Physicochemical, Phytochemical and Antioxidant Study of Pyrus Malus

Research Article | DOI: https://doi.org/10.31579/2639-4162/256

Physicochemical, Phytochemical and Antioxidant Study of Pyrus Malus

  • Rakhi Mishra
  • Binit Dwivedi

 DDPRC CRI(H), Under CCRH, Ministry of AYUSH, Government of India, A 1/1, Sector 24, Noida, UP 201301, India.

*Corresponding Author: Rakhi Mishra, DDPRC CRI(H), Under CCRH, Ministry of AYUSH, Government of India, A 1/1, Sector 24, Noida, UP 201301, India.

Citation: Rakhi Mishra, Binit Dwivedi, (2025), Physicochemical, Phytochemical and Antioxidant Study of Pyrus Malus, J. General Medicine and Clinical Practice, 8(5); DOI:10.31579/2639-4162/256

Copyright: © 2025, Rakhi Mishra. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Received: 17 February 2025 | Accepted: 20 May 2025 | Published: 29 May 2025

Keywords: HPTLC; UV; physicochemical; pyrus malus; homoeopathy; antioxidant study

Abstract

Background/Aim: Pyrus malus (apple), commonly utilized in traditional medicine, is gaining interest for its therapeutic potential. However, standardized information on its mother tincture and raw drug material is limited. This study aimed to establish a comprehensive profile of Pyrus malus through physicochemical, phytochemical, and chromatographic analyses to support its pharmaceutical standardization.

Materials and Methods: The mother tincture and raw drug material of Pyrus malus leaves were analyzed using standard pharmacopoeial procedures. Physicochemical parameters such as pH, alcohol content, total solids, loss on drying, water and alcohol soluble extractive values were measured. Preliminary Phytochemical screening was performed to detect the presence of various bioactive compounds. High Performance Thin Layer Chromatography (HPTLC) was conducted using chloroform: methanol (9:1, v/v) as the mobile phase, with spot visualization under UV light at 254nm and 366nm and after derivation with anisaldehyde sulphuric acid reagent.

Results: The mother tincture showed a pH of 4.58, alcohol content of 41% v/v, and total solid of 2.05 v/v. The raw drug material exhibited a 4.90% loss on drying and an 18.60% water-soluble extractive value. Phytochemical screening revealed the presence of tannins, flavonoids, alkaloids, glycosides, and phenolic compounds. HPTLC analysis provided distinct spot patterns, indicating multiple bioactive constituents. The optimized mobile phase demonstrated effective separation, confirming the chemical complexity of the extract.

Conclusions: The study successfully established a physicochemical and phytochemical profile of Pyrus malus mother tincture and raw drug material. Chromatographic profile further confirmed the presence of diverse bioactive compounds. These findings support the standardization of Pyrus malus for pharmaceutical applications and encourage pharmacological properties and expand its medicinal applications.

1. Introduction

Pyrus malus also known as apple belongs to the family of Rosaceae[1] and comprises more than 300 species. It is an important remunerative crop grown across the temperature regions of the world. Pyrus malus is native to South-West Asia[2]. Pyrus malus are rich in vitamins and minerals[3]consuming these fruits aids in weight loss enhances brain health, provides antioxidants, reduces the risk of metabolic syndrome and prevents some forms of cancer[2]. Pyrus malus is reported to contain triterpenoids, flavonoids, organic acids and sterols[4]. Pharmacologically Pyrus malus is reported to have antioxidants, antiobesity, anticholesterol, anticancer, enzyme inhibitor and antimicrobial activity[5]. Pyrus malus is beneficial for general skin health with topical application. Pyrus malus strengthens the immune system and prevents diseases. Pyrus malus leaves and fruits are rich in vitamins and minerals, trace elements, amino acids and flavonoids[5]. The fruits of Pyrus malus contain malic acid, which has tonic, keratolytic and antiseptic properties. The astringent principle of Pyrus malus includes tannins, tannin derivatives and colouring materials(flavones)[6]. Due to its rich vitamin and mineral content, it helps in skin degeneration, which causes premature ageing. Its soothing antiseptic and keratolytic agents help loosen dead skin fragrant. Pyrus malus has low antigen content which is useful in managing immune-mediated diseases[7]. Pyrus malus is believed to be a good source of antioxidants[8]. Previous research reported that Pyrus malus fruit contains epicatechin, procyanidins, gallic acid, protocatechuic, chlorogenic, ferulic and p-coumaric, quercetin, and myricetin[9]. Pyrus malus plant has high levels of antioxidants and anti-inflammatory activities, which help to reduce age-associated diseases such as cardiovascular disease, hypertension, diabetes, CVD associated with obesity, and stroke and improve bone health[10] Homoeopathy, Pyrus malus leaves are used. The major characteristics of a plant are affected by its genetic background and the environment where it is grown.  Pyrus malus, commonly known as the apple tree, has long been a subject of interest in homoeopathy and phytochemistry due to its medicinal properties. While there's limited specific research on the homoeopathic mother tincture of Pyrus malus regarding its phytochemical composition, antioxidant activity, and HPTLC profiling, addressing these factors is crucial for authenticating and enhancing its medicinal potential. Homoeopathic mother tinctures are primarily alcoholic extracts, retaining a variety of secondary metabolites such as flavonoids, phenolics, tannins, and alkaloids. These compounds are often responsible for antioxidant and therapeutic properties. Comprehensive phytochemical screening involves determining the presence of bioactive constituents commonly found in Pyrus malus leaves homoeopathic mother tincture. The antioxidant potential of Pyrus malus homoeopathic mother tinctures can be evaluated using assays like DPPH[11], ABTS[12], or FRAP[13]. These studies help quantify the free radical scavenging capacity of the tincture, which is a significant indicator of its therapeutic efficacy. High-Performance Thin Layer Chromatography (HPTLC)[14] is a standard method for identifying and quantifying specific phytochemicals in herbal tinctures. HPTLC can be employed to fingerprint phenolic acids and flavonoids, ensuring the consistency and authenticity of the homoeopathic preparation. The quality and composition of phytochemicals [15] in Pyrus malus mother tincture depend significantly on climatic conditions, soil type, altitude, and agricultural practices. Plants grown in temperate regions with optimal sunlight and humidity are likely to yield higher levels of bioactive compounds. Documenting and standardising raw material sources is crucial for ensuring batch-to-batch consistency in the tincture. Homoeopathic mother tinctures should be prepared following strict pharmacopoeia guidelines, provided the raw drug is fresh, authentic, and contamination-free. The origin of the raw drug must be authenticated through botanical identification and traceability to reputable sources. Studies focusing on standardization, bioactivity, and clinical efficacy of Pyrus malus leaves homoeopathic mother tincture are needed to establish its scientific basis. Comparative studies using different Pyrus malus leaves mother tinctures might reveal variations in effectiveness and bioactivity, emphasizing the importance of genetic and environmental factors. Standardization parameters were determined by the guidelines given by the World Health Organization (WHO)[15]. Parameters including extractive values, ash values and loss on drying were determined. Preliminary phytochemical test, antioxidant stud and chromatographic profiling were performed for the identification and standardization of Pyrus malus leaves. Pyrus malus leaves are a rich source of antioxidant compounds Antioxidant components are micro constituents that inhibit lipid oxidation by inhibiting the initiation or propagation of oxidizing chain reactions and are involved in scavenging free radicals[16]. In view of that, we designed the study to evaluate the antioxidant potential of Pyrus malus by various assay methods. The present study is helpful for the determination and quantification of antioxidant compounds which are useful for producing Pyrus malus safer drugs for the treatment of various ailments of human beings.

