Development of an eco-friendly HPLC method for the stability indicating assay of binary mixture of ibuprofen and phenylephrine

doi:10.21203/rs.3.rs-2635611/v1

Download PDF

Research Article

Development of an eco-friendly HPLC method for the stability indicating assay of binary mixture of ibuprofen and phenylephrine

https://doi.org/10.21203/rs.3.rs-2635611/v1

This work is licensed under a CC BY 4.0 License

Journal Publication

published 24 Oct, 2023

Read the published version in BMC Chemistry →

You are reading this latest preprint version

The development and validation of the stability indicating HPLC technique has contributed to the understanding of the stability profile of ibuprofen (IBU) and phenylephrine (PHE). Stability profile was achieved for PHE, the drug was found to be liable to be influenced by stress oxidative conditions. Two oxidative degradates were formed and their structures were confirmed using IR and mass spectrometry. The drugs and degradation products were successfully separated using a a gradient elution method on YMC-C8 column with 0.1% hexanesulfonic acid and acetonitrile as solvents at pH 6.6. The flow rate was 1.0 mL.min^− 1, and a diode array detector operating at 220 nm was used for UV detection. The proposed technique has been validated using ICH guidelines and its greenness was assessed according to Analytical Eco Scale metric.

Degradation

Green analytical chemistry

HPLC

Ibuprofen

Phenylephrine

Stability indicating assay

Stability studies can help to identify the probable degradation pathways under various stress conditions ¹. Also, they are absolutely necessary for shelf life calculation of pharmaceutical products and confirmation of quality, safety and efficiency ². Different techniques of analytical chemistry can be used for development of stability-indicating assay methods such as spectrophotometry ³ and electrochemistry ⁴ with HPLC representing the main technique of choice for this application ⁵.

Ibuprofen (IBU), (2RS)-2-[4-(2-Methylpropyl)phenyl]propanoic acid ⁶ (Fig. 1a) is used to relieve pain, fever and inflammation as it is a nonsteroidal anti-inflammatory medicine. It acts mainly as an anti-inflammatory, antipyretic and analgesic drug through inhibiting the cyclooxygenase-2 enzyme ⁷. Phenylephrine (PHE), (1R)-1-(3-Hydroxyphenyl)-2-(methylamino)ethanol ⁶ (Fig. 1b) is a medicine mainly utilized as a decongestant, to enlarge the pupil, to raise the blood pressure, and to treat hemorrhoids ⁸. The combination dosage form of IBU and PHE is indicated to treat the symptoms of the common cold and flu ⁹, such as stuffy nose, nasal congestion, headache, fever and moderate aches and pains. It is available in the market in the form of coated tablets under trade name Grippostad®.

Scientific reports showed several methods for assay of IBU ¹⁰ and PHE ¹¹ in pharmaceuticals using different techniques for their simultaneous determination in different mixtures. Literature survey revealed few techniques for assaying IBU and PHE in their binary combination simultaneously such as spectrophotometry ¹², spectrofluorimetry ¹³ and HPLC ¹⁴. The two drugs were subjected to stability studies in their mixtures with other drugs ^{11d, 15}, but no method was developed for the stability indicating assay of this binary mixture.

Green analytical chemistry (GAC) is advanced field which is connected with developing analytical methods that eliminate or minimize the harmful effects of various organic solvents, on health as well as on the environment ¹⁶. Nowadays, the environment protection and the personal health and safety are considered in the field of chemistry, that led to increasing number of published articles about how to work according to green principles and guidelines to obtain better clean handling of chemistry ¹⁷. GAC has been considered in different techniques such as spectrophotometry ¹⁸, spectrofluorimetry ¹⁹, electrochemistry ²⁰ and chromatography ²¹. The methods for evaluating greenness of an analytical method should be quantitative, and consistent evaluating methods. Examples of these methods are the National Environmental Methods Index (NEMI), that is regarded as the most traditional evaluating method ²², Analytical Eco Scale (AES)²³, the Green Analytical Procedure Index (GAPI)²⁴ and Analytical Greenness metric (AGREE)²⁵.

