INTRODUCTION:

Psoriasis is a most common long lasting chronic autoimmune inflammatory skin disease. Psoriasis is affecting approximately 2% to 3% of world population. The word psoriasis is derived from Greek word “Psora” means itch and “iasis” means condition.

It characterized papules or erythematous plaques and skin lesions covered with silvery scales. The most commonly skin affected areas elbows, knees, breasts, scalp, hand, feet, palms, fingers and toes to treatment of complete body. In case, darker skin the patches may be purple in color.^(1,2)

The symptoms observed psoriasis skin patches are irritation, dryness, pain, redness and itching on skin. For detailed studied psoriasis, it categorized in different psoriasis such as invasive psoriasis, plaque psoriasis, psoriatic arthritis, nail psoriasis, scalp psoriasis and several types psoriasis. According to IFPA (International Federation of Psoriasis Association), the psoriasis like symptoms and disease was found to be in 125 millon people across the world. Around 10 million cases every year in India. Therefore, we need to do more efforts on psoriasis treatment.⁽³⁾

Nowadays, many allopathic treatments are available to reduce the symptoms of psoriasis but they produce some side effects. So people is moving towards the use of crude drugs and herbals drug for the treatment with lesser adverse effects. So now days various herbal formulations was prepared for the treatment of psoriasis with lesser side effects. For the local treatment of psoriasis we will use hydrogel as dosage forms.⁽⁴⁾

Hydrogels is a three-dimensional hydrophilic polymer network chains. Due to their individual cross-linking chain with chemical or physical polymer. Hydrogels are highly absorbent natural and synthetic polymeric networks. It contains three phases: polymeric network matrix solid phase, interstitial fluid phase, and ionic phase. Hydrogel was being used for the treatment of various skin diseases due to better patient compliance and good permeation properties as compared to conventional creams and ointment. Further we can use various permeation enhancer for the good permeability of the drug in to skin. Permeation enhancers are promote absorption of drug through skin temporary by transient enhancing skin permeability. They employed transfer delivery of drugs which ionizable to maintain drug level in blood.⁽⁵⁾

The berberine hydrochloride obtained from heart-leaved moonseed obtained from the plant of Tinospora cordifolia belonging to the family Menispermaceae. Berberine HCl is a compound that can be extracted from a number of herbs and plants. They inhibits proliferative A549 cells, the cells was treated with variety of concentration. They suppressed growth of cells and A549 cells. They shows the antipsoriatic activity and anti-inflammatory activity.^(6,7) Nimisha et al., 2017 they reported that berberine used for the treatment of antipsoriatic and anti-inflammatory activity.⁽⁷⁾

In the present paper we focus to prepare the hydrogel of Berberine Hydrochloride for the treatment of psoriasis. Further we evaluated the effect of permeation enhancer on the skin for the treatment of psoriasis.

MATERIAL AND METHODS:

Berberine hydrochloride was purchased from Yucca Chemical, Mumbai. The carbopol 934 was purchased from SDFCL Mumbai, gylcerine, propylene glycol, propyl paraben, Transcutol P and triethanolamine was purchased from Central drug house (P) Ltd. New Delhi, Methanol was purchased from HiMedia Laboratories Pvt. Ltd. 23 Badhani Ind. Est. LBS Marg Mumbai and methyl paraben was purchased from Qualikems fine chemicals Pvt. Ltd.