                                                                                                        Figure 1: Image of Pyrus malus leaf

2. Materials and Methods

The leaves of Pyrus malus were collected from the flora of the Nilgiri District, Tamil Nadu by Center of Medicinal Plants Research in Homoeopathy (CMPRH), Emerald, Tamil Nadu, India and was authenticated by the staff of the Center of Medicinal Plants Research in Homoeopathy (CMPRH), Emerald, Ooty. The voucher specimen has been deposited in the herbarium and the laboratory of DDPR Central Research Institute for Homoeopathy, Noida, Uttar Pradesh, India, for future reference. Authentic plant material was used to prepare the alcoholic extract. The solvents ethanol, HPLC water, methanol and chloroform were of analytical grade purity (Merck Ltd., India).

2.1. Physicochemical studies of raw drug standardization

2.1.1. Determination of Loss on Drying

Loss on drying method used for determination of moisture content as per methods recommended in Homoeopathic Pharmacopeia of India [17]. Percentage loss on drying was calculated.

2.1.2. Determination of Foreign matter

For foreign matter determination, 100 g of plant raw material has been taken and outspread in the form of a thin-layer. The sample was examined by a 6x lens or with an unaided eye, and the foreign organic matter was picked manually. The ratio of total foreign matter weighed, and the weight of the drug taken gave the % of foreign matter.

2.1.3. Determination of Total Ash value

In drug, the impurity present in the form of organic matter was determined with the help of total Ash value. For its determination, 2 g of the dried raw drug was weighed in powdered form in a pre-weighed silica crucible[18]. The sample was incinerated in silica crucible by gradually increasing the temperature up to 450°C for 4 hours or until it became carbon free. The crucible was cooled and weighed until constant weight was obtained. Percent of total ash value was then calculated by taking the ratio of loss in weight to weight of sample taken.

2.1.4. Determination of Acid Insoluble Ash value

After total cash value determination, 25 mL of 5 M hydrochloric acid was added to the dried ash and boiled in a water bath for 10 minutes. The solution was concentrated till its colour changed to yellow. Acid insoluble matter was filtered using ashless Whatman paper number 1 followed by washing with distilled water. The paper was again ignited in a crucible at a temperature not more than 450°C for 4 hours[18], after which the crucible was kept in a desiccator, cooled and weighed. With reference to the originally taken air-dried powdered drug, % of acid insoluble ash value was calculated.

2.1.5. Determination of Water-soluble extractive value

The extractive value determines the number of active constituents in a given amount of plant raw drug material when extracted with a solvent. These values indicate the extent of polar and non-polar components present in the plant material[19]. For determination of water extractive value, 2 g of sample was accurately weighed, air dried powdered drug was put in a conical flask with 100 mL water added in it. The solution was allowed to stand for 24 hours with intermittent shaking of the flask after every 4 hours. The water-soluble extractive was filtered using Whatman filter paper. 25 mL of this filtrate was completely dried on a pre-weighed petri plate at 105°C[20]. The increase in weight of the petri dish was noted to calculate the water-soluble extractive value determination. With reference to the originally taken air-dried powdered drug, the % of water-soluble extractive value was calculated.

2.1.6. Determination of Alcohol-soluble extractive value

For determination of alcohol soluble extractive value accurately weighed 2 g air dried powdered drug was put in a conical flask and 100 mL absolute alcohol added to it. The whole solution was left for 24 hours for complete extraction at room temperature. The solution was shaken vigorously for a few minutes after every 6 hours. The extract was filtered with the help of Whatman filter paper taking precautions to avoid evaporation and loss of alcohol from the extract. Weighed the empty flat-bottomed petri dish[20]. The petri dish with 25 mL of filtrate was heated at 105°C[21] in an electric oven then cooled in a desiccator and weighed. With reference to the originally taken air-dried powdered % of alcohol-soluble extractive value calculated and recorded in Table 2.

2.1.7. Preparation of crude extract/Inhouse mother tincture

100 g of coarsely dried powdered Pyrus malus leaves were taken in which 600 ml distilled water and 430 ml strong alcohol (95%) was added to make one thousand millilitres of the mother tincture using the percolation method (as per Homoeopathic Pharmacopoeia of India)[17]. This tincture was transferred to a tightly packed amber glass container and stored for further study. 

