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Abstract

To develop and optimize UV- Spectrophotometric method for Berberine Hydrochloride API. To validate newly developed UV- Spectrophotometric method using various parameter according to ICH guidelines. Materials and method: UV-Spectrophotometric method was developed using Phosphate Buffer pH 7.4 the developed method was standardized in terms of validation parameter such as simple, sensitive, precise, linear, accurate, robust, reproducible as per ICH Q2 (R1) guidelines for estimation of Berberine Hydrochloride in marketed formulation this newly developed method was successfully applied. Results: Berberine Hydrochloride exhibits ? max at 275nm and in the concentration range 50 to 250 ?g/ml beers law was obeyed. The Limit of Detection was found to be 42.83 ?g/ml and Limit of quantification was found to be 142.79 ?g/ml. Recovery of Berbeshine in Marketed formulation was obtained in range of 102-115%, All the precision and repeatability results were observed within the acceptance range i.e. less than 2%. Assay of Berbeshine was found to be in range of 95-97%. Conclusion: The method was found to be simple accurate and precise reproducible and marketed formulation of Berberine Hydrochloride was estimated.

Keywords

Berberine Hydrochloride, Beer’s law, Phosphate Buffer pH 7.4, UV-Spectrophotometer, Validation

Introduction

1.1 Berberine Hydrochloride:1,2,3

Berberine Hydrochloride is an Isoquinoline alkaloid found in a variety of medicinal plants, mainly in the Berberis genus and the family is Berberidaceae [Berberis vulgaris]. It is Bright yellow in colour and is widely used in traditional medicines. Charka Samhita prescribed that the extract of the plant be taken internally for the treatment of haemorrhage, piles, pruritus and alopecia. Sushruta Samhita described it is being useful internally in indigestion deficiency of breast milk and in uterine and vaginal disorders. It also possesses activity against diabetes, High cholesterol, Hypertension, Anti-microbial.

Drug Profile:4

Table no.1 Drug profile of Berberine Hydrochloride

 

Drug Name

Berberine Hydrochloride

Structures:

2D Str

3D Str

 

Iupac Name

5,6-dihydro-9,10-dimethoxybenzo[g]-1,3-benz0dioxolo[5,6a]-quinolizinium

Solubility

Phosphate Buffer pH 7.4

Mechanism Of Action

Berberine Hydrochloride inhibits pseudomonas aeruginosa biofilms growth by down- regulating pslA and pelA.  It inhibits inflammation in mice.

 

Molecular Formula

C20H18C1NO4

Molecular Weight

371.81gm/ml

Category

Diabeties, High cholesterol, Hypertension, Anti microbial

Colour

Bright yellow colour

Odour And Taste

Charecteristics odour and taste

Melting Point

204-206

State

Solid

Storage

Store in dark place

Part Used

Root, Rizhome and Bark

 

 

1.2 UV-Spectroscopy

Basic Principles of spectroscopy5

Spectroscopy deals with the production, measurements and interpretation of spectra arising from the interaction of electromagnetic radiation with matter. There are many different spectroscopic methods available for solving a wide range of analytical problems. The methods differ with respect to species to be analysed (e.g. molecular or atomic spectroscopy), the type of radiation matter interaction to be monitored (e.g. absorption, emission or diffraction) and the region of the electromagnetic spectrum used in the analysis. Spectroscopic methods are very informative and widely used for the both quantitative and qualitative analyses. Spectroscopic method based on the absorption or emission of radiation in the ultraviolet (UV), visible(vis), infrared (IR) and radio (nuclear magnetic resonance NMR) frequency ranges are most commonly encountered in traditional food analysis laboratories. Each of these methods is distinct in that it monitors different types of molecular or aromatic transitions.

Spectroscopy Method:6,7

It is the branch of science dealing with the study of interaction between Electromagnetic radiation and matter. It is a most powerful tool available for the study of atomic and molecular structure/s and is used in the analysis of wide range of samples o [tical spectroscopy includes the region on electromagnetic spectrum between 100 A0 and 400 μ m. The region of electromagnetic spectrum.

