INTRODUCTION:

Despite the fact that corticosteroids have been shown to reduce mortality in severe COVID-19-related illnesses, precise therapy data that can prevent the disease, reduce hospitalisations, and reduce deaths are lacking. Ivermectin has been evaluated as a treatment for and preventative strategy against COVID-19 infection in both observational and randomised studies since the start of the SARS-CoV-2 epidemic.^1,2,3

IVR, Chemical moiety given in Figure 1, is an anthelmintic and semisynthetic drug. IVR chemically (22,23-dihydroavermectin B_1a+ 22,23-dihydroavermectin B_1b) is a member of the Avermectins, a category that comprises natural chemicals generated by fermentation of the soil-dwelling actinomycete Streptomyces avermitilis.⁴

Ivermectin, a macrocyclotic lactone, is a well-known, effective, and safe antiparasitic drug. It is frequently used to treat scabies, ascariasis, onchocerciasis, trichuriasis, strongyloidiasis, and enterobiasis because it works by opening -aminobutyric acid (GABA) channel-I.^4,5 Attention has been drawn to ivermectin's additional mode of action, which involves preventing certain positive single-stranded RNA viruses from replicating in vitro, In particular, recent studies confirm the potential effect of ivermectin in treating Covid-19 infected patients.^6,7,8,9,10

Ivermectin was determined using a variety of analytical techniques, including mass spectrometry or liquid chromatography^11,12, electrospray ionization mass spectrometry¹³, high-performance liquid chromatography^14,15,16,17, and spectrophotometry^18,19,20. However, they were all conducted using unsafe, environmentally harmful, and expensive solvents, compared to our research, which used water as the base solution.

A European Pharmacopoeia monograph was developed to evaluate and identify various compounds in samples of active medicinal components.²¹

Because of the lengthy chromatographic study duration, quality control laboratories do not favor the United States Pharmacopoeia approach ²². And as a result, a sensitive, effective, and reliable analytical approach is still desperately needed for the regular analysis of ivermectin in injectables and other end products.

Here, we created and verified a novel spectrophotometric approach for the ivermectin test, which includes figuring out and recognizing ivermectin in pharmaceutical products. These analytical techniques are robust, linear, accurate, selective, and exact.

Ivermectin dissolves in methanol, ethanol, propylene glycol, and other solutions, but it does not dissolve in water. Therefore, the use of a surfactant Sodium Lauryl Sulfate (SLS) can help in improving its solubility in water, and thus providing the possibility of using an inexpensive, harmless, and readily available solution.

Consequently, quality control laboratories would find these recently developed and verified procedures suitable for analyzing ivermectin in pharmaceutical products.

Figure 1: Chemical structure of Ivermectin

MATERIAL AND METHOD:

Instrument and Apparatus:

All absorbance measurements for the investigations were taken with a double beam UV/Visible spectrophotometer (Rigol 3660) and a pair of 10 mm matched quartz cells. The UV-Probe system software obtained the spectra automatically. All weights were collected using an analytical balance with a precision of 0.0001 g (Precisa), the ultrasonic bath model (JENEK / PS-80 A). Pipettes, burettes, and a beaker were utilized.

Chemicals:

IVR Pure Standards ingredients generously given by Barakat Pharmaceutical Industries with 99.5% purity, as specified by the supplier. All of the chemicals utilized have medicinal or analytical properties. Double distilled water was filtered with a 0.45 μ membrane.

Reagents:

SLS solution (0.5%) was made by dissolving 2.5 gr of SLS powder in a volumetric flask (500 mL) filled with distilled water. The material was sonicated for 15 minutes to get the final concentration of 500 μg mL^-1.

Pharmaceutical Formulations:

Ivermectin tablets, Barakat Pharmaceutical Industries (Piramectin*), Aleppo, Syria, labeled 3 mg of Ivermectin.