Preformulation studies of Berberine hydrochloride:

Various kinds of physicochemical, chemical characterization were being done for the drugs and excipients by physically characterized on basis of color, odour and taste. The melting point of the drug was estimated using capillary tube fusion method. The obtained melting point was compared with standard.⁽⁸⁾

FTIR Analysis:

The compound was estimated by FTIR to determine the functional group of Berberine hydrochloride. It was studied by FTIR (Shimadzu Affinity-1) spectrophotometry. The potassium bromide was ground into fine powder by using mortar pestle and then compressed into KBr disc in a hydraulic press at a 75Kg/cm pressure. The KBr disc was scanned 20 times at a 2cm resolution. The characteristic peaks were recorded. The mixture of drug and potassium bromide was mesh in fine powder by using mortar pestle and then compressed in to a KBr discs in a hydraulic press at a pressure of 75Kg/cm. The each potassium bromide (KBr) disc was scanned 20 times at a resolution of 2cm. The characteristic peaks were recorded. To identify the characteristics peaks of excipients used in the formulations with combination of Berberine hydrochloride, the combined IR spectroscopy was performed with the help of above method. The compatibility of the drug and excipients was studied on the basis of interpretation of characteristics peaks of drugs and specific excipients.^(9,10)

UV-Visible Spectrophotometry:

An accurately weight of Berberine hydrochloride (25 mg) dissolved in methanol as cosolvent and volume made up with pH 6.8 phosphate buffer (25mL) volumetric flask. 10mL solution was pipetted out and transferred to 100mL volumetric flask was made up to 100mL and volume with phosphate buffer. Then, further diluted 10mL of solution was pipette out and transferred to 100mL in a volumetric flask and volume was made up to 100mL with phosphate buffer and series of concentration 1, 2, and 5µg/mL was diluted and scanned 359nm at UV Spectrophotometer.⁽¹⁰⁾

Table 1: Formulation study of hydrogels

Ingredients (%age)	F1	F2	F3	F4	F5	F6
Drug	0.5	0.5	0.5	0.5	0.5	0.5
Carbopol 934	0.5	1	1.5	2	1.5	1.5
Glycerine	5	5	5	5	5	5
Propyl glycol	2.5	2.5	2.5	2.5	2.5	2.5
Methanol	2.5	2.5	2.5	2.5	2.5	2.5
Transcutol P	-	-	-	-	0.1	0.2
Methyl paraben	0.2	0.2	0.2	0.2	0.2	0.2
Propyl paraben	0.1	0.1	0.1	0.1	0.1	0.1
Triethanolamine	QS	QS	QS	QS	QS	QS
Water	QS	QS	QS	QS	QS	QS

Preparation of hydrogels:

The hydrogels formulation was prepared by using drug in 0.5% concentration with or without permeation enhancers. The Carbopol 934 was mixed with distilled water and kept for overnight. Then methyl paraben, propyl paraben were added along with other excipients like glycerin, propyl glycol, methanol and triethanolamine was added for maintain pH upto 5.5. The formulation table of hydrogel was shown in table 1.

Evaluation of hydrogels:

The prepared hydrogels was evaluated for physical appearance, drug content, viscosity, pH, grittiness, spreadability, homogeneity and in-vitro release study.

Physical Appearance:

The physical appearance of prepared gel formulations was done with the help of visual as well as with the help of microscope. The sample was placed on the glass slide and then cover slip was placed. Then slide was observed under microscope and physical appearances of prepared gel formulation was studied.^(10,11)

Drug content:

1gm of gel from each formulations containing drug was taken in 10mL volumetric flask dissolved in methanol and volume was made up to 10mL with methanol. Then, the solutions was filtered and samples was analyzed using UV spectrophotometer after suitable dilutions at 359nm.⁽¹¹⁾

Viscosity:

Viscosity of the formulations was determined using Brookfield viscometer. The gel was filled in sufficient quantity of sample holder separately. The gel height was filled in the sample holder should sufficiently allow to dip the spindle. The rpm of the spindle was adjusted. The viscosities of the formulations was recorded.⁽¹²⁾

Grittiness:

All gel formulations were evaluated microscopically and were observed for any significant particulate matter. The gel preparation was also self-determined from any kind of grittiness.⁽¹³⁾

Microscopy evaluation:

The prepared formulations of gel were microscopically evaluated to determine the presence of lumps. This was determine by taking few amount of gel and kept it on a glass slide and observes under microscope.⁽¹⁴⁾

Spreadability:

The spreadability of the gel formulations was determined at 24 hours after permeation, by measuring the spreading diameter of 1gm gel between two horizontal plates (20 cm × 20cm) after one min. The time taken for both plates travel distance 7.5cm and separated away from lower slide under the weight was recorded.^(11,15) The spreadability of gel was calculated.