  1. 2.1.8. Qualitative Phytochemical screening

Phytochemical tests were performed on crude extract for qualitative estimation of phytochemicals present in in-house mother tincture of Pyrus malus with all respective testing procedures as described in the textbook of Harborne JB (eds.)[22] (Table 3). 

  1. 2.1.9. Standardization of mother tincture

Standardization of mother tincture was conducted to identify the organoleptic and physicochemical properties of mother tincture [23]. Organoleptic properties measurement was done for colour, odour and clarity of solution. The samples were tested for various physicochemical properties like sediments, pH, total solids, Wt/mL and total alcohol content.

  1. 2.2. Preparation of chloroform extract

25 mL of Mother Tincture was taken in a 50 mL beaker. To remove the ethanol, the solution was evaporated in a water bath and extracted three times with 20 mL chloroform. Combined and concentrated chloroform extract up to 2 mL volume. Carried out TLC[24] of chloroform extract of mother tincture on silica gel 60 F254 pre-coated plate [25]. 

2.3. HPTLC fingerprinting profile study

HPTLC fingerprinting study [26] was carried out by following the methodology of High-Performance Thin Layer Chromatography for the analysis of Medicinal plant. A densitometric HPTLC Camag Linomat 5 instrument was used for the study. As a sample applicator Camag Linomat 5 was used for spotting TLC plate. Spots were made on silica gel 60 F254 pre-coated plate (Merck) 10 × 10 cm plate with an aid of sampling machine and solvent front was run up to 70mm height [27]. For development of mobile phase, a saturating chamber Camag Twin Trough glass chamber was used. Camag TLC Scanner and software vision CATS were used for scanning purpose [28]. HPLC grade solvents were used for all the extracts solution. Volume applied for sample 2 µL. For detection of active constituents’ various mobile phase was used. TLC spots were visualized after illumination at 254 nm, 366 nm and after derivatization.

2.4. Study of Antioxidant Potential

2.4.1. Determination of Total Phenolic content

The total phenolic content of the extracts was determined by Folin-Ciocalteu’s reagent procedure reported by Singleton [29]. The total phenol content was estimated in Pyrus malus mother tincture. Ascorbic acid was used as reference standard. Different concentration (0.2661-8.517 mM) of ascorbic acid were prepared and analysed at 736 nm and calibration curve was plotted as absorbance versus concentration. Total Phenolic content was estimated by using Ascorbic acid as standard approximately 50 µl of the mother tincture was mixed with 5 mL 10 % Folin-Ciocalteu’s [30] (phenol reagent) and 4 mL of sodium carbonate. The mixture was allowed to stand for one hour in dark. After one hour the colour changed from yellow to blue. The absorbance of the solutions was measured at λmax 736 nm using a UV-VIS spectrophotometer. The total phenolic content was calculated from the calibration curve and in results total phenolic content of Pyrus malus mother tincture was calculated as the ascorbic acid equivalents using standard ascorbic acid (Y= 0.0753x +0.0256, R2 0.9995) curve standardised in the lab for the calculation of ascorbic acid equivalent. Total Phenolic content was expressed in mM concentration of ascorbic acid equivalent[31]. 

2.4.2. Determination of DPPH Radical Scavenging Assay

The free radical scavenging activity of Pyrus malus sample in-house mother tincture(A) and market sample (B, C and D) were measured by 2,2-diphenyl-1-picryl-ydraxyl (DPPH) radical scavenging assay[32]. The standard solution of DPPH was prepared by dissolving 0.025 g in 25 mL methanol and different concentration of standards/mother tincture sample (100µl) was mixed with 4 mL methanol and 1 mL of DPPH standard. The mixture was allowed to stand for one hour in dark, after which the absorbance was measured at 517 nm using a UV-VIS spectrophotometer. The percentage inhibition was determined by comparing the test result and the control (methanol used as solvent blank). Percentage degradation was calculated by the formula:

Where,

A= absorbance of sample

B= absorbance of control

The inhibiting effects of mother tincture showed varied levels of DPPH radical scavenging activity, expressed as percentage degradation [29].

2.4.3. Determination of Total Flavonoid content

Total flavonoid content was determined by aluminium chloride colorimetric assay method. Total flavonoid content using rutin hydrate as standard. An aliquot of 1 mL of the mother tincture mixed with 4 mL distilled water add 300 µl (5%) sodium nitrite in it. After five minutes add 300 µl (10%) aluminium chloride then after five minutes add 2 mL methanol and 2 mL (1M) sodium hydroxide then add 2.4 mL distilled water to make up the volume up to 10 mL. The mixture was shaken vigorously and left to stand in dark at room temperature. The resulting mixture colour change yellow to pink. The absorbance of the reaction mixture was measured at λ max 510 nm with a spectrophotometer. A standard calibration curve was constructed using rutin hydrate standard solutions of (31.25- 500 µg/mL) of each standard was treated in the same manner as the samples above to generate calibration curve. The total flavonoid content of mother tincture was determined from the curve and the results recalculated and expressed as the rutin hydrate equivalents.

2.4.5. Determination of ABTS Assay

Free radical scavenging activity of Pyrus malus mother tincture was determined by ABTS (2, 2-azino-bis-(3-ethylbenzothiazoline-6-sulfonate) [33] radical cation decolourization assay. ABTS. + cation radical was produced by the reaction between 7mM ABTS in water and 2.45 mM potassium persulfate (1:1) stored in the dark at room temperature for 16 hours [34] before use. ABTS. + solution was then diluted with methanol to obtain an absorbance of 0.700 at 746 nm. After the addition of 10 µl of mother tincture/standard in 2 mL of diluted ABTS. + solution, the absorbance was measured at 5 min after the initial mixing. An appropriate solvent blank (methanol) was run in each assay.