The molecular dynamic simulation technique (MDS) is an important computational tool for understanding the system's dynamic development, determining the physical foundation of the structure, assessing the stability of interactions at the molecular level, and verifying practical work outcomes. As a result, the primary goal of using MDS, which was employed for the first time in chromatographic approaches, was to evaluate the introduced drug's interaction with the stationary phase and to confirm the results of the proposed method by revealing which drug will be less retained in the stationary phase and which one will be more retained ²⁶.

The aim of this developed method is to prepare, separate and detect the oxidative degradates of PHE and to develop an eco-friendly HPLC method for assay the binary mix with the degradates of PHE. MDS was used to analyse the elution behavior of the intact drugs and degradates and describe their interaction with the stationary phase. AES metric was used for calculating of the greenness of this method and comparison to the published method.

2.1. Instrumentation

HPLC system (Aglient 1100 Model G1315A) with a photodiode array detector (DAD) and 100 µL injection loop. YMC-C8 column (100 × 4.6 mm, 3 µm) had been used. Auto sampler system was used to inject the sample.

2.2. Materials and reagents

2.2.1. Pure standard

IBU and PHE were kindly supplied by NODCAR. Their purities were found to be 99.62 ± 1.11% and 100.02 ± 1.14% for IBU and PHE, respectively, in accordance to the BP methods ⁶.

2.2.2. Pharmaceutical dosage form

Grippostad® film coated tablets produced by ES, Laboratorio Stada. Each tablet contains 200 mg of ibuprofen and 5 mg of phenylephrine.

2.2.3. Chemicals and reagents

Acetonitrile of HPLC grade (Merck, Darmstadt), hexansulfonic acid (Merck, Darmstadt).

2.2.4. Solutions

Standard solutions of of 1.0 mg mL^− 1 of IBU and PHE were prepared using methanol.

Laboratory-prepared mixtures containing various proportions of IBU and PHE and its degradates were equipped by moving various volumes from their stock solutions into volumetric flasks of 10-mL. The mobile phase for HPLC method was used to complete each volume to the mark.

2.3. chromatographic conditions

At 35 c^o, gradient separation was carried out on YMC-C8 column (100 × 4.6 mm, 3 µm) using mobile phase consists of 0.1% hexanesulfonic acid and acetonitrile. The gradient starts with 0.1% hexanesulfonic acid and acetonitrile at ratio 80: 20, and after 3 minutes the ratio changed into 60: 40 till the end of the run. The mobile phase was pumped at a flow rate of 1 ml/min and de-gassed. 20 µL of solution had been injected and detected at 220 nm.

2.4. Procedures

2.4.1. Preparation of PHE degradates

Oxidative degradation was performed by refluxing 100.0 mg of PHE powder with 25 mL of 3% H₂O₂ for 3 hr and protected from light and then get rid of excess H₂O₂ by evaporation. The solution was cooled, moving into volumetric flask of 25-mL and completed to volume using methanol, after that confirming for degradation completely through TLC. Silica gel 60 F₂₅₄ plates were used to check the disappearance of intact drug spot at R_f 0.12 and appearance of two spots of degradation products at R_f 0.45 and 0.55 using ethyl acetate-methanol-30% ammonia solution (8:2:0.1, by volume) as the developing mobile phase. We obtain the degradation products by scratching the separated spots on TLC individually and dissolving it in methanol. The obtained degradation products Fig. 2 were separated via preparative TLC, purified and were identified using infra-red Fig. 3 and mass spectrometry Fig. 4.

2.4.2. Construction of the calibration curves

Aliquots equivalent to 100–1000 µg of IBU and 3-100 µg of PHE were precisely moved from their stock solutions into two separate sets of volumetric flasks of 10-mL and the volumes were adjusted using the mobile phase. Membrane filter 0.45 µm was used to filter the resulting solutions and 20 µL aliquots has been injected into the column under the previous chromatographic conditions. Plotting the integrated peak area against the corresponding concentrations to construct calibration curves.

2.4.3. Assay of laboratory prepared mixtures

Various combinations of IBU, PHE and its degradates were prepared and mixed in various prpportions, similar to the solutions section. The prepared solutions were then analyzed by the developed method as described under construction of calibration curve.

