Research Article

Absorption enhancing activity of seven herbal extracts on ciprofloxacin using everted sac method

Vedamurthy Joshi1,2*, Shivakumar Inamdar3, Rajesh Kowti1, Sachin A.B.1, Ankit Acharya1

1Sri adichunchananagiri College of Pharmacy, B G Nagara, Karnataka, India

2Research scholar, PRIST University, Vallam, Thanjavur, Tamil Nadu, India

3HKES College of Pharmacy, Gulbarga, Karnataka, India

*For correspondence

Vedamurthy Joshi,

Asst. Professor,

Department of Pharmaceutics, S.A.C. College of Pharmacy,

B.G. Nagar, Karnataka, India.

E.mail: vedamurthyjoshi @gmail.com

 

 

 

 

 

 

 

 

 

Received: 05 July 2016

Accepted: 27 July 2016

ABSTRACT

Objective: Ciprofloxacin is a broad-spectrum fluoroquinolone antibiotic with poor permeability. The present study aimed to investigate the influence of seven alcoholic herbal extracts on the in-vitro absorption of ciprofloxacin using everted sac method.

Methods: The herbal extracts include Curcuma longa (turmeric), Piper nigrum (black piper), Moringa oleifera (drum stick), Piper betle (betel leaf), Trachyspermum ammi (ajwain seeds), Murraya koenigii (curry leaves), Plumbago zeylanica. Influence of Verapamil, a known P-gp inhibitor on ciprofloxacin absorption was also studied. Drug-extract compatibility studies were carried with FTIR. Histological study was carried to detect the influence of the extract on the isolated intestine.

Results: The results indicated significant improvement in the apparent permeability coefficient (Paa) of ciprofloxacin when co-incubated with plant extracts. Amongst all herbal extracts, Piper nigrum extract showed the highest percentage of ciprofloxacin permeability i.e. Paa increases from 15.245 cm/sec to 18.012 cm/sec, which was 18.24% higher than ciprofloxacin alone as control. Paa of ciprofloxacin (3.0 mM) was found increased in the following order ranging from highest to lowest. Piper nigrum >Curcuma longa >Piper betle >Murraya koenigii >Moringa oleifera >Plumbago zeylanica >Trachyspermum ammi.

Conclusions: In conclusion, the permeability of Ciprofloxacin can be increased with the co incubation of the above extracts.

Keywords: Ciprofloxacin, Everted sac method, Permeability coefficient, Bioenhancers, Piper nigrum, Moringa oleifera, Piper betle, Trachyspermum ammi, Murraya koenigii, Plumbago zeylanica

 

Introduction

Herbs and their products are globally used for treatment of huge number of pathological conditions. According to a UNAIDS survey, one-third of the people in developed countries and over 80% of people from Africa depend on herbal medicines to treat common ailments such as cold, cough, inflammatory disorders, heart disease, diabetes, and central nervous system diseases.1 There is also much evidence that large number of people use herbal medicines along with chemical drugs. Although herbal medicine is natural, it contains a complex mixture of several active and inactive chemical constituents and involved directly and indirectly in drug metabolism and transport. To confirm patient safety and efficacy, large number of research is being conducted to determine possible harmful interactions between these herbal products and prescribed medications.2

The oral bioavailability of ciprofloxacin is 50–80% and at least 10% of ciprofloxacin is eliminated via intestinal secretion, of which <1% is due to biliary excretion. Further the resistance of the drug is also increasing in the population.3 A number of studies have reported that ciprofloxacin is a substrate for P-gp and also the substrate of absorptive transporters.4 In this context ciprofloxacin is chosen as a model candidate for P-gp.

