Back to Journals » Drug Design, Development and Therapy » Volume 12

Berberine nanoparticles with enhanced in vitro bioavailability: characterization and antimicrobial activity

Authors Sahibzada MUK, Sadiq A, Faidah HS, Khurram M, Amin MU, Haseeb A, Kakar M

Received 9 November 2017

Accepted for publication 27 December 2017

Published 14 February 2018 Volume 2018:12 Pages 303—312


Checked for plagiarism Yes

Review by Single-blind

Peer reviewers approved by Dr Palas Chanda

Peer reviewer comments 2

Editor who approved publication: Dr Anastasios Lymperopoulos

Muhammad Umar Khayam Sahibzada,1,2 Abdul Sadiq,2 Hani S Faidah,3 Muhammad Khurram,4 Muhammad Usman Amin,4 Abdul Haseeb,5,6 Maria Kakar4

1Department of Pharmacy, Sarhad University of Science and Information Technology, Peshawar, Pakistan; 2Department of Pharmacy, University of Malakand, Chakdara, Pakistan; 3Department of Microbiology, Faculty of Medicine, Umm Al Qura University, Makkah, Saudi Arabia; 4Department of Pharmacy, Abasyn University, Peshawar, Pakistan; 5Discipline of Social and Administrative Pharmacy, School of Pharmaceutical Science, Universiti Sains Malaysia, Peneng, Malaysia; 6Department of Clinical Pharmacy, College of Pharmacy, Umm Al Qura University, Makkah, Saudi Arabia

Background: Berberine is an isoquinoline alkaloid widely used in Ayurveda and traditional Chinese medicine to treat illnesses such as hypertension and inflammatory conditions, and as an anticancer and hepato-protective agent. Berberine has low oral bioavailability due to poor aqueous solubility and insufficient dissolution rate, which can reduce the efficacy of drugs taken orally. In this study, evaporative precipitation of nanosuspension (EPN) and anti-solvent precipitation with a syringe pump (APSP) were used to address the problems of solubility, dissolution rate and bioavailability of berberine.
Methods: Semi-crystalline nanoparticles (NPs) of 90–110 nm diameter for APSP and 65–75 nm diameter for EPN were prepared and then characterized using differential scanning calorimetry (DSC) and X-ray powder diffractometry (XRD). Thereafter, drug content solubility and dissolution studies were undertaken. Berberine and its NPs were evaluated for their antibacterial activity.
Results: The results indicate that the NPs have significantly increased solubility and dissolution rate due to conversion of the crystalline structure to a semi-crystalline form.
Conclusion: Berberine NPs produced by both APSP and EPN methods have shown promising activities against Gram-positive and Gram-negative bacteria, and yeasts, with NPs prepared through the EPN method showing superior results compared to those made with the APSP method and the unprocessed drug.

Keywords: berberine, EPN, APSP, bioavailability, dissolution, antibacterial activity, precipitation method


In pharmaceuticals, solubility is one of the main factors that keeps many potential drug molecules from the market. Poor aqueous solubility affects bioavailability, due to reduced dissolution of the drug in the body, which leads to poor drug absorption, and thus the desired plasma concentration is not achieved to cause pharmacological action.1 Problems with solubility result in increased costs, as a much higher dose is needed to reach the required plasma concentration level. Moreover, higher doses result in undesired pharmacological responses, such as more adverse effects and poor patient compliance, when the outcome is not what a patient expects.2,3

Solid dosage forms like tablets and capsules, when administered orally, first undergo dissolution in gastrointestinal fluids prior to absorption. For less soluble drugs, the dissolution rate limits the bioavailability. To develop a suitable dosage form, many difficulties arise due to the poor water solubility of drugs, as the therapeutic efficacy of a drug depends upon the solubility of drug molecules.4

Many approaches have been adopted to solve the problems of poor solubility and decreased bioavailability. To improve solubility, some of the techniques that have been used so far are particle-size reduction, solid dispersion, and presentation of a drug in the form of nanoparticles (NPs). NPs are smaller in size than conventional drug particles, and so there is increased drug surface area.57

Drug nanocrystals can be produced by several techniques, including “top-down” and “bottom-up” approaches.8,9 Top-down methods are used frequently in the pharmaceutical industry, and include milling and high-pressure homogenization,10,11 while bottom-up approaches, eg, supercritical fluid technology, antisolvent precipitation, and spray freezing into liquid, are used less commonly. Among the bottom-up approaches, the antisolvent precipitation method is an easy, simple and cost-effective way to achieve a nano scale.1214 If a drug is soluble in an organic solvent, precipitation would be a feasible method.10 Evaporative precipitation of nanosuspension (EPN) and anti-solvent precipitation with a syringe pump (APSP) are among the approaches that are used to address the problems of solubility, dissolution rate and bioavailability.11,15 Fessi et al first developed and patented a solvent displacement method for the simple and rapid preparation of a nanosuspension as presented in Bilati et al.12

