ LnC₀ with high correlation cofficient r² values (r² = 0.9984) (Supplementary data file, Figure S3) indicating a first order kinetic mechanism of the degradation of cisplatin with natural laboratory light. The rate constant values (κ) corresponded to the slope of this linear plot was equal to κ_25°C = 0.0815 Day⁻¹ The corresponding half time of this first-order degradation of cisplatin determined by the equation t_1/2 = Ln2/k was t_1/2 = 8.5 Day at 25°C.

Validation procedure of the cisplatin HILIC stability indicating assay method : linearity, LOD and LOQ, precision, accuracy, robustness

For concentrations ranging from 0.05 mg/mL to 1.0 mg/mL, and for three replicates of the CS, the mean linear regression was equal to peak area = m₁*x (mg/mL) +m₀ where m₁ = 14740.79 and m₀ = −97.43 for cisplatin with an excellent mean determination coefficient r² equal to 0.9998. The LOD and LOQ values of cisplatin were 2 and 11 µg/mL, respectively, indicating that the developed method was sensitive enough to detect cisplatin at low concentrations. The precision of this HILIC method was performed by analyzing intra-day and inter-day analysis, which demonstrated excellent precision with high repeatability and reproducibility as expressed by the RSD values lower than 2% (Table 1). In terms of accuracy, the results revealed that the percentage recovery of cisplatin was between 99.93 ± 1.10% and 101.40 ± 1.00% (Table 2) with RSD % of each level of the test concentration less than 2% indicating that our HILIC method was highly accurate. Concerning the robustness testing, slight adjustments of the phase phase A (water + 0.05% H₃PO₄) have been made following the optimal chromatographic conditions and the retention time (RT) change was studied. For the variations in phase A% (9, 10, 11), the corresponding RSD value of the cisplatin retention time change RT=6.72 ± 0.09 min (n=3) was always lower than 2% confirming the excellent robustness of our HILIC method.

Table 1.

Validation of precision of the HILIC method for cisplatin.

Theoretical concentration (mg/mL)	Intra-day^a (n = 3)		Inter-day^b (n=9)
Theoretical concentration (mg/mL)	Measured concentration (mg/mL)	RSD (%)^c	Measured concentration (mg/mL)	RSD (%)^c
0.06	0.057 ± 0.001	1.70	0.062 ± 0.001	1.60
0.20	0.194 ± 0.003	1.50	0.207 ± 0.003	1.40
0.40	0.394 ± 0.004	1.00	0.411 ± 0.005	1.20
0.50	0.523± 0.009	1.70	0.545 ± 0.007	1.30
0.80	0.793 ± 0.011	1.30	0.813 ± 0.012	1.50

All values are mean ± SD as obtained by triplicate analyses in a day.

All values are mean ± SD, obtained by triplicate analyses per day over 3 days.

The relative standard deviation (RSD) = SD/mean × 100%.

Table 2.

Validation of accuracy of the HILIC method for cisplatin.

Test concentration (mg/mL)	Recovery (%)^a
Test concentration (mg/mL)	L1	L2	L3
0.06	99.98(0.98)^b	99.93(1.10)	101.4(0.98)
0.80	101.20(1.09)	101.30(0.95)	100.56(1.03)

All accuracy values obtained by triplicate analyses for each test concentration level (80%(L1), 100%(L2) and 120%(L3)). In parentheses the relative standard deviation (RSD) = SD/mean × 100%.

Physical stability: Visual inspection, particulate contamination, turbidity, lightness and chromaticity measurements

For each concentration studied the solutions of cisplatin stayed limpid. There was no appearance of any visible particle matter, haziness, or gas development. Freshy prepared solutions were clear without any visible particles. Before storage, (D₀) the mean (SD) number of particles >10 µm was 0.3 (0.1), 0.3 (0.1) and 0.2 (0.1) per mL respectively for 0.0590 mg/mL, 0.3970 mg/mL and 0.7874 mg/mL infusion bags, while the mean (SD) number of particles of >25 µm was 0.0 (0.0) per mL for 0.0590 mg/mL, 0.3970 mg/mL and 0.7874 mg/mL infusion bags. On D₁₅₀, the mean (SD) of particles of >10 µm was 0.4 (0.1), 0.3 (0.1) and 0.3 (0.1) per mL respectively for 0.0590 mg/mL, 0.3970 mg/mL and 0.7874 mg/mL infusion bags while the mean (SD) number of particles of > 25 µm was 0.0 (0.0) per mL for 0.0590 mg/mL, 0.3970 mg/mL and 0.7874 mg/mL infusion bags. All these values were less than the acceptable limits. No elevation of the absorbance value measured at 350 nm 410 nm or 550 nm was over 0.004 ± 0.002 and were thus less than our acceptable limit 0.010 AU throughout the study for all the cisplatin infusion bags of 30 mg, 200 mg and 400 mg (see values in supplementary data file Table S2). Turbidity analysis thus did not reveal the formation of any additional particles. The evolution of the lightness (L*) and chromaticity (a*, b*) measurements of the cisplatin solutions throughout the study was presented in supplementary data file Table S2. The L* and a*, b* measurements remained stable over the study period at all the concentrations studied (in all comparative conditions (n=3), p > 0.80 for L* and p > 0.70 for a* and b*). These results demonstrated that the color of the solutions did not change over time and confirmed our visual examination