P-glycoprotein (P-gp) is the ATP-binding cassette (ABC) transporter primarily found in epithelial cells lining the colon, small intestine, pancreatic ductules, bile ductules, kidney proximal tubules, adrenal gland also in the endothelial cells of the blood-brain barrier. It acts as a physiological barrier and protects these susceptible organs from toxic compounds, preventing them to enter the cytosol and extrude them to the exterior, thus enhances the secretion of metabolites and xenobiotics into bile, urine, and the lumen of gastrointestinal tract5. P-gp shows greater impact on limiting cellular uptake of many hydrophobic drugs (anticancer agents, immunosuppressants, steroid hormones, calcium-channel blockers, beta-adrenoreceptor blockers, cardiac glycosides, antibiotics) from the intestinal lumen into epithelial cells than on enhancing the excretion of drugs out of hepatocytes and renal tubules into the adjacent luminal space.6 Therefore, P-gp can potentially reduce the oral absorption and bioavailability of these drugs and thus decrease the retention time of a number of drugs.7 To minimize these problems, various chemicals inhibitors have been used to overcome P-gp mediated efflux, but unfortunately currently used P-gp inhibitors did not show the promising results. So, consequently, research on herbal P-gp inhibitors identification has been performed.8 Herbal components form the ideal P-gp inhibitor as they have been consumed since long with a view to improve health status of humans. The potential of herbal constituents to act as P-gp inhibitors, enabling increased absorption, brain penetration, inhibition of excretion and modulation of multi-drug resistance. However, the detail studies on herbal products and their role in P-gp inhibitions need to be performed for better efficacy of the drug.9 Ciprofloxacin belongs to class III drug of Biopharmaceutical classification system characterized by low permeability and high solubility.10 Drug is also known to secrete in the intestine may be by P gp efflux.11

This work focuses on influence of alcoholic extracts of Curcuma longa (turmeric), Piper nigrum (black pepper), Moringa oleifera (drum stick), Piper betle (betel leaf), Trachyspermum ammi (ajwain seeds), Murraya koenigii (curry leaves) and roots of Plumbago zeylanica on the absorption of ciprofloxacin using everted sac method. Further to study the influence of Verapamil, a known P-gp inhibitor on the in-vitro absorption of ciprofloxacin.

Materials and Methods

Materials

Ciprofloxacin was obtained as gift sample from Biocon Lab, Bangalore. Turmeric, drum stick leaves, betel leaf, ajwain seeds and black pepper were procured from the local market (Bellur, B.G. Nagara, Karnataka). Ethanol was procured from SD Fine Chemicals, Mumbai. All other reagents used were analytical grade. Double distilled water was used throughout experiment.

Animal used

Male albino rats weighing 170-220 g on standard feed and tap water were used throughout study and were maintained under standard lab conditions (22±3oC) with 12 h light and 12 h dark exposure. Animals were fasted overnight before scarification.

Ethical approval

The study protocol was approved by Institutional Animal Ethics Committee (IAEC), Sri Adichunchanagiri College of Pharmacy, B.G. Nagara-571448, Karnataka, India. Reg. no.SACCP/IAEC/271(c)/2014-15.

Methods

Plant extraction

Roots of Curcuma longa and Plumbago zeylanica; seeds of Trachyspermum ammi) and Piper nigrumwere purchased from the local market of Bellur, dried and powdered using a mixer (Philips 750 watts). Fresh leaves of Murraya koenigii, Moringa oleifera and Piper betle were collected locally, dried in a cool place about 4 days at the laboratory further kept at 40oC at for 4 h in hot air oven (Kadavil electro mechanical Ind., Kerala) finely powdered and packed in the self-sealing plastic cover until further use. Dried powders were extracted with 99% ethanol using a Soxhlet extraction apparatus. The 100 g power was put in Soxhlet thimble and put into a Soxhlet thimble tube. 1000 ml of ethanol was added to a Soxhlet flask, and then extracted at 60°C until the extract was not clear. The ethanol was removed under pressure using a rotator evaporator (Buchi Rota evaporator, Singapore). Then dried residual extract were stored in amber coloured bottle in refrigerator (Samsung) at 4°C and next step was evaluation of chemical constituents.12

Phytochemical evaluation

Qualitative estimation

Phytoconstituents such as carbohydrates, proteins, Alkaloids, glycosides, terpenes, steroids, flavanoids, tannins and saponins were determined using crude extracts. Methods for phytochemical evaluation were employed as per Kokate CK et al13 and Harborne JB et al14. Methods for determination of chemical constituents were described briefly.

Carbohydrates

The extract was dissolved in 10 ml of distilled water and filtered through Whatmann filter paper and the filtrate is subjected to various tests for carbohydrates.

Molish test

Place 2 ml of solution in a test tube. 1 drop of Molish reagent was added to the same test tube. Then 2 ml of conc. HCL was added from the sides of the test tube. The test tube was observed for formation of a violet ring at the junction of the two liquids. If violet ring appears at the junction of the two liquids, it confirms the presence of carbohydrates.