Phospholipid carriers have become an attractive tool to address the issue of poorly water-soluble active pharmaceutical ingredients.16 Phytosomes have emerged as a new technology to incorporate phytoconstituents into phospholipid complexes, with subsequent improvement in bioavailability and absorption of poorly soluble compounds.17

Berberine (BBR) is an isoquinoline alkaloid found in the stem bark and roots of Berberis aristata (family Berberidaceae), commonly known as “Daru haldi” in Urdu. BBR formulations are widely used in Ayurveda and traditional Chinese medicine18 to treat illnesses like hypertension19 and inflammatory conditions.20,21 BBR has also been reported to have a number of pharmacological actions including anti-malarial,22 anti-arrhythmic,23,24 anti-hyperglycemic,19 anticancer,2527 hepato-protective,28 antioxidant,29 and antimicrobial.30,31 However, the poor water solubility of BBR impacts its dissolution rate and oral bioavailability, thus limiting its clinical use.15 BBR appears to be a hydrophilic compound and has a log P-value of −1.5,32 which makes BBR a class III drug in the biopharmaceutical classification system (BCS). Drugs included in this class are lipophobic and have poor membrane permeability, and the absorption of the drugs is mostly limited to the paracellular pathway. This limits intestinal absorption and leads to low bioavailability.33

BBR NPs were prepared using APSP and EPN methods in order to improve the bioavailability of the drug. The prepared NPs were investigated for parameters including dissolution, solubility, and antimicrobial properties.

Materials and methods


All the chemicals utilized in this study were of analytical grade from Sigma-Aldrich Co. (St Louis, MO, USA). Unprocessed BBR powder was received as a kind gift from Dr Javed Ali, Department of Microbiology, Pakistan Council of Scientific and Industrial Research (PCSIR) Laboratories, Peshawar, Pakistan. Ethanol, n-hexane, potassium phosphate monobasic, sodium hydroxide, HCl (36.5% w/w), hydroxy propyl methyl cellulose (HPMC), and propylene glycol (PG) were also from Sigma-Aldrich Co, while the deionized double distilled water used in this work was obtained using the Millipore Q® system (EMD Millipore, Billerica, MA, USA).


Preparation of BBR NPs

BBR NPs were prepared through previously described EPN and APSP methods with slight modification for this study.11 Briefly, in the APSP method, a solvent (ethanol) was used to prepare a saturated solution of BBR which was rapidly injected into a particular volume of deionized water (anti-solvent) at a fixed flow rate of 1 mL/min under mechanical stirring (3,000 rpm), with the help of a syringe. The same procedure was used while incorporating different volumes of the deionized water with the same volume of saturated drug solution (1:10, 1:15, 1:20 v/v). After stirring, the resulting mixture, which was in the form of a turbid/opaque suspension, was evaporated quickly in a vacuum using a rotary evaporator to obtain nano-sized drug particles.11

In EPN, a pure drug saturated solution was prepared in ethanol and then, as a result of the quick addition of hexane (anti-solvent) to the prepared drug solution, a nanosuspension was formed. Different solvent:anti-solvent ratios (1:10, 1:15, 1:20 v/v) were used. Nano-sized drug particles from a nanosuspension were obtained by rapid evaporation of the solvent and anti-solvent in a vacuum.11 HPMC and PG at a concentration of 1% were used as stabilizers for NP preparation by both methods.


The prepared NPs were characterized using Fourier-transform infrared spectroscopy (FTIR) for structure confirmation, scanning electron microscopy (SEM) for surface morphology and particle size, X-ray diffraction (XRD) for the determination of structure lattice, and differential scanning calorimetry (DSC) to study the thermal behavior of the sample.

BBR NPs were characterized to determine their particle size and associated polydispersity index (PDI) by dynamic light scattering (Zetasizer® NanoS, Malvern Instruments, Malvern, UK). All the samples were analyzed in triplicate (n=3), and results were presented as mean ± SD.

FTIR was performed in the range of 4,400–200 cm−1 using an infrared spectrophotometer (IR Prestige-21 Fourier-transform infrared spectrophotometer, Shimadzu, Kyoto, Japan) in the range of 200–4,400 cm−1.

