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Heba Elmansi Pharmaceutical Analytical Chemistry Department, Faculty of Pharmacy, Mansoura University, Mansoura, 35516, Egypt

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Mohamed I. El-Awady Pharmaceutical Analytical Chemistry Department, Faculty of Pharmacy, Mansoura University, Mansoura, 35516, Egypt
Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Delta University for Science and Technology, International Coastal Road, Gamasa, 11152, Egypt

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Sona Barghash Pharmaceutical Analytical Chemistry Department, Faculty of Pharmacy (Girls), Al-Azhar University, Cairo, Egypt

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Sawsan Abd El-Razeq Pharmaceutical Analytical Chemistry Department, Faculty of Pharmacy (Girls), Al-Azhar University, Cairo, Egypt

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Fathalla Belal Pharmaceutical Analytical Chemistry Department, Faculty of Pharmacy, Mansoura University, Mansoura, 35516, Egypt

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Abstract

A fast reliable micellar electrokinetic methodology was investigated for the concurrent quantitation of six antimicrobial and anti-inflammatory drugs, namely, ciprofloxacin, dexamethasone, metronidazole, ornidazole, spiramycin and tinidazole. The method has the merits of rapidity, precision, and sensitivity. The separation was carried out in less than 7 min by applying a basic background electrolyte consisting of 25 mM disodium tetraborate buffer, pH 9 containing 50 mM SDS at 25 kV using photodiode array detector at 230 and 315 nm. The internal standard used during analysis was cromolyn sodium and validation was carried out following ICH guidelines. The proposed method showed linear response over the range from 0.5 to 10.0 μg mL−1 reaching limits of detection and limits of quantitation in the ranges of 0.09–0.2 μg mL−1 and 0. 27–0.6 respectively. The method's greenness was estimated using the GAPI tool where excellent greenness was concluded. Co-formulated or single-ingredient commercial preparations were investigated and the results were statistically evaluated.

Abstract

A fast reliable micellar electrokinetic methodology was investigated for the concurrent quantitation of six antimicrobial and anti-inflammatory drugs, namely, ciprofloxacin, dexamethasone, metronidazole, ornidazole, spiramycin and tinidazole. The method has the merits of rapidity, precision, and sensitivity. The separation was carried out in less than 7 min by applying a basic background electrolyte consisting of 25 mM disodium tetraborate buffer, pH 9 containing 50 mM SDS at 25 kV using photodiode array detector at 230 and 315 nm. The internal standard used during analysis was cromolyn sodium and validation was carried out following ICH guidelines. The proposed method showed linear response over the range from 0.5 to 10.0 μg mL−1 reaching limits of detection and limits of quantitation in the ranges of 0.09–0.2 μg mL−1 and 0. 27–0.6 respectively. The method's greenness was estimated using the GAPI tool where excellent greenness was concluded. Co-formulated or single-ingredient commercial preparations were investigated and the results were statistically evaluated.

1 Introduction

Ciprofloxacin (CIP) is a broad spectrum antibacterial agent belonging to the group of fluoroquinolones [1]. CIP is used with an antimicrobial medication that kills anaerobes, like metronidazole (MNZ), to treat mixed aerobic/anaerobic infections [2]. On the other hand, a combination of CIP and ornidazole (ORZ) is used in the treatment of bacterial & parasitic infections. It is used to treat diarrhoea, gynaecological and pelvic infections. It also helps treat mixed infections of teeth and gums. CIP works by preventing the bacterial cells from dividing and repairing, thereby killing the bacteria. ORZ kills parasites and anaerobic bacteria that cause infections by damaging their DNA [3]. At the same time, combined CIP and tinidazole (TNZ) was found to be effective in the cases of pelvic inflammatory disease [4] while, CIP and dexamethasone (DXM) otic drops are used to treat outer ear infections in adults and children and acute (suddenly occurring) middle ear infections in children with ear tubes. The combination of CIP and DXM works by killing the bacteria that cause infections and reducing swelling in the ear. Additional uses have been described in children with Acute Otitis Media and Otorrhea (AOMT) [5], topical otic therapy with CIP/DXM is superior to CIP alone and leads in a faster clinical remission, furthermore a combination dosage form of spiramycin (SPI) and metronidazole (MNZ) was found to be effective in the treatment of active periodontitis [6]. This combination is beneficial in reducing brain cysts caused by toxoplasma gondii [7]. As different combinations present in the market containing two or more of the six cited drugs, we aimed to develop a fast reliable capillary electromigration separation method for the quantitation of these antimicrobial and anti-inflammatory drugs in their different dosage forms, based on MEKC. This will help in quality control testing without the need for sample preparation and costly solvents. Additionally, simultaneous estimation plays a very important role in the pharmaceutical analysis as it is very feasible and time saving in quality control laboratories.

Figure 1 illustrates the chemical structure of the studied analytes. Various methods have been developed for the analysis of these compounds in the literature. A good guide for the work reported for CIP up to 2018 is found in the critical review written by Kawas et al. [8]. Another review article covering the reported methods for ORZ up to 2014 was prepared by Zameeruddin et al. [9]. Regarding TNZ, Sebastian [10] reviewed its literature up to 2017. The recent articles concerning the analysis of CIP include spectrophotometric [11, 12], High performance liquid chromatography (HPLC) [13, 14] and capillary electrophoresis (CE) [15, 16] methods. Regarding DXM, the recent methods include spectrophotometric [17], TLC [18], HPLC [19, 20] and CE [21] methods. For MNZ, the reported methods include spectrophotometric [22, 23] electrochemical [24, 25], TLC [26, 27] HPLC [28, 29] and CE [30, 31] methods. Determination of ORZ was carried out using spectrophotometric [32, 33] and HPLC [34, 35] methods. The literature for SPI determination includes spectrophotometric [36], HPLC [3738], UHPLC [39, 40] and CE [41] methods. As for TNZ, the recent methods include spectrophotometry [42, 43], electrochemical determination [44], TLC [45] and HPLC [46, 47].

Fig. 1.
Fig. 1.

Chemical structure of: (A) Ciprofloxacin HCL; (B) Dexamethasone; (C) Metronidazole; (D) Ornidazole; (E) Spiramycin; (F) Tinidazole

Citation: Acta Chromatographica 2022; 10.1556/1326.2022.01057

Micellar Electrokinetic Chromatography (MEKC) is a technique that relied on adding surfactants to the background electrolytes [48]. This technique permits the simultaneous separation of both neutral and charged species. The surfactant is added to the background electrolyte (BGE) above its critical micelle concentration (CMC) where individual surfactant molecules aggregate to form micelles. The micelles are dynamic structures with hydrophobic tails positioned in the center away from the hydrophilic buffer and charged heads located outside and towards the aqueous buffer. As the micelles are charged, they act as a pseudo-stationary phase for the analytes and the partitioning of the analytes occur between the moving micellar and aqueous phases which affects the separation process. Micelles and solutes interactions are electrostatic and/or hydrophobic in nature. During electrophoresis micelles also show a differential migration due to their effective mobility. For a neutral species, the interactions are essentially hydrophobic whereas for a charged species, they are a mixture of electrostatic and hydrophobic interactions [49]. As polarity plays an important role in the analysis of pharmaceuticals, the introduction of MEKC resulted in versatility of applications in this area [50–52].

The Society of Analytical Chemistry is recently following the applications of Eco-friendly methods that exclude or reduce toxic and caustic waste [53]. Assessment of the greenness of analytical methods is now an important issue in the development stage. The small scales in which MEKC separations are performed make this family of techniques a greener alternative to chromatography. These approaches consume less amounts of solvents and samples. Using micellar biodegradable system is an additional merit. Among the new methods used for greenness evaluation is GAPI [54]. It is superior to other green assessment tools because it is based on the evaluation of the green character of an entire analytical methodology, from sample collection to final determination. We, therefore, evaluated the greenness using GAPI method.

2 Materials and methods

2.1 Materials

Pure CIP, TNZ and SPI were supplied by the National Organization of Drug Control and Research (NODCAR), Cairo, Egypt.

Pure ORZ was provided by Pharonia Pharmaceuticals (Pharo Pharma), Borg EL-ARAB, Alexandria, Egypt.

Pure MNZ was provided by Amriya Pharmaceutical Industries, Alexandria, Egypt.

Pure DXM was provided by Egyptian International Pharmaceutical Industries Company (EIPICO), 10th of Ramadan City, Egypt.

Cromolyn Sodium was provided by Sigma Pharmaceutical Company, 6th October City, Egypt, used as the internal standard (IS).

Pharmaceutical preparations were bought from the local Egyptian market. These include

Ciprobay® tablets; containing 500 mg of CIP, product of Hikma Pharma S.A.E. Egypt.

Ornidaz® tablets; containing 500 mg of ORZ, product of Chemipharm Pharmaceutical Industries S.A.E. 6th October City, Egypt.

Protozole tablets; containing 500 mg of TNZ, product of Medical Union Pharmaceuticals, Ismaelia, Egypt.

Flagyl ® tablets; containing 500 mg of MNZ, product of Sanofi Aventis. Cairo, Egypt.

Dexamethasone-MUP ampoules Batch No.20246, each ampoule contains 8 mg dexamethasone phosphate produced by Medical Union Pharmaceuticals, Abu-Sultan, Ismailia, Egypt.

Spirex® tablets; product of Medical Union Pharmaceuticals, Ismaelia, Egypt.

Tinifloxacin tablets, containing 500 mg of CIP and 600 mg of TNZ, produced by Organopharma. Cairo, Egypt.

Ciprodiazole tablets, contain 500 mg CIP and 500 mg MNZ/tablet, by Minapharm for Pharmaceuticals and Chemical Industries 10th of Ramadan City- Egypt.

Co-formulated tablets Ciplox-OZ [55] were prepared in laboratory by mixing 10 mg CIP and 10 mg ORZ fine powder together. The powder was compacted into tablets with common diluents and fillers.

Spirazole forte tablet, containing 1.5 M.I.U spiramycin and 250 mg metronidazole; produced by Pharonia Pharmaceuticals (Pharo Pharma), Borg EL-ARAB, Alexandria, Egypt.

Zonacip otic drops, containing 3 mg CIP and 1 mg DXM/mL; produced by Pharonia Pharmaceuticals (Pharo Pharma), Borg EL-ARAB, Alexandria, Egypt.

2.2 Reagents

Analytical grade NaH2PO4, Na2B4O7.10 H2O (borax), H3PO4 and NaOH were from Merck, Darmstadt, Germany.

Sodium dodecyl sulfate (SDS) was bought from Fluka, Buchs, Switzerland.

0.45 μm Syringe filters (Minisart RC25) were bought from Sartorius-Stedim (Gottingen, Germany) were used for filtration of samples and BGE.

Deionized water was used throughout the study.

2.3 Instruments

  • ➢ Agilent 7100 Capillary Electrophoresis system (Agilent Technologies, Waldbronn, Germany) with a diode-array detector was used. Data were recorded with Agilent Open LAB CDS software. Fused-silica capillaries (50 μm I.D.) were from Agilent Technologies (Waldbronn, Germany) with a total length of 48.8 cm and effective length: 40.5 cm the generated electric current was around 35 μA under the optimized conditions,

  • ➢ For pH measurements, a Consort NV P-901 pH–Meter (Belgium) was used.

  • ➢ Ultrasonic bath BHA-180 T (Abbotta, USA).