Fig No.1 UV- Spectrophotometer

Instrumentation of UV- Spectrophotometry8

UV-Visible spectrophotometry is one of the most frequently employed technique in pharmaceutical analysis. It involves measuring the amount of ultraviolet or visible radiation absorbed by a substance in solution. Instrument which measures the ratio, or function of ratio, of the intensity of two beams of light in the UV-Visible region are called Ultraviolet-Visible spectrophotometers. In qualitative analysis, organic compounds can be identified by use of spectrophotometer, if any recorded data is available, and quantitative spectrophotometric analysis is used to ascertain the quantity of molecular species absorbing the radiation. Spectrophotometric technique is simple, rapid, moderately specific and applicable to small quantities of compounds. The fundamental law that governs the quantitative spectrophotometric analysis is the Beer -Lambert law. Beer’s law: It states that the intensity of a beam of parallel monochromatic radiation decreases exponentially with the number of absorbing molecules. In other words, absorbance is proportional to the concentration.

Lambert’s law: It states that the intensity of a beam of parallel monochromatic radiation decreases exponentially as it passes through a medium of homogeneous thickness. combination of these two laws yields the Beer-Lambert law.

Beer-Lambert law: When beam of light is passed through a transparent cell containing a solution of an absorbing substance, reduction of the intensity of light may occur.

Mathematically, Beer Lambert law is expressed as

A=a b c

Where,

  • A-Absorbance or optical density
  • a-absorptivity or extinction coefficient
  • b-path length of radiation through sample (cm)
  • c- concentration of solute in solution.

Both b and a are constants so a is directly proportional to the concentration c when c in gm/100, then the constant is called A (1%, 1cm)

A=A1%/1cm’bc

Quantification of medicinal substance using spectrophotometer may carried out by preparing solution in transparent solvent and measuring its absorbance at suitable wavelength. The wavelength normally selected is wavelength of maximum absorption (λ  max), where small error in setting the wavelength scale has little effect on measured absorbance. Ideally, concentration should be adjusted to give an absorbance of approximately 0.9 around which the accuracy and precision of the measurements are optimal. The assay of single component sample, which contains other absorbing substances, is then calculated from the measured absorbance by using one of three principal procedures.

Fig No.2 Schematic instrumentation of UV spectrophotometer

  1. METHODOLOGY:
    1. Materials and methods:

The materials required to carry out the work includes reagents, chemicals and analytical instruments as given below:

Chemicals and Apparatus:

Chemicals:  Drug sample Berberine Hydrochloride API was procured from Zyrex Ayurveda India for method development of Berberine Hydrochloride. Methanol, Ethanol, Phosphate Buffer6,8,7,7.4 and Distilled water. For Method Validation: Phosphate Buffer 7.4, Berberine Hydrochloride extract, Distilled water, tablet marketed formulation. Apparatus: 100ml volumetric flask, 10ml of volumetric flask, beakers, funnels, micro pipette, measuring cylinders, cuvette, soft tissue papers, aluminium foil. Instruments: Shimadzu UV 1800 Spectrophotometer.

    1. Method Development
  1. Solubility: In estimation of Berberine Hydrochloride drug it is necessary to know the solubility of the drug. So, the solvent selection was done by performing Gravimetric method in different solvents.

Gravimetric Method10: Take 10 ml of each solvent and add 100mg of drug to it. Stir continuously for 24hours.filter the solution and evaporate the solvent using hot air oven at 400 c. After drying weigh the residue. Calculate the solubility by using the formula.

Table No.2 Solubility

Solvents

W1

W2

W3=W2-W1

Ethanol

1.08

1.108

0.028

Methanol

1.08

1.103

0.023

PH 7

1.08

1.141

0.061

PH 7.4

1.08

1.108

0.028

PH 6.8

1.08

1.136

0.056

Distilled water

1.08

1.12

0.043

Formula: W3= W2- W1

By performing above gravimetric analysis we have come to the conclusion that we should use Phosphate Buffer pH 7.4 should be used.

Calculation:

  • W1=weight of empty filter paper.
  • W2=weight of filter paper with residue after drying.
  • W3=undissolved residue.

Note:

Methodology for UV-spectroscopy:

First select a suitable solvent then selection of maximum absorbance range and wavelength and determination of wavelength then determination of UV- Spectrophotometric method.

 Determination of Maximum Wavelength (λ max):

Take 0.1gm of Berberine Hydrochloride powder API is dissolved in 100ml of Phosphate Buffer pH 7.4 transfer 1ml of above solution into 10ml of vol flask containing 10ml of Phosphate Buffer pH 7.4 then the solution was scanned against black for the entire UV visible wavelength 200-400nm. Based on the spectrum λ max of 275nm was selected for further analysis.