2.4.4. Application to tablet dosage form

Ten of Grippostad® film coated tablets (labelled to contain 200.0 mg of IBU and 5.0 mg PHE per tablet) were precisely weighed, the tablets were first stripped of the film before being finely powdered. A precisely weighed portion containing 200 mg of IBU and 5 mg of PHE was sonicated in 30 mLof methanol for 10 min before being filtered into a 100-mL volumetric flask. The residues were rinsed multiple times with 10 mL of methanol, and the solutions were adjusted to the mark by the same solvent. An aliquot was then diluted in order to prepare a solution containing 40.0 and 1.0 µg mL^− 1 of IBU and PHE, respectively.

3.1. Preparation and structure elucidation of degradation products of PHE

We should study the possible degradation pathways and products for these drugs at similar stress conditions, for developing analytical method indicating stability. PHE was reported to be more stable toward acidic, alkaline, thermal and photodegradation conditions ^{11d, 15a−c}. While it is more susceptible to oxidative conditions producing 2 degradation products ^11d. IBU was reported to be susceptible to strong oxidative and thermal degradation ^15d−f.

In our work, IBU and PHE were subjected to the same oxidative stress conditions (refluxing with 3% H₂O₂ for 3 hr), and the degradation was monitored using TLC. While two main resolved peaks at R_f 0.45 and 0.55 appeared for the PHE degradation solution, IBU degradation solution showed only the intact drug spot. This result agrees to the study that concluded IBU is susceptible only to strong oxidation conditions ^15d. The two degradation products of PHE were separated via preparative TLC, purified and their mass and IR spectra were obtained to confirm their structures.

The IR spectra of the degradation products (Fig. 3) show appearance of aromatic ketone bands around 1640 cm^− 1 and oxide band in deg2 spectrum at 1453 cm^− 1. Mass spectra of deg1 and deg2 (Fig. 4) indicated peaks at m/z 184 and 195 consistent with the molecular weight of their proposed structures. The proposed pathway for PHE degradation is shown in Fig. 2.

3.2. Method development and optimization

The main goal of this article is to provide a sensitive, accurate and selective HPLC method for assaying IBU, PHE and its oxidative degradates simultaneously in their quaternary mixture and in dosage forms. Important factors affecting the efficiency of the HPLC technique were studied carefully. Levels of a presented factor were changed while the other chromatographic conditions remained unchanged.

Many trials have been done on several sorts of columns to achieve optimum separation, e.g., XTerra MS (100 x 4.6 mm, 5 µm), C18 X-bridge shield RP and YMC-C8 column (100 × 4.6 mm, 3 µm). The C18 columns showed very bad retention and the peaks were highly overlapped, while C8 one showed best retention and resolution. Different elution modes and mobile phases with various compositions and ratios have been applied for ideal separation. By using isocratic elution and a mobile phase of 0.1% hexanesulfonic acid sodium salt monohydrate and methanol (80:20%), IBU could not be separated. We used also buffer: methanol in different ratios (70:30 and 30:70), but the peaks were severely overlapped. All solvent compositions and ratios failed to separate four compounds using isocratic elution, so gradient elution was utilized. Finally, YMC-C8 column was used and gradient elution using 0.1% hexanesulfonic acid sodium salt monohydrate and acetonitrile as solvents. The optimum gradient started with hexanesulfonic acid sodium salt monohydrate and acetonitrile at ratio 80: 20, and after 3 minutes the ratio changed into 60: 40 till the end of the run. Various flow rates 0.8, 0.9, 1 and 1.2 mL.min^− 1 were tested and 1 mL.min^− 1 was the best regarding separation and resolution. The best UV wavelength used for detection was 220 nm regarding the sensitivity of the two drugs. Different temperatures were applied to investigate best column temperature for separation and 35 C^° was the optimum. Good chromatographic separation was achieved with tR of 2.0, 2.2, 3.2 and 7.0 min for deg1, deg2, IBU and PHEN, respectively (Fig. 5). The system suitability parameters have been calculated for the method where acceptable values were got in the recommended ranges as shown in Table 1.

Table 1

System suitability parameters of the proposed HPLC method
	PHE	IBU	Reference values
Retention time (t_R) [min]	3.71	6.28	NA
Resolution (R_s)	7.86		R_{s > 1.5}
Selectivity factor (α)	1.71		α > 1
Tailing factor (T)	0.97	1.71	T < 2
Capacity factor (K’)	2.73	5.30	1 < K’<10
Column efficiency (N)	2196.32	2080.36	N > 2000
Height equivalent to theoretical Plate (HETP) [mm]	0.0198	0.0455	NA

3.3. Method validation

Validation of the proposed technique was made in accordance with the ICH guidelines ²⁷.