2.4 Preparation of background electrolyte

Borate buffer stock (100 mM, pH 9.0) was prepared by dissolving 3.714 g of disodium tetraborate (Na2B4O7.10H2O) in 100 mL of deionized water and the final pH was adjusted to pH 9.0 with 0.1 M NaOH. Accurately weighted 5.778 g SDS was dissolved in 100 mL of deionized water to obtain a stock SDS solution (200 mM) and the solution was kept in the refrigerator and used within one week. These stock solutions used for the preparation of the final optimized BGE which is a 20 mM borate buffer with pH of 9. 0 and 50 mM SDS. The BGE was degassed in an ultrasonic bath for 10 min after being filtered using a 0.45 mm syringe filter.

2.5 Standard solutions

Freshly prepared stock solutions containing 1.0 mg mL−1 of each of TNZ, MNZ, ORZ, CIP, DXM, SPM and Cromolyn sodium (I.S) were prepared separately in deionized water. Working solutions were prepared by further dilution with the same solvent to obtain concentration of 0.1 mg mL−1.

2.6 Electrophoretic procedure

  • ➢ For the first usage, the capillary was conditioned by flushing with 1.0 M NaOH for 30 min, then with water for 15 min, followed by 5 min of drying by purging air through the capillary.

  • ➢ The capillary was rinsed for 30 min with 0.2 M NaOH, 10 min with water, and then BGE for 30 min at the start of each working day.

  • ➢ To keep optimal run-to-run injection repeatability, the capillary was preconditioned with 0.2 M NaOH for 2 min, water for 1 min and BGE for 3 min before each injection. .

  • ➢ Photodiode array (PDA) detector was set at 230 and 315 nm with a bandwidth of 10 nm.

  • ➢ Samples were introduced at hydrodynamic pressure of 50 mbar for 10 s,

  • ➢ The temperature of the capillary was kept constant at 25 °C and a voltage of 25 kV was applied (positive polarity).

3 Procedures

3.1 Construction of Calibration Graphs

In a set of 10-mL volumetric flasks, aliquots of the working standard solutions of the six drugs (0.1 mg mL−1) were transferred and a constant volume of 10 μL cromolyn sodium stock solution (1.0 mg mL−1) was added. The flasks were completed to volume with deionized water to reach final concentrations in the range of 0.5–5.0 μg mL−1 for TNZ and ORZ, 1.0–10.0 μg mL−1 for MNZ, CIP. The peaks of the corresponding drugs were detected at 315 nm while DXM and SPI were detected at 230 nm for treplicate and average responses were calculated. The samples were then analyzed under the above optimum conditions. The corrected peak area ratio was plotted accordingly versus the final concentration of each drug in μg mL−1 and the corresponding regression equation was derived.

3.2 Pharmaceutical applications

3.2.1 For single-component tablets

Ten Ciprobay®, Ornidaz®, Flagyl®, Protozole or Spirex ® tablets were weighed and finely powdered. An amount equivalent to 10.0 mg of each drug was transferred in to separate 100-mL volumetric flasks and about 50 mL of deionized water was added. The flasks were then sonicated for 20 min, completed with deionized water to obtain a solution containing 0.1 mg mL−1 for each drug. The solutions were then filtered through a syringe filter.

3.2.2 For DXM in injections

Into a 100-mL volumetric flask, accurately measured volumes of freshly mixed five Dexamethasone ® ampoules equivalent to 10 mg DXM were transferred. About 75.0 mL of deionized water were added, mixed, and completed to obtain a solution containing 0.1 mg mL−1 DXM.

3.2.3 For multi-component tablets

Ten tablets of Ciplox-OZ [55], Ciprodiazole, Tinifloxacin and Spirazole forte ® tablets were grinded then transferred into four separate 100-mL volumetric flasks and about 50.0 mL of deionized water were added and mixed well then sonicated for 20 min. Volumes were completed to the mark and filtered.

3.2.4 Zonacip otic drops

The contents of five Zonacip otic bottles were mixed and a volume of the freshly mixed suspension equivalent to 30 mg of CIP and 10 mg of DXM was transferred to 100-mL volumetric flask. The flask was completed to the mark with deionized water, then filtered using a syringe filter.

As illustrated under “Construction of Calibration Graphs,” further dilution with deionized water was made as suitable for each preparation. Each analyte's corrected peak area ratio (analyte/I.S.) was calculated and the concentration was estimated using the corresponding regression equation.

4 Results and discussion

Various combinations in dosage forms are present in the pharmaceutical market. These multi component formulations need new methods for their analysis. There is increasing interest in the use of capillary electrophoresis for applications in analytical and biochemical fields. Two problems were faced while attempting to separate the studied analytes: The first problem is the different pka range between the analytes. The second one is the structural similarity of the three studied nitroimidazole, which need to be separated by high resolution power. MEKC has thought to be a promising method for separating charged and neutral molecules with high resolving power, small sample volumes, and less time compared with HPLC. The designated method permitted the separation of the six studied drugs in a single run (less than 7 min) with good resolution. Figure 2A and B show typical electropherograms for the studied drugs under the described conditions at 315 and 230 nm. The residence times for TNZ, MNZ, ORZ, CIP, DXM and SPI were 2.5, 2.6, 2.8, 3.6, 4.0 and 6.8 min, respectively.

Fig. 2.
Fig. 2.

(A) Electropherogram for the studied analytes using the optimized experimental conditions at 315 nm; (analyte concentration: 2.0 µg mL−1 for TNZ and ORZ, 5.0 µg mL−1 for MNZ and CIP and 1.0 µg mL−1 for Cromolyn Sodium as I.S.; A: TNZ (2.5 min), B: MNZ (2.6 min), C: ORZ (2.8 min), D: CIP (3.6 min) and I.S: (4.2 min). (B) Electropherogram for the studied analytes using the optimized experimental conditions at 230 nm; (analyte concentration: 5.0 μg mL−1 for DXM and SPI; D: CIP (3.6 min), E: DXM (4.0 min), I.S: (4.2 min), F: SPI (6.8 min)

Citation: Acta Chromatographica 2022; 10.1556/1326.2022.01057

4.1 Optimization of capillary electrophoretic conditions

All parameters influencing the efficiency and selectivity of the proposed method were studied and optimized.

4.2 Detection wavelength

Depending on the absorbance properties of each compound Diode array detector was set at different wavelengths. At wavelengths of 200, 220, 230, 270, and 315 nm, different electropherograms were run. TNZ, MNZ, ORZ, and CIP electropherograms were found to be the best at 315 nm while the best electropherograms were obtained at 230 nm for DXM and SPI, so the two wavelengths were used for detection to enhance the sensitivity the method.

4.3 Rinsing of the capillary between two runs

Alkaline conditioning to remove adsorbates and refresh the surface by deprotonation of the silanol groups is most commonly employed.

In preliminary studies the capillary was rinsed with 0.1 M NaOH for one minute, deionized-water for one minute and buffer solution for three minutes. The steps were performed at a pressure of 50 mbar. Also 0.1 M NaOH for 2 min, water for 1 min and BGE for 3 min was tried but a delay in the migration time was noted. This led to the use of 0.2 M NaOH. Different durations of the washing steps were investigated and 0.2 M NaOH for 2 min, water for 1 min and BGE for 3 min was the best rinsing procedure that maintained appropriate injection repeatability from run to run.

4.4 Effect of pH

It is an important notice to state that the pka values of TNZ, MEZ, ORZ, CIP, DXM and SPI are 4.7,2.62,2.4,6.09,1.89 and 8.0 respectively [56], so alkaline pH would be an excellent choice for separation. The selection of alkaline pH instead of neutral or acidic was also relied on the fact that the alkaline medium holds the capillary tube's inner silica ionized [57] thus reducing the migration time.

The effect of pH was examined over the pH range 6–10, using 20 mM buffer solutions prepared at various pH values. As the pH increased, the migration times were shown to decrease. Lower pH values resulted in an increase in the migration time with accompanying peak broadening of TNZ which is co-eluted with solvent front. The optimum pH was 9.0 as it resulted in the highest efficiency, well resolved peaks, best peak shape, and sensitivity (Fig. S1).

4.5 Buffer type and concentration

Two running buffers phosphate and borate buffers at different pHs, were tested for the best resolution. In borate buffer, better resolved peaks with low background current were achieved.

Borate buffer concentration was changed from 10 to 40 mM. The EOF is reduced by increasing the buffer concentration, and vice versa. Moreover, Joule heating will occur upon using a high concentration of the buffer. 20 mM concentration of borate buffer was chosen as the optimum BGE to keep better peak shape within a reasonable migration time (Fig. S2).

4.6 SDS concentration

The concentration of SDS was investigated at concentrations ranging from 30 to 80 mM. The increase of SDS concentration from 50 to 80 mM, resulted in an improvement of separation pattern between analytes. At the same time, using 80 mM SDS leads to a significant increase in the current and the migration time. While a co-elution of the first eluting peaks was found at 10 mM SDS with a significant broadening peak. Therefore, 50 mM SDS was found to be the best concentration, providing moderate generated currents and reasonable analysis times (Fig. S3).

4.7 Applied voltage

The applied voltage affects the pseudo effective electrophoretic mobility, the electroosmotic mobility, the efficiency and hence the resolution of the analytes [58]. Under the optimum BGE conditions selected above, the influence of voltage (20–30 kV) on the migration time was studied. The EOF is increased by increasing the applied voltage, resulting in shorter analysis times and higher efficiency. Voltage of 25 kV was chosen as the optimal, as it maintains the separation within a reasonable time (Fig. S4).

4.8 Injection volume

The optimum injection volume was studied in order to gain highest sensitivity and improve the signal to-noise ratio [59]. For selection of the optimum injection parameters, samples were injected hydrodynamically at 50 mbar while injection times ranged from 5 to 20 s. After 10 s, the peak shapes were deformed and the resolution between peaks also decreased, thus 50 mbar for 10 s were the optimum injection parameters and corresponded to the highest sensitivity without compromising separation efficiency (Fig. S5).

4.9 Capillary cartridge temperature

Only a narrow range of temperatures can be examined [60]. Temperature effects were studied at 20, 25, and 30 °C. 25 °C was chosen because it produced the best resolution, peak shape, reasonable migration time and the generated current was not so high, about 35 μA. Increasing the temperature causing overlapped analytes peaks in addition to slight deformation.

4.10 Sample solvent

Different sample matrices were used with the same BGE composition (50 mM SDS and 20 mM disodium tetraborate buffer, pH 9.0). The matrices: 10 mM phosphoric acid (pH 2.15), borate buffer and water were used as the three sample matrices. The most favorable results were obtained upon dilution of stock solutions with water (Fig. S6).

4.11 Selection of the Internal Standard (I.S.)

Internal standard can enhance the method's precision significantly [61]. Atorvastatin, ezetimibe, saxagliptin and cromolyn sodium were tested. Cromolyn sodium was chosen as the I.S. because it provided higher resolution and better peak shape. To reduce major integration errors, it is preferred to employ a high concentration of the I.S. with a minimum signal-to-noise ratio of 30 [60]. The concentration of cromolyn sodium was kept constant at 1.0 μg mL−1 throughout the developed method.

5 Method validation

The designed MEKC method was validated in accordance with ICH guidelines [62].