Percent inhibition of absorbance at 746 nm was calculated using the formula:

ABTS ion scavenging effect (%) = [(AB-AA)×100] ÷AB

Where,

AB is absorbance of ABTS radical+ methanol

AA is absorbance of ABTS radical + sample/standard

Trolox was used as standard substance [34]..

3. Results

3.1. Physiochemical and Phytochemical studies

The physicochemical properties of the Pyrus malus mother tincture for parameters like sediments, pH, total solids, alcohol content and weight per mL were analysed and tabulated in Table 1. The results obtained for various physiochemical studies of Pyrus malus raw drug are tabulated in Table 2. Phytochemical tests performed on the crude extract of whole plant of Pyrus malus showed positive results for various tests as mentioned in Table 3. Organoleptic observations of the prepared in-house mother tincture indicated formation of a clear green solution with characteristic tincture odors.

S. No.ParameterIn-house sample
1SedimentsNil
2pH4.58
3Total solid2.05% w/v
4Wt/mL0.94g
5Alcohol content41.0% v/v

                                               Table 1: Result of physicochemical properties of In-house sample of Pyrus malus mother tinctures.

S. No.Name of testResults %
1Foreign matter2.00
2Loss on drying4.90
3Total ash value3.69
4Acid Insoluble ash value1.20
5Water-soluble extractive value18.60
6Alcohol-soluble extractive value5.65

                                                             Table 2: Result of physicochemical properties of Pyrus malus raw drug material.

S. No.Name of Phytochemical testResults  
1Tannins (lead acetate test)Positive
2Flavonoids (alkaline reagent test)Positive
3AlkaloidsPositive
 Mayer’s test 
 Hager’s test 
 Wagner’s test 
 Dragendorff’s test 
4Phenolic compounds (ferric chloride testPositive
5Glycoside (Sodium hydroxide reagent test)Positive

Table 3: Result of Phytochemical tests for screening various phytochemicals present in the mother tincture of Pyrus malus.

3.2. Results of HPTLC study

Pyrus malus chloroform extract was prepared in the laboratory. Based on extensive literature reviews, various combinations of solvent systems were studied to have an appropriate mobile phase composition for the best and most efficient HPTLC chromatographic separation of active constituents in Pyrus malus chloroform extract. Among all the mobile phase combinations studied, chloroform: methanol (9:1, v/v) was finalized as the ideal. Thus, the most appropriate mobile phase composition for the entire HPTLC method development study was finalized. The profile of Chromatographic separation was scanned at UV 254nm and 366nm wavelength. At white light nine spots appear at Rf. 0.26(yellow), 0.33(yellow), 0.40(green), 0.47(green), 0.55(green) and 0.79(light green) Figure 2. At UV 254nm six brown spots appear at Rf. 0.26, 0.32, 0.40, 0.46, 0.55 and 0.82 Figure 3. At UV 366nm eight spots appear at Rf. 0.24(green), 0.33(red), 0.38(red), 0.46(red), 0.55(red), 0.67(red),0.79 (red) and 0.84(red) Figure 4.

Figure 2: High-performance thin-layer chromatography fingerprints of Pyrus malus at white light. Track 1-4 Pyrus malus mother tincture.

Figure 3: High-performance thin-layer chromatography fingerprints of Pyrus malus at UV 254nm. Track 1-4 Pyrus malus mother tincture.

Figure 4: High-performance thin-layer chromatography fingerprints of Pyrus malus at UV 366nm. Track 1-4 Pyrus malus mother tincture.

3.3. Results of Antioxidant study

3.3.1. Result of Total phenol content

In the present study the total phenolic content of Pyrus malus in-house mother tincture was determined by Folin–Ciocalteu method and reported as ascorbic acid equivalents. Study reveals total phenolic content found in Pyrus malus in-house sample (75µL) is 17.47 mM AAE (Ascorbic acid equivalents) Table 4.

Sample.Concentration in (mM) of AAE Absorbance
Pyrus malus mother tincture17.472.3633
Control0.100.0151

                                                                                Table 4: Clinical Studies on Sugar Intake and Cognitive Decline.

3.3.2. Result of DPPH Assay

In the present study the DPPH assay of Pyrus malus in-house mother tincture was determined by DPPH radical scavenging assay method and reported as ascorbic acid equivalents (AAE). Study reveals Pyrus malus in-house sample was able to decolorize DPPH free radical, the DPPH scavenging increased with the concentration of the extract. The percentage of inhibition found in 100 µL volume of Pyrus malus in-house mother tincture was 86.18% Table 5.

Table 5: Result of DPPH Assay in Pyrus malus mother tincture.

In DPPH assay a significant correlation coefficient (R, 0.9944) was found between the antioxidant activity of alcoholic extracts (mother tinctures) of Pyrus malus in-house sample. The hydrogen radical scavenging action is known to be one of the important mechanisms for measuring antioxidant activity. This assay determines the scavenging of stable radical species DPPH by antioxidant compounds present in the mother tincture. Our study indicates that the Pyrus malus mother tincture exhibits a high content of Phenolic compound i.e.,17.47 mM which was significantly correlated with DPPH radical scavenging activity % i.e., 86.18% Table 4 & 5. 

3.3.3. Result of Total flavonoid content 

In the present study the total flavonoid content of Pyrus malus mother tincture was determined by AlCl3 method and reported as rutin hydrate equivalents. Study reveals Pyrus malus mother tinctures (1ml) contain 3293.75 µg/ml amount of flavonoid compound Table 6.

Table 6: Result of Total Flavonoid content in Pyrus malus mother tincture.

3.3.4. Result of ABTS assay 

In the present study the ABTS assay of Pyrus malus in-house sample was determined by ABTS assay method and reported in terms of Trolox equivalents. A significant correlation coefficient (R, 0.9901) was found between the antioxidant activity of alcoholic extracts (mother tinctures) of Pyrus malus in-house sample. Study reveals Pyrus malus in-house mother tincture was able to decolorize ABTS+ free radical, the ABTS radical cation scavenging activity increased with the concentration of the extract. The result showed the percentage of inhibition found in 10 µL volume of Pyrus malus in-house mother tincture was 99.44% Table 7

Table 7: Result of ABTS radical cation scavenging activity against Pyrus malus mother tincture.