  1. Standard Preparation:

0.1gm of Berberine Hydrochloride API was weighed accurately then dissolve in 100ml 0f Phosphate Buffer pH7.4 transfer 0.5, 1, 1.5, 2, 2.5 ml of above solution into 10ml vol. flask make up the mark with phosphate Buffer pH 7.4 to make 50, 100, 150, 200, 250 μ g/ml solutions then take absorbance of these standard measured at λ max of 275nm. The standard curve was drawn by plotting concentration v/s absorbance.

  1. Instruments:

Table No:3 Analytical Instruments

Instruments

UV- Spectroscopy

Model

UV1800

Software

UV probe

Company

Shimadzu

Solvent

Phosphate Buffer pH 7.4

Scanning wavelength range

400-200nm

Maximum absorbance of Berberine Hydrochloride

275nm

  1. Method validation: The method was validated according to ICH guidelines (Q2R 1). Method validation was   done by using following parameters like Linearity and Range, Precision, LOD, LOQ, Ruggedness, Robustness, Assay, Accuracy.
  1. Linearity and range: From stock solution transfer 50, 100, 150, 200, 250 μ g/ml of standard solution in 10ml vol. flask make up the volume with phosphate buffer pH 7.4 up to the mark then scanned at 275nm and calibration curve was plotted by taking concentration X axis and absorbance on Y axis then correlation coefficient was calculated and it was found to be 0.999 calibration was shown in fig No.4 results are in Table No. 4
  2. Precision: Precision was performed in two ways that is Intraday and interday precision.

Intraday precision:  For intraday precision prepare triplicates of 50 μ g/ml, 150 μ g/ml, 250 μ g/ml from stock solution in 10ml vol. flask make up the volume with phosphate buffer pH 7.4 up to the mark then scanned at 275nmand % RSD was found to be less than 2%. The results are shown in Table No. 5

Interday Precision: For interday precision prepare triplicates of 50 μ g/ml, 150 μ g/ml, 250 μ g/ml from stock solution in 10ml vol. flask make up the volume with phosphate buffer pH 7.4 up to the mark and scanned at 275nm and %RSD was found to be less than 2%. The results are shown in Table. No 6

  1. Ruggedness: The ruggedness was performed change in analyst or change in instrument by preparing the triplicates of upper, middle and lower concentration i.e. 50 μ g/ml, 150 μ g/ml, 250 μ g/ml respectively. The reproducibility was checked which showed %RSD less than 2% and this indicates that the method developed is rugged. The results are shown in Table No.7
  2. Robustness: Robustness is done by changing the wavelength by preparing the triplicates of upper, middle and lower concentration i.e. 50 μg/ml, 150 μg/ml, 250 μg/ml respectively. The reproducibility was checked which showed %RSD less than 2% and this indicates that the method developed is robust. The results are shown in Table No. 8
  3. LOD: The limit of detection was calculated using equation: LOD=3.3×σ /S

Where,

σ  – Standard deviation of Y intercept of calibration curve

S - Slope of regression equation, the results are shown Table No. 9

  1. LOQ: The limit of quantification was calculated using equation: LOQ: 3.3 σ /S

Where,

 σ  – Standard deviation of Y intercept of calibration curve

S - Slope of regression equation, the results are shown Table No.10

Assay: 10 tablets of Berbeshine were weighed that contains 500mg of Berberine Hydrochloride. Average weight of tablet is calculated then calculate the theoretical drug content. Prepare stock solution of Berberine Hydrochloride and stock solution of Berbeshine and filter the solution through Whatman filter paper then 100 μ g/ml concentration sample solution and standard solution scanned at 275nm then calculate the practical yield.

Formula:

%Purity=Practical drug content Theoretical drug content× 100

Average weight of 10 tablets = 842mg

Lable claim = 500mg Berberine Hydrochloride

(Limit for % purity of drug is 90-120%)

The results are shown in Table No. 10

  1. Accuracy:
  1. Stock solution of Berberine Hydrochloride: 0.1gm of Berberine Hydrochloride powder was weighed accurately. Dissolved in 100ml vol. flask containing 100ml Phosphate Buffer pH 7.4
  2. Sample solution of Berbeshine: 0.1gm of Berbeshine powder was weighed accurately. Dissolved in 100ml vol. flask in 100ml Phosphate Buffer pH 7.4.