Electron photomicrographs of NPs prepared by APSP and EPN were obtained by SEM (JEOL JSM-5910, Tokyo, Japan), which was operated at 20 kV using the standard procedure for sample preparation.

The XRD patterns of unprocessed BBR and prepared NPs were recorded using an X’Pert PRO X-ray diffractometer (PANalytical, Almelo, the Netherlands). The operating voltage was 40 kV, operating current was 30 mA, the start angle 2θ was 5° and the finishing angle was 60°.

DSC studies were done using Mettler-Toledo 822e (Greifensee, Switzerland). The procedure involved using ~3–6 mg of sealed sample in one pan while an empty pan served as a standard. Heat flow to both pans was provided at a rate of 10°C/min under nitrogen gas flow. Any change due to thermal effect in the pans was recorded.

In vitro analysis

Solubility studies

Solubility is the maximum amount of a compound/material in a solution for a particular solvent. The solubility of large particles (micrometers or more) is generally not dependent on particle size. However, the solubility of NPs mainly depends on particle size, with an increase of solubility as the particle size decreases.34

For the solubility study, a surplus amount (equivalent to 200 mg) of unprocessed BBR and the prepared NPs were placed in separate vials. Ten mL of distilled water was added to each vial and shaken vigorously in an orbital shaker (HS501 orbital shaker, IKA GmbH, Staufen im Breisgau, Germany) at 25°C (room temperature) for 72 hours at 3,000 rpm. After mixing, samples were centrifuged at 3,000 rpm and filtered through a Whatman filter paper no 1 (Thermo Fisher Scientific, Waltham, MA, USA). For determination of solubility, the filtered portion was diluted and analyzed at 263 nm using a UV-visible spectrophotometer (PharmaSpec 1,700 UV-visible spectrophotometer, Shimadzu). The same procedure was adopted for PBS pH 6.8 and 0.1 M HCl as solvents to determine the solubility. All tests were run in triplicate. The data were evaluated to determine their significance by applying statistical analysis (one-way ANOVA followed by Dunnett’s post hoc test).


Three different dissolution media – distilled water, 0.1 M HCl, and PBS (pH 6.8) – were used in the dissolution studies in accordance with United States Pharmacopeia (USP) method II (paddle method) as reported previously, with slight modifications.3537 The volume used for each medium was 900 mL at 37°C±0.5°C at a rotation speed of 100 rpm. Unprocessed BBR (100 mg) and prepared NPs were subjected to the dissolution vessels. Five mL aliquots were drawn at predetermined intervals (15, 30, 45, 60, 90, and 120 minutes) and filtered through Whatman filter paper no 1. To maintain the sink conditions, the same volume of medium was replaced.38 Filtered samples were suitably diluted and observed spectrophotometrically using a double-beam spectrophotometer (Agilent 8453 UV/visible spectrophotometer, Agilent Technologies, Santa Clara, CA, USA) at a maximum wavelength of 263 nm. All tests were conducted in triplicate.

Antimicrobial study

The NPs prepared by APSP and EPN methods were tested for their in vitro antimicrobial potentials against four bacteria – two Gram-positive (Staphylococcus aureus and Bacillus subtilis) and two Gram-negative (Escherichia coli and Pseudomonas aeruginosa) – as well as two yeasts (Candida albicans and Candida glabrata) using 96-well microtest plate methods in accordance with the Clinical and Laboratory Standards Institute guidelines39 and a previously reported method.40 The lowest concentration of test NPs that totally inhibited the growth of bacteria and yeasts was considered as the minimal inhibitory concentration (MIC; μg/mL) of the respective test NPs. Positive controls included norfloxacin (for Gram-negative bacteria), clarithromycin (for Gram-positive bacteria), and miconazole (for yeasts), while PBS was used as a vehicle for NP solutions from the respective methods.

Results and discussion

Characterization of the NPs prepared by APSP and EPN

NPs of BBR prepared by APSP and EPN methods were characterized by the following analytical techniques: SEM, Zetasizer, FTIR, DSC and XRD.

Surface morphology

SEM studies were carried out for unprocessed BBR, and its NPs were prepared by APSP (Figure 1) and EPN (Figure 2). The white patches in the figures show the formation of NPs with diameters of approximately 90–110 nm for APSP and 65–75 nm for EPN. The high surface area due to particle size reduction enhanced solubility, dissolution rate and the bioavailability of the NPs.

Figure 1 Scanning electron microscope images of berberine nanoparticles prepared by anti-solvent precipitation with a syringe pump.
Abbreviation: CRL UOP, Central Resource Lab, University of Peshawar.