3.3.1. Linearity and range

The linearity for each drug was estimated by investigating various concentrations of IBU (10–100 µg mL^− 1) and PHE (0.3–10 µg mL^− 1). The regression equations results and their corresponding relative standard errors accompanied by the LOD and LOQ were shown in Table 2.

Table 2

Assay parameters and validation sheet for determination of ibuprofen and phenylephrine by the proposed HPLC method
	PHE	IBU
Range	0.3–10 µg mL^− 1	10–100 µg mL^− 1
Slope	989994	1611.40
Intercept	528735	-11978
SE of the slope	14376.11	18.4132
SE of the intercept	68432.38	1143.568
Correlation coefficient (r)	0.9995	0.9997
LOD*	0.4399	2.75
LOQ**	1.333	8.33
Accuracy	99.44%	99.45%
Repeatability	1.34	1.09
Intermediate precision	1.47	1.23
Specificity (mean ± SD)	99.67 ± 1.67	100.75 ± 1.44
Robustness	1.94	1.19
* LOD = 3.3σ/S

**LOQ = 10σ/S.

3.3.2. Accuracy

Accuracy was assessed by applying the recently stated methods on pure samples with various concentrations, where satisfactory recoveries were attained as shown in Table 2.

3.3.3. Precision

3.3.3.1. Repeatability

On the same day, three different concentrations for IBU (10, 40, 60 µg mL^− 1) and PHE (0.3, 1.0 and 3.0 µg mL^− 1) were assessed three times. Relative standard deviations (RSD%) values indicated low deviations and high repeatability as shown in Table 2.

3.3.3.2. Intermediate precision

On three different days, the previous procedures were performed for the analysis of previously selected concentrations. RSD% showed high precision as shown in Table 2.

3.3.4. Robustness

Some experimental parameters were intentionally modified to calculate the robustness of the method. This time, organic solvent ratio in the mobile phase (20 ± 1.0%) and flow rate (1.0 ± 0.1 mL min^− 1)were considered, and reasonable RSD% were obtained (Table 2).

3.3.5. Specificity

Various laboratory-prepared combinations with different proportions of the investigated components with the degradates were analyzed. The recovery percentages and SD are shown in Table 2.

3.4. Application the proposed method to Grippostad® tablets and statistical comparison

The developed method was successfully applied for assaying IBU and PHE in Grippostad® tablets. The standard addition technique has been applied and good recoveries were obtained as shown in Table 3.

The results obtained from IBU and PHE analysis in pure powder and the official method ⁶ of analysis were compared statistically. T and F tests confirmed no significant change between the developed method and the official one as shown in Table 4.

Table 3

Determination of ibuprofen and phenylephrine in Grippostad® tablets and application of standard addition technique using the proposed method
	Grippostad® tablets	Standard Addition Technique
	Mean^* ± SD	Taken (µg mL ^− 1)	Added (µg mL^− 1)	Recovery%*
PHE	99.44 ± 1.38	0.5	0.25	99.20
			0.5	101.15
			1	98.90
		Mean^a ± SD		99.75 ± 1.22
IBU	99.78 ± 1.27	20	10	99.80
			20	99.82
			40	101.34
		Mean^a ± SD		100.32 ± 0.88

Table 4

Statistical comparison for the results obtained by the proposed HPLC method and the official method
Parameter	Proposed method		Official method [10]
	PHE	IBU	PHE	IBU
Mean of recoveries	99.44	99.78	100.02	99.57
SD	1.38	1.27	0.85	1.09
Variance	1.92	0.13	0.73	1.19
N	5	5	5	5
Student’s t-test (2.30)*	0.78	0.24	NA
F-test (6.38)*	2.60	0.11	NA

* the values in parentheses are the tabulated values

3.5. Assessment of the greenness of the developed method

The greenness profile of the proposed method was assessed and ranked according to AES tool. AES is proposed as a new comprehensive approach for greenness assessment of analytical methodology. The ease of the calculation of this method makes more advantages over other methods. The AES is determined by assigning penalty points to each analytical procedure factor that does not agree with or match with perfect green analysis. An Eco-Scale score of 100 is considered ideal for eco-friendly analysis, excellent if more than75, acceptable if more than50, and inadequate if less than50. The AES value of the developed method was calculated as shown in Table 5. In accordance to Table 5, the method we developed has high score of 86 and is considered an excellent green analytical method.