For 50%: From the above standard stock solution pipette out 0.5ml standard solution and 0.25ml of sample solution transferred into 10 vol. flask and the volume is made up to the mark using phosphate buffer pH 7.4. solvent three replicates were prepared to performed the study and calculate % recovery.

For 100%: From the above standard stock solution pipette out 0.5ml standard solution and 0.5ml of sample solution transferred into 10 vol. flask and the volume is made up to the mark using phosphate buffer pH 7.4. solvent three replicates were prepared to performed the study and calculate % recovery.

For 150%: From the above standard stock solution pipette out 0.5ml standard solution and 0.75ml of sample solution transferred into 10 vol. flask and the volume is made up to the mark using phosphate buffer pH 7.4. solvent three replicates were prepared to performed the study and calculate % recovery.

(Limit for % recovery of drug is 90-120%)

The results were shown in Table No. 11

  1. RESULTS AND DISCUSSION:

Discussion: The UV- Spectroscopic method was developed for estimation of Berberine Hydrochloride using phosphate buffer pH 7.4 at 275nm. Validation was performed as per ICH guidelines.

UV-spectroscopic method development and validation: UV method was developed by using Shimadzu UV 1800 spectrophotometer instrument using a solvent system phosphate buffer pH 7.4 at 275nm.

Fig No. 3: UV spectra of Berberine Hydrochloride

Linearity:  The linearity range of Berberine Hydrochloride was in the range of 50-250 μ g/ml correlation coefficient :0.999

Table No. 4

Sr No

Concentration

Absorbance

1

0

0

2

50

0.184

3

100

0.371

4

150

0.522

5

200

0.761

6

250

0.898

 

r2

0.999

SLOPE

0.0036

LOD

42.83

LOQ

142.79

Report: The results of UV spectroscopic method were within the acceptance limit. The calibration curve of the drug gives correlation coefficient value 0.999

Specificity and selectivity: Solvent used in method developed was interfered with the drug hence the method was found to be specific.

Precision: Precision was implemented intraday and interday Precision. The % RSD was found within acceptance criteria

Table No. 5: Intraday precision of Berberine Hydrochloride

Sample no

Conc μ g/ml

Absorbance

%RSD

Average %RSD

 

 

Morning

Afternoon

Evening

 

 

0.49%

1

50

0.252

0.269

0.263

0.75%

2

150

0.514

0.521

0.523

0.48%

3

250

0.749

0.752

0.753

0.26%

Table No. 6: Interday precision of Berberine Hydrochloride

Sample no

Conc μ g/ml

Absorbance

%RSD

Average %RSD

 

 

Day 1

Day 2

Day 3

 

 

0.4%

1

50

0.253

0.266

0.268

0.8%

2

150

0.515

0.518

0.523

0.4%

3

250

0.750

0.755

0.755

0.2%

Ruggedness: : By change in analyst ruggedness was performed. %RSD was found within acceptance criteria.

Table No.7: Ruggedness Study

Analysts

Sample no.

Conc μ g/ml

Absorbance

SD

%RSD

Analyst 1

1

50

0.189

0.0031

1.65%

Analyst 2

2

150

0.487

0.001

0.21%

Analyst 3

3

250

0.774

0.0015

0.20%

Robustness: : By change in wavelength the robustness was performed. The %RSD was found within acceptance criteria

Table No.8

Sample no

Conc μ g/ml

Wavelength

%RSD

Average %RSD

 

 

273nm

275nm

277nm

 

0.66%

1

50

0.148

0.149

0.147

1.34

2

150

0.469

0.474

0.472

0.44

3

250

0.725

0.733

0.731

0.21

Sensitivity: LOD and LOQ data of Berberine Hydrochloride:

Table No.9

Drug

LOD μ g/ml

LOQ μ g/ml

Berberine Hydrochloride

42.83

142.79

Accuracy: By taking the absorbance of Standard solution and Marketed solution Accuracy was performed. %Recovery was within acceptance criteria.

Table No. 10 Accuracy of Berberine Hydrochloride

Total conc

μ g/ml,

Std conc of BH

Sample conc of BH

Absorbance

275nm

Conc

Y=mx+c

Sample conc difference

μ g/ml,

% Recovery

std

sample

50%

25

25

0.184

0.186

50.83

25.83

103.3%

150%

25

75

0.371

0.373

102.7

77.7

103.65

250%

25

125

0.522

0.540

149.1

124.1

99.25%

Assay:

Table No:11Assay of Berberine Hydrochloride

Sr no

Sample

Absorbance

%Assay

1

Standard solution

0.512

105.5%

2

Marketed solution

0.229

CONCLUSION: It was revealed that the newly developed UV-Spectrophotometer method for determining the Berberine Hydrochloride in its pharmaceutical dosage form was simple, specific and cost effective. Using the ICH criteria all validation parameters were found to be acceptable ranges.