5.1 Linearity and range

Each concentration was analyzed for three times and average peak area were calculated. Plotting the corrected peak area ratio (peak area of the drug/peak area of cromolyn sodium) alongside the drug concentration in μg mL−1 yielded a linear relationship. Table 1 summarizes the results of the statistical analysis [63] of the data and shows the linear range for each analyte. The correlation coefficients (r) are high enough (0.9998–0.9999) indicating the linearity of the calibration curves. Limits of quantitation and limits of detection (LOQ and LOD) were calculated mathematically according to ICH guidelines [62] and abridged in Table 1. The minimum concentration at which each drug could be detected was established practically by calculating signal to noise ratio 3:1. Similarly, the quantitation limit was obtained for each of the three drugs by calculating signal to noise ratio 10:1. The mathematical equations are represented as follows: LOD = 3.3 Sa/b, LOQ = 10 Sa/b, Where, Sa is the standard deviation of the intercept of a regression line and b is the slope.

Table 1.

Analytical performance data for the determination of the studied drugs by the proposed MEKC method

Parameter TNZ (at 315nm) ORZ (at 315nm) MNZ (at 315nm) CIP (at 315nm) DXM (at 230nm) SPI (at 230 nm)
Linearity range (µg mL−1) 0.5–5.0 0.5–5.0 1.0–10.0 1.0–10.0 1.0–10.0 1.0–10.0
Intercept ± S.D. (Sa) 0.33 ± 0.02 −0.04 ± 0.03 −6.7x10−3 ± 0.04 3x10−4 ± 0.018 0.0174 ± 0.02 0.0153 ± 2.9x10−2
Slope ± S.D. (Sb) 8.3 ± 0.74 × 10−3 0.0102 ± 0.96 × 10−2 7.6 ± 0.69 × 10−3 4.5 ± 0.29 × 10−3 5.9 ± 3.3 × 10−4 8.2 ± 0.44 × 10−3
Correlation coefficient (r) 0.9998 0.9998 0.9998 0.9999 0.9999 0.9999
S.D. of residuals (Sy/x) 0.03 0.038 0.05 2.29 × 10−2 0.026 0.03
LOD (µg mL−1) 0.09 0.095 0.20 0.13 0.12 0.12
LOQ (µg mL−1) 0.27 0.3 0.60 0.40 0.37 0.35

5.2 Accuracy

The data of analysis of the six cited drugs in their raw material were compared to those obtained by the previously reported methods [37, 64–67] to demonstrate the proposed method's accuracy. The data represented non-significant difference using Students' t-test and the variance ratio F-test [63] as shown in Tables 2–4.

Table 2.

Determination of TNZ and MNZ in raw material by the proposed MECK and reported methods, [66, 67]

Ranges Proposed method Reported method, [66, 67]
Taken (µg mL−1) Found* (µg mL−1) % Recovery Taken (µg mL−1) Found (µg mL−1) % Recovery
TNZ 0.5 0.49 97.48 2.0 1.95 97.56
1.0 1.02 101.85 5.0 5.08 101.52
2.0 2.02 101.01 10.0 9.97 99.67
3.0 2.99 99.52
4.0 3.95 98.78
5 5.04 100.75
Mean % ± S.D. 99.90 ± 1.62 99.58 ± 1.98
t-test 0.26 (2.36)
F-test 1.51 (5.79)
MNZ 1.0 1.02 101.84 10.0 9.65 96.45
3.0 2.92 97.32 100.0 100.07 100.67
5.0 5.06 101.13 200.0 199.68 99.84
7.0 7.07 101.06
8.0 7.95 99.34
10.0 9.99 99.85
Mean % ± S.D. 99.89 ± 0.92 98.99 ± 2.24
t-test 0.85 (2.36)
F-test 1.87 (5.79)

*Average of three separate determinations.

The values between parentheses are the tabulated t and F values at P = 0.05 [63].

Table 3.

Determination of ORZ and CIP in raw material by the proposed MEKC and reported methods [64, 65]

Ranges Proposed method Reported method [64, 65]
Taken (µg mL−1) Found (µg mL−1) % Recovery Taken (µg mL−1) Found (µg mL−1) % Recovery
ORZ 0.5 0.51 102.02 3.0 2.95 98.50
1.0 1.02 101.86 10.0 10.07 100.66
2.0 1.99 99.71 18.0 17.94 99.69
3.0 2.97 99.0
4.0 3.96 98.98
5.0 5.05 100.98
Mean % ± S.D. 100.43 ± 1.38 99.62 ± 1.08
t-test 0.88 (2.36)
F-test 1.63 (19.296)
CIP 1.0 0.99 99.98 10.0 9.81 98.08
2.0 1.97 98.46 30.0 30.38 101.28
4.0 3.99 99.59 50.0 49.81 99.62
6.0 6.09 101.44
8.0 8.00 100.04
10.0 9.96 99.60
Mean % ± S.D. 99.85 ± 0.96 99.66 ± 1.60
t-test 0.23 (2.36)
F-test 2.76 (5.79)
Table 4.

Determination of DXM and SPI at 230 nm in raw material by the proposed MEKC and reported methods [37, 64]

Ranges Proposed method Reported method [37, 64]
Taken (µg mL−1) Found (µg mL−1) % Recovery Taken (µg mL−1) Found (µg mL−1) % Recovery
DXM 1.0 0.98 98.05 5.0 5.14 102.85
3.0 2.99 99.97 10.0 9.79 97.88
5.0 5.02 100.35 20.0 20.07 100.37
7.0 7.05 100.75
9.0 8.94 99.30
10.0 10.01 100.13
Mean %± S.D. 99.76 ± 0.96 100.37 ± 2.49
t-test 0.55 (2.36)
F-test 6.65 (5.79)
SPI 1.0 0.99 99.18 1.0 1.03 102.73
3.0 2.96 98.80 10.0 9.95 99.49
5.0 5.03 100.67 20.0 20.03 100.13
7.0 7.05 100.78
9.0 8.99 99.89
10 9.96 99.64
Mean %± S.D. 99.83 ± 0.79 100.78 ± 1.72
t-test 1.19 (2.36)
F-test 4.7 (5.79)

The values between parentheses are the tabulated t and F values at P = 0.05 [63].

5.3 Precision

Interday and Intraday precision of the proposed MEKC method were assessed and %RSD ranged from 0.37 to 2.07% (Table S1). These data were obtained by analysis of three concentrations within the linear range in the same day and in three successive days respectively.

5.4 Selectivity

The method's specificity was determined by examining any interference from the formulation's ingredients mentioned in the information leaflet. There was no interference from any additives, indicating that the suggested method is sufficiently selective.

5.5 Applications

5.5.1 Pharmaceutical formulations

By determining the investigated analytes in different co-formulated and single component formulations, the applicability of the new developed method was approved (Figs 3 and 4). As shown in Tables 5 & S2, the % recoveries for the studied drugs were in the range of 97.63%–102.56%. Moreover, the statistical tests revealed that the results obtained were in good agreement with those obtained using the comparison methods [37, 64–67].

Fig. 3.
Fig. 3.

Electropherograms for the analysis of: (A) Protozole 500 mg tablets; (B) Flagyl ® 500 mg tablets; (C) Ornidaz®500 mg tablets; (D) Ciprobay® 500 mg tablets; (E) Dexamethasone-MUP ampoules; (F) Spirex tablets

Citation: Acta Chromatographica 2022; 10.1556/1326.2022.01057

Fig. 4.
Fig. 4.

Electropherograms for the analysis of: (A) Tinifloxacin tablets; (B) Ciprodiazole tablets; (C) Laboratory prepared Co-formulated tablets Ciplox-OZ; (D) Zonacip otic drops

Citation: Acta Chromatographica 2022; 10.1556/1326.2022.01057

Table 5.

Assay results for the determination of the studied drugs in their co-formulated tablets by the developed MEKC method and the comparison methods [37, 64–67]

Dosage form Proposed method Reference methods [37, 64–67]
Amount taken (µg mL−1) % Founda Amount taken (µg mL−1) % Founda
Tinifloxacin® 500 mg tablets (500 mg CIP + 600 mg TNZ per tablet) CIP TNZ CIP TNZ CIP TNZ CIP TNZ
1.0 1.2 101.13 97.77 10.0 2.0 102.88 101.90
2.0 2.4 99.15 101.68 30.0 5.0 98.08 98.80
4.0 4.8 100.14 99.72 50.0 10.0 100.58 100.25
Mean % 100.14 99.72 100.51 100.32
± S.D. 0.99 1.96 2.40 1.55
t-testb 0.25 (2.776) 0.41 (2.776)
F-testb 5.88 (19) 1.59 (19)
Ciprodiazole® tablets (500 mg CIP + 500 mg MNZ per tablet) CIP MNZ CIP MNZ CIP MNZ CIP MNZ
2.0 2.0 98.97 102.13 10.0 10.0 102.88 97.65
5.0 5.0 100.66 98.63 30.0 100.0 98.08 100.45
10.0 10.0 99.88 100.25 50.0 200.0 100.58 99.89
Mean % 99.84 100.34 100.51 99.33
± S.D. 0.85 1.75 2.40 1.48
t-testb 0.46 (2.776) 0.76 (2.776)
F-testb 8.06 (19) 1.39 (19)
Laboratory prepared tablets

CIP, ORZ [55]
CIP ORZ CIP ORZ CIP ORZ CIP ORZ
1.0 1.0 98.33 101.97 10.0 3.0 102.88 96.18
3.0 3.0 101.11 98.68 30.0 10.0 98.08 102.11
5.0 5.0 99.67 100.39 50.0 18.0 100.58 99.43
Mean % 99.70 100.35 100.51 99.24
± S.D. 1.39 1.65 2.40 2.97
t-testb 0.51 (2.776) 0.56 (2.776)
F-testb 2.98 (19) 3.26 (19)
Zonacip otic drops each 1 mL contains (3.0 mg CIP and 1.0 mg DXM). CIP DXM CIP DXM CIP DXM CIP DXM
3.0 1.0 98.12 99.06 10.0 5.0 102.88 101.90
6.0 3.0 101.86 100.94 30.0 10.0 98.08 98.57
9.0 5.0 99.37 99.69 50.0 20.0 100.58 100.24
Mean % 99.78 99.90 100.51 100.24
± S.D. 1.90 0.96 2.40 1.67
t-testb 0.41 (2.776) 0.31 (2.776)
F-testb 1.59 (19) 3.03 (19)
Spirazole forte® tablets (379mgSPI+250 mg MNZ SPI MEZ SPI MNZ SPI MNZ SPI MNZ
1.52 1.0 98.36 101.04 1.0 10.0 97.44 97.65
4.56 3.0 100.91 99.43 10.0 100.0 100.48 100.45
9.12 6.0 99.82 100.12 20.0 200.0 99.88 99.89
Mean % 99.70 100.20 99.33
± S.D. 1.28 0.81 1.48
t-testb 0.36 (2.776) 1.12 (2.776)
F-testb 1.58 (19) 3.04 (19)

aeach result is average of three readings. bthe figures between parentheses are the tabulated t and F values at P = 0.05.

6 Assessment of the greenness using GAPI tool

The assumptions about green chemistry's major purpose is to make both the environment and the operator safer. In today's world, determining a method's greenness and environmental impact is a vital stage in any established analytical method [68]. Green Analytical Procedure Index (GAPI) was used to investigate the suggested method's eco-friendly properties. To assess the greenness, it is necessary to cover the whole procedure items in 15 parameters [54]. Fifteen colored zones are represented by the GAPI pictogram (Fig. 5). In the proposed method; we don't use any organic solvent in the BGE to improve greenness. The procedure was also carried out without any previous derivatization, heating, or the inclusion of any other chemicals. Further dilutions were made with water. It might be viewed as a sensitive, environmentally friendly alternative to the previously reported methods.

Fig. 5.
Fig. 5.

