Synergistic Detection, Quantification and Standardization of Oral Dental Preparation Comprising Curcumin and Eugenol By RP-HPLC Method

Dhrumi Prajapati, Pinak Patel, Pratik Patel, Krunal Detholia,

doi:10.5281/zenodo.14898157

Research Paper | Open Access
Volume 03 | Issue 02 | Article Id IJPS/250302159

Synergistic Detection, Quantification and Standardization of Oral Dental Preparation Comprising Curcumin and Eugenol By RP-HPLC Method
Dhrumi Prajapati* Pinak Patel Pratik Patel Krunal Detholia
Department of Pharmaceutical Quality Assurance, Smt. S. M. Shah Pharmacy College, Mahemdabad, Gujarat.

Abstract

In the current study, a reverse phase high performance liquid chromatographic method has been developed and validated for standardization of dental preparation on the basis of Curcumin and Eugenol. Both the active components were determined and quantified using Agilent 1260 Infinity II, with the mobile phase consisted of 40 parts acetonitrile, 30 parts methanol, and 30 parts water. The separation was performed using a Supphirus C18 HP Classic Column (250*4.6 mm, 5 µm). In order to obtain the chromatogram, 240 nm was set as analytical wavelength, the flow rate was kept constant at 1 ml/min. At 4.692 and 5.985 minutes, respectively, the chromatogram showed the elution of Curcumin and Eugenol. Linearity showed up for both Eugenol and Curcumin comprising the range of 0.5 to 5 µg/mL. For Curcumin and Eugenol, the values of R2 were determined to be 0.996 and 0.994, respectively. It was discovered that the LOD for Eugenol and Curcumin was 0.01 and 0.03 µg/mL, respectively. For Eugenol and Curcumin, the LOQ was found to be 0.03 and 0.11 µg/mL, respectively. The ICH Q2 (R2) guideline was successfully followed in the validation of the optimized method.

Keywords

Curcumin (CUR), Eugenol (EUG), Reverse Phase High Performance Liquid Chromatography (RP-HPLC), Analytical Method Development (AMD), Analytical Method Validation (AMV), System Suitability Parameters (SSTs).

Introduction

Curcumin (CUR), the primary active component of turmeric, has long been used for a number of health advantages. The yellow, crystalline powder known as CUR is a hydrophobic polyphenolic derivative that is extracted from the roots and rhizomes of Curcuma longa (family: Zingiberaceae). Scientific research has demonstrated that it benefits in numerous ways, such as producing anti-oxidant, anti-inflammatory, anti-proliferative, antibacterial, antiviral and antifungal properties. ^[1-4]Tulsi extract (Ocimum sanctum), clove oil (Syzygium aromaticum), tea tree oil (Melaleuca alternifolia) and eucalyptus oil (Eucalyptus globulus) are sources of Eugenol (EUG). EUG has many beneficial pharmacological qualities that can help alleviate tooth pain and improve oral health, such as its analgesic, antiseptic, and antibacterial qualities. ^[5-8]

Figure 1: Structure of Curcumin

Figure 2: Structure of Eugenol

The above drug combination is marketed as an oral liquid rinse with 0.1% turmeric, 0.05% Tulsi, 0.09% Eucalyptus, 0.05% Clove oil, 0.1% Tea tree oil, and 0.15% Mint per 100 milliliters. The quantitative analysis of CUR and EUG has been reported using a variety of methods ^[8–21], either alone or in combination with other ingredients. However as of right now, the only method that can instantly quantify CUR and EUG from their combined marketed formulation is the HPTLC method ^[22].

MATERIALS AND METHODS

Instrumentation and reagent:

Sample processing was performed using an Agilent technologies (1260 infinite II) HPLC equipment with a UV-VIS detector that operates in binary gradient mode. Chromatographic analysis was conducted utilizing OpenLab software for processing and integration. Column employed in the operation was a Supphirus C₁₈ HP classic (250 × 4.6mm, 5 µm). Precise material quantities were determined using a Mettler Toledo weighing scale with a sensitivity of 0.1 mg. pH adjustments were made using the Labman LMPH 10 pH meter. Oxford Lab Fine Chem LLP, Mumbai, India, supplied the active pharmaceutical ingredients as complimentary samples. Additionally, chemicals such as methanol, acetonitrile, and water, all meeting HPLC quality standards, were procured from Merck Life Sciences Private Limited (Mumbai, India). Apart from this AR grade methanol was purchased from Anacham Laboratories for the experimental purpose. Finar provided the HPLC grade methanol, acetonitrile, and water that were used in the experimental work.