Summary: Method was developed by using Phosphate buffer pH 7.4 as solvent at 275nm.method was validated by using phosphate buffer pH 7.4 as solvent and accuracy was done by using market formulation (Berbeshine). By using ICH guidelines, the validation was performed

ACKOWLEDGMENT: The completion of our project work has been an intense learning period for us, not only in scientific arena, but also a personal level. In this acknowledgement we take privilege to express sincere gratitude towards the fine individuals for their inspiration guidance and support that we received. First and foremost, we thank the almighty who blessed us with this opportunity. We would like to express our gratitude to our guide Kiran patil Lecturer of Dept of pharmaceutical chemistry, Rani chennamma college of pharmacy. Her valuable guidance encouragement timely suggestions throughout our work will always remembered. We wish to express our thanks to principal of Rani chennamma college of pharmacy, Belagavi. We express  our sincere gratitude to Udaykumar Bolmal sir and all teachers for their valuable Feedback and profound co-operation offered during the time of research work. Thanks to one all……

REFERENCES

  1. Tavade S, Patil K, Kurangi B, Suryawanshi S. Development and validation of UV-spectrophotometric method for estimation of berberine hydrochloride in marketed formulation and poly lactic co-glycolic acid nanoparticles. Indian J Pharm Educ Res. 2022 Jul 1;56(3):873-880.
  2. Pasrija A, Singh R, Katiyar CK. Validated HPLC-UV method for the determination of berberine in raw herb Daruharidra (Berberis aristata DC), its extract, and in commercially marketed ayurvedic dosage forms. International Journal of Ayurveda Research. 2010 Oct;1(4):243.
  3. Majumder KK, Kumar MA, Pahwa RA, Lamba AK, Shankar RA, Tiwari AB, Tiwari VA, Rishabh. Formulation and Characterization of Floating Tablet Dosage Form of Dual Delivery of Drug Curcumin and Berberine Hydrochloride Using Simultaneous Estimation by Uv Spectroscopy. International Journal of Applied Pharmaceutics. 2021 Sep 7:306-310.
  4. IUPAC Chemical Nomenclature and Structure Representation Division (2013). "P-73.3.1". In Favre HA, Powell WH (eds.). Nomenclature of Organic Chemistry: IUPAC Recommendations and Preferred Names 2013. IUPAC–RSC. ISBN 978-0-85404-182-
  5. Penner MH. Basic principles of spectroscopy. Food analysis. 2017:79-88.
  6. Willard HH, Merritt Jr LL, Dean JA, Settle Jr FA. Instrumental methods of analysis.
  7. Chatwal Gr, Anand KS. Instrumental methods of chemical analysis, 5th Edn Himalaya publishing House. Mumbai 2002:2-149.
  8. Davidson AG, Beckett AH, Stenlake JB. Practical pharmaceutical chemistry.
  9. Behera S, Ghanty S, Ahmad F, Santra S , Banerjee S .Analytical and Bioanalytical techniques UV – visible spectrophotometric method development and validation of assay of paracetamol tablet formulation. Journal of Analytical and Bioanalytical Techniques. 2012;3(6).
  10. Qureshi A, Dr. Vyas J, Dr. Upadhyay UM. Determination of solubility by gravimetric method: A brief review National journal of pharmaceutical sciences. 2022;2(1):1-3.
  11. Zanwar AS, Sen DB, Memon M, Sanathra R, Patel D, Maheshwari RA, Sen AK. Spectrophotometric Methods for the Analysis of Berberine Hydrochloride and Eugenol in Formulated Emulgel. Journal of Natural Remedies. 2022 Jul 30;22(3):440-488.
  12. Lalka S, Momin M, Sherje A. simultaneous estimation of berberine hydrochloride and indomethacin for in vitro diffusion profile by spectroscopy. Indian Drugs. 2021 Feb 1;58(2):
  13. Chauhan I, Singh V, Majhi S, Verma M, Yasir M. Development and Validation of UV-Spectrophotometric Method for Berberine Quantification. Journal of Applied Pharmaceutical Sciences and Research. 2024 Aug 1;7(2):28-33.
  14. Mujtaba MA. Development and validation of UV-spectrophotometric methods for the determination of berberine in polymeric nanoparticles. Advanced Science, Engineering and Medicine. 2019 Dec 1;11(12):1273-1278.
  15. Latif A, Siddiqui N, Razique A, Sukul RR. Spectrophotometric estimation of berberine and TLC fingerprint profile of a Unani eye drop formulation. IJPRD. 2010;2(7):5.
  16. Lamichhane J, Lamichhane T, Patil SB, Balasubramanian J, Joshi J, Tripathi P, Manikyam HK. Microwave Assisted Extraction of Berberine and Preparation of Berberine Hydrochloride from Berberis Aristata Variety of Nepal, and Quantification using RP-HPLC and HPTLC Methods. European Journal of Medicinal Plants. 2021 Dec 14;32(12):46-53.
  17. Alam SD, Beg MA, Bagadi M, Locatelli M, ALOthman ZA, Mustaqeem M, Ali I. Facile extraction of berberine from different plants, separation, and identification by thin?layer chromatography, high?performance liquid chromatography, and biological evaluation against Leishmaniosis. Journal of Separation Science. 2023 Nov;46(21):2300582.
  18. Chauhan S, Tyagi S, Maan P. Simultaneous UV Spectrophotometric Method for Estimation of Berberine and Withanolide in Polyherbal Immunity Booster Dosage Form. International Journal of Newgen Research in Pharmacy & Healthcare. 2023 Dec 30:26-31.
  19. Ali SA, Parveen N, Ali AS. Promoting Melanocyte Regeneration Using Different Plants and Their Constituents. Cancer Therapy; Bentham Science: Sharjah, United Arab Emirates. 2019 Sep 2;3:247-276.
  20. Mulsa N, Sanghvi G, Purohit P, Sheth N, Vaishnav D. Development of the UV spectroscopic method of phenytoin sodium in API and stress degradation studies. Inventi Rapid: Pharma Analysis & Quality Assurance. 2013:1-5.