Table 5

Comparison of the greenness profiles of proposed and reported methods using ESA tool.
Reagents and instruments	Proposed method	Reported method [26]
Chloroform	0	2
Methanol	0	6
Ammonia	0	2
0.1%hexanesulfonic acid	0	0
Acetonitrile	6	0
Waste	3	3
Occupational hazards	3	3
Total penalty points	13	16
Total score	86	84

3.6. Molecular Dynamic Simulation

Before conducting the simulation, dielectric constant (DEC) was adjusted first at 21.58 and then adjusted at 25.56 as the proposed method is gradient. DEC is equivalent to the total ratios of the mobile phase, as we cannot introduce the ratios of the solvent as it will give inaccurate simulation results. Here, the composition of the column plays a serious role in the interaction of the drugs with the stationary phase. The Reversed-phase C₈ had only an octa chain which could provide hydrophobic interactions; however, using YMC-C8, which offered the same interaction as reversed-phase C₈, beside it will provide hydrogen bond donor owing to the embedded (Si-OH) group Fig. 6 in the bonded phase ligand. Thus, in this work, we used an YMC-C8 column where the interactions between the drugs and the stationary phase were conducted, and the binding energies were calculated. In the case of oxidative degradation of PHE, the resulting degradates and the intact drug were simulated under the mentioned conditions, and after a thorough examination of the resulting trajectories, we found that both degradates and the intact drug interact with the stationary phase in several different modes. The interactions between drugs and the column depend on polarity as the least polar drug will be the most retained with stationary phase and the most polar drug will be the least retained with stationary phase. For deg 2, there is positive and negative charge on N-oxide group which increase polarity of the component and also quinone group make tautomerism into its corresponding phenolic groups and that make this component the most polar between four components, so it gets first with binding energy of -1.18 Kcal/mol, as seen in Fig. 7. Deg 1 shows less polarity as there is no positive and negative charge, so get second with binding energy of -3.16 Kcal/mol as shown in Fig. 8. The intact drug molecules were found to form hydrophobic van der Waals interactions. For IBU, there is free carboxylic group despite methyl group which make weak hydrophobic van der waals interaction with stationary phase, so it gets third with binding energy of -3.35 Kcal/mol as in Fig. 9. which make component less polar than degradate 1 and there is For PHE, shows strong van der waals interaction with stationary phase due to phenyl group which make the drug the highest retained component with binding energy of -6.67 Kcal/mol as in Fig. 10. Due to the diversity of the interactions between the analyzed components and the stationary phase, these observations were not enough to prove the arrangement of the separated components regarding their binding to the stationary phase. So, after conducting the molecular dynamics simulation, the solvation energies were calculated from the emerging trajectories. A calibration plot for each component was generated by plotting the simulation time versus its solvation energy. Figure 11 arrange the oxidative degradations and the intact drug according to their solvation energies throughout the whole simulation time. The data extracted from the calculated solvation energies were in line with the binding energy data and experimental data from both the proposed chromatographic methods.

An eco-friendly HPLC method has been developed for the determination of IBU, PHE and its oxidative degradates simultaneously. The proposed approach is simple, accurate and precise and can be utilized in the analysis of binary Mixture of IBU and PHEN in presence of the oxidative degradates in pure form and in dosage form. The method has been proved to be green with respect to AES metrics.

Ethics approval and consent to participate

Not applicable

Consent for publication

Not applicable

Competing interests

The authors declare no conflict of interest

Availability of data and materials

All data generated or analyzed during this study are included in this published article and its supplementary information files listed below.

Funding

Not applicable. This study did not receive any funds from any source.

Author contributions

KMK:research idea conceptualization;supervised manuscript writing,revision and editing. YMF:support and supervision. AMA: support and supervision. RAF: conducted the lab work, data collecting and presentation and writing first drift of the manuscript.SAH: supervised the study; data analysis and manuscript writing. All authors read and approved the final manuscript.