Preparation of Standard Solution:

The procured CUR and EUG had purities of 95% and 99%, respectively. CUR (105 mg) and EUG (101 mg) were precisely weighed and then transferred to 100 mL volumetric flasks. The levels were raised with acetonitrile to produce a solution containing 100 ?g/mL CUR and EUG. The working standards containing 2.5 ?g/ml CUR and EUG were obtained by diluting 0.5 ml aliquots from the stock solutions of CUR and EUG in 10 ml volumetric flasks of acetonitrile.

Selection of Detection Wavelength:

CUR (10 mg) and EUG (20 mg) solutions were prepared in methanol with the aim of determining an analytical wavelength. The solution was scanned in the UV wavelength range of 200-800 nm, where the observed spectra overlapped. 240 nm wavelength was chosen due to significant absorption.

Optimization of separation conditions:

Initially, the testing was conducted using a 50:50 v/v mixture of methyl nitrile and water. In that case, CUR and EUG were separated with an early retention time; However, the detection was carried out at 254 nm. The subsequent trial's detection was conducted at 240 nm with methanol and water (50:50) as the mobile phase due to the early retention period; But the peak symmetry was questionable. Considering the previously mentioned problems, the composition of mobile phase was again changed in the next trial. Both components were separated, but still there was an issue with their resolution. Eventually, at 240 nm using Acetonitrile: Methanol: water (40:40:20 on volume basis) as the mobile phase, both components were separated with adequate resolution.

Table 1 Optimized Chromatographic Condition

Stationary phase	Sapphirus C18 (250×4.6 mm, 5 µm)
Mobile phase	Acetonitrile: Methanol: Water (40:30:30)
Column temperature	Ambient
Flow rate	1 ml/ min
Detection wavelength	PDA detection (240 nm)
Injection volume	10 µL
Run time	10 min
Retention time	CUR (5.98 min), EUG (4.69 min)

Figure 3 Chromatogram of Optimized Condition

RESULTS AND DISCUSSION

System Suitability:

In order to determine the suitability of the technique for the given parameters, including theoretical plates and tailing factor, a representative biphasic mixture containing 50µg/mL been examined and evaluated for total five time. And the mentioned parameters in Table 2 were statistically studied to check whether they fall within prescribed limits or not. Which later found that all of the parameters fell within the acceptable ranges.

Table 2 System Suitability Parameters for CUR and EUG

Parameters	CUR (50µg/mL)	EUG (50µg/mL)
Retention time	5.98 ± 0.01 %RSD = 0.22	4.69 ± 0.01 %RSD = 0.26
Tailing factor	1.00 ± 0.01 %RSD = 1.48	0.97 ± 0.02 %RSD = 1.98
Number of theoretical plates	2546.42 ± 38.01 %RSD = 1.49	2140.74 ± 28.92 %RSD = 1.35

Validation of Optimized method:

Linearity

Accurately weighed 105 mg of CUR and 101 mg of EUG were placed into a 100 ml volumetric flask, and the volume was adjusted with 1000 µg/ml of acetonitrile. Of the resultant solution, 0.5 ml was taken and diluted to 10 ml with acetonitrile, and a range of different strengths were produced, with 0.5+0.5 µg/mL, 1.25+1.25 µg/mL, 2.5+2.5 µg/mL, 3.75+3.75 µg/mL, and 5+5 µg/mL, as the respective amounts for CUR and EUG. Three injections of each concentration were made.

Table 3 Linearity data for CUR

Sr no.	Concentration (µg/mL)	Mean Area ± SD	%RSD
1	0.5	16010.40 ± 301.21	1.88
2	1.25	30717.47 ± 416.95	1.36
3	2.5	75749.28 ± 1004.19	1.33
4	3.75	119942.89 ± 1152.55	0.96
5	5	156677.44 ± 1065.54	0.68
Linearity Regression Equation		y = 32008x + 2789.2
Regression Coefficient		0.996

Figure 4 Linearity Curve for CUR

Table 4 Linearity data for EUG

Sr no.	Concentration (µg/mL)	Mean Area ± SD	%RSD
1	0.5	22472.50 ± 432.27	1.92
2	1.25	37445.65 ± 671	1.79
3	2.5	66425.53 ± 849.07	1.28
4	3.75	98908.75 ± 921.10	0.93
5	5	125143.17 ± 485.82	0.39
Linearity Regression Equation		y = 24330x + 5510.3
Regression Coefficient		0.995

Figure 5 Linearity Curve for EUG

Figure 6 Overlain chromatogram of CUR and EUG from Linearity studies

Repeatability

Five injections of each concentration from the range were made, and the relative standard deviation (RSD) of each level was determined to assess repeatability.