GAPI assessment tool for the proposed MEKC method

Citation: Acta Chromatographica 2022; 10.1556/1326.2022.01057

7 Comparison with previously published methods

In Table 5, we compared the analysis of different tablet dosage forms using previous and current approaches. The statistical evaluation indicated no significant difference regarding accuracy and precision. However, when comparing the previous published methods [37, 64–67] with the proposed MEKC method, the current approach displays some advantages. Firstly, it is the first method for the concurrent determination of the six drugs. Moreover, the HPLC methods include the usage of organic modifiers as acetonitrile [37, 65, 66] or methanol [67]. Accordingly, the proposed method is a green alternative using lower sample volumes, micellar system and low time of analysis.

8 Conclusion

Simultaneous estimation of the drugs plays a very important role in pharmaceutical world and quality control. Therefore, a green, novel, precise and accurate MEKC method was investigated for the simultaneous estimation of TNZ, ORZ, MNZ, CIP, DXM and SPI in pharmaceutical dosage forms. The proposed approach is considered a green alternative due to the low sample volume, low solvent consumption and reduced analysis time, which accordingly decreases the generated waste. GAPI tool was used to assess the greenness of the method. In comparison to some previously published reports for the analytes, the suggested method is repeatable, fast, cost effective in addition to being direct sample injection without pretreatment.

Conflict of interest

The authors declared they have no conflict of interest.

Supplementary materials

Supplementary data to this article can be found online at https://doi.org/10.1556/1326.2022.01057.

References

  • 1.

    Nelson, J. M. ; Chiller, T. M. ; Powers, J. H. ; Angulo, F. J. Fluoroquinolone-resistant Campylobacter species and the withdrawal of fluoroquinolones from use in poultry: a public health success story. J. Clin. Infect. Dis. 2007, 44(7), 977980. https://doi.org/10.1086/512369.

    • Search Google Scholar
    • Export Citation
  • 2.

    Werk, R. ; Schneider, L. Ciprofloxacin in combination with metronidazole. Infect. J 1988, 16(4), 257260. https://doi.org/10.1007/BF01650774.

    • Search Google Scholar
    • Export Citation
  • 3.

    Boeckh, M. ; Lode, H. ; Deppermann, K. M. ; Grineisen, S. ; Shokry, F. ; Held, R. ; et al. Pharmacokinetics and serum bactericidal activities of quinolones in combination with clindamycin, metronidazole, and ornidazole. J. Antimicro. Agents Chemother. 1990, 34(12), 24072414. https://doi.org/10.1128/aac.34.12.2407.

    • Search Google Scholar
    • Export Citation
  • 4.

    Malhotra, M. ; Sharma, J. B. ; Batra, S. ; Arora, R. ; Sharma, S. Ciprofloxacin-tinidazole combination, fluconazole- azithromicin-secnidazole-kit and doxycycline- metronidazole combination therapy in syndromic management of pelvic inflammatory disease: a prospective randomized controlled trial. Indian J. Med. Sci. 2003, 57(12), 549555, PMID: 14701947.

    • Search Google Scholar
    • Export Citation
  • 5.

    Roland, P. S. ; Anon, J. B. ; Moe, R. D. ; Conroy, P. J. ; Wall, G. M. ; Dupre, S. J. ; et al. Topical ciprofloxacin/dexamethasone is superior to ciprofloxacin alone in pediatric patients with acute otitis media and otorrhea through tympanostomy tubes. J. Laryngoscope. 2003, 113(12), 21162122. https://doi.org/10.1097/00005537-200312000-00011.

    • Search Google Scholar
    • Export Citation
  • 6.

    Poulet, P-P. ; Duffaut, D. ; Barthet, P. ; Brumpt, I. Concentrations and in vivo antibacterial activity of spiramycin and metronidazole in patients with periodontitis treated with high-dose metronidazole and the spiramycin/metronidazole combination. J. Antimicrob. Chemother. 2005, 55(3), 347351. https://doi.org/10.1093/jac/dki013.

    • Search Google Scholar
    • Export Citation
  • 7.

    Chew, W. K. ; Segarra, I. ; Ambu, S. ; Mak, J. W Significant reduction of brain cysts caused by Toxoplasma gondii after treatment with spiramycin coadministered with metronidazole in a mouse model of chronic toxoplasmosis. J. Antimicro. Agents Chemother. 2012, 56(4), 17621768. https://dx.doi.org/10.1128%2FAAC.05183-11.

    • Search Google Scholar
    • Export Citation
  • 8.

    Kawas, G. ; Marouf, M. ; Mansour, O. ; Sakur, A. A. Analytical methods of ciprofloxacin and its combinations review. Res. J. Pharm. Technol. 2018, 11(5), 21392148. https://doi.org/10.5958/0974-360X.2018.00396.7.

    • Search Google Scholar
    • Export Citation
  • 9.

    Zameeruddin, M. S. ; Kalyankar, S. S. ; Jadhav, S. B. ; Kadam, V. S. ; Bharkad, V. B. Review on analytical method validation of nitroimidazoles. World J.Pharm. Pharm. Sci. 2014, 3(3), 557577.

    • Search Google Scholar
    • Export Citation
  • 10.

    Sebastian, M. P. ; Krishnakumar, K. A review of analytical methods for estimation of amoxicillin trihydrate and tinidazole in pharmaceutical formulations. Asian J. Res. Pharm. Sci. Biotech. 2017, 5(1), 15.

    • Search Google Scholar
    • Export Citation
  • 11.

    Nguyen, T. D. ; Le, H. B. ; Dong, T. O. ; Pham, T. D. Determination of fluoroquinolones in pharmaceutical formulations by extractive spectrophotometric methods using ion-pair complex formation with bromothymol blue. J. Anal. Methods Chem. 2018, 2018, 111. https://doi.org/10.1155/2018/8436948.

    • Search Google Scholar
    • Export Citation
  • 12.

    Obaydo, R. H. ; Sakur, A. A. Spectrophotometric strategies for the analysis of binary combinations with minor component based on isoabsorptive point's leveling effect: an application on ciprofloxacin and fluocinolone acetonide in their recently delivered co-formulation. Spectrochim. Acta A. Mol. Biomol. Spectrosc. J. 2019, 219, 186194. https://doi.org/10.1016/j.saa.2019.04.036.

    • Search Google Scholar
    • Export Citation
  • 13.

    Eman, I. ; Khamis, E. F. ; Belal, S. F. ; Moneim, M. M. A Sensitive inexpensive chromatographic determination of an antimicrobial combination in human plasma and its pharmacokinetic application. J. Chromatogr. B. 2018, 1097, 94100. https://doi.org/10.1016/j.jchromb.2018.09.008.

    • Search Google Scholar
    • Export Citation
  • 14.

    da Silva, D. C. ; Oliveira, C. C. Development of micellar HPLC-UV method for determination of pharmaceuticals in water samples. J. Anal. Methods Chem. 2018, 2018(2), 112. https://doi.org/10.1155/2018/9143730.

    • Search Google Scholar
    • Export Citation
  • 15.

    He, T. ; Xu, Z. ; Ren, J. Pressure-assisted electrokinetic injection stacking for seven typical antibiotics in waters to achieve μg/L level analysis by capillary electrophoresis with UV detection. Microchem. J. 2019, 146, 12951300. https://doi.org/10.1016/j.microc.2019.02.057.

    • Search Google Scholar
    • Export Citation
  • 16.

    Díaz-Quiroz, C. A. ; Hernandez-Chavez, J. F. ; Ulloa-Mercado, G. ; Gortáres-Moroyoqui, P. ; Martínez-Macías, R. ; Meza-Escalante, E. ; et al. Simultaneous quantification of antibiotics in wastewater from pig farms by capillary electrophoresis. J. Chromatogr. B. 2018, 1092, 386393. https://doi.org/10.1016/j.jchromb.2018.06.017.

    • Search Google Scholar
    • Export Citation
  • 17.

    Sversut, R. A. ; Vieira, J. C. ; Rosa, A. M. ; Singh, A. K. ; do Amaral, M. S. ; Kassab, N. M. Improved UV spectrophotometric method for precise, efficient and selective determination of dexamethasone in pharmaceutical dosage forms, orbital: electron. J. Chem. 2015, 7(1), 59. http://dx.doi.org/10.17807/orbital.v7i1.630.

    • Search Google Scholar
    • Export Citation
  • 18.

    Saad, M. N. ; Essam, H. M. ; Elzanfaly, E. S. ; Amer, S. M. Economic chromatographic methods for simultaneous quantitation of some fluoroquinolones and corticosteroids present in different binary ophthalmic formulations. J. Liq. Chromatogr. Relat. Technol. 2020, 43(7–8), 271281. https://doi.org/10.1080/10826076.2020.1725041.

    • Search Google Scholar
    • Export Citation
  • 19.

    Ibrahim, F. A. ; Elmansi, H. ; Fathy, M. E. Green RP-HPLC method for simultaneous determination of moxifloxacin combinations: investigation of the greenness for the proposed method. Microchem. J. 2019148, 151161. https://doi.org/10.1016/j.microc.2019.04.074.

    • Search Google Scholar
    • Export Citation
  • 20.

    Saad, M. N. ; Essam, H. M. ; Elzanfaly, E. S. ; Amer, S. M. A two-step optimization approach: validated RP-HPLC method for determination of gatifloxacin and dexamethasone in ophthalmic formulation. J. Chromatogr. Sci. 2020, 58(6), 504510. https://doi.org/10.1093/chromsci/bmaa013.

    • Search Google Scholar
    • Export Citation
  • 21.

    Essam, H. M. ; Saad, M. N. ; Elzanfaly, E. S. ; Amer, S. M. Stepwise optimization and sensitivity improvement of green micellar electrokinetic chromatography method to simultaneously determine some fluoroquinolones and glucocorticoids present in various binary ophthalmic formulations. J. Biomed.Chromatogr. 2020, 34(11), e4941. https://doi.org/10.1002/bmc.4941.

    • Search Google Scholar
    • Export Citation
  • 22.

    Abdelwahab, N. S. ; Mohamed, M. A. Three new methods for resolving ternary mixture with overlapping spectra: comparative study. J.Chem. Pharm. Bull. 2017, 65(6), 558565. https://doi.org/10.1248/cpb.c17-00132.

    • Search Google Scholar
    • Export Citation
  • 23.

    Masood, Z. ; Ansari, M. T. ; Adnan, S. ; Saeed, M. ; Farooq, M. ; Ahmad, M. Development and application of spectrophotometric method for quantitative determination of Metronidazole in pure and tablet formulations. Pak. J. Pharm. Res. 2016, 2(1), 2832.

    • Search Google Scholar
    • Export Citation
  • 24.

    Ranganathan, P. ; Mutharani, B. ; Chen, S-M. ; Sireesha, P. Biocompatible chitosan-pectin polyelectrolyte complex for simultaneous electrochemical determination of metronidazole and metribuzin. Carbohydr. Polym. J. 2019, 214, 317327. https://doi.org/10.1016/j.carbpol.2019.03.053.

    • Search Google Scholar
    • Export Citation
  • 25.

    Yang, M. ; Guo, M. ; Feng, Y. ; Lei, Y. ; Cao, Y. ; Zhu, D. ; et al. Sensitive voltammetric detection of metronidazole based on three-dimensional graphene-like carbon architecture/polythionine modified glassy carbon electrode. J. Electrochem. Soc. 2018, 165(11), B530. http://dx.doi.org/10.1149/2.1311811jes.

    • Search Google Scholar
    • Export Citation
  • 26.