Reference

  1. Tavade S, Patil K, Kurangi B, Suryawanshi S. Development and validation of UV-spectrophotometric method for estimation of berberine hydrochloride in marketed formulation and poly lactic co-glycolic acid nanoparticles. Indian J Pharm Educ Res. 2022 Jul 1;56(3):873-880.
  2. Pasrija A, Singh R, Katiyar CK. Validated HPLC-UV method for the determination of berberine in raw herb Daruharidra (Berberis aristata DC), its extract, and in commercially marketed ayurvedic dosage forms. International Journal of Ayurveda Research. 2010 Oct;1(4):243.
  3. Majumder KK, Kumar MA, Pahwa RA, Lamba AK, Shankar RA, Tiwari AB, Tiwari VA, Rishabh. Formulation and Characterization of Floating Tablet Dosage Form of Dual Delivery of Drug Curcumin and Berberine Hydrochloride Using Simultaneous Estimation by Uv Spectroscopy. International Journal of Applied Pharmaceutics. 2021 Sep 7:306-310.
  4. IUPAC Chemical Nomenclature and Structure Representation Division (2013). "P-73.3.1". In Favre HA, Powell WH (eds.). Nomenclature of Organic Chemistry: IUPAC Recommendations and Preferred Names 2013. IUPAC–RSC. ISBN 978-0-85404-182-
  5. Penner MH. Basic principles of spectroscopy. Food analysis. 2017:79-88.
  6. Willard HH, Merritt Jr LL, Dean JA, Settle Jr FA. Instrumental methods of analysis.
  7. Chatwal Gr, Anand KS. Instrumental methods of chemical analysis, 5th Edn Himalaya publishing House. Mumbai 2002:2-149.
  8. Davidson AG, Beckett AH, Stenlake JB. Practical pharmaceutical chemistry.
  9. Behera S, Ghanty S, Ahmad F, Santra S , Banerjee S .Analytical and Bioanalytical techniques UV – visible spectrophotometric method development and validation of assay of paracetamol tablet formulation. Journal of Analytical and Bioanalytical Techniques. 2012;3(6).
  10. Qureshi A, Dr. Vyas J, Dr. Upadhyay UM. Determination of solubility by gravimetric method: A brief review National journal of pharmaceutical sciences. 2022;2(1):1-3.
  11. Zanwar AS, Sen DB, Memon M, Sanathra R, Patel D, Maheshwari RA, Sen AK. Spectrophotometric Methods for the Analysis of Berberine Hydrochloride and Eugenol in Formulated Emulgel. Journal of Natural Remedies. 2022 Jul 30;22(3):440-488.
  12. Lalka S, Momin M, Sherje A. simultaneous estimation of berberine hydrochloride and indomethacin for in vitro diffusion profile by spectroscopy. Indian Drugs. 2021 Feb 1;58(2):
  13. Chauhan I, Singh V, Majhi S, Verma M, Yasir M. Development and Validation of UV-Spectrophotometric Method for Berberine Quantification. Journal of Applied Pharmaceutical Sciences and Research. 2024 Aug 1;7(2):28-33.
  14. Mujtaba MA. Development and validation of UV-spectrophotometric methods for the determination of berberine in polymeric nanoparticles. Advanced Science, Engineering and Medicine. 2019 Dec 1;11(12):1273-1278.
  15. Latif A, Siddiqui N, Razique A, Sukul RR. Spectrophotometric estimation of berberine and TLC fingerprint profile of a Unani eye drop formulation. IJPRD. 2010;2(7):5.