Table 5 Repeatability of CUR

Area	Concentration (µg/mL) CUR
Area	0.5	1.25	2.5	3.75	5
1	15977.79	30472.40	75449.58	119951.30	157523.70
2	15758.52	30575.58	74525.75	120858.36	156236.00
3	15928.85	30254.99	76958.78	120585.85	157998.67
4	16528.66	31284.52	76589.75	120349.95	156258.96
5	15858.20	30999.88	75222.52	117969.01	155369.89
Mean	16010.40	30717.47	75749.28	119942.89	156677.44
SD	301.21	416.95	1004.19	1152.55	1065.54
%SD	1.88	1.36	1.33	0.96	0.68

Table 6 Repeatability for EUG

Area	Concentration (µg/mL) EUG
Area	0.5	1.25	2.5	3.75	5
1	22228.05	37145.00	66172.95	99059.96	124748.66
2	22586.20	37258.00	66599.10	98751.20	124954.64
3	22852.52	36589.00	67258.89	99213.89	124919.57
4	21848.23	38254.25	66999.85	100025.89	125112.02
5	22847.51	37982.02	65096.85	97492.81	125980.98
Mean	22472.50	37445.65	66425.53	98908.75	125143.17
SD	432.27	671.00	849.07	921.10	485.82
%SD	1.92	1.79	1.28	0.93	0.39

Limit of detection and quantitation

Two methods, known as statistical calculation and visual detection technique, were used to determine LOD and LOQ. Data from repeatability regression were used to statistically compute the detection and quantification limits. Using the average of slope (S) and standard deviation of standard error (?), LOD and LOQ were computed.

Table 7 LOD and LOQ of CUR and EUG

API	LOD	LOQ
API	By Statistical calculation	By Statistical calculation
Curcumin	0.03	0.11
Eugenol	0.01	0.03

Accuracy

In order to determine the percentage of analyte recovery, the sample was spiked with standard at 50%, 100%, and 150% of the target concentration (Table 8). The technique employed for spiking was blank with standard. Using linear regression analysis, the amount of area observed at each spiking level was used to quantify the % recovery after each concentration was injected three times.

Table 8 Spiking of samples for accuracy study of CUR and EUG

Level of Spiking	Volume taken by	Volume made up to	Total concentration
Un-Spiked	-	-	-
50 % Spiked	0.25 ml	10 ml	1.25 µg/ml
100 % Spiked	0.50 ml	10 ml	2.5 µg/ml
150 % Spiked	0.75 ml	10 ml	3.75 µg/ml

For all components, the accuracy of the optimized technique has been established, and recovery tests have been performed at documented levels (Table 9 and 10). The obtained findings were found to fall within 98 and 102%.

Table 9 Accuracy for CUR

Level of Spiking	Amount of drug added (µg/mL)	Amount of drug recovered (µg/mL)	% Mean recovery
Un-Spiked	-	-	-
50 % Spiked	1.5	1.23 ± 0.01	98.40 ± 0.80
100 % Spiked	2.5	2.48 ± 0.01	99.20 ± 0.40
150 % Spiked	3.75	3.72 ± 0.02	99.20 ± 0.53

Table 10 Accuracy for EUG

Level of Spiking	Amount of drug added (µg/mL)	Amount of drug recovered (µg/mL)	% Mean recovery
Un-Spiked	-	-	-
50 % Spiked	1.5	1.22 ± 0.01	97.33 ± 0.46
100 % Spiked	2.5	2.47 ± 0.02	98.67 ± 0.61
150 % Spiked	3.75	3.72 ± 0.02	99.11 ± 0.56

Robustness

Operational parameters like the mobile phase's composition and flow rate were purposefully changed within an acceptable range in order to examine the effects on system suitability parameters like retention time and assay percentage. Table 11 demonstrate that minor adjustments to the method parameters had no discernible impact on the outcomes, indicating the method's robustness.