Chemical stability

The evolution of pH and osmolality of the cisplatin solutions throughout the study were presented in Table 3. The pH and osmolality (osm) (mOsm/kg) measurements remained stable over the study period at all the concentration studied (in all comparative conditions(n=3), p > 0.60). The HILIC chromatograms showed no sign of the presence of degradation products for the long - term stability study. Figure S4 in supplementary data material showed the purity angle of the cisplatin peak was always less than the threshold indicating that the cisplatin peak remained over time spectrally homogeneous and not subject to co-elution. The results for measured cisplatin concentrations for infusion bags at the three concentrations are summarized in Table 3. To demonstrate that cisplatin concentrations remained well within the 95–105% concentration range for the long-term stability analysis, Figure S5 in supplementary data material showed the estimated regression line, and the one-sided lower 95% confidence limits at the three concentrations. For the 0.0590 mg/mL, 0.3970 mg/mL and 0.7874 mg/mL infusion bags, as the slope of the regression line is negative with p < 0.006, the estimated stability time given by the intersection of the lower one-sided 95% confidence limit with the acceptance criterion of 5% were respectively equal to 180 days, 190 days and 200 days.

Table 3.

Evolution for cisplatin infusion bags of pH, osmolality and concentration over time during the long-term stability. All data are representative of three independent experiments performed in triplicate.

(A)*Expressed in percentage of the cisplatin initial concentration: 0.0590 mg/mL
Day	pH	Osmolality	Cisplatin (%) *
0	5.74±0.06	274± 3	100
8	5.67±0.05	278 ± 2	104.78 ± 0.99
32	5.88±0.04	284 ± 2	99.87 ± 1.14
57	5.95±0.05	284 ± 3	103.56 ± 0.98
99	6.06±0.09	280± 2	101.88 ± 1.03
143	5.78±0.05	281 ± 2	97.89 ± 1.00
165	5.93±0.05	285 ± 3	99.44 ± 1.49
220	5.65±0.06	280 ± 3	100.37 ± 1.51

(B)* Expressed in percentage of the cisplatin initial concentration: 0.3970 mg/mL
Day	pH	Osmolality	Cisplatin (%) *
0	4.05±0.08	277± 3	100
8	4.09±0.06	283 ± 2	99.81 ± 1.24
32	4.17±0.05	284 ± 3	100.35 ± 1.56
57	4.19±0.07	284 ± 3	99.44 ± 1.34
99	4.66±0.08	280± 3	99.84 ± 1.28
143	4.52±0.09	284 ± 3	97.39 ± 1.00
165	4.51±0.07	284 ± 2	96.90 ± 0.84
220	4.54±0.08	283 ± 3	97.99 ± 1.00

(C)* Expressed in percentage of the cisplatin initial concentration: 0.7874 mg/mL
Day	pH	Osmolality	Cisplatin (%)
0	3.69±0.07	280± 2	100
8	3.74±0.05	284 ± 3	99.91 ± 1.05
32	3.70±0.06	284 ± 2	100.12 ± 0.99
57	3.72±0.08	286 ± 2	100.46 ± 1.05
99	3.64±0.07	280± 2	100.28 ± 1.00
143	3.83±0.06	288 ± 3	99.42 ± 1.23
165	3.91±0.05	285 ± 2	98.11 ± 1.34
220	3.70±0.07	288 ± 2	98.64 ± 1.30