  16. Lamichhane J, Lamichhane T, Patil SB, Balasubramanian J, Joshi J, Tripathi P, Manikyam HK. Microwave Assisted Extraction of Berberine and Preparation of Berberine Hydrochloride from Berberis Aristata Variety of Nepal, and Quantification using RP-HPLC and HPTLC Methods. European Journal of Medicinal Plants. 2021 Dec 14;32(12):46-53.
  17. Alam SD, Beg MA, Bagadi M, Locatelli M, ALOthman ZA, Mustaqeem M, Ali I. Facile extraction of berberine from different plants, separation, and identification by thin?layer chromatography, high?performance liquid chromatography, and biological evaluation against Leishmaniosis. Journal of Separation Science. 2023 Nov;46(21):2300582.
  18. Chauhan S, Tyagi S, Maan P. Simultaneous UV Spectrophotometric Method for Estimation of Berberine and Withanolide in Polyherbal Immunity Booster Dosage Form. International Journal of Newgen Research in Pharmacy & Healthcare. 2023 Dec 30:26-31.
  19. Ali SA, Parveen N, Ali AS. Promoting Melanocyte Regeneration Using Different Plants and Their Constituents. Cancer Therapy; Bentham Science: Sharjah, United Arab Emirates. 2019 Sep 2;3:247-276.
  20. Mulsa N, Sanghvi G, Purohit P, Sheth N, Vaishnav D. Development of the UV spectroscopic method of phenytoin sodium in API and stress degradation studies. Inventi Rapid: Pharma Analysis & Quality Assurance. 2013:1-5.

Photo
Kiran Patil
Corresponding author

Department of Pharmaceutical Chemistry, Rani Chennamma College of Pharmacy, Rajiv Gandhi University of Health Science, Belagavi 590001, Karnatak, India.

Photo
Abishek Shedabal
Co-author

Department of Pharmaceutical Chemistry, Rani Chennamma College of Pharmacy, Rajiv Gandhi University of Health Science, Belagavi 590001, Karnatak, India

Photo
Kalpana Gulappagol
Co-author

Department of Pharmaceutical Chemistry, Rani Chennamma College of Pharmacy, Rajiv Gandhi University of Health Science, Belagavi 590001, Karnatak, India

Photo
MD Umar Gokak
Co-author

Department of Pharmaceutical Chemistry, Rani Chennamma College of Pharmacy, Rajiv Gandhi University of Health Science, Belagavi 590001, Karnatak, India

Photo
Pushpa Kadalagi
Co-author

Department of Pharmaceutical Chemistry, Rani Chennamma College of Pharmacy, Rajiv Gandhi University of Health Science, Belagavi 590001, Karnatak, India

Photo
Vidya Bellivari
Co-author

Department of Pharmaceutical Chemistry, Rani Chennamma College of Pharmacy, Rajiv Gandhi University of Health Science, Belagavi 590001, Karnatak, India

Kiran Patil*, Abishek Shedabal, Kalpana Gulappagol, MD Umar Gokak, Pushpa Kadalagi, Vidya Bellivari, UV-Spectrophotometric Method Development and Validation of Berberine Hydrochloride, Int. J. of Pharm. Sci., 2025, Vol 3, Issue 6, 2825-2836. https://doi.org/10.5281/zenodo.15704448

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