Table 11 Robustness of CUR and EUG

Parameters	Level of changes	CUR		EUG
Parameters	Level of changes	Mean Area ± SD	%RSD	Mean Area ± SD	%RSD
Flow rate of mobile phase	0.8mL/min	75463 ± 58.39	0.08	66164.20 ± 175.29	0.26
	1 mL/min	75464.89 ± 42.02	0.06	66147.00 ± 115.01	0.17
	1.2 mL/min	75778.57 ± 51.41	0.07	66747.29 ± 84.10	0.13
Composition of mobile phase	38:32:30v/v	75835.64 ± 51.41	0.07	66224.34 ± 201.26	0.30
	40:30:30v/v	75457.46 ± 60.23	0.08	66385.69 ± 300.36	0.45
	40:28:32v/v	75084.64 ± 627.13	0.84	66639.73 ± 188.69	0.28
	42:30:28 v/v	75273.79 ± 236.42	0.31	66799.89 ± 447.65	0.67

(n=5 determinations)

Method precision

The method precision was calculated employing both Intraday and Interday precision. For intraday precision, the whole range (0.5–5 µg/ml) of a combination of CUR and EUG was assessed on the same day at various time intervals, and for interday precision, on a different day. (At each level, n = 3 considerations)

Table 12 Intraday and Interday precision for CUR

Concentration µg/mL CUR	Intraday		Interday
Concentration µg/mL CUR	Mean Area ± SD	%RSD	Mean Area ± SD	%RSD
0.5	15876.93 ± 93.29	0.59	15813.01 ± 145.50	0.92
2.5	75640.64 ± 297.13	0.39	75412.75 ± 433.29	0.57
5	157493.82 ± 34.20	0.02	157504.84 ± 17.46	0.01

Table 13 Intraday and Interday Precision for EUG

Concentration µg/mL EUG	Intraday		Interday
Concentration µg/mL EUG	Mean Area ± SD	%RSD	Mean Area ± SD	%RSD
0.5	22356.61 ± 180.94	0.81	22290.91 ± 83.20	0.37
2.5	66156.08 ± 119.10	0.18	66140.64 ± 124.67	0.19
5	124819.15 ± 74.0	0.06	124691.45 ± 62.07	0.05

Sample analysis (Standardization)

To begin the sample analysis, 5 ml of the formulation was diluted to 100 ml using methanol solution, sonicated for five minutes, and then filtered through 0.45 µ filter paper. Using mobile phase, 1 ml, 2 ml and 0.5 ml of solution was further diluted to 10 ml in separate volumetric flasks. (dilution factor 200, 100 and 50, respectively), the resulting solutions were chromatographed under optimized chromatographic conditions and the peak area was observed for EUG and CUR. The obtained area was subjected to statistical transformation on the basis of regression analysis and the amount of EUG and CUR were derived in mg/100 ml. At dilution factor 200, CUR was not detected; just EUG was. Therefore, it was reasonable to lower the dilution factor from 200 to 100, and as a result, CUR was detected with very less intensity, however was not quantified. Finally, when it was spiked with 10 ?g/ml CUR, the CUR and EUG were separated with significant intensity. Three separate chromatograms of the solution having dilution factor of 200 were taken. The total percentage of CUR and EUG was then determined by statistically transforming the mean observed area using a linear regression equation. (Table 14) (Figure 7, 8, 9)

Table 14 Standardization of Sample Formulation

Formulation

Amount of Formulation taken

Amount Found (µg/ml)

Amount Found (mg)

CUR

EUG

CUR

EUG

Cur Q Fresh

(contains Curcumin and Eugenol)

5 ml

110 µg/ml

11 mg

Figure 7 Sample analysis (Dilution factor:200)

Figure 9 Sample analysis (Spiked with 10 µg/mL CUR, Dilution factor:50)

Table 15 Summary and Conclusion of CUR and EUG

Parameters	Limit	Result		Inference
Parameters	Limit	CUR	EUG	Inference
Linearity	R²> 0.995	0.996	0.994	Method was linear
Repeatability	% RSD < 2>	0.68 – 1.88	0.39 – 1.92	Method was repeatable
LOD	-	0.03	0.01	-
LOQ	-	0.11	0.03	-
Intraday precision	% RSD < 2>	0.02 – 0.59	0.06 – 0.81	Method was precise
Inter-day precision	% RSD < 2>	0.01 – 0.92	0.05 – 0.37	Method was precise
Robustness	% RSD < 2>	-	-	Method was robust