Discussion

The retrospective analysis of the cisplatin preparations was carried out on data retrieved from preparation software (BPC Chimio^®, France) used in our chemotherapy preparation unit. Three most frequently produced doses (30 mg, 200 mg, 400 mg) were identified. During a six months period following DB implementation, 546 preparations amounting to 15 different doses were prepared. Among those 546 preparations: 530 (97%) were standard doses, 11 (2%) preparations were under the lowest standard dose (30mg) and 5 (1%) were above the highest standard dose 400mg. These results confirmed the interest of implementing these banding dose and studying the corresponding stability. For this, an Eco-Friendly HILIC optimized stability indicating assay coupled to a DAD detector was developed. For the cisplatin degradation process analysis, the number of stability studies^7–9 suffered from shortcomings due to the lack of research into optimal degradation conditions to detect the greatest number of degradation products on the chromatogram. For a good stability indicating chromatographic method, it was necessary that the stress chromatograms gave efficient retentions of the active pharmaceutical ingredients i.e., cisplatin and its degradation products (DPs). The optimal chromatographic retention of a molecule with stationary phase inside the column was usually considered optimal when the retention factor k of this molecule was in the range²⁸ 2–10. If k is below 2 (below 1), there is insufficient (no) retention between the analyte and the stationary phase, whilst above 10, the run times can be excessive, and the chromatographic peak height will decrease as the bandwidth of the peaks increases. In our case, with our HILIC method conditions described above, the column dead time was 1.70 min, the retention time (RT) of cisplatin was RT=6.82 min corresponding to a retention factor k_Cisplatin ≈ 3.00. This value confirmed an excellent optimal interaction of cisplatin with the HILIC HPLC column. If ACN was used instead of ethanol and with the same other chromatographic conditions, the retention time (RT) of cisplatin was RT=3.01 min corresponding to a dramatic drop in the retention factor k_Cisplatine ≈ 0.80 < 1. This last result demonstrated that with ACN, cisplatin was not retained on the HILIC stationary phase and thus confirmed the great advantage to use ethanol as a green and sustainable organic solvent. With our HILIC method, the cisplatin peak appeared well on the chromatogram and was separated from the peak of DPs (Rs >1.5) in accordance with the optimal chromatographic conditions. Our mass balance correlates well with this affirmation. As well, detection performed at 210 nm associated with the HILIC retention mechanism allow the detection and separation of the highest polar DPs of cisplatin i.e., MHC. For the basic stress chromatogram, the DPs appeared as 2 uniform peaks with retention times (RTs) equal to 3.34 min and 8.98 min corresponding to TriCAP and MHC, this last molecule being the main DPs usually obtained in strong basic conditions ([OH⁻] ⩾ 0.01 M).²⁹ For the acid stress chromatogram, the DP appeared as 1 uniform peak with a retention time (RT) equal to 5.30 min corresponding to TetraCP the main stable DP in HCl medium.³⁰ For the photodegradation stress, our data demonstrated that a nine day exposure of the POF bags to normal laboratory light leads to a 50% reduction in cisplatin concentration. Therefore, for a potential exposure of the cisplatin infusion bags into the laboratory natural light, the time to fall to 95% (90%) of the initial concentration would be estimated et eight minutes (sixteen minutes). This last result confirmed it was necessary to prevent such exposure during patient administration.¹⁰ This is an other strong point of this work. In most stability studies, color changes were analyzed only by a simple visual examination as described in the European pharmacopeia.³¹ The US Pharmacopeia³² described a method utilizing a UV-spectrometer to measure luminosity and color difference of a drug formulation. Despite these recommendations, most published stability studies fail to adequately integrate this notion, and still rely on visual examination to find a color change. Our results demonstrated that during the long-term stability study of the cisplatin infusion bags at 25°C and protected from daylight, the luminosity (L*) and chromaticity (a*, b*) parameters did not vary significantly throughout the study confirming no color change during the visual examination. This present work was the first report including this spectrophotometric analysis for the study of cisplatin stability and this point must be also highlighted. As well, this study was, to our knowledge, the first to evaluate the long-term stability of cisplatin in POF infusion bags for three dose banding and up to three months. In the literature,^7–9 stability was studied only for a maximum of 30 days. None of these studies pursued analysis beyond the end date originally envisaged.

Conclusion

Cisplatin in 0.9% NaCl at standardized 30 mg, 200 mg and 400 mg rounded doses for POF bags is physically, and chemically stable at 25°C ± 3°C and 60% ± 5% RH for at least 25 weeks in the dark. The time to fall to 95% (90%) of the initial concentration would be estimated at eight minutes (sixteen minutes) for a potential excursion of the cisplatin infusion bags into the natural light. Thanks to this long-term stability, the preparation of ready-to-use solutions by a hospital pharmacy is compatible with routine practice and should help to decrease the risks involved in multiples dilutions. As well, we showed that HILIC analytical methods are easy to implement using classical HPLC material in a pharmaceutical department and thus opens up exciting prospects for Eco-Friendly control of pharmaceutical preparations.

Supplemental Material

sj-docx-1-opp-10.1177_10781552261475929 - Supplemental material for Long term stability study of cisplatin in polyolefin infusion bags in dose banding conditions using eco-friendly hydrophilic interaction chromatography and colorimetry methods

Supplemental material, sj-docx-1-opp-10.1177_10781552261475929 for Long term stability study of cisplatin in polyolefin infusion bags in dose banding conditions using eco-friendly hydrophilic interaction chromatography and colorimetry methods by Yves Claude Guillaume, Lydie Lethier, Maurad Cabut, Cédric Peudepiece, Christine Fagnoni-Legat and Claire ANDRE in Journal of Oncology Pharmacy Practice

Footnotes

ORCID iD

Yves Claude Guillaume

Ethics approval statement

Not applicable

Author contributors

YG, CA and LL conceptualized this work, designed the experiments. YG, CA wrote the manuscript. All authors approved the final version of the manuscript for submission.

Funding

The authors have not declared a specific grant for this research for any funding agency in the public, commercial or not for profit sectors.

Funding

The authors received no financial support for the research, authorship, and/or publication of this article.

Declaration of conflicting interests

The authors declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

Data availability statement

The authors confirm that the data supporting the findings of this study are available within the article [and/or] its supplementary materials.

Supplemental material

Supplemental material for this article is available online.

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