Figure 2 Scanning electron microscope images of berberine nanoparticles prepared by evaporative precipitation of nanosuspension.
Abbreviation: CRL UOP, Central Resource Lab, University of Peshawar.

Particle size measurement

The mean particle sizes of BBR NPs prepared through APSP and EPN methods are represented in Figure 3A and 3B, respectively. Both figures represent mean particle size ranges from 50±5 nm to 170±7 nm. The mean particle size and PDI values of NPs prepared by the EPN method was 71.53 nm, while the NPs prepared by APSP method had a mean particle size of 102.62 nm. These values show that both NPs have narrow size distribution.

Figure 3 Particle size of the nanocrystals and associated polydispersity index of berberine nanoparticles prepared by (A) anti-solvent precipitation with a syringe pump; and (B) evaporative precipitation of nanosuspension.

FTIR studies

FTIR spectra of unprocessed BBR (Figure 4A) and its NPs prepared by APSP (Figure 4B) and EPN (Figure 4C) indicate sharp peaks with proper intensities as the vibrational changes play a significant role in the intermolecular interactions in solid materials. In the FTIR spectra, the characteristic peaks are as follows: 700–1,300 cm−1 (skeletal C−C vibrations), 1,103.28 cm−1 (C−O), 1,597.06 cm−1, and 1,504.48 cm−1 (aromatic C=C stretching), 1,504.48 cm−1 (skeleton vibration of aromatic C=C ring stretching), 1,386.82 cm−1 and 1,361.74 cm−1 (C=C stretching), 1,276.88 cm−1 (C−O−C stretching), and 1,035.77–1,184.29 cm–1 (in plane = C−H bending).41

Figure 4 Fourier-transform infrared spectroscopy spectra of (A) unprocessed berberine; and nanoparticles prepared by (B) anti-solvent precipitation with a syringe pump (APSP), and (C) evaporative precipitation of nanosuspension (EPN).
Abbreviations: % T, percentage transmission; MB, material berberine; MB EPN, berberine nanoparticles prepared by EPN method; MB APSP, berberine nanoparticles prepared by APSP method.

X-ray diffractometry

The XRD pattern of unprocessed BBR (Figure 5) shows sharp and intense diffraction peaks at 2θ of 8.6°, 9.1°, 12.9°, 16.2°, 20.9°, 25.4°, and 30.1°, which indicates that unprocessed BBR is crystalline in nature. The NPs prepared by the APSP and EPN methods show diffraction peaks with less intensity, which is indicative of a change in the crystalline nature of the material (Figure 5). Less crystalline (semi-crystalline) and amorphous materials have greater free energy compared to their corresponding crystalline forms. Therefore, less crystalline or amorphous forms of the drugs can be more easily solubilized and have enhanced dissolution rates compared to their respective crystalline forms.4144 Thus, modification in the crystalline nature through nano-sizing may be an ideal approach for enhancement of solubility and dissolution rates of drug molecules, which will further improve bioavailability.41

Figure 5 X-ray diffractograms of unprocessed berberine and nanoparticles prepared by anti-solvent precipitation with a syringe pump (APSP), and evaporative precipitation of nanosuspension (EPN).
Abbreviations: MB, material berberine; MB EPN, berberine nanoparticles prepared by EPN method; MB APSP, berberine nanoparticles prepared by APSP method.

Differential scanning calorimetry

NPs prepared by the APSP and EPN methods were further characterized by DSC to determine the effect of crystal structure during melting (Tm) on the heat enthalpy of unprocessed BBR and the prepared NPs (Figure 6). In the present study, unprocessed BBR gave a sharp endothermic peak showing melting of BBR at 145°C. Prepared NPs had almost the same melting point as that of the unprocessed drug, but the melting endothermic peak was less intense, with an enthalpy of heat fusion lower than the unprocessed drug. The reduction in the enthalpy values is an indication of reduction in crystallinity due to decrease in particle size.4143,45

Figure 6 Differential scanning calorimetry results of unprocessed berberine (original berberine) and nanoparticles prepared through anti-solvent precipitation with a syringe pump (APSP) and evaporative precipitation of nanosuspension (EPN).
Abbreviation: MB, material berberine.

In vitro evaluation

Solubility studies

Next, the solubility of unprocessed BBR (Figure 7) and the prepared NPs in distilled water, 0.1 M HCl and PBS (pH 6.8) were studied. The results clearly indicate that solubility of BBR increases when converted to the nano form in all three solvents. The enhancement of the solubility may be attributed to the changes in the crystalline nature of BBR to a semi-crystalline or less crystalline form.15,42,4649 The nano form of BBR has more free energy compared to the micro form which further helps in improving the solubility of the NPs. It is evident from Figure 7 that the solubility of BBR in distilled water is very close to that of PBS (pH 6.8). The solubility of BBR in 0.1 M HCl was lower than that in distilled water and PBS (pH 6.8). The decrease in solubility in 0.1 M HCl solution may be due to conversion of BBR to berberine chloride, which is less soluble than BBR.