R²= Regression Coefficient

RSD = Relative Standard Coefficient

LOD = Limit of Detection

LOQ = Limit of Quantification

CONCLUSION

For the standardization of formulation on the basis of CUR and EUG, there wasn't any technique developed yet apart from planner chromatography. The recommended RP-HPLC technique precisely isolated and measured the constituents from their combination. In all aspects, including accuracy and reproducibility, the developed, optimized, and validated method was found to be considerably superior to the documented HPTLC method. It was possible to separate both of the active components with this approach. Both of the active components could be successfully analyzed and quantified when the formulation was subjected to the optimized procedure. However, when method was practiced on the formulation, the quality of the formulation was found to be substandard on the basis of CUR as the former component was not detected while Eugenol was successfully quantified. Upon spiking the formulation with standard CUR, the peak corresponding to the same was detected with quite ease.

ACKNOWLEDGEMENTS

The authors would like to thank, Oxford Lab Fine Chem LLP, Mumbai, India for donating gift samples (API) for the development of the technique. The management of Smt. S.M. Shah Pharmacy College, Amsaran, Mahemdabad, Gujarat, India, and Navjyoti Analytics and Research Laboratory has provided the valuable guidance and facilities to the writers to carry out the research work.

Financial Support

For the work accomplished, funding has neither been disclosed nor received.

Conflict Of Interest

No conflicts of interest, either financial or otherwise, have been revealed by the authors.

REFRENCES

Indian Pharmacopoeia. Government of India: Ministry of Health and family welfare Indian Pharmacopoeia Commission, Ghaziabad, Edition 8, 2018: 3, 3799-3800.
Gupta AK: Quality Standard of Indian Medicinal Plants. Indian Council of Medicinal Research, New Delhi, 2003: 8, 138-148.
Martin D: The Complete Drug Reference. Pharmaceutical Press, London, Edition 35, 2007: 1326.
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Ulanowska M and Olas B: Biological Properties and Prospects for the Application of Eugenol—A Review. International journal of molecular sciences 2021; 22: 3671.
Nikhat P and Amber J: Eugenol- A review of a versatile molecule with remarkable pharmacological Properties. Journal of advanced scientific research 2021; 12(8):8-12.
Muhammad FN, Mahnoor K, Muhammad R, Jinyin C, Yali Y and Chunpeng CW: Pharmacological Properties and Health Benefits of Eugenol: A comprehensive Review. Hindawi Oxidative Medicine and Cellular Longevity 2021; 1-7.
Suchitra P and Rajashree H: A new stability-indicating RP-HPLC method for determination of curcumin: An application to nanoparticulate formulation. International Journal of Pharmacy and Pharmacology 2016; 8(12):149-155.
Haroon KS, Kai BL, Gabriel OK and Kok KP: Stability indicating HPLC–UV method for detection of curcumin in Curcuma longa extract and emulsion formulation. Food Chemistry 2015; 170:321-326.
Sultana S and Vandana J: Development and validation of a RP-HPLC method for the simultaneous determination of curcumin, piperine and camphor in an ayurvedic formulation. International Journal of Pharmacy and Pharmaceutical Sciences 2018; 10:115-21.
Ankita K, Srinivas B and Rajashree H: Development and validation of RP-HPLC method for simultaneous estimation of curcumin and piperine. International Journal of Applied Pharmaceutics 2018; 10(5):43-8.
Neha D, Munira M, Upasana S, Tabassum K and Atul S: Analytical method development and validation for simultaneous estimation of curcumin and cyclosporine by RP-HPLC. International Journal of Pharmacy and Pharmaceutical Sciences 2019; 11:26-33.
Rubiya K, Sachin KS, Bhupinder K, Monica G, Sheetu W, Saurabh G, Prateek P, Deep K, Kamal D, Sharath LM, Babu UV, Manish S, Hardik S and Vijay K: Development and validation of RP-HPLC method for simultaneous determination of curcumin and quercetin in extracts, marketed formulations, and self-nanoemulsifying drug delivery system. Regenerative Medicine: Genomics Open 2021; 1(1):43-52.
Reolon JB, Maicon B, Sandra EH, Eduardo AB, Marcelo DM and Leticia MC: Development and validation of high-performance liquid chromatography method for simultaneous determination of acyclovir and curcumin in polymeric microparticles. Journal of Applied Pharmaceutical Science 2018; 8(1):136-141.
Ankalgi DA, Nitin KC, Pooja K and Mahendra AS: Method development and validation for simultaneous estimation of methotrexate and curcumin in Bulk Drug by using RP-HPLC. Journal of Drug Delivery and Therapeutics 2019; 9.
Duygu E, Mustafa C, Derya D and Sacide A: RP-HPLC Method Development for Determination of Curcumin in Commercial Turmeric Capsules. Hacettepe University Journal of the Faculty of Pharmacy 2024; 44.1,1-8.
Sagar S, Sasikumar M, Sunita S and Priyanka P: Validated RP-HPLC method to estimate eugenol from commercial formulations like Caturjata Churna, Lavangadi Vati, Jatiphaladi Churna, Sitopaladi Churna and clove oil. Journal of Pharmaceutical Research 2013; 6(1):53-60.
Zeynep A, Gülcemal Y, Ece MY and Hassan YA: Determination of eugenol in plants and pharmaceutical form by HPLC with amperometric detection at graphene-modified carbon paste electrode. Graphene Tech S.L. 2018; 3, 1-9.
Kannissery P, Ura KI, Yoonuskunju TK, Sayeed A, Shahid HA and Javed A: Development and validation of RP-HPLC-PDA method for the quantification of eugenol in developed nanoemulsion gel and nanoparticles. Journal of Analytical Science and Technology 2013; 4, 1-6.
Rana IS, Aarti SR and Ram CR: Evaluation of antifungal activity in essential oil of the Syzygium aromaticum (L.) by extraction, purification and analysis of its main component eugenol. Brazilian Journal of Microbiology 2011; 42(4):1269-77.
Balkrishna A, Sharma P, Joshi M, Srivastava J and Varshney A: Development and validation of a rapid high?performance thin?layer chromatographic method for quantification of gallic acid, cinnamic acid, piperine, eugenol, and glycyrrhizin in Divya?Swasari?Vati, an ayurvedic medicine for respiratory ailments. Journal of Separation Science 2021; 44(16):3146-1357