Guideline IHT. International Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human Use 2003.
Yasmeen A, Sofi G. J Phytopharmacology. 2019;8:147–51.
aDarwish HW, Hassan SA, Salem MY, El-Zeany BA. 2016, 154, 58–66; bHassan, S. A.; Nashat, N. W.; Elghobashy, M. R.; Abbas, S. S.; Moustafa, A. A., Spectrochim. Acta A 2022, 267, 120576.
Hassan SA, Nashat NW, Elghobashy MR, Abbas SS, Moustafa AA, Mahmoud AM. Microchem J. 2022;178:107323.
aBasha MA, Abdelrahman MK, Bebawy L, Mostafa A, Hassan SA, cGad M, Hassan SA, Zaazaa HE, Amer SM. 2020, 243, 118756; bWeshahy, S. A.; Yehia, A. M.; Helmy, A. H.; Youssef, N. F.; Hassan, S. A., Microchem. J. 2020, 157, 105047; cGad M, Hassan SA, Zaazaa HE, Amer SM, Chromatographia 2019, 82 (11), 1641–1652; dHassan, S. A.; Nashat, N. W.; Elghobashy, M. R.; Abbas, S. S.; Moustafa, A. A., J. Chromatogr. Sci. 2020, 58 (8), 747–758.
Medicines. and Healthcare Products Regulatory Agency (MHRA): London, 2019.
Sweetman SC. 2005.
Esteve-Taboada JJ, Águila-Carrasco D, Antonio J, Bernal-Molina P, Ferrer-Blasco T, López-Gil N, Montés-Micó R. Journal of Ophthalmology 2016, 2016.
Farrer F. Prof Nurs Today. 2010;14(2):8–12.
aCunha RR, Chaves SC, Ribeiro MM, Torres LM, Muñoz RA, Santos WTD, Richter EM, cPeikova L, Georgieva M, Tsvetkova B, Suliman SM. 2015, 38 (10), 1657–1662; bPadró, J. M.; Osorio-Grisales, J.; Arancibia, J. A.; Olivieri, A. C.; Castells, C., J. Chromatogr. A 2016, 1467, 255–260; cPeikova L, Georgieva M, Tsvetkova B, Pharmacia 2014, 61, 3–6; dSave, T. K.; Parmar, D.; Devarajan, P. V., journal of chromatography b: biomedical sciences and applications 1997, 690 (1–2), 315–319; eAl-Kindy, Suliman SM, F. E. O., Luminescence: The journal of biological and chemical luminescence 2007, 22 (4), 294–301.
aPalabiyik IM, Onur F, Hassan FA, Khamis EM, Ragab EF, Hamdy MA, Elsayed NM, Hassan GM. 2010, 26 (8), 853–859; bElfatatry, H. M.; Mabrouk, M. M.; Hammad, S. F.; Mansour, F. R.; Kamal, A. H.; Alahmad, S., J. AOAC Int. 2016, 99 (5), 1247–1251; cEl Yazbi, Hassan FA, Khamis EM, Ragab EF, Hamdy MA, M. M., Journal of chromatographic science 2016, 54 (5), 819–828; dRezk, M. R.; Fayed, A. S.; Marzouk, H. M.; Abbas, S. S., J. AOAC Int. 2017, 100 (2), 434–444; eMostafa, Elsayed NM, Hassan GM, N. Y.; El Mously, D. A., Journal of AOAC International 2017, 100 (6), 1761–1770.
Patel M, Patel B, Parmar S. Journal of Spectroscopy 2013, 2013.
Salem YA, Hammouda ME, El-Enin MAA, El-Ashry SM J. S. A. P. A. M.;, Spectroscopy B. 2019, 210, 387–397.
aRagab MA, Abdel-Hay MH, Ahmed HM, Mohyeldin SM. 2019, 57 (7), 592–599; bVemula, V. R. B.; Sharma, P. K., Tropical Journal of Pharmaceutical Research 2014, 13 (6), 967–974.
aRagab MA, El Yazbi FA, Hassan EM, Khamis EF, Hamdy MM, cPatel KB, Thula KC, Maheshwari DG, eAhirrao V, Pawar R. 2018, 8 (3), 331–347; bPatel, B. S.; Nayak, P. P.; Parmar, R. R.; Shah, D. A., Pharm Anal. Qual. Assur. 2014, 3, 1–7; cPatel KB, Thula KC, Maheshwari DG, Pharmacophore 2014, 5 (2), 262–272; dCaviglioli, G.; Valeria, P.; Brunella, P.; Sergio, C.; Attilia, A.; Gaetano, B., J. Pharm. Biomed. Anal. 2002, 30 (3), 499–509; eAhirrao V, Pawar R, Research Journal of Pharmaceutical Sciences 2013, 2319, 555X; fFarmer, S.; Anderson, P.; Burns, P.; Velagaleti, R., Pharm. Technol. 2002, 28 (42), 13.
Płotka-Wasylka J, Mohamed HM, Kurowska-Susdorf A, Dewani R, Fares MY, Andruch V. Curr Opin Green Sustainable Chem. 2021;31:100508.
Anastas P, Eghbali N. Chem Soc Rev. 2010;39(1):301–12.
ElDin NB, Abd El-Rahman MK, Zaazaa HE, Moustafa AA, Hassan SA.Microchem. J.2021,106419.
aIbrahim N, Elzanfaly ES, El Gendy AE, Hassan SA. 2021, 14 (4), 1880–1886; bAbdelaal, S. H.; El Azab, N. F.; Hassan, S. A.; El-Kosasy, A. M., Spectrochim. Acta A 2021, 120032.
Elhassan MM, Mahmoud AM, Hegazy MA, Mowaka SJCP. 2021, 75 (8),4165–4176.
aHassan SA, Ibrahim N, Elzanfaly ES, El Gendy AE. 2021, 1–7; bPeleshok, K.; Piponski, M.; Ajie, E. A.; Poliak, O.; Zarivna, N.; Denefil, O.; Logoyda, L. J. P., 2021, 68, 43.
Keith LH, Brass HJ, Sullivan DJ, Boiani JA, Alben KT, editors, editors. Vol.;ACS Publications, 2005.
Gałuszka A, Migaszewski ZM, Konieczka P, Namieśnik J. TRAC Trends Anal Chem. 2012;37:61–72.
Płotka-Wasylka J. Talanta 2018, 181,204–209.
Pena-Pereira F, Wojnowski W, Tobiszewski M. Anal Chem. 2020;92(14):10076–82.
Jameson CJ, Wang X, Murad S.AIChE Journal2021, 67 (3), e17143.
In International Conference on Harmonization (ICH)ed., Vol. 62, 2005.