4. Discussion

The present study provides a comprehensive analysis of the antioxidant potential of Pyrus malus in-house mother tincture through the quantification of total phenolic and flavonoid contents, and its evaluation via DPPH and ABTS radical scavenging assays. The findings strongly support the hypothesis that Pyrus malus contains bioactive compounds capable of exerting significant antioxidant effects. The total phenolic content was found to be 17.47 mM AAE at 75 µL concentration, indicating a high level of polyphenolic compounds. Phenolic compounds are well-known for their capacity to donate hydrogen atoms or electrons, thereby neutralizing free radicals and contributing to the plant’s antioxidant defense mechanism. Similarly, the flavonoid content, measured as 3293.75 µg/mL of rutin hydrate equivalents, further confirms the richness of Pyrus malus in phytochemicals with antioxidant properties. Flavonoids act as potent scavengers of reactive oxygen species (ROS) and are associated with various health benefits, including anti-inflammatory and cardioprotective effects. The strong radical scavenging activity observed in both DPPH and ABTS assays provides functional evidence of antioxidant capability. The DPPH assay demonstrated 86.18% inhibition at 100 µL concentration, while the ABTS assay showed even higher scavenging activity with 99.44% inhibition at only 10 µL. These values are significant, especially considering the small volumes used, and suggest a high potency of the mother tincture. Importantly, strong correlation coefficients were found between phytochemical content and antioxidant activity (R = 0.9944 for DPPH and R = 0.9901 for ABTS), supporting a direct association between phenolic/flavonoid presence and free radical neutralization. These findings align with previous studies highlighting the antioxidant potential of apple derived products and extracts, and further suggest that the preparation method of the mother tincture retains or possibly concentrates these bioactive compounds. The results not only validate the traditional use of Pyrus malus in herbal medicine but also open new avenues for its application in pharmaceutical formulations aimed at oxidative stress-related disorders.

5. Conclusion

This study demonstrated that the alcoholic extract (mother tincture) of Pyrus malus leaves possesses favorable physicochemical properties and significant antioxidant potential. HPTLC fingerprinting confirmed the presence of bioactive compounds in the chloroform extract, supporting the chemical richness of the formulation. The antioxidant activity, evaluated through total phenol content (17.47 mM AAE), total flavonoid content (3293.75 µg/mL), DPPH assay (86.18% inhibition), and ABTS assay (99.44% inhibition), showed a strong correlation between polyphenol concentration and radical scavenging capacity. These findings suggest that the potent antioxidant effects of Pyrus malus may contribute to its therapeutic efficacy in homoeopathic applications. The study establishes a foundational framework for the standardization and scientific validation of Pyrus malus mother tincture in alternative medicine. Future research should aim at the quantitative estimation and pharmacological evaluation of additional phytoconstituents to further elucidate the mechanisms behind its medicinal properties and broaden its clinical applications.

References

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Hao Jiang

As an author who has recently published in the journal "Brain and Neurological Disorders". I am delighted to provide a testimonial on the peer review process, editorial office support, and the overall quality of the journal. The peer review process at Brain and Neurological Disorders is rigorous and meticulous, ensuring that only high-quality, evidence-based research is published. The reviewers are experts in their fields, and their comments and suggestions were constructive and helped improve the quality of my manuscript. The review process was timely and efficient, with clear communication from the editorial office at each stage. The support from the editorial office was exceptional throughout the entire process. The editorial staff was responsive, professional, and always willing to help. They provided valuable guidance on formatting, structure, and ethical considerations, making the submission process seamless. Moreover, they kept me informed about the status of my manuscript and provided timely updates, which made the process less stressful. The journal Brain and Neurological Disorders is of the highest quality, with a strong focus on publishing cutting-edge research in the field of neurology. The articles published in this journal are well-researched, rigorously peer-reviewed, and written by experts in the field. The journal maintains high standards, ensuring that readers are provided with the most up-to-date and reliable information on brain and neurological disorders. In conclusion, I had a wonderful experience publishing in Brain and Neurological Disorders. The peer review process was thorough, the editorial office provided exceptional support, and the journal's quality is second to none. I would highly recommend this journal to any researcher working in the field of neurology and brain disorders.

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Dr Shiming Tang

Dear Agrippa Hilda, Journal of Neuroscience and Neurological Surgery, Editorial Coordinator, I trust this message finds you well. I want to extend my appreciation for considering my article for publication in your esteemed journal. I am pleased to provide a testimonial regarding the peer review process and the support received from your editorial office. The peer review process for my paper was carried out in a highly professional and thorough manner. The feedback and comments provided by the authors were constructive and very useful in improving the quality of the manuscript. This rigorous assessment process undoubtedly contributes to the high standards maintained by your journal.

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Raed Mualem

International Journal of Clinical Case Reports and Reviews. I strongly recommend to consider submitting your work to this high-quality journal. The support and availability of the Editorial staff is outstanding and the review process was both efficient and rigorous.

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Andreas Filippaios

Thank you very much for publishing my Research Article titled “Comparing Treatment Outcome Of Allergic Rhinitis Patients After Using Fluticasone Nasal Spray And Nasal Douching" in the Journal of Clinical Otorhinolaryngology. As Medical Professionals we are immensely benefited from study of various informative Articles and Papers published in this high quality Journal. I look forward to enriching my knowledge by regular study of the Journal and contribute my future work in the field of ENT through the Journal for use by the medical fraternity. The support from the Editorial office was excellent and very prompt. I also welcome the comments received from the readers of my Research Article.

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Dr Suramya Dhamija

Dear Erica Kelsey, Editorial Coordinator of Cancer Research and Cellular Therapeutics Our team is very satisfied with the processing of our paper by your journal. That was fast, efficient, rigorous, but without unnecessary complications. We appreciated the very short time between the submission of the paper and its publication on line on your site.