Figure 7 Solubility studies of unprocessed berberine and nanoparticles prepared by anti-solvent precipitation with a syringe pump (APSP) and evaporative precipitation of nanosuspension (EPN) methods.
Notes: ***P<0.001 as compared to unprocessed berberine. One-way ANOVA was used, followed by post-hoc Dunnett’s test.
Abbreviations: ANOVA, analysis of variance; D/water, distilled water.

It was found that the solubility of the NPs was significantly higher (P<0.001) than the unprocessed BBR in the distilled water, 0.1 M HCl and PBS.

Dissolution studies

Dissolution studies of unprocessed BBR and of the NPs prepared by the APSP and EPN methods were carried out in distilled water (Figure 8A), PBS pH 6.8 (Figure 8B) and 0.1 M HCl (Figure 8C) over a period of 120 minutes. Samples were drawn at different time intervals (15, 30, 45, 60, 90, and 120 minutes). Dissolution rate analysis clearly shows that unprocessed BBR dissolution was very low (<30%) in distilled water, 0.1 M HCl, and in PBS (pH 6.8), but NPs prepared by APSP and EPN methods showed an improved dissolution rate compared to that of raw BBR over the same time range (120 minutes). NPs made by both methods showed more than 70% dissolution within 15 minutes in all three dissolution media, which clearly indicates an enhanced dissolution rate. The enhanced dissolution rate of NPs can be ascribed to certain factors such as increased surface area, conversion to amorphous form or reduction in the crystallinity, good dispersibility, decrease in agglomeration and aggregation between the hydrophobic drug particles.45

Figure 8 In vitro dissolution profiles of unprocessed berberine and nanoparticles prepared through anti-solvent precipitation with a syringe pump (APSP) and evaporative precipitation of nanosuspension (EPN) methods in (A) distilled water; (B) PBS pH 6.8; and (C) 0.1 M HCl.
Abbreviation: D/water, distilled water.

NPs prepared by the EPN method showed a superior dissolution rate compared to those prepared by the APSP method.

Antimicrobial assays

BBR NPs prepared by APSP and EPN methods had better antibacterial and antifungal activities than BBR (unprocessed) and respective standard antimicrobial drugs (Table 1). The antibacterial activity of BBR NPs produced by the EPN method was increased by three- to four-fold against Gram-positive bacteria. However, BBR NPs prepared by the APSP method showed a two- to three-fold increase in activity against Gram-positive bacteria as shown by a reduction in MICs given in Table 1. The results indicate a significant increase in antibacterial activity of BBR NPs as compared to unprocessed BBR. BBR NPs prepared by the EPN method were found to be more effective than miconazole, ie, BBR had a MIC value of 64 μg/mL against C. glabrata while the MIC of miconazole was 128 μg/mL.

Table 1 In vitro antibacterial and antifungal activities of berberine and its nanoparticles (NPs) prepared by anti-solvent precipitation with a syringe pump (APSP) and evaporative precipitation of nanosuspension (EPN) methods
Note: Minimum inhibitory concentration values shown as μg/mL.
Abbreviations: S. aureus, Staphylococcus aureus; B. subtilis, Bacillus subtilis; E. coli, Escherichia coli; P. aeruginosa, Pseudomonas aeruginosa; C. albicans, Candida albicans; C. glabrata, Candida glabrata.

Infections caused by pathogens such as P. aeruginosa, S. aureus, E. coli, and C. albicans have a high global prevalence, incidence rates and very significant clinical implications, especially in developing countries like Pakistan. Factors involved in the increasing incidence rates include insufficient supply of antimicrobials, patient compliance issues, and self-medication, especially in poorer countries, and occurrence of antibiotic resistance. Plants and their derivatives, considered to be natural remedies, have thus been increasingly used not only in developing and poorer countries but also in developed countries, in which herbal medicines are currently gaining popularity. This use is not confined to a single domain but includes treatment of all ailments including infections caused by microorganisms.50