Reference

Indian Pharmacopoeia. Government of India: Ministry of Health and family welfare Indian Pharmacopoeia Commission, Ghaziabad, Edition 8, 2018: 3, 3799-3800.
Gupta AK: Quality Standard of Indian Medicinal Plants. Indian Council of Medicinal Research, New Delhi, 2003: 8, 138-148.
Martin D: The Complete Drug Reference. Pharmaceutical Press, London, Edition 35, 2007: 1326.
The Merck Index: An Encyclopedia of Chemicals, Drugs, and Biologicals. Merck Research Laboratory, U.S.A., Edition 14, 2006: 1687.
Indian Pharmacopoeia. Government of India: Ministry of Health and family welfare Indian Pharmacopoeia Commission, Ghaziabad, Edition 8, 2018: 3, 3771-3772.
Ulanowska M and Olas B: Biological Properties and Prospects for the Application of Eugenol—A Review. International journal of molecular sciences 2021; 22: 3671.
Nikhat P and Amber J: Eugenol- A review of a versatile molecule with remarkable pharmacological Properties. Journal of advanced scientific research 2021; 12(8):8-12.
Muhammad FN, Mahnoor K, Muhammad R, Jinyin C, Yali Y and Chunpeng CW: Pharmacological Properties and Health Benefits of Eugenol: A comprehensive Review. Hindawi Oxidative Medicine and Cellular Longevity 2021; 1-7.
Suchitra P and Rajashree H: A new stability-indicating RP-HPLC method for determination of curcumin: An application to nanoparticulate formulation. International Journal of Pharmacy and Pharmacology 2016; 8(12):149-155.
Haroon KS, Kai BL, Gabriel OK and Kok KP: Stability indicating HPLC–UV method for detection of curcumin in Curcuma longa extract and emulsion formulation. Food Chemistry 2015; 170:321-326.
Sultana S and Vandana J: Development and validation of a RP-HPLC method for the simultaneous determination of curcumin, piperine and camphor in an ayurvedic formulation. International Journal of Pharmacy and Pharmaceutical Sciences 2018; 10:115-21.
Ankita K, Srinivas B and Rajashree H: Development and validation of RP-HPLC method for simultaneous estimation of curcumin and piperine. International Journal of Applied Pharmaceutics 2018; 10(5):43-8.
Neha D, Munira M, Upasana S, Tabassum K and Atul S: Analytical method development and validation for simultaneous estimation of curcumin and cyclosporine by RP-HPLC. International Journal of Pharmacy and Pharmaceutical Sciences 2019; 11:26-33.
Rubiya K, Sachin KS, Bhupinder K, Monica G, Sheetu W, Saurabh G, Prateek P, Deep K, Kamal D, Sharath LM, Babu UV, Manish S, Hardik S and Vijay K: Development and validation of RP-HPLC method for simultaneous determination of curcumin and quercetin in extracts, marketed formulations, and self-nanoemulsifying drug delivery system. Regenerative Medicine: Genomics Open 2021; 1(1):43-52.
Reolon JB, Maicon B, Sandra EH, Eduardo AB, Marcelo DM and Leticia MC: Development and validation of high-performance liquid chromatography method for simultaneous determination of acyclovir and curcumin in polymeric microparticles. Journal of Applied Pharmaceutical Science 2018; 8(1):136-141.