    Moussa, B. A. ; El-Kady, E. F. ; Mohamed, M. F. ; Youssef, N. F. Greener thin-layer chromatographic solvents for the determination of pantoprazole sodium sesquihydrate, metronidazole and clarithromycin in pharmaceutical formulations used as triple therapy in Helicobacter infection. J. Planar Chromatogr. 2017, 30(6), 481487. https://doi.org/10.1556/1006.2017.30.6.4.

    • Search Google Scholar
    • Export Citation
  • 27.

    Morcoss, M. ; Abdelwahab, N. S. ; Ali, N. W. ; Elsaady, M. T. Different chromatographic methods for simultaneous determination of diloxanide furoate, metronidazole and its toxic impurity. J. Iran. Chem. Soc. 2016, 13(9), 16431651. https://doi.org/10.1007/s13738-016-0881-3.

    • Search Google Scholar
    • Export Citation
  • 28.

    Dorn, C. ; Kratzer, A. ; Schießer, S. ; Kees, F. ; Wrigge, H. ; Simon, P. Determination of total or free cefazolin and metronidazole in human plasma or interstitial fluid by HPLC-UV for pharmacokinetic studies in man. J. Chromatogr. B 2019, 1118, 5154. https://doi.org/10.1016/j.jchromb.2019.04.025.

    • Search Google Scholar
    • Export Citation
  • 29.

    Maslarska, V. ; Tsvetkova, B. ; Peikova, L. ; Bozhanov, S. HPLC method for simultaneous determination of metronidazole and preservatives in vaginal gel formulation. J. Acta Chromatogr. 2018, 30(2), 127130.

    • Search Google Scholar
    • Export Citation
  • 30.

    Airado-Rodríguez, D. ; Hernández-Mesa, M. ; García-Campaña, A. M. ; Cruces-Blanco, C. Evaluation of the combination of micellar electrokinetic capillary chromatography with sweeping and cation selective exhaustive injection for the determination of 5-nitroimidazoles in egg samples. J. Food Chem. 2016, 1213, 215222. https://doi.org/10.1016/j.foodchem.2016.06.056.

    • Search Google Scholar
    • Export Citation
  • 31.

    Bol’shakov, D. ; Amelin, V. ; Nikeshina, T. Identification and determination of antibacterial substances in drugs by capillary electrophoresis. J. Anal. Chem. 2016, 71(1), 94101. https://doi.org/10.1134/S1061934815110039.

    • Search Google Scholar
    • Export Citation
  • 32.

    Gauncar, F. L. ; Kudchadkar, S. S. Development and validation of UV spectrophotometric method for determination of ofloxacin and ornidazole in combined dosage form using simultaneous equation method. World J. Pharm. Res. 2017, 6(8), 10261039ADSX. https://doi.org/10.20959/wjpr20178-8873.

    • Search Google Scholar
    • Export Citation
  • 33.

    Patel, B. H. P. ; Satish, A. Development and validation of analytical method for the estimation of ornidazole in pharmaceutical formulation. Int. Res. J. .Pharm. 2017, 8(11), 115119.

    • Search Google Scholar
    • Export Citation
  • 34.

    Sharma, B. R. ; Shah, C. N. Analytical method validation and method development for simultaneous estimation for ornidazole and diloxanide furoate in pharmaceutical solid dosage form. Pharma. Sci. Monit. 2017, 8(1), 7588.

    • Search Google Scholar
    • Export Citation
  • 35.

    Gauncar, F. L. ; Kudchadkar, S. S. Development and validation of RP-HPLC method for simultaneous estimation of ofloxacin and ornidazole in presence of ornidazole impurity in combined pharmaceutical dosage form. World J. Pharm. Res. 2017, 6(9Spec.Iss.), 604620.

    • Search Google Scholar
    • Export Citation
  • 36.

    El Demerdash, A. O. ; Razeq, S. A. A. ; Fouad, M. M. ; El Sanabary, H. F. Densitometric and UV-spectrophotometric methods for simultaneous determination of spiramycin adipate in binary mixture with oxytetracycline-HCl or tetracycline-HCl. Int. Res. J. Pure Appl. Chem. 2018, 17(1), 121. https://doi.org/10.9734/IRJPAC/2018/44345.

    • Search Google Scholar
    • Export Citation
  • 37.

    Mahmoudi, A. Efficient and simple HPLC method for spiramycin determination in urine samples and in pharmaceutical tablets. J. Sep. Sci.plus 2018, 1(4), 253260. https://doi.org/10.1002/sscp.201800014.

    • Search Google Scholar
    • Export Citation
  • 38.

    Katsidzira, R. M. ; Wessels, A. ; Aucamp, M. A novel RP-HPLC method for the detection and quantification of clarithromycin or spiramycin in bulk drug samples and dosage forms. Int. J. Pharm. Pharm. Sci. 2016, 8(12), 310313. https://doi.org/10.22159/ijpps.2016v8i12.15058.

    • Search Google Scholar
    • Export Citation
  • 39.

    Gbylik-Sikorska, M. ; Gajda, A. ; Nowacka-Kozak, E. ; Posyniak, A. Simultaneous determination of 45 antibacterial compounds in mushrooms-Agaricus bisporus by ultra-high performance liquid chromatography-tandem mass spectrometry. J. Chromatogr. A. 2019, 1587, 111118. https://doi.org/10.1016/j.chroma.2018.12.013.

    • Search Google Scholar
    • Export Citation
  • 40.

    Susakate, S. ; Poapolathep, S. ; Chokejaroenrat, C. ; Tanhan, P. ; Hajslova, J. ; Giorgi, M. ; et al. Multiclass analysis of antimicrobial drugs in shrimp muscle by ultra high performance liquid chromatography-tandem mass spectrometry. J. Food Drug Anal. 2019, 27(1), 118134. https://doi.org/10.1016/j.jfda.2018.06.003.

    • Search Google Scholar
    • Export Citation
  • 41.

    Kowalski, P. ; Olędzka, I. ; Plenis, A. ; Miękus, N. ; Pieckowski, M. ; Bączek, T. Combination of field amplified sample injection and hydrophobic interaction electrokinetic chromatography (FASI-HIEKC) as a signal amplification method for the determination of selected macrocyclic antibiotics. J. Anal. Chim. Acta 2019, 1046, 192198.

    • Search Google Scholar
    • Export Citation
  • 42.

    Kamal, A. H. ; El-Malla, S. F. Factor space analysis and dual wavelength spectrophotometric methods for simultaneous determination of norfloxacin and tinidazole in tablet. J. Anal. Chem. Lett. 2018, 8(3), 393404. https://doi.org/10.1080/22297928.2018.1470941.

    • Search Google Scholar
    • Export Citation
  • 43.

    Fraihat, S. M. Green methods for the determination of nitrite in water samples based on a novel diazo coupling reaction. J.Green Process. Synth. 2017, 6(2), 245248. https://doi.org/10.1515/gps-2016-0005.

    • Search Google Scholar
    • Export Citation
  • 44.

    Nikodimos, Y. ; Hagos, B. Electrochemical behaviour of tinidazole at 1, 4-benzoquinone modified carbon paste electrode and its direct determination in pharmaceutical tablets and urine by differential pulse voltammetry. J. Anal. Methods Chem. 2017, 2017, 110. https://doi.org/10.1155/2017/8518707.

    • Search Google Scholar
    • Export Citation
  • 45.

    Meshram, D. B. ; Priyanka, M. ; Desai, S. D. ; Tajne, M. R. Simultaneous determination of fluconazole and tinidazole in combined dose tablet using high performance thin layer chromatography. J. Chemica Sinica 2017, 8(1), 133137.

    • Search Google Scholar
    • Export Citation
  • 46.

    Mamatha, M. ; Karnakar, N. ; Sunil, R. ; Middha, A. RP-HPLC method development and validation for simultaneous estimation of fluconazole and tinidazole in tablet dosage form. Eur. J. Biomed. Pharm. Sci. 2018, 5(8), 17.

    • Search Google Scholar
    • Export Citation
  • 47.

    Ibrahim, A. M. ; Hendawy, H. A. M. ; Hassan, W. S. ; El-sayed, H. M. ; Shalaby, A. Response surface and tolerance analysis approach for optimizing HPLC method. Microchem. J. 2019, 146, 220226. https://doi.org/10.1016/j.microc.2019.01.007.

    • Search Google Scholar
    • Export Citation
  • 48.

    Riekkola, M. L. ; Joensson, J. A. ; Smith, R. M. Terminology for analytical capillary electromigration techniques (IUPAC Recommendations 2003). Pure Appl. Chem. 2004, 76(2), 443451. https://doi.org/10.1351/pac200476020443.

    • Search Google Scholar
    • Export Citation
  • 49.

    Terabe, S. Electrokinetic chromatography: an interface between electrophoresis and chromatography. Trends Anal. Chem. 1989, 8, 129134. https://doi.org/10.1016/0165-9936(89)85022-8.

    • Search Google Scholar
    • Export Citation
  • 50.

    El-Awady, M. ; Belal, F. ; Pyell, U. Robust analysis of the hydrophobic basic analytes loratadine and desloratadine in pharmaceutical preparations and biological fluids by sweeping—cyclodextrin-modified micellar electrokinetic chromatography. J. Chromatogr. A. 2013, 1309, 6475. https://doi.org/10.1016/j.chroma.2013.08.020.

    • Search Google Scholar
    • Export Citation
  • 51.

    Belal, F. ; El-Din, M. S. ; Tolba, M. ; El-Awady, M. ; Elmansi, H. Analysis of four antimigraine drugs in two ternary mixtures by sweeping-micellar electrokinetic chromatography with retention factor gradient effect and dynamic pH junction. Microchem. J. 2016, 127, 1121. https://doi.org/10.1016/j.microc.2016.01.009.

    • Search Google Scholar
    • Export Citation
  • 52.

    Belal, F. ; El-Razeq, S. A. ; Fouad, M. ; Zayed, S. ; Fouad, F. Simultaneous determination of five coccidiostats in veterinary powders, feed premixes, and baby food by micellar electrokinetic chromatography: application to chicken tissues and liver. Food Anal. Methods 2018, 11(12), 35313541. https://doi.org/10.1007/s12161-018-1330-y.

    • Search Google Scholar
    • Export Citation
  • 53.

    Armenta, S. ; Garrigues, S. ; de la Guardia, M. Green analytical chemistry. TrAC - Trends Anal. Chem. J. 2008, 27(6), 497511. https://doi.org/10.1016/j.trac.2008.05.003.

    • Search Google Scholar
    • Export Citation
  • 54.

    Płotka-Wasylka, J. J. T. A new tool for the evaluation of the analytical procedure: green Analytical Procedure Index. Talanta 2018, 181, 204209. https://doi.org/10.1016/j.talanta.2018.01.013.

    • Search Google Scholar
    • Export Citation
  • 55.

    CIPLOX-OZ tablets (ciprofloxacin + ornidazole). https://ciplamed.com/content/ciplox-oz-tablets,2019(Accessed July 2019).

  • 56.

    Wishart, D. S. ; Feunang, Y. D. ; Guo, A. C. ; Lo, E. J. ; Marcu, A. ; Grant, J. R. ; et al. DrugBank 5.0: a major update to the DrugBank database for 2018. J.Nucleic Acids Res. 2018, 46(D1), D1074D1082. https://doi.org/10.1093/nar/gkx1037.

    • Search Google Scholar
    • Export Citation
  • 57.

    Goyon, A. ; Francois, Y. N. ; Colas, O. ; Beck, A. ; Veuthey, J. L. ; Guillarme, D. High‐resolution separation of monoclonal antibodies mixtures and their charge variants by an alternative and generic CZE method. Electrophoresis 2018, 39(16), 20832090. https://doi.org/10.1002/elps.201800131.