BBR has been demonstrated to reduce the infectivity of bacteria, fungi, and protozoa in both animals and humans.5053 Previous studies have shown that BBR has negligible activity against Gram-positive bacteria,50 with Zhang et al40 reporting the antibacterial activity of BBR against both Gram-positive and Gram-negative bacteria, with activities equal to or greater than 512 μg/mL against S. aureus and E. coli, respectively. In the present study, BBR NPs prepared by APSP and EPN methods have MIC values of 128 and 64 μg/mL respectively, which indicates a substantial (300%–400%) increase in antibacterial activity against Gram-positive bacteria. Similarly, a profound increase in antibacterial activity by BBR NPs prepared by the EPN method was found against E. coli, ie, MIC 32 μg/mL. The anticandidal activity of BBR NPs was shown to be better than unprocessed BBR. Against C. albicans and C. glabrata, the activity was measured as 64, 128, and 256 μg/mL, respectively, for NPs prepared by the EPN method, NPs prepared by the APSP method, and unprocessed BBR.


The dissolution of NPs prepared by EPN and APSP methods in aqueous medium were 76.8% and 74.1%, respectively, while the solubilities were 1.992 mg/mL and 1.847 mg/mL, respectively. Thus, NPs prepared by the EPN method showed better results than those prepared by the APSP method in terms of solubility and dissolution rate. Moreover, enhanced solubility and dissolution rate in turn increase the bioavailability of the respective NPs. BBR NPs produced by both APSP and EPN methods have shown promising activities against Gram-positive and Gram-negative bacteria, and yeasts, with NPs prepared by the EPN method showing superior results compared to those made with the APSP method, and the unprocessed drug.


The authors would like to thank Mr Javed Ali, Senior Scientific Officer, Pakistan Council of Scientific and Industrial Research (PCSIR) Laboratories, Peshawar, KPK, Pakistan for the provision of berberine raw material for the study. No grant was received from any external source to carry out the research.

Author contributions

All authors contributed toward data analysis, drafting and critically revising the paper and agree to be accountable for all aspects of the work.


The authors report no conflicts of interest in this work.



Mullauer FB, van Bloois L, Daalhuisen JB, et al. Betulinic acid delivered in liposomes reduces growth of human lung and colon cancers in mice without causing systemic toxicity. Anticancer Drugs. 2011;22(3):223–233.


Cheng Z, Chen A-F, Wu F, et al. 8,8-Dimethyldihydroberberine with improved bioavailability and oral efficacy on obese and diabetic mouse models. Bioorg Med Chem. 2010;18(16):5915–5924.


Wang S, Song B, Li K. [Determination of berberine in decocted liquid from shenshu granules with water by reversed-phase liquid chromatography]. Se Pu. 2000;18(3):261–262. Chinese [with English abstract].


Hua W, Ding L, Chen Y, Gong B, He J, Xu G. Determination of berberine in human plasma by liquid chromatography–electrospray ionization–mass spectrometry. J Pharm Biomed Anal. 2007;44(4):931–937.


Zuo F, Nakamura N, Akao T, Hattori M. Pharmacokinetics of berberine and its main metabolites in conventional and pseudo germ-free rats determined by liquid chromatography/ion trap mass spectrometry. Drug Metab Dispos. 2006;34(12):2064–2072.


Müller R, Peters K, Becker R, Kruss B. Nanosuspensions – a novel formulation for the iv administration of poorly soluble drugs. Paper presented at: 1st World Meeting on Pharmaceutics, Biopharmaceutics, Pharmaceutical Technology, APGI 7th International Conference on Pharmaceutical Technology, 41st Annual Congress of APV; May 9–11, 1995; Budapest.


Müller R, Peters K, Becker R, Kruss B. Nanosuspensions for the iv administration of poorly soluble drugs-stability during sterilization and long-term storage. Paper presented at: Proceedings of the 22nd International Symposium on Controlled Release of Bioactive Materials; July 30–August 2, 1995.


Keck CM, Müller RH. Drug nanocrystals of poorly soluble drugs produced by high pressure homogenisation. Eur J Pharm Biopharm. 2006;62(1):3–16.


Pattekari P, Zheng Z, Zhang X, Levchenko T, Torchilin V, Lvov Y. Top-down and bottom-up approaches in production of aqueous nanocolloids of low solubility drug paclitaxel. Phys Chem Chem Phys. 2011;13(19):9014–9019.


Jacobs C, Müller RH. Production and characterization of a budesonide nanosuspension for pulmonary administration. Pharm Res. 2002;19(2):189–194.


Kakran M, Sahoo NG, Tan I-L, Li L. Preparation of nanoparticles of poorly water-soluble antioxidant curcumin by antisolvent precipitation methods. J Nanopart Res. 2012;14(3):1–11.


Bilati U, Allémann E, Doelker E. Development of a nanoprecipitation method intended for the entrapment of hydrophilic drugs into nanoparticles. Eur J Pharm Sci. 2005;24(1):67–75.