Ankalgi DA, Nitin KC, Pooja K and Mahendra AS: Method development and validation for simultaneous estimation of methotrexate and curcumin in Bulk Drug by using RP-HPLC. Journal of Drug Delivery and Therapeutics 2019; 9.
Duygu E, Mustafa C, Derya D and Sacide A: RP-HPLC Method Development for Determination of Curcumin in Commercial Turmeric Capsules. Hacettepe University Journal of the Faculty of Pharmacy 2024; 44.1,1-8.
Sagar S, Sasikumar M, Sunita S and Priyanka P: Validated RP-HPLC method to estimate eugenol from commercial formulations like Caturjata Churna, Lavangadi Vati, Jatiphaladi Churna, Sitopaladi Churna and clove oil. Journal of Pharmaceutical Research 2013; 6(1):53-60.
Zeynep A, Gülcemal Y, Ece MY and Hassan YA: Determination of eugenol in plants and pharmaceutical form by HPLC with amperometric detection at graphene-modified carbon paste electrode. Graphene Tech S.L. 2018; 3, 1-9.
Kannissery P, Ura KI, Yoonuskunju TK, Sayeed A, Shahid HA and Javed A: Development and validation of RP-HPLC-PDA method for the quantification of eugenol in developed nanoemulsion gel and nanoparticles. Journal of Analytical Science and Technology 2013; 4, 1-6.
Rana IS, Aarti SR and Ram CR: Evaluation of antifungal activity in essential oil of the Syzygium aromaticum (L.) by extraction, purification and analysis of its main component eugenol. Brazilian Journal of Microbiology 2011; 42(4):1269-77.
Balkrishna A, Sharma P, Joshi M, Srivastava J and Varshney A: Development and validation of a rapid high?performance thin?layer chromatographic method for quantification of gallic acid, cinnamic acid, piperine, eugenol, and glycyrrhizin in Divya?Swasari?Vati, an ayurvedic medicine for respiratory ailments. Journal of Separation Science 2021; 44(16):3146-1357

Dhrumi Prajapati

Corresponding author

Department of Pharmaceutical Quality Assurance, Smt. S. M. Shah Pharmacy College, Mahemdabad, Gujarat

Pinak Patel

Co-author

Department of Pharmaceutical Quality Assurance, Smt. S. M. Shah Pharmacy College, Mahemdabad, Gujarat

Pratik Patel

Co-author

Department of Pharmaceutical Quality Assurance, Smt. S. M. Shah Pharmacy College, Mahemdabad, Gujarat

Krunal Detholia

Co-author

Department of Pharmaceutical Quality Assurance, Smt. S. M. Shah Pharmacy College, Mahemdabad, Gujarat

Dhrumi Prajapati*, Pinak Patel, Pratik Patel, Krunal Detholia, Synergistic Detection, Quantification and Standardization of Oral Dental Preparation Comprising Curcumin and Eugenol By RP-HPLC Method, Int. J. of Pharm. Sci., 2025, Vol 3, Issue 2, 1673-1682. https://doi.org/10.5281/zenodo.14898157

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Synergistic Detection, Quantification and Standardization of Oral Dental Preparation Comprising Curcumin and Eugenol By RP-HPLC Method

Abstract

Keywords

Introduction

Reference

Dhrumi Prajapati

Pinak Patel

Pratik Patel

Krunal Detholia

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