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Bruno Chauffert

I am very glad to say that the peer review process is very successful and fast and support from the Editorial Office. Therefore, I would like to continue our scientific relationship for a long time. And I especially thank you for your kindly attention towards my article. Have a good day!

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Baheci Selen

"We recently published an article entitled “Influence of beta-Cyclodextrins upon the Degradation of Carbofuran Derivatives under Alkaline Conditions" in the Journal of “Pesticides and Biofertilizers” to show that the cyclodextrins protect the carbamates increasing their half-life time in the presence of basic conditions This will be very helpful to understand carbofuran behaviour in the analytical, agro-environmental and food areas. We greatly appreciated the interaction with the editor and the editorial team; we were particularly well accompanied during the course of the revision process, since all various steps towards publication were short and without delay".

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Jesus Simal-Gandara

I would like to express my gratitude towards you process of article review and submission. I found this to be very fair and expedient. Your follow up has been excellent. I have many publications in national and international journal and your process has been one of the best so far. Keep up the great work.

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Douglas Miyazaki

We are grateful for this opportunity to provide a glowing recommendation to the Journal of Psychiatry and Psychotherapy. We found that the editorial team were very supportive, helpful, kept us abreast of timelines and over all very professional in nature. The peer review process was rigorous, efficient and constructive that really enhanced our article submission. The experience with this journal remains one of our best ever and we look forward to providing future submissions in the near future.

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Dr Griffith

I am very pleased to serve as EBM of the journal, I hope many years of my experience in stem cells can help the journal from one way or another. As we know, stem cells hold great potential for regenerative medicine, which are mostly used to promote the repair response of diseased, dysfunctional or injured tissue using stem cells or their derivatives. I think Stem Cell Research and Therapeutics International is a great platform to publish and share the understanding towards the biology and translational or clinical application of stem cells.

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Dr Tong Ming Liu

I would like to give my testimony in the support I have got by the peer review process and to support the editorial office where they were of asset to support young author like me to be encouraged to publish their work in your respected journal and globalize and share knowledge across the globe. I really give my great gratitude to your journal and the peer review including the editorial office.

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Husain Taha Radhi

I am delighted to publish our manuscript entitled "A Perspective on Cocaine Induced Stroke - Its Mechanisms and Management" in the Journal of Neuroscience and Neurological Surgery. The peer review process, support from the editorial office, and quality of the journal are excellent. The manuscripts published are of high quality and of excellent scientific value. I recommend this journal very much to colleagues.

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S Munshi

Dr.Tania Muñoz, My experience as researcher and author of a review article in The Journal Clinical Cardiology and Interventions has been very enriching and stimulating. The editorial team is excellent, performs its work with absolute responsibility and delivery. They are proactive, dynamic and receptive to all proposals. Supporting at all times the vast universe of authors who choose them as an option for publication. The team of review specialists, members of the editorial board, are brilliant professionals, with remarkable performance in medical research and scientific methodology. Together they form a frontline team that consolidates the JCCI as a magnificent option for the publication and review of high-level medical articles and broad collective interest. I am honored to be able to share my review article and open to receive all your comments.

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Tania Munoz

“The peer review process of JPMHC is quick and effective. Authors are benefited by good and professional reviewers with huge experience in the field of psychology and mental health. The support from the editorial office is very professional. People to contact to are friendly and happy to help and assist any query authors might have. Quality of the Journal is scientific and publishes ground-breaking research on mental health that is useful for other professionals in the field”.

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George Varvatsoulias

Dear editorial department: On behalf of our team, I hereby certify the reliability and superiority of the International Journal of Clinical Case Reports and Reviews in the peer review process, editorial support, and journal quality. Firstly, the peer review process of the International Journal of Clinical Case Reports and Reviews is rigorous, fair, transparent, fast, and of high quality. The editorial department invites experts from relevant fields as anonymous reviewers to review all submitted manuscripts. These experts have rich academic backgrounds and experience, and can accurately evaluate the academic quality, originality, and suitability of manuscripts. The editorial department is committed to ensuring the rigor of the peer review process, while also making every effort to ensure a fast review cycle to meet the needs of authors and the academic community. Secondly, the editorial team of the International Journal of Clinical Case Reports and Reviews is composed of a group of senior scholars and professionals with rich experience and professional knowledge in related fields. The editorial department is committed to assisting authors in improving their manuscripts, ensuring their academic accuracy, clarity, and completeness. Editors actively collaborate with authors, providing useful suggestions and feedback to promote the improvement and development of the manuscript. We believe that the support of the editorial department is one of the key factors in ensuring the quality of the journal. Finally, the International Journal of Clinical Case Reports and Reviews is renowned for its high- quality articles and strict academic standards. The editorial department is committed to publishing innovative and academically valuable research results to promote the development and progress of related fields. The International Journal of Clinical Case Reports and Reviews is reasonably priced and ensures excellent service and quality ratio, allowing authors to obtain high-level academic publishing opportunities in an affordable manner. I hereby solemnly declare that the International Journal of Clinical Case Reports and Reviews has a high level of credibility and superiority in terms of peer review process, editorial support, reasonable fees, and journal quality. Sincerely, Rui Tao.

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Rui Tao

Clinical Cardiology and Cardiovascular Interventions I testity the covering of the peer review process, support from the editorial office, and quality of the journal.

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Khurram Arshad

Clinical Cardiology and Cardiovascular Interventions, we deeply appreciate the interest shown in our work and its publication. It has been a true pleasure to collaborate with you. The peer review process, as well as the support provided by the editorial office, have been exceptional, and the quality of the journal is very high, which was a determining factor in our decision to publish with you.

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Gomez Barriga Maria Dolores

The peer reviewers process is quick and effective, the supports from editorial office is excellent, the quality of journal is high. I would like to collabroate with Internatioanl journal of Clinical Case Reports and Reviews journal clinically in the future time.