Preparation of Sample and Standard Solutions:

Preparation of Standard Stock Solutions: It was dissolved by adding 12.5 mg of ivermectin reference standard and 25 mL of SLS solution (0.5%) to dissolve into a 25 mL volumetric tube by sonication and was obtained final concentration 500 μg ml^-1 of stock standard of Ivermectin.

Preparation of Working Standard Solution: From the standard stock solution, appropriate aliquots were pipetted out into a series of 25 mL volumetric flasks. SLS solution (0.5%) was added to the volume to achieve the desired set of solutions with concentration ranges of 2.5, 5, 10, 15, 20, and 25 μg mL^-1 for IVR. Plotting the calibration curve of absorbance versus concentration was done after the absorbances of the aforementioned solutions were measured at 245 nm.

Ivermectin sample solutions: An average of twenty pills were ingested, annihilated into a powder, and then transferred to a 50 mL volumetric flask. 30 mL of SLS solution (0.5%) was then added and dissolved. SLS solution (0.5%) was used to get the total volume up to 50 mL. In a 10 mL measuring flask, 1 milliliter of this solution was collected and diluted with SLS solution (0.5%) to give 6 μg mL^-1 of ivermectin as the theoretical concentration of the stock solution.

RESULTS AND DISCUSSION:

Analytical Instruments and Conditions:

The use of the SLS solution in determining and measuring ivermectin is a simple, inexpensive, harmless and practical method, as water was used as the basic solution. Thus, the problem of poor solubility of ivermectin in water was solved.

The developed method based on UV Spectrophotometric analysis at Zero-order without any separation step.²³ To get the λmax value in the 200–400 nm region, the standard solution was scanned in a UV spectrophotometer. Measurements were then made blindly against the 0.5% SLS solution. At a wavelength of 245 nm, the absorbance values of the reference solution were measured and recorded, together with the quantity of ivermectin. Ivermectin absorbance readings at this wavelength have been demonstrated to be proportionate to standard mixture concentrations. (Figure 2)

Figure 2: Overlay spectrum of ivermectin standard solutions (2.5-25 μg mL^-1)

Validation:

The ICH recommendations' protocols are followed in the complete validation of the optimized spectrophotometric technique. Q2(R1) to confirm analytical techniques.^{24,25,26,27,28}

Linearity:

To assess the linearity of the UV spectrophotometer, a set of standard solutions with six distinct concentrations were analyzed. The studies were carried out in triplicate. For IVR, it was discovered that the procedure was linear throughout a concentration range of (2.5–25 μg ml^-1) (table 1). As shown in Figure 3, the regression equation for IVR was y = 0.0187x + 0.0225. The values of the correlation coefficients and the standard deviations around the slope and the intercept (Table 2) make it abundantly obvious that the calibration curves are linear. As a result, the new approach is linear (table 2)

Table 1: Linearity data of Ivermectin

S. No.	Conc. of IVR (μg mL^-1)	Abs at λmax of IVR (245nm)
1	2.5	0.084
2	5	0.099
3	10	0.211
4	15	0.299
5	20	0.401
6	25	0.49

Table 2: Regression Characteristics

Parameters	IVR
Slope	0.0187
Intercept	0.022455
Correlation coefficient	0.996

Figure 3: Linearity of Ivermectin

Accuracy:

The mean % recovery of the triple determination for IVR at three concentrations within the linearity range was computed in order to complete the calculation. The findings obtained are summarized in Table 3. The mean percentage recoveries were found to be 99.5±0.89, which were within the acceptable range (98-102%), indicating that the approach is accurate and feasible for detecting IVR.

Table 3: Evaluation of accuracy for the determination of IVR

Theoretical concentration (μg mL^-1)	Actual concentration (μg mL^-1)	Recovery %	Mean Rec ± S.D.
5	4.96	99.2	99.5±0.89
10	9.88	98.8
15	15.08	100.5

Precision:

Repeatability (intra-day precision): was assessed by computing the relative standard deviations (%RSD) for three separate IVR test concentrations that were determined in triplicate within the linearity range on the same day (table 4)

Intermediate (inter-day): The Relative Standard Deviations (%RSD) of three distinct IVR test concentrations within the linearity range on three separate days were computed in order to assess accuracy (table 5).