S = M × ––

S = Spreadability

M = Weight tied to upper slides

L = Length moved to glass slide

T =Time taken in sec

In- vitro drug release study:

The diffusion studies was performed of prepared gels was carried out in Franz diffusion cell. Cellophane membrane was soaked in pH 6.8 phosphate buffer overnight. 1gm gel was taken in cellophane membrane and diffusion study was carried out at 37± 0.5^oC using phosphate buffer of pH 6.8 as dissolution medium. 5 ml of sample was withdrawn from the beaker at 1, 2, 3, 4, 5, 6, 7 and 8 hours and sample was replaced with equal volume of phosphate buffer. The sample was analyzed by using spectrophotometrically method.⁽¹²⁾

Skin Permeation Study:

The skin permeation study was performed on Franz diffusion cells using skin of goat skin as the permeation barrier. Harvested skin of goat was freed of adhering fat layers and mounted on diffusion cells with a cross-sectional area of 3.142cm² and the receptor volume was made about 30mL. Diffusion medium placed in receptor compartment was composed of phosphate buffer solution of pH 6.8. Diffusion cell was maintained at normal body temperature 37±2°C while placed medium was continuously stirred. The optimized formulations containing Berberine hydrochloride (0.5% w/w) was applied onto the mice skin at donor compartment and 5mL sample was withdrawn at suitable time intervals (0, 1, 2, 3, 4 and 6 hours). After the sampling an equal volume of fresh diffusion medium was replaced in receptor compartment.^(14,15)

RESULTS AND DISCUSSION:

Preformulation study Berberine hydrochloride:

Physically Berberine hydrochloride was observed to be crystalline yellow powder. The melting point of berberine hydrochloride was determined by using digital melting point method was found to be 143-148°C.⁽¹⁶⁾ Thin layer chromatography of berberine hydrochloride was measured using thin layer chromatography and compared with standard Rf value 0.2 and the calculated Rf values shown in Table 2.^(17,18)

FTIR Analysis of berberine hydrochloride:

FTIR spectra of Berberine hydrochloride was measured at the range 4000-400cm^-1. The presence of characterstics peaks was identified to find the availability of peaks with their functional groups. In the FTIR spectra of Berberine hydrochloride the identified peaks was 1212.14, 1227.74, 1231.60, 1263.43 due to C-O, 1670.43, 1678.14, 1686.82 die to C=N and 1299.11, 1279.89, 1322.26 due to C-N.

The various functional groups showed the presence of characterstics peaks of Berberine in comparison with standard peaks, which indicates the purity of Berberine hydrochloride was observed in Figure 1 (a). The FTIR spectra of Berberine hydrochloride with excipients used in formulations showed the presence of characterstics peaks of each functional group in Figure 1 (b). So, we examined that there was no degradation of peaks in the FTIR spectra of drug with excipients used in the formulation. All the frequency with their respective functional groups was present.

UV-Visible Spectrophotometry Analysis of berberine hydrochloride:

The UV-Visible spectra of Berberine hydrochloride was made in phosphate buffer of 6.8 pH. Berberine hydrochloride was dissolved in methanol (co-solvent) and then the volume was maintained with buffer solution. The spectrum of Berberine hydrochloride was examined at selected wavelength 359 nm. The dilutions were prepared in the concentration range 1, 2, 3, 4 and 5µg/mL. The result of linearity curve of Berberine hydrochloride was found to be 0.9974 R² value showed in Figure 2.