    • Search Google Scholar
    • Export Citation
  • 58.

    Issaq, H. J. ; Atamna, I. Z. ; Muschik, G. M. ; Janini, G. M. The effect of electric field strength, buffer type and concentration on separation parameters in capillary zone electrophoresis. Chromatographia 1991, 32(3), 155161. https://doi.org/10.1007/BF02325019.

    • Search Google Scholar
    • Export Citation
  • 59.

    Williams, S. J. ; Goodall, D. M. ; Evans, K. P. Analysis of anthraquinone sulphonates: comparison of capillary electrophoresis with high-performance liquid chromatography. J. Chromatogr. A. 1993, 629(2), 379384. https://doi.org/10.1016/0021-9673(93)87052-N.

    • Search Google Scholar
    • Export Citation
  • 60.

    Waetzig, H. ; Degenhardt, M. ; Kunkel, A. Strategies for capillary electrophoresis. Method development and validation for pharmaceutical and biological applications. Electrophoresis 1998, 19, 26952752. https://doi.org/10.1002/elps.1150191603.

    • Search Google Scholar
    • Export Citation
  • 61.

    Altria, K. ; Fabre, H. Approaches to optimisation of precision in capillary electrophoresis. J. Chromatographia 1995, 40(5–6), 313320.

    • Search Google Scholar
    • Export Citation
  • 62.

    ICH Technical Requirements for the Registration of Pharmaceutical for Human Use, Validation of Analytical Procedures: Text and Methodology Q2(R1), IFPMA, Geneva, Switzerland, November, 2005; pp 113.

    • Search Google Scholar
    • Export Citation
  • 63.

    Miller, J. N. ; Miller, J. C. Statistics and Chemometrics for Analytical Chemistry, 5th ed.; Pearson Education Limited: Harlow, England, 2005.

    • Search Google Scholar
    • Export Citation
  • 64.

    The United States Pharmacopoeia 34, the National Formulary 29, the US Pharmacopoeial Convention: Rockville, MD, USA, 2011.

  • 65.

    Krishnaiah, Y. S. R. ; Bhaskar, Y. M. P. ; Shyale, S. Development and validation of a reversed-phase HPLC method for the analysis of ornidazole in pharmaceutical dosage forms. Asian J. Chem. 2003, 15(2), 925929.

    • Search Google Scholar
    • Export Citation
  • 66.

    Rani, N. U. ; Rao, J. V. L. N. S. Etimation of tinidazole in tablets by RP–HPLC method. Int. J. Chem. Sci. 2010, 8(4), 23252330.

  • 67.

    Oltean, E. G. ; Nica, A. Development and validation of A RP-HPLC method for the quantization studies of metronidazole in tablets and powders dosage forms. Vet.Drug 2011, 5, 7173.

    • Search Google Scholar
    • Export Citation
  • 68.

    Płotka-Wasylka, J. ; Namieśnik, J. Green Analytical Chemistry: Past, Present and Perspectives, Green Chemistry and Sustainable Technology; Springer: Berlin, 2019.

    • Search Google Scholar
    • Export Citation

Supplementary Materials

  • 1.

    Nelson, J. M. ; Chiller, T. M. ; Powers, J. H. ; Angulo, F. J. Fluoroquinolone-resistant Campylobacter species and the withdrawal of fluoroquinolones from use in poultry: a public health success story. J. Clin. Infect. Dis. 2007, 44(7), 977980. https://doi.org/10.1086/512369.

    • Search Google Scholar
    • Export Citation
  • 2.

    Werk, R. ; Schneider, L. Ciprofloxacin in combination with metronidazole. Infect. J 1988, 16(4), 257260. https://doi.org/10.1007/BF01650774.

    • Search Google Scholar
    • Export Citation
  • 3.

    Boeckh, M. ; Lode, H. ; Deppermann, K. M. ; Grineisen, S. ; Shokry, F. ; Held, R. ; et al. Pharmacokinetics and serum bactericidal activities of quinolones in combination with clindamycin, metronidazole, and ornidazole. J. Antimicro. Agents Chemother. 1990, 34(12), 24072414. https://doi.org/10.1128/aac.34.12.2407.

    • Search Google Scholar
    • Export Citation
  • 4.

    Malhotra, M. ; Sharma, J. B. ; Batra, S. ; Arora, R. ; Sharma, S. Ciprofloxacin-tinidazole combination, fluconazole- azithromicin-secnidazole-kit and doxycycline- metronidazole combination therapy in syndromic management of pelvic inflammatory disease: a prospective randomized controlled trial. Indian J. Med. Sci. 2003, 57(12), 549555, PMID: 14701947.

    • Search Google Scholar
    • Export Citation
  • 5.

    Roland, P. S. ; Anon, J. B. ; Moe, R. D. ; Conroy, P. J. ; Wall, G. M. ; Dupre, S. J. ; et al. Topical ciprofloxacin/dexamethasone is superior to ciprofloxacin alone in pediatric patients with acute otitis media and otorrhea through tympanostomy tubes. J. Laryngoscope. 2003, 113(12), 21162122. https://doi.org/10.1097/00005537-200312000-00011.

    • Search Google Scholar
    • Export Citation
  • 6.

    Poulet, P-P. ; Duffaut, D. ; Barthet, P. ; Brumpt, I. Concentrations and in vivo antibacterial activity of spiramycin and metronidazole in patients with periodontitis treated with high-dose metronidazole and the spiramycin/metronidazole combination. J. Antimicrob. Chemother. 2005, 55(3), 347351. https://doi.org/10.1093/jac/dki013.

    • Search Google Scholar
    • Export Citation
  • 7.

    Chew, W. K. ; Segarra, I. ; Ambu, S. ; Mak, J. W Significant reduction of brain cysts caused by Toxoplasma gondii after treatment with spiramycin coadministered with metronidazole in a mouse model of chronic toxoplasmosis. J. Antimicro. Agents Chemother. 2012, 56(4), 17621768. https://dx.doi.org/10.1128%2FAAC.05183-11.

    • Search Google Scholar
    • Export Citation
  • 8.

    Kawas, G. ; Marouf, M. ; Mansour, O. ; Sakur, A. A. Analytical methods of ciprofloxacin and its combinations review. Res. J. Pharm. Technol. 2018, 11(5), 21392148. https://doi.org/10.5958/0974-360X.2018.00396.7.

    • Search Google Scholar
    • Export Citation
  • 9.

    Zameeruddin, M. S. ; Kalyankar, S. S. ; Jadhav, S. B. ; Kadam, V. S. ; Bharkad, V. B. Review on analytical method validation of nitroimidazoles. World J.Pharm. Pharm. Sci. 2014, 3(3), 557577.

    • Search Google Scholar
    • Export Citation
  • 10.

    Sebastian, M. P. ; Krishnakumar, K. A review of analytical methods for estimation of amoxicillin trihydrate and tinidazole in pharmaceutical formulations. Asian J. Res. Pharm. Sci. Biotech. 2017, 5(1), 15.

    • Search Google Scholar
    • Export Citation
  • 11.

    Nguyen, T. D. ; Le, H. B. ; Dong, T. O. ; Pham, T. D. Determination of fluoroquinolones in pharmaceutical formulations by extractive spectrophotometric methods using ion-pair complex formation with bromothymol blue. J. Anal. Methods Chem. 2018, 2018, 111. https://doi.org/10.1155/2018/8436948.

    • Search Google Scholar
    • Export Citation
  • 12.

    Obaydo, R. H. ; Sakur, A. A. Spectrophotometric strategies for the analysis of binary combinations with minor component based on isoabsorptive point's leveling effect: an application on ciprofloxacin and fluocinolone acetonide in their recently delivered co-formulation. Spectrochim. Acta A. Mol. Biomol. Spectrosc. J. 2019, 219, 186194. https://doi.org/10.1016/j.saa.2019.04.036.

    • Search Google Scholar
    • Export Citation
  • 13.

    Eman, I. ; Khamis, E. F. ; Belal, S. F. ; Moneim, M. M. A Sensitive inexpensive chromatographic determination of an antimicrobial combination in human plasma and its pharmacokinetic application. J. Chromatogr. B. 2018, 1097, 94100. https://doi.org/10.1016/j.jchromb.2018.09.008.

    • Search Google Scholar
    • Export Citation
  • 14.

    da Silva, D. C. ; Oliveira, C. C. Development of micellar HPLC-UV method for determination of pharmaceuticals in water samples. J. Anal. Methods Chem. 2018, 2018(2), 112. https://doi.org/10.1155/2018/9143730.

    • Search Google Scholar
    • Export Citation
  • 15.

    He, T. ; Xu, Z. ; Ren, J. Pressure-assisted electrokinetic injection stacking for seven typical antibiotics in waters to achieve μg/L level analysis by capillary electrophoresis with UV detection. Microchem. J. 2019, 146, 12951300. https://doi.org/10.1016/j.microc.2019.02.057.

    • Search Google Scholar
    • Export Citation
  • 16.

    Díaz-Quiroz, C. A. ; Hernandez-Chavez, J. F. ; Ulloa-Mercado, G. ; Gortáres-Moroyoqui, P. ; Martínez-Macías, R. ; Meza-Escalante, E. ; et al. Simultaneous quantification of antibiotics in wastewater from pig farms by capillary electrophoresis. J. Chromatogr. B. 2018, 1092, 386393. https://doi.org/10.1016/j.jchromb.2018.06.017.

    • Search Google Scholar
    • Export Citation
  • 17.

    Sversut, R. A. ; Vieira, J. C. ; Rosa, A. M. ; Singh, A. K. ; do Amaral, M. S. ; Kassab, N. M. Improved UV spectrophotometric method for precise, efficient and selective determination of dexamethasone in pharmaceutical dosage forms, orbital: electron. J. Chem. 2015, 7(1), 59. http://dx.doi.org/10.17807/orbital.v7i1.630.

    • Search Google Scholar
    • Export Citation
  • 18.

    Saad, M. N. ; Essam, H. M. ; Elzanfaly, E. S. ; Amer, S. M. Economic chromatographic methods for simultaneous quantitation of some fluoroquinolones and corticosteroids present in different binary ophthalmic formulations. J. Liq. Chromatogr. Relat. Technol. 2020, 43(7–8), 271281. https://doi.org/10.1080/10826076.2020.1725041.

    • Search Google Scholar
    • Export Citation
  • 19.

    Ibrahim, F. A. ; Elmansi, H. ; Fathy, M. E. Green RP-HPLC method for simultaneous determination of moxifloxacin combinations: investigation of the greenness for the proposed method. Microchem. J. 2019148, 151161. https://doi.org/10.1016/j.microc.2019.04.074.

    • Search Google Scholar
    • Export Citation
  • 20.

    Saad, M. N. ; Essam, H. M. ; Elzanfaly, E. S. ; Amer, S. M. A two-step optimization approach: validated RP-HPLC method for determination of gatifloxacin and dexamethasone in ophthalmic formulation. J. Chromatogr. Sci. 2020, 58(6), 504510. https://doi.org/10.1093/chromsci/bmaa013.

    • Search Google Scholar
    • Export Citation
  • 21.

    Essam, H. M. ; Saad, M. N. ; Elzanfaly, E. S. ; Amer, S. M. Stepwise optimization and sensitivity improvement of green micellar electrokinetic chromatography method to simultaneously determine some fluoroquinolones and glucocorticoids present in various binary ophthalmic formulations. J. Biomed.Chromatogr. 2020, 34(11), e4941. https://doi.org/10.1002/bmc.4941.