Horn D, Rieger J. Organic nanoparticles in the aqueous phase – theory, experiment, and use. Angew Chem Int Ed Eng. 2001;40(23):4330–4361.


Rogers TL, Gillespie IB, Hitt JE, et al. Development and characterization of a scalable controlled precipitation process to enhance the dissolution of poorly water-soluble drugs. Pharm Res. 2004;21(11):2048–2057.


Sahibzada MUK, Sadiq A, Khan S, Faidah HS. Fabrication, characterization and in vitro evaluation of silibinin nanoparticles: an attempt to enhance its oral bioavailability. Drug Des Devel Ther. 2017;11:1453–1464.


Pattni BS, Chupin VV, Torchilin VP. New developments in liposomal drug delivery. Chem Rev. 2015;115(19):10938–10966.


Bhattacharya S, Ghosh A. Phytosomes: the emerging technology for enhancement of bioavailability of botanicals and nutraceuticals. Internet J Aesthet Antiaging Med. 2009;2(1):141–153.


Taylor CT, Winter DC, Skelly MM, et al. Berberine inhibits ion transport in human colonic epithelia. Eur J Pharmacol. 1999;368(1):111–118.


Pan G-Y, Huang Z-J, Wang G-J, et al. The antihyperglycaemic activity of berberine arises from a decrease of glucose absorption. Planta Med. 2003;69(07):632–636.


Küpeli E, Koşar M, Yeşilada E, Başer KHC. A comparative study on the anti-inflammatory, antinociceptive and antipyretic effects of isoquinoline alkaloids from the roots of Turkish Berberis species. Life Sci. 2002;72(6):645–657.


Yeşilada E, Küpeli E. Berberis crataegina DC. root exhibits potent anti-inflammatory, analgesic and febrifuge effects in mice and rats. J Ethnopharmacol. 2002;79(2):237–248.


Le Tran Q, Tezuka Y, Ueda J-Y, et al. In vitro antiplasmodial activity of antimalarial medicinal plants used in Vietnamese traditional medicine. J Ethnopharmacol. 2003;86(2):249–252.


Sánchez-Chapula J. Increase in action potential duration and inhibition of the delayed rectifier outward current IK by berberine in cat ventricular myocytes. Br J Pharmacol. 1996;117(7):1427–1434.


Tsai P-L, Tsai T-H. Hepatobiliary excretion of berberine. Drug Metab Dispos. 2004;32(4):405–412.


Gao S, Basu S, Yang G, Deb A, Hu M. Oral bioavailability challenges of natural products used in cancer chemoprevention. Prog Chem. 2013;25(9):1553–1574.


Jantová S, Cipák L, Cernáková M, Košťálová D. Effect of berberine on proliferation, cell cycle and apoptosis in HeLa and L1210 cells. J Pharm Pharmacol. 2003;55(8):1143–1149.


Kettmann V, Košťálová D, Jantova S, Čerňáková M, Drímal J. In vitro cytotoxicity of berberine against HeLa and L1210 cancer cell lines. Pharmazie. 2004;59(7):548–551.


Teodoro JS, Duarte FV, Gomes AP, et al. Berberine reverts hepatic mitochondrial dysfunction in high-fat fed rats: a possible role for SirT3 activation. Mitochondrion. 2013;13(6):637–646.


Mišík V, Bezáková L, Máleková L, Košťálová D. Lipoxygenase inhibition and antioxidant properties of protoberberine and aporphine alkaloids isolated from Mahonia aquifolium. Planta Med. 1995;61(4):372–373.


Hayashi K, Minoda K, Nagaoka Y, Hayashi T, Uesato S. Antiviral activity of berberine and related compounds against human cytomegalovirus. Bioorg Med Chem Lett. 2007;17(6):1562–1564.


Birdsall TC, Kelly GS. Berberine therapeutic potential of an alkaloid found in several medicinal plants. Alt Med Rev. 1997;2(2):94–103.


Battu SK, Repka MA, Maddineni S, Chittiboyina AG, Avery MA, Majumdar S. Physicochemical characterization of berberine chloride: a perspective in the development of a solution dosage form for oral delivery. AAPS PharmSciTech. 2010;11(3):1466–1475.


Madara JL. Loosening tight junctions. Lessons from the intestine. J Clin Invest. 1989;83(4):1089–1094.


Kwok PC, Chan H-K. Nanotechnology versus other techniques in improving drug dissolution. Curr Pharm Des. 2014;20(3):474–482.