Characterization of hydrogel formulation:

The prepared formulations of hydrogels was further evaluated for their physical appearance, spreadability, viscosity, drug content, grittiness, homogeneity, microscopical evaluation and in-vitro drug release study. The various results of physical parameter were given in the table no. 3. According to this table we find that at the lower concentration of carbopol the F1 formulation becomes like thick liquid only not as semisolid. When we had increased the concentration of carbopol from F1 to F4, 0.5 to 2% respectively It was found to with the increased concentration of polymer leads to better gel properties but at higher concentration gel becomes thick and non homogenious with poor spreadability. So from this observation F3 formulation was selected as it showed good results. So, further transcutol P in the concentration of 0.1 and 0.2 percent in F5 and F6 formulations was added to evaluate the permeation properties.

In vitro drug release study:

In-vitro release study was evaluated by release kinetic models (zero order, first order, higuchi, korsmeyer). The in vitro release profile of prepared topical herbal gel formulations was represented graphically in Figure 3.

Drug release effect on formulations without permeation enhancers:

The effect of Berberine hydrochloride release in hydrogels was examined. The cumulative percentage drug Release was found to be from the observed release data in Table 4, it was found to be formulation F3, F5 and F6 showed 91.2, 96.34 and 99.54% release at the end of 8 hrs study graphically represented in Figure 3.

Effect of Transcutol P on drug release:

Transcutol P was used in formulations (F5, F6) in two concentration ranges (0.1-0.2%). The effect of Transcutol P on drug release is better at 0.2% and it will also give the larger effect on release as compare to rest of two permeation enhancers. The increase in the concentration of Transcutol P revealed the increasing effect on release of drug across the skin. It is a potent penetration enhancer which is miscible with drug molecules. The increase in drug release data at 0.2% of Transcutol P in formulations (F5 and F6) was improved and provides better topical application of drugs.

Table 3: Evaluation parameter of Hydrogel

Formulation	F1	F2	F3	F4	F5	F6
Color	Yellow	Yellow	Yellow	Yellow	Yellow	Yellow
Physical appearance	Liquid form	Liquid to semi solid	Semi-solid	Semi-solid	Semi-solid	Semi-solid
Drug content (%age)	95.4	95.6	96.8	93.1	96.9	96.1
Grittiness	No	No	No	No	No	No
Homogeneity	Poor	Good	Good	Poor	Good	Good
Microscopy evaluation	No lumps / liquid form	No lumps	No lumps	Lumps	No lumps	No lumps
Spreadability (g.cm/sec)	28.09±0.06	23.02±0.05	21.09±0.08	14.06±0.04	21.06±0.05	21.02±0.03
Viscosity (Centipoises)	2590	3795	4209	4494	4209	4211

Fig 3: In-Vitro Drug Release Profile of Berberine hydrochloride with or without Permeation Enhancer

Table 5: Kinetics of In-Vitro Release from hydrogel

Formulation	Zero order	First order	Higuchi	Korsemeyer
Code	R²	R²	R²	N	R²
F3	0.9753	0.9576	0.9543	1	0.9812
F5	0.9902	0.9157	0.9427	1.0	0.9348
F6	0.9861	0.9322	0.9511	1.0	0.9249

For the release mechanism of the drug release data were explored by kinetic models. The kinetic model seems to fit best was noticed by observing the R² value of each formulation. The best fit highest determination R² coefficient was observed by first order and higuchi model followed zero order which indicate drug release.

The n value was found to be 0.8 to 1.0 these values shows the non-fickian drug release mechanism shown in table 5. Therefore, the hydrogels with permeation enhancer (F5 and F6) to increase the release rate as compared to without permeation enhancer (F3).

In research study six formulations of hydrogel (F1-F6) was prepared. In which four formulations (F1, F2, F3 and F4) was prepared without permeation enhancer and rest of the two formulations was prepared with permeation enhancers with two concentration 0.1-0.2%. By performing the in vitro drug release study, formulation (F6) revealed the highest release of drug in 8 hrs as compare to the formulation (F3). The best mechanism of drug release is showed in the formulation (F6) with 0.2% Transcutol P is well suited for drug delivery through skin. By comparing the release data of hydrogel formulations, can find that formulation F3without permeation show low release and formulation (F5 and F6) with permeation enhancers show highest release of drug.