    • Search Google Scholar
    • Export Citation
  • 22.

    Abdelwahab, N. S. ; Mohamed, M. A. Three new methods for resolving ternary mixture with overlapping spectra: comparative study. J.Chem. Pharm. Bull. 2017, 65(6), 558565. https://doi.org/10.1248/cpb.c17-00132.

    • Search Google Scholar
    • Export Citation
  • 23.

    Masood, Z. ; Ansari, M. T. ; Adnan, S. ; Saeed, M. ; Farooq, M. ; Ahmad, M. Development and application of spectrophotometric method for quantitative determination of Metronidazole in pure and tablet formulations. Pak. J. Pharm. Res. 2016, 2(1), 2832.

    • Search Google Scholar
    • Export Citation
  • 24.

    Ranganathan, P. ; Mutharani, B. ; Chen, S-M. ; Sireesha, P. Biocompatible chitosan-pectin polyelectrolyte complex for simultaneous electrochemical determination of metronidazole and metribuzin. Carbohydr. Polym. J. 2019, 214, 317327. https://doi.org/10.1016/j.carbpol.2019.03.053.

    • Search Google Scholar
    • Export Citation
  • 25.

    Yang, M. ; Guo, M. ; Feng, Y. ; Lei, Y. ; Cao, Y. ; Zhu, D. ; et al. Sensitive voltammetric detection of metronidazole based on three-dimensional graphene-like carbon architecture/polythionine modified glassy carbon electrode. J. Electrochem. Soc. 2018, 165(11), B530. http://dx.doi.org/10.1149/2.1311811jes.

    • Search Google Scholar
    • Export Citation
  • 26.

    Moussa, B. A. ; El-Kady, E. F. ; Mohamed, M. F. ; Youssef, N. F. Greener thin-layer chromatographic solvents for the determination of pantoprazole sodium sesquihydrate, metronidazole and clarithromycin in pharmaceutical formulations used as triple therapy in Helicobacter infection. J. Planar Chromatogr. 2017, 30(6), 481487. https://doi.org/10.1556/1006.2017.30.6.4.

    • Search Google Scholar
    • Export Citation
  • 27.

    Morcoss, M. ; Abdelwahab, N. S. ; Ali, N. W. ; Elsaady, M. T. Different chromatographic methods for simultaneous determination of diloxanide furoate, metronidazole and its toxic impurity. J. Iran. Chem. Soc. 2016, 13(9), 16431651. https://doi.org/10.1007/s13738-016-0881-3.

    • Search Google Scholar
    • Export Citation
  • 28.

    Dorn, C. ; Kratzer, A. ; Schießer, S. ; Kees, F. ; Wrigge, H. ; Simon, P. Determination of total or free cefazolin and metronidazole in human plasma or interstitial fluid by HPLC-UV for pharmacokinetic studies in man. J. Chromatogr. B 2019, 1118, 5154. https://doi.org/10.1016/j.jchromb.2019.04.025.

    • Search Google Scholar
    • Export Citation
  • 29.

    Maslarska, V. ; Tsvetkova, B. ; Peikova, L. ; Bozhanov, S. HPLC method for simultaneous determination of metronidazole and preservatives in vaginal gel formulation. J. Acta Chromatogr. 2018, 30(2), 127130.

    • Search Google Scholar
    • Export Citation
  • 30.

    Airado-Rodríguez, D. ; Hernández-Mesa, M. ; García-Campaña, A. M. ; Cruces-Blanco, C. Evaluation of the combination of micellar electrokinetic capillary chromatography with sweeping and cation selective exhaustive injection for the determination of 5-nitroimidazoles in egg samples. J. Food Chem. 2016, 1213, 215222. https://doi.org/10.1016/j.foodchem.2016.06.056.

    • Search Google Scholar
    • Export Citation
  • 31.

    Bol’shakov, D. ; Amelin, V. ; Nikeshina, T. Identification and determination of antibacterial substances in drugs by capillary electrophoresis. J. Anal. Chem. 2016, 71(1), 94101. https://doi.org/10.1134/S1061934815110039.

    • Search Google Scholar
    • Export Citation
  • 32.

    Gauncar, F. L. ; Kudchadkar, S. S. Development and validation of UV spectrophotometric method for determination of ofloxacin and ornidazole in combined dosage form using simultaneous equation method. World J. Pharm. Res. 2017, 6(8), 10261039ADSX. https://doi.org/10.20959/wjpr20178-8873.

    • Search Google Scholar
    • Export Citation
  • 33.

    Patel, B. H. P. ; Satish, A. Development and validation of analytical method for the estimation of ornidazole in pharmaceutical formulation. Int. Res. J. .Pharm. 2017, 8(11), 115119.

    • Search Google Scholar
    • Export Citation
  • 34.

    Sharma, B. R. ; Shah, C. N. Analytical method validation and method development for simultaneous estimation for ornidazole and diloxanide furoate in pharmaceutical solid dosage form. Pharma. Sci. Monit. 2017, 8(1), 7588.

    • Search Google Scholar
    • Export Citation
  • 35.

    Gauncar, F. L. ; Kudchadkar, S. S. Development and validation of RP-HPLC method for simultaneous estimation of ofloxacin and ornidazole in presence of ornidazole impurity in combined pharmaceutical dosage form. World J. Pharm. Res. 2017, 6(9Spec.Iss.), 604620.

    • Search Google Scholar
    • Export Citation
  • 36.

    El Demerdash, A. O. ; Razeq, S. A. A. ; Fouad, M. M. ; El Sanabary, H. F. Densitometric and UV-spectrophotometric methods for simultaneous determination of spiramycin adipate in binary mixture with oxytetracycline-HCl or tetracycline-HCl. Int. Res. J. Pure Appl. Chem. 2018, 17(1), 121. https://doi.org/10.9734/IRJPAC/2018/44345.

    • Search Google Scholar
    • Export Citation
  • 37.

    Mahmoudi, A. Efficient and simple HPLC method for spiramycin determination in urine samples and in pharmaceutical tablets. J. Sep. Sci.plus 2018, 1(4), 253260. https://doi.org/10.1002/sscp.201800014.

    • Search Google Scholar
    • Export Citation
  • 38.

    Katsidzira, R. M. ; Wessels, A. ; Aucamp, M. A novel RP-HPLC method for the detection and quantification of clarithromycin or spiramycin in bulk drug samples and dosage forms. Int. J. Pharm. Pharm. Sci. 2016, 8(12), 310313. https://doi.org/10.22159/ijpps.2016v8i12.15058.

    • Search Google Scholar
    • Export Citation
  • 39.

    Gbylik-Sikorska, M. ; Gajda, A. ; Nowacka-Kozak, E. ; Posyniak, A. Simultaneous determination of 45 antibacterial compounds in mushrooms-Agaricus bisporus by ultra-high performance liquid chromatography-tandem mass spectrometry. J. Chromatogr. A. 2019, 1587, 111118. https://doi.org/10.1016/j.chroma.2018.12.013.

    • Search Google Scholar
    • Export Citation
  • 40.

    Susakate, S. ; Poapolathep, S. ; Chokejaroenrat, C. ; Tanhan, P. ; Hajslova, J. ; Giorgi, M. ; et al. Multiclass analysis of antimicrobial drugs in shrimp muscle by ultra high performance liquid chromatography-tandem mass spectrometry. J. Food Drug Anal. 2019, 27(1), 118134. https://doi.org/10.1016/j.jfda.2018.06.003.

    • Search Google Scholar
    • Export Citation
  • 41.

    Kowalski, P. ; Olędzka, I. ; Plenis, A. ; Miękus, N. ; Pieckowski, M. ; Bączek, T. Combination of field amplified sample injection and hydrophobic interaction electrokinetic chromatography (FASI-HIEKC) as a signal amplification method for the determination of selected macrocyclic antibiotics. J. Anal. Chim. Acta 2019, 1046, 192198.

    • Search Google Scholar
    • Export Citation
  • 42.

    Kamal, A. H. ; El-Malla, S. F. Factor space analysis and dual wavelength spectrophotometric methods for simultaneous determination of norfloxacin and tinidazole in tablet. J. Anal. Chem. Lett. 2018, 8(3), 393404. https://doi.org/10.1080/22297928.2018.1470941.

    • Search Google Scholar
    • Export Citation
  • 43.

    Fraihat, S. M. Green methods for the determination of nitrite in water samples based on a novel diazo coupling reaction. J.Green Process. Synth. 2017, 6(2), 245248. https://doi.org/10.1515/gps-2016-0005.

    • Search Google Scholar
    • Export Citation
  • 44.

    Nikodimos, Y. ; Hagos, B. Electrochemical behaviour of tinidazole at 1, 4-benzoquinone modified carbon paste electrode and its direct determination in pharmaceutical tablets and urine by differential pulse voltammetry. J. Anal. Methods Chem. 2017, 2017, 110. https://doi.org/10.1155/2017/8518707.

    • Search Google Scholar
    • Export Citation
  • 45.

    Meshram, D. B. ; Priyanka, M. ; Desai, S. D. ; Tajne, M. R. Simultaneous determination of fluconazole and tinidazole in combined dose tablet using high performance thin layer chromatography. J. Chemica Sinica 2017, 8(1), 133137.

    • Search Google Scholar
    • Export Citation
  • 46.

    Mamatha, M. ; Karnakar, N. ; Sunil, R. ; Middha, A. RP-HPLC method development and validation for simultaneous estimation of fluconazole and tinidazole in tablet dosage form. Eur. J. Biomed. Pharm. Sci. 2018, 5(8), 17.

    • Search Google Scholar
    • Export Citation
  • 47.

    Ibrahim, A. M. ; Hendawy, H. A. M. ; Hassan, W. S. ; El-sayed, H. M. ; Shalaby, A. Response surface and tolerance analysis approach for optimizing HPLC method. Microchem. J. 2019, 146, 220226. https://doi.org/10.1016/j.microc.2019.01.007.

    • Search Google Scholar
    • Export Citation
  • 48.

    Riekkola, M. L. ; Joensson, J. A. ; Smith, R. M. Terminology for analytical capillary electromigration techniques (IUPAC Recommendations 2003). Pure Appl. Chem. 2004, 76(2), 443451. https://doi.org/10.1351/pac200476020443.

    • Search Google Scholar
    • Export Citation
  • 49.

    Terabe, S. Electrokinetic chromatography: an interface between electrophoresis and chromatography. Trends Anal. Chem. 1989, 8, 129134. https://doi.org/10.1016/0165-9936(89)85022-8.

    • Search Google Scholar
    • Export Citation
  • 50.

    El-Awady, M. ; Belal, F. ; Pyell, U. Robust analysis of the hydrophobic basic analytes loratadine and desloratadine in pharmaceutical preparations and biological fluids by sweeping—cyclodextrin-modified micellar electrokinetic chromatography. J. Chromatogr. A. 2013, 1309, 6475. https://doi.org/10.1016/j.chroma.2013.08.020.

    • Search Google Scholar
    • Export Citation
  • 51.

    Belal, F. ; El-Din, M. S. ; Tolba, M. ; El-Awady, M. ; Elmansi, H. Analysis of four antimigraine drugs in two ternary mixtures by sweeping-micellar electrokinetic chromatography with retention factor gradient effect and dynamic pH junction. Microchem. J. 2016, 127, 1121. https://doi.org/10.1016/j.microc.2016.01.009.

    • Search Google Scholar
    • Export Citation
  • 52.