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Lin Shaw Chin

Clinical Cardiology and Cardiovascular Interventions, I would like to express my sincerest gratitude for the trust placed in our team for the publication in your journal. It has been a true pleasure to collaborate with you on this project. I am pleased to inform you that both the peer review process and the attention from the editorial coordination have been excellent. Your team has worked with dedication and professionalism to ensure that your publication meets the highest standards of quality. We are confident that this collaboration will result in mutual success, and we are eager to see the fruits of this shared effort.

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Maria Dolores Gomez Barriga

Dear Dr. Jessica Magne, Editorial Coordinator 0f Clinical Cardiology and Cardiovascular Interventions, I hope this message finds you well. I want to express my utmost gratitude for your excellent work and for the dedication and speed in the publication process of my article titled "Navigating Innovation: Qualitative Insights on Using Technology for Health Education in Acute Coronary Syndrome Patients." I am very satisfied with the peer review process, the support from the editorial office, and the quality of the journal. I hope we can maintain our scientific relationship in the long term.

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Dr Maria Dolores Gomez Barriga

Dear Monica Gissare, - Editorial Coordinator of Nutrition and Food Processing. ¨My testimony with you is truly professional, with a positive response regarding the follow-up of the article and its review, you took into account my qualities and the importance of the topic¨.

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Dr Maria Regina Penchyna Nieto

Dear Dr. Jessica Magne, Editorial Coordinator 0f Clinical Cardiology and Cardiovascular Interventions, The review process for the article “The Handling of Anti-aggregants and Anticoagulants in the Oncologic Heart Patient Submitted to Surgery” was extremely rigorous and detailed. From the initial submission to the final acceptance, the editorial team at the “Journal of Clinical Cardiology and Cardiovascular Interventions” demonstrated a high level of professionalism and dedication. The reviewers provided constructive and detailed feedback, which was essential for improving the quality of our work. Communication was always clear and efficient, ensuring that all our questions were promptly addressed. The quality of the “Journal of Clinical Cardiology and Cardiovascular Interventions” is undeniable. It is a peer-reviewed, open-access publication dedicated exclusively to disseminating high-quality research in the field of clinical cardiology and cardiovascular interventions. The journal's impact factor is currently under evaluation, and it is indexed in reputable databases, which further reinforces its credibility and relevance in the scientific field. I highly recommend this journal to researchers looking for a reputable platform to publish their studies.

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Dr Marcelo Flavio Gomes Jardim Filho

Dear Editorial Coordinator of the Journal of Nutrition and Food Processing! "I would like to thank the Journal of Nutrition and Food Processing for including and publishing my article. The peer review process was very quick, movement and precise. The Editorial Board has done an extremely conscientious job with much help, valuable comments and advices. I find the journal very valuable from a professional point of view, thank you very much for allowing me to be part of it and I would like to participate in the future!”

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Zsuzsanna Bene

Dealing with The Journal of Neurology and Neurological Surgery was very smooth and comprehensive. The office staff took time to address my needs and the response from editors and the office was prompt and fair. I certainly hope to publish with this journal again.Their professionalism is apparent and more than satisfactory. Susan Weiner

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Dr Susan Weiner

My Testimonial Covering as fellowing: Lin-Show Chin. The peer reviewers process is quick and effective, the supports from editorial office is excellent, the quality of journal is high. I would like to collabroate with Internatioanl journal of Clinical Case Reports and Reviews.

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Lin-Show Chin

My experience publishing in Psychology and Mental Health Care was exceptional. The peer review process was rigorous and constructive, with reviewers providing valuable insights that helped enhance the quality of our work. The editorial team was highly supportive and responsive, making the submission process smooth and efficient. The journal's commitment to high standards and academic rigor makes it a respected platform for quality research. I am grateful for the opportunity to publish in such a reputable journal.

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Sonila Qirko

My experience publishing in International Journal of Clinical Case Reports and Reviews was exceptional. I Come forth to Provide a Testimonial Covering the Peer Review Process and the editorial office for the Professional and Impartial Evaluation of the Manuscript.

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Luiz Sellmann

I would like to offer my testimony in the support. I have received through the peer review process and support the editorial office where they are to support young authors like me, encourage them to publish their work in your esteemed journals, and globalize and share knowledge globally. I really appreciate your journal, peer review, and editorial office.

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Zhao Jia

Dear Agrippa Hilda- Editorial Coordinator of Journal of Neuroscience and Neurological Surgery, "The peer review process was very quick and of high quality, which can also be seen in the articles in the journal. The collaboration with the editorial office was very good."

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Thomas Urban

I would like to express my sincere gratitude for the support and efficiency provided by the editorial office throughout the publication process of my article, “Delayed Vulvar Metastases from Rectal Carcinoma: A Case Report.” I greatly appreciate the assistance and guidance I received from your team, which made the entire process smooth and efficient. The peer review process was thorough and constructive, contributing to the overall quality of the final article. I am very grateful for the high level of professionalism and commitment shown by the editorial staff, and I look forward to maintaining a long-term collaboration with the International Journal of Clinical Case Reports and Reviews.

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Cristina Berriozabal

To Dear Erin Aust, I would like to express my heartfelt appreciation for the opportunity to have my work published in this esteemed journal. The entire publication process was smooth and well-organized, and I am extremely satisfied with the final result. The Editorial Team demonstrated the utmost professionalism, providing prompt and insightful feedback throughout the review process. Their clear communication and constructive suggestions were invaluable in enhancing my manuscript, and their meticulous attention to detail and dedication to quality are truly commendable. Additionally, the support from the Editorial Office was exceptional. From the initial submission to the final publication, I was guided through every step of the process with great care and professionalism. The team's responsiveness and assistance made the entire experience both easy and stress-free. I am also deeply impressed by the quality and reputation of the journal. It is an honor to have my research featured in such a respected publication, and I am confident that it will make a meaningful contribution to the field.