The relative standard deviations were found to be less than 2% for IVR.²⁹

Table 4: Intra-day precision

Theoretical concentration (μg mL^-1)	Actual concentration (μg mL^-1)	Recovery %	SD	RSD
5	4.9	98	0.05	1.020408
10	9.98	99.8	0.072111	0.722555
15	14.79	98.6	0.255016	1.723472

Table 5: Inter-day precision

Theoretical concentration (μg mL^-1)	Actual concentration (μg mL^-1)	Recovery %	SD	RSD
5	5.01	100.17	0.01	0.199601
10	9.93	99.8	0.051316	0.516604
15	14.86333	98.6	0.066583	0.44797

Detection and quantitation limits:

The LOQ and LOD were used to evaluate the sensitivity of spectrophotometric methods (table 6).

Limit of Detection:

It is the lowest concentration of analyte in a sample that, under the given experimental circumstances, may be detected but may not necessarily be quantified. According to ICH rules, the limit of detection may be computed using the following formula.

LOD = 3.3 ×σ/S

Where, σ is the standard deviation of response of the drug and S is the slope of the corresponding calibration curve.

Limit of Quantification:

It is the lowest analyte concentration in a sample that, under the specified experimental circumstances, may be determined with a reasonable level of precision and accuracy. According to ICH, the limit of quantification may be computed using the following formula.

LOQ = 10× σ /S

Where, σ is the standard deviation of response of the drug and S is the slope of the corresponding calibration curve.³⁰

Table 6: Data of LOD and LOQ

	LOD (μg/ml)	LOQ (μg mL^-1)
IVR	3.9626	12

Assay of IVR in tablet dosage form:

The IVR in tablet dose form was effectively determined using the suggested validated approach, and the quantitative analysis results were shown in Table 7. The sample solution's absorbance was measured at 245 nm in comparison to the 0.5% SLS solution (figure 4). There were three duplicates found. The concentrations of all the tested formulations met the criteria, which states that IVR tablets should contain between 90% and 110% of the quantity listed on the label.

Table 7: The results obtained from the analysis of dosage form

Parameters	IVR
Drug Content %	91.35%
SD	0.134904
RSD %	0.02527232

Figure 4: Spectrum of IVR tablet solution (6 μg mL^-1)

The spectrophotometric approach was found to be linear, precise, and accurate, and the recovery percentage fell between the predetermined ranges (98–102%). The suggested method's analysis of the pharmaceutical formulation also proved to be highly repeatable, reliable, and in good accordance with the drug's label claim. You can use this technique in place of the current ones. The usual additives and excipients did not interfere with the analysis of real samples containing IVR.

Because it is less costly, has fewer steps and processes, and requires less time than the LC chromatographic technique, the UV spectrophotometric method is more favorable than the latter.

CONCLUSION:

The devised approach was proven to be accurate, exact, simple, sensitive, and quick and can typically be utilised for measurement of Ivermectin in their dose form. The devised approach was validated at par with ICH principles and all metrics met the requirements.

CONFLICT OF INTEREST:

The authors declare that they have no conflicts of interest.

ACKNOWLEDGMENTS:

The current study was supported by Al-Watanyia Private University.

DATA AVAILABILITY:

All data generated or analyzed during this study are included in this article, and the datasets used or analyzed during the current study are available from the corresponding author with reasonable request.

FUNDING STATEMENT:

The study described in this article was conducted without any external financial support or funding from any specific organization or grant. The research was self-funded by the authors.

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Received on 27.03.2025 Revised on 12.09.2025

Accepted on 25.11.2025 Published on 13.01.2026

Available online from January 17, 2026

Research J. Pharmacy and Technology. 2026;19(1):278-282.

DOI: 10.52711/0974-360X.2026.00039

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