    Belal, F. ; El-Razeq, S. A. ; Fouad, M. ; Zayed, S. ; Fouad, F. Simultaneous determination of five coccidiostats in veterinary powders, feed premixes, and baby food by micellar electrokinetic chromatography: application to chicken tissues and liver. Food Anal. Methods 2018, 11(12), 35313541. https://doi.org/10.1007/s12161-018-1330-y.

    • Search Google Scholar
    • Export Citation
  • 53.

    Armenta, S. ; Garrigues, S. ; de la Guardia, M. Green analytical chemistry. TrAC - Trends Anal. Chem. J. 2008, 27(6), 497511. https://doi.org/10.1016/j.trac.2008.05.003.

    • Search Google Scholar
    • Export Citation
  • 54.

    Płotka-Wasylka, J. J. T. A new tool for the evaluation of the analytical procedure: green Analytical Procedure Index. Talanta 2018, 181, 204209. https://doi.org/10.1016/j.talanta.2018.01.013.

    • Search Google Scholar
    • Export Citation
  • 55.

    CIPLOX-OZ tablets (ciprofloxacin + ornidazole). https://ciplamed.com/content/ciplox-oz-tablets,2019(Accessed July 2019).

  • 56.

    Wishart, D. S. ; Feunang, Y. D. ; Guo, A. C. ; Lo, E. J. ; Marcu, A. ; Grant, J. R. ; et al. DrugBank 5.0: a major update to the DrugBank database for 2018. J.Nucleic Acids Res. 2018, 46(D1), D1074D1082. https://doi.org/10.1093/nar/gkx1037.

    • Search Google Scholar
    • Export Citation
  • 57.

    Goyon, A. ; Francois, Y. N. ; Colas, O. ; Beck, A. ; Veuthey, J. L. ; Guillarme, D. High‐resolution separation of monoclonal antibodies mixtures and their charge variants by an alternative and generic CZE method. Electrophoresis 2018, 39(16), 20832090. https://doi.org/10.1002/elps.201800131.

    • Search Google Scholar
    • Export Citation
  • 58.

    Issaq, H. J. ; Atamna, I. Z. ; Muschik, G. M. ; Janini, G. M. The effect of electric field strength, buffer type and concentration on separation parameters in capillary zone electrophoresis. Chromatographia 1991, 32(3), 155161. https://doi.org/10.1007/BF02325019.

    • Search Google Scholar
    • Export Citation
  • 59.

    Williams, S. J. ; Goodall, D. M. ; Evans, K. P. Analysis of anthraquinone sulphonates: comparison of capillary electrophoresis with high-performance liquid chromatography. J. Chromatogr. A. 1993, 629(2), 379384. https://doi.org/10.1016/0021-9673(93)87052-N.

    • Search Google Scholar
    • Export Citation
  • 60.

    Waetzig, H. ; Degenhardt, M. ; Kunkel, A. Strategies for capillary electrophoresis. Method development and validation for pharmaceutical and biological applications. Electrophoresis 1998, 19, 26952752. https://doi.org/10.1002/elps.1150191603.

    • Search Google Scholar
    • Export Citation
  • 61.

    Altria, K. ; Fabre, H. Approaches to optimisation of precision in capillary electrophoresis. J. Chromatographia 1995, 40(5–6), 313320.

    • Search Google Scholar
    • Export Citation
  • 62.

    ICH Technical Requirements for the Registration of Pharmaceutical for Human Use, Validation of Analytical Procedures: Text and Methodology Q2(R1), IFPMA, Geneva, Switzerland, November, 2005; pp 113.

    • Search Google Scholar
    • Export Citation
  • 63.

    Miller, J. N. ; Miller, J. C. Statistics and Chemometrics for Analytical Chemistry, 5th ed.; Pearson Education Limited: Harlow, England, 2005.

    • Search Google Scholar
    • Export Citation
  • 64.

    The United States Pharmacopoeia 34, the National Formulary 29, the US Pharmacopoeial Convention: Rockville, MD, USA, 2011.

  • 65.

    Krishnaiah, Y. S. R. ; Bhaskar, Y. M. P. ; Shyale, S. Development and validation of a reversed-phase HPLC method for the analysis of ornidazole in pharmaceutical dosage forms. Asian J. Chem. 2003, 15(2), 925929.

    • Search Google Scholar
    • Export Citation
  • 66.

    Rani, N. U. ; Rao, J. V. L. N. S. Etimation of tinidazole in tablets by RP–HPLC method. Int. J. Chem. Sci. 2010, 8(4), 23252330.

  • 67.

    Oltean, E. G. ; Nica, A. Development and validation of A RP-HPLC method for the quantization studies of metronidazole in tablets and powders dosage forms. Vet.Drug 2011, 5, 7173.

    • Search Google Scholar
    • Export Citation
  • 68.

    Płotka-Wasylka, J. ; Namieśnik, J. Green Analytical Chemistry: Past, Present and Perspectives, Green Chemistry and Sustainable Technology; Springer: Berlin, 2019.

    • Search Google Scholar
    • Export Citation
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Senior editors

Editor(s)-in-Chief: Kowalska, Teresa

Editor(s)-in-Chief: Sajewicz, Mieczyslaw

Editors(s)

  • Danica Agbaba (University of Belgrade, Belgrade, Serbia)
  • Ivana Stanimirova-Daszykowska (University of Silesia, Katowice, Poland)
  • Monika Waksmundzka-Hajnos (Medical University of Lublin, Lublin, Poland)

Editorial Board

  • R. Bhushan (The Indian Institute of Technology, Roorkee, India)
  • J. Bojarski (Jagiellonian University, Kraków, Poland)
  • B. Chankvetadze (State University of Tbilisi, Tbilisi, Georgia)
  • M. Daszykowski (University of Silesia, Katowice, Poland)
  • T.H. Dzido (Medical University of Lublin, Lublin, Poland)
  • A. Felinger (University of Pécs, Pécs, Hungary)
  • K. Glowniak (Medical University of Lublin, Lublin, Poland)
  • B. Glód (Siedlce University of Natural Sciences and Humanities, Siedlce, Poland)
  • A. Gumieniczek (Medical University of Lublin, Lublin, Poland)
  • U. Hubicka (Jagiellonian University, Kraków, Poland)
  • K. Kaczmarski (Rzeszow University of Technology, Rzeszów, Poland)
  • H. Kalász (Semmelweis University, Budapest, Hungary)
  • K. Karljiković Rajić (University of Belgrade, Belgrade, Serbia)
  • I. Klebovich (Semmelweis University, Budapest, Hungary)
  • A. Koch (Private Pharmacy, Hamburg, Germany)
  • Ł. Komsta (Medical University of Lublin, Lublin, Poland)
  • P. Kus (Univerity of Silesia, Katowice, Poland)
  • D. Mangelings (Free University of Brussels, Brussels, Belgium)
  • E. Mincsovics (Corvinus University of Budapest, Budapest, Hungary)
  • Á. M. Móricz (Centre for Agricultural Research, Budapest, Hungary)
  • G. Morlock (Giessen University, Giessen, Germany)
  • A. Petruczynik (Medical University of Lublin, Lublin, Poland)
  • R. Skibiński (Medical University of Lublin, Lublin, Poland)
  • B. Spangenberg (Offenburg University of Applied Sciences, Germany)
  • T. Tuzimski (Medical University of Lublin, Lublin, Poland)
  • Y. Vander Heyden (Free University of Brussels, Brussels, Belgium)
  • A. Voelkel (Poznań University of Technology, Poznań, Poland)
  • B. Walczak (University of Silesia, Katowice, Poland)
  • W. Wasiak (Adam Mickiewicz University, Poznań, Poland)
  • I.G. Zenkevich (St. Petersburg State University, St. Petersburg, Russian Federation)

 

KOWALSKA, TERESA
E-mail: kowalska@us.edu.pl

SAJEWICZ, MIECZYSLAW
E-mail:msajewic@us.edu.pl

Indexing and Abstracting Services:

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2021  
Web of Science  
Total Cites
WoS
652
Journal Impact Factor 2,011
Rank by Impact Factor Chemistry, Analytical 66/87
Impact Factor
without
Journal Self Cites
1,789
5 Year
Impact Factor
1,350
Journal Citation Indicator 0,40
Rank by Journal Citation Indicator Chemistry, Analytical 72/99
Scimago  
Scimago
H-index
29
Scimago
Journal Rank
0,27
Scimago Quartile Score Chemistry (miscellaneous) (Q3)
Scopus  
Scopus
Cite Score
2,8
Scopus
CIte Score Rank
General Chemistry 210/409 (Q3)
Scopus
SNIP
0,586

2020
 
Total Cites
650
WoS
Journal
Impact Factor
1,639
Rank by
Chemistry, Analytical 71/83 (Q4)
Impact Factor
 
Impact Factor
1,412
without
Journal Self Cites
5 Year
1,301
Impact Factor
Journal
0,34
Citation Indicator
 
Rank by Journal
Chemistry, Analytical 75/93 (Q4)
Citation Indicator
 
Citable
45
Items
Total
43
Articles
Total
2
Reviews
Scimago
28
H-index
Scimago
0,316
Journal Rank
Scimago
Chemistry (miscellaneous) Q3
Quartile Score
 
Scopus
393/181=2,2
Scite Score
 
Scopus
General Chemistry 215/398 (Q3)
Scite Score Rank
 
Scopus
0,560
SNIP
 
Days from
58
submission
 
to acceptance
 
Days from
68
acceptance
 
to publication
 
Acceptance
51%
Rate

2019  
Total Cites
WoS
495
Impact Factor 1,418
Impact Factor
without
Journal Self Cites
1,374
5 Year
Impact Factor
0,936
Immediacy
Index
0,460
Citable
Items
50
Total
Articles
50
Total
Reviews
0
Cited
Half-Life
6,2
Citing
Half-Life
8,3
Eigenfactor
Score
0,00048
Article Influence
Score
0,164
% Articles
in
Citable Items
100,00
Normalized
Eigenfactor
0,05895
Average
IF
Percentile
20,349
Scimago
H-index
26
Scimago
Journal Rank
0,255
Scopus
Scite Score
226/167=1,4
Scopus
Scite Score Rank
Chemistry (miscellaneous) 240/398 (Q3)
Scopus
SNIP
0,494
Acceptance
Rate
41%

 

Acta Chromatographica
Publication Model Online only
Gold Open Access
Submission Fee none
Article Processing Charge 400 EUR/article
Regional discounts on country of the funding agency World Bank Lower-middle-income economies: 50%
World Bank Low-income economies: 100%
Further Discounts Editorial Board / Advisory Board members: 50%
Corresponding authors, affiliated to an EISZ member institution subscribing to the journal package of Akadémiai Kiadó: 100%
Subscription Information Gold Open Access
Purchase per Title  

Acta Chromatographica
Language English
Size A4
Year of
Foundation
1992
Volumes
per Year
1
Issues
per Year
4
Founder Institute of Chemistry, University of Silesia
Founder's
Address
PL-40-007 Katowice, Poland, Bankowa 12
Publisher Akadémiai Kiadó
Publisher's
Address
H-1117 Budapest, Hungary 1516 Budapest, PO Box 245.
Responsible
Publisher
Chief Executive Officer, Akadémiai Kiadó
ISSN 2083-5736 (Online)

Monthly Content Usage

Abstract Views Full Text Views PDF Downloads
Apr 2022 0 0 0
May 2022 0 0 0
Jun 2022 0 0 0
Jul 2022 0 0 0
Aug 2022 0 100 47
Sep 2022 0 66 30
Oct 2022 0 0 0