Ma S, Wang Y, Shang X, Yan F. Formulation of berberine hydrochloride and hydroxypropyl-β-cyclodextrin inclusion complex with enhanced dissolution and reduced bitterness. Trop J Pharm Res. 2012;11(6):871–877.


Shi C, Tong Q, Fang J, Wang C, Wu J, Wang W. Preparation, characterization and in vivo studies of amorphous solid dispersion of berberine with hydrogenated phosphatidylcholine. Eur J Pharm Sci. 2015;74:11–17.


Zhaojie M, Ming Z, Shengnan W, et al. Amorphous solid dispersion of berberine with absorption enhancer demonstrates a remarkable hypoglycemic effect via improving its bioavailability. Int J Pharm. 2014;467(1–2):50–59.


Mutalik S, Anju P, Manoj K, Usha AN. Enhancement of dissolution rate and bioavailability of aceclofenac: a chitosan-based solvent change approach. Int J Pharm. 2008;350(1):279–290.


Clinical and Laboratory Standards Institute. M02-A12: Performance standards for antimicrobial disc susceptibility testing. 2015. Available from:


Zhang SL, Chang JJ, Damu GL, et al. Novel berberine triazoles: synthesis, antimicrobial evaluation and competitive interactions with metal ions to human serum albumin. Bioorg Med Chem Lett. 2013;23(4):1008–1012.


Patel RP, Patel MM. Solid-state characterization and dissolution properties of lovastatin hydroxypropyl-β-cyclodextrin inclusion complex. Pharm Tech. 2007;31(2):72–81.


Kakran M, Sahoo N, Li L, et al. Fabrication of drug nanoparticles by evaporative precipitation of nanosuspension. Int J Pharm. 2010;383(1):285–292.


Shid RL, Dhole SN, Kulkarni N, Shid SL. Formulation and evaluation of nanosuspension delivery system for simvastatin. Int J Pharm Sci Nanotechnol. 2014;7:2459–2476.


Jiang T, Han N, Zhao B, Xie Y, Wang S. Enhanced dissolution rate and oral bioavailability of simvastatin nanocrystal prepared by sonoprecipitation. Drug Dev Ind Pharm. 2012;38(10):1230–1239.


Leuner C, Dressman J. Improving drug solubility for oral delivery using solid dispersions. Eur J Pharm Biopharm. 2000;50(1):47–60.


Hancock BC, Parks M. What is the true solubility advantage for amorphous pharmaceuticals? Pharm Res. 2000;17(4):397–404.


Murdande SB, Pikal MJ, Shanker RM, Bogner RH. Solubility advantage of amorphous pharmaceuticals: I. A thermodynamic analysis. J Pharm Sci. 2010;99(3):1254–1264.


Murdande SB, Pikal MJ, Shanker RM, Bogner RH. Solubility advantage of amorphous pharmaceuticals: II. Application of quantitative thermodynamic relationships for prediction of solubility enhancement in structurally diverse insoluble pharmaceuticals. Pharm Res. 2010;27(12):2704–2714.


Müller RH, Junghanns J. Drug nanocrystals/nanosuspensions for the delivery of poorly soluble drugs. In: Torchilin VP, editor. Nanoparticulates as Drug Carriers. London: Imperial College Press; 2006:307–328.


de Oliveira DR, Tintino SR, Braga MF, et al. In vitro antimicrobial and modulatory activity of the natural products silymarin and silibinin. Biomed Res Int. 2015;2015:292797.


Sun D, Abraham SN, Beachey EH. Influence of berberine sulfate on synthesis and expression of Pap fimbrial adhesin in uropathogenic Escherichia coli. Antimicrob Agents Chemother. 1988;32(8):1274–1277.


Park K-S, Kang K-C, Kim J-H, Adams DJ, Johng T-N, Paik Y-K. Differential inhibitory effects of protoberberines on sterol and chitin biosyntheses in Candida albicans. J Antimicrob Chemother. 1999;43(5):667–674.


Quan H, Cao Y-Y, Xu Z, et al. Potent in vitro synergism of fluconazole and berberine chloride against clinical isolates of Candida albicans resistant to fluconazole. Antimicrob Agents Chemother. 2006;50(3):1096–1099.

Creative Commons License This work is published and licensed by Dove Medical Press Limited. The full terms of this license are available at and incorporate the Creative Commons Attribution - Non Commercial (unported, v3.0) License. By accessing the work you hereby accept the Terms. Non-commercial uses of the work are permitted without any further permission from Dove Medical Press Limited, provided the work is properly attributed. For permission for commercial use of this work, please see paragraphs 4.2 and 5 of our Terms.

Download Article [PDF]