Skin Permeability Study:

The best two formulations F5 and F6 with higher release of Berberine hydrochloride was selected for skin permeation study. The permeability on mice skin was studied for 6 hours respectively. The permeability on mice skin was graphically represented with percent cumulative amount of drug released vs. Time (hr) in Figure 4.

Table 4: Skin Permeation Study of Berberine hydrochloride from Selected Formulation F5, F6

Time (hr’s)	Percentage Cumulative Amount of Drug Permeated
Time (hr’s)	F5	F6
0	0	0
1	3.28±0.09	4.28±0.33
2	7.08± 0.28	9.08±0.65
3	10.09±0.56	11.44±0.21
4	15.48±0.42	14.56±0.08
6	17.09±0.22	19.98±0.02

Fig 4: Skin Permeation study of Berberine hydrochloride from selected formulation F5, F6

Skin permeation study showed that the topical preparation do not remain for more than 6h on skin. The result of skin permeation was recorded for 6 hrs on mice skin. They usually get removed by sweat or eliminate by contact of skin. The above data of permeability indicating that amount of drug released from F6 on skin was more than formulation F5. It was found to formulation F6 is better as compared to F5 in skin permeation studies.

CONCLUSION:

Psoriasis is a most common long lasting chronic autoimmune inflammatory skin disease. The topical approaches are the best way to cure such type of inflammatory diseases. In this research work, six formulations of hydrogel from F1 to F6 were prepared with carbopol 934, methanol, propyl glycol, glycerine, Transcutol P, methyl paraben, propyl paraben and triethanolamine. In which four formulations of hydrogels F1, F2, F3 and F4 was prepared without permeation enhancer and rest of the two formulations was prepared with permeation enhancer in two concentration ranges from 0.1-0.2%. All the formulation was evaluated for their physical appearance, grittiness, viscosity, spreadability, drug content, pH and microscopical evaluation. By evaluating the formulation with above parameters further we can find the in vitro drug release study of all the formulation. The in vitro release profile of formulation F6 revealed the highest release of drug in 8 hrs as compare to the formulation F3. The best mechanism of drug release is showed in the formulation F6 with 0.2% Transcutol P suited for skin by comparing the release data of hydrogel formulation F3 without permeation showed less release in comparison with formulation F5 and F6. As per the kinetics study of hydrogels all the formulations followed non fickian diffusion mechanism with the n value between 0.8 to 1.0. From the above release we can find that hydrogels with permeation enhancers was effective to increase the rate of drug release. After that the formulation F5 and F6 with higher release data was selected for skin permeation study and it was found to be formulation F6 with 0.2% concentration of permeation enhancer showed higher permeability in 6 hr’s as compare to F5 with 0.1% permeation enhancer. The research showed that, permeability of skin was enhanced with the use of penetration enhancers. The effective rate of permeability was showed that the hydrogel formulation F6 was good and useful to penetrate the drug through mice’s skin. So, we had concluded that the present research work confirm the development of hydrogel with best topical penetration properties.

ACKNOWLEDGEMENT:

The author wishes to acknowledge Laureate Institute of Pharmacy, Jawalamukhi, Himachal Pradesh (176031), for providing their support and other required facilities in the preparation of this research article.

CONFLICT OF INTEREST:

None.

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Received on 29.08.2020 Modified on 13.10.2020

Res. J. Pharma. Dosage Forms and Tech.2021; 13(1):12-18.

DOI: 10.5958/0975-4377.2021.00003.3

Drug	Stationary phase	Mobile phase	Spraying reagent	Color of spot	Calculated Rf value
Berberine hydrochloride	Silica gel-G	n-butanol: ethyl acetate: formic acid: distilled water (30:50:10:10)	dragendroff’s reagent	Yellow	0.292