No competing interests reported.

Download PDF

Journal Publication

published 24 Oct, 2023

Read the published version in BMC Chemistry →

Editorial decision: Major revision
12 Jun, 2023
Reviews received at journal
12 Jun, 2023
Reviews received at journal
08 Jun, 2023
Reviewers agreed at journal
27 May, 2023
Reviewers invited by journal
27 May, 2023
Editor invited by journal
25 May, 2023
Editor assigned by journal
15 Mar, 2023
Submission checks completed at journal
06 Mar, 2023
First submitted to journal
27 Feb, 2023

You are reading this latest preprint version

Development of an eco-friendly HPLC method for the stability indicating assay of binary mixture of ibuprofen and phenylephrine

Status:

Journal Publication

Version 1

Abstract

Figures

1. Introduction

2. Experimental

2.1. Instrumentation

2.2. Materials and reagents

2.2.1. Pure standard

2.2.2. Pharmaceutical dosage form

2.2.3. Chemicals and reagents

2.2.4. Solutions

2.3. chromatographic conditions

2.4. Procedures

2.4.1. Preparation of PHE degradates

2.4.2. Construction of the calibration curves

2.4.3. Assay of laboratory prepared mixtures

2.4.4. Application to tablet dosage form

3. Results And Discussion

3.1. Preparation and structure elucidation of degradation products of PHE

3.2. Method development and optimization

3.3. Method validation

3.3.1. Linearity and range

3.3.2. Accuracy

3.3.3. Precision

3.3.3.1. Repeatability

3.3.3.2. Intermediate precision

3.3.4. Robustness

3.3.5. Specificity

3.4. Application the proposed method to Grippostad® tablets and statistical comparison

3.5. Assessment of the greenness of the developed method

3.6. Molecular Dynamic Simulation

Conclusion

Declarations

References

Additional Declarations

Status:

Journal Publication

Version 1