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Dr Tewodros Kassahun Tarekegn

"I am grateful for the opportunity of contributing to [International Journal of Clinical Case Reports and Reviews] and for the rigorous review process that enhances the quality of research published in your esteemed journal. I sincerely appreciate the time and effort of your team who have dedicatedly helped me in improvising changes and modifying my manuscript. The insightful comments and constructive feedback provided have been invaluable in refining and strengthening my work".

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Dr Shweta Tiwari

I thank the ‘Journal of Clinical Research and Reports’ for accepting this article for publication. This is a rigorously peer reviewed journal which is on all major global scientific data bases. I note the review process was prompt, thorough and professionally critical. It gave us an insight into a number of important scientific/statistical issues. The review prompted us to review the relevant literature again and look at the limitations of the study. The peer reviewers were open, clear in the instructions and the editorial team was very prompt in their communication. This journal certainly publishes quality research articles. I would recommend the journal for any future publications.

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Dr Farooq Wandroo

Dear Jessica Magne, with gratitude for the joint work. Fast process of receiving and processing the submitted scientific materials in “Clinical Cardiology and Cardiovascular Interventions”. High level of competence of the editors with clear and correct recommendations and ideas for enriching the article.

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Dr Anyuta Ivanova

We found the peer review process quick and positive in its input. The support from the editorial officer has been very agile, always with the intention of improving the article and taking into account our subsequent corrections.

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Dr David Vinyes

My article, titled 'No Way Out of the Smartphone Epidemic Without Considering the Insights of Brain Research,' has been republished in the International Journal of Clinical Case Reports and Reviews. The review process was seamless and professional, with the editors being both friendly and supportive. I am deeply grateful for their efforts.

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Gertraud Teuchert-Noodt

To Dear Erin Aust – Editorial Coordinator of Journal of General Medicine and Clinical Practice! I declare that I am absolutely satisfied with your work carried out with great competence in following the manuscript during the various stages from its receipt, during the revision process to the final acceptance for publication. Thank Prof. Elvira Farina

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Dr Elvira Farina

Dear Jessica, and the super professional team of the ‘Clinical Cardiology and Cardiovascular Interventions’ I am sincerely grateful to the coordinated work of the journal team for the no problem with the submission of my manuscript: “Cardiometabolic Disorders in A Pregnant Woman with Severe Preeclampsia on the Background of Morbid Obesity (Case Report).” The review process by 5 experts was fast, and the comments were professional, which made it more specific and academic, and the process of publication and presentation of the article was excellent. I recommend that my colleagues publish articles in this journal, and I am interested in further scientific cooperation. Sincerely and best wishes, Dr. Oleg Golyanovskiy.

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Dr Oleg Golyanovski

Dear Ashley Rosa, Editorial Coordinator of the journal - Psychology and Mental Health Care. " The process of obtaining publication of my article in the Psychology and Mental Health Journal was positive in all areas. The peer review process resulted in a number of valuable comments, the editorial process was collaborative and timely, and the quality of this journal has been quickly noticed, resulting in alternative journals contacting me to publish with them." Warm regards, Susan Anne Smith, PhD. Australian Breastfeeding Association.

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Dr Susan Anne Smith

Dear Jessica Magne, Editorial Coordinator, Clinical Cardiology and Cardiovascular Interventions, Auctores Publishing LLC. I appreciate the journal (JCCI) editorial office support, the entire team leads were always ready to help, not only on technical front but also on thorough process. Also, I should thank dear reviewers’ attention to detail and creative approach to teach me and bring new insights by their comments. Surely, more discussions and introduction of other hemodynamic devices would provide better prevention and management of shock states. Your efforts and dedication in presenting educational materials in this journal are commendable. Best wishes from, Farahnaz Fallahian.

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Dr Farahnaz Fallahian

Dear Maria Emerson, Editorial Coordinator, International Journal of Clinical Case Reports and Reviews, Auctores Publishing LLC. I am delighted to have published our manuscript, "Acute Colonic Pseudo-Obstruction (ACPO): A rare but serious complication following caesarean section." I want to thank the editorial team, especially Maria Emerson, for their prompt review of the manuscript, quick responses to queries, and overall support. Yours sincerely Dr. Victor Olagundoye.

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Dr Victor Olagundoye

Dear Ashley Rosa, Editorial Coordinator, International Journal of Clinical Case Reports and Reviews. Many thanks for publishing this manuscript after I lost confidence the editors were most helpful, more than other journals Best wishes from, Susan Anne Smith, PhD. Australian Breastfeeding Association.

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Dr Susan Anne Smith

Dear Agrippa Hilda, Editorial Coordinator, Journal of Neuroscience and Neurological Surgery. The entire process including article submission, review, revision, and publication was extremely easy. The journal editor was prompt and helpful, and the reviewers contributed to the quality of the paper. Thank you so much! Eric Nussbaum, MD

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Dr Eric S Nussbaum

Dr Hala Al Shaikh This is to acknowledge that the peer review process for the article ’ A Novel Gnrh1 Gene Mutation in Four Omani Male Siblings, Presentation and Management ’ sent to the International Journal of Clinical Case Reports and Reviews was quick and smooth. The editorial office was prompt with easy communication.

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Hala Al Shaikh

Dear Erin Aust, Editorial Coordinator, Journal of General Medicine and Clinical Practice. We are pleased to share our experience with the “Journal of General Medicine and Clinical Practice”, following the successful publication of our article. The peer review process was thorough and constructive, helping to improve the clarity and quality of the manuscript. We are especially thankful to Ms. Erin Aust, the Editorial Coordinator, for her prompt communication and continuous support throughout the process. Her professionalism ensured a smooth and efficient publication experience. The journal upholds high editorial standards, and we highly recommend it to fellow researchers seeking a credible platform for their work. Best wishes By, Dr. Rakhi Mishra.

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Dr Rakhi Mishra