Protein

The crude extract was dissolved in 8 ml of distilled water and filtered through Whatmann filter paper, and the filtrate was subjected to tests for proteins and amino acids.

Millons test

To 2 ml of filtrate, few drops of Millon's reagent are added. A white precipitates was obtained, which indicate presences of proteins.

Biuret test

An aliquot of 2 ml of filtrate was treated with drop of 2% copper sulphate solution. To this, 1 ml of ethanol (95%) was added, followed by excess of potassium hydroxide pellets. The pink color in ethanol layer indicated presences of proteins.

Alkaloid

About 50 mg of dried extract was stirred with 3 ml of dil. HCl and then filtered thoroughly. The filtrate was utilized for various tests for alkaloid.

Mayer's test

To a 1 ml of filtrate, few drops of Mayer's reagent are added by the side of the test tube. The white or creamy precipitate indicated test as positive.

Wagner's test

To a 1 ml of filtrate, few drops of Wagner's reagent are added by the side of the test tube. The color change was observed. Appearance of reddish-brown precipitates confirms the test as positive.

Dragendorff's test

To 1 ml of filtrate, 2 ml of Dragendorff's reagent was added.

Appearance of the prominent yellow precipitate confirms the presence of Alkaloid.

Glycosides

Borntrager's Test

Crude extract was boiled with dilute sulphuric acid and filtered. Filtrate was shaken with chloroform. The organic layer was separated to which ammonia is added slowly. If pink color appears, it indicates the presence of glycoside 15.

Legal Test

The crude extract was dissolved in pyridine and then adds sodium nitroprusside. Appearance of pink or red color indicates the presence of glycoside.

Terpenoid and steroid

4 mg of crude extract was treated with 0.5 ml of acetic anhydride and 0.5 ml of chloroform. Then conc. sulphuric acid was added slowly. Appearance of red violet color indicates the presence ofterpenoid and green bluish color indicates steroids.

Flavonoid

4 mg of crude extract was treated with 1.5 ml of 50% methanol solution. The solution was warmed and metal magnesium was added. To this solution, 5-6 drops of conc. HCl was added. Appearance of red color indicates presence of flavonoids and orange color indicates flavones.

Tannins

0.5 ml of crude extract was taken in 1 ml of water and 1-2 drops of ferric chloride solution was added to it. Appearance of blue color indicates the presence of gallic tannins and green black indicates catecholic tannins.

Saponins

About 200 mg of the crude extract was shaken with 5 ml of RO water and then heated to boil. Appearance of creamy miss of small bubbles shows the presence of saponins.

Quantitative estimation

Ethanolic extracts of Curcuma longa, Plumbago zeylanica, Trachyspermum ammi,Piper nigrum, Murraya koenigii, Moringaoleifera and Piper betle were subjected to quantitative evaluation like moisture content, ash values, extractive values and pH (three determinations for each parameter).

Screening of P-Glycoprotein (Pgp) modulation activity

Composition and preparation of Krebs-Ringer's solution (KRS)

Krebs-Ringer's solution of composition (g/L): NaCl 6.9 g, KCl 0.35 g, MgSO4.7H2O 0.29 g, KH2PO4 0.16 g, NaHCO3 2.1 g, CaCl2 0.28 g and 0.2% glucose (pH 7.3). Except CaCl2, all ingredients were dissolved in 600 ml of purified water in 1 litre volumetric flask. In a separate 250 ml beaker containing 100 ml purified water, CaCl2 was dissolved. The CaCl2 solution was added to the above solution and made up to 1000 ml using purified water.

Everted rat intestine apparatus

The experiment setup is designed as per Dixit P et al.16 The apparatus consists of two cylindrical glass tubes; one (110×17 mm) joined to other (48×17 mm) via J-shaped tapering end (Figure 1 (a and b). Both the tubes are held together by a glass joint on the upper end. On the lower ends of both tubes, a bulge is given for proper mounting of tissue. The dimensions of the apparatus (110×49×17 mm) are such that it can be conveniently set up in a 250 ml glass beaker. After mounting the everted intestinal segment on the apparatus and setting it in the beaker; the inside of the glass tubes serves as the mucosal compartment and the beaker serves as the serosal compartment.17

Preparation of everted intestinal sac

The everted intestinal sac technique is a suitable and universal in-vitro model for the study of intestinal transference of the drugs. After being fasted overnight with free access to water, threats were anesthetized by chloroform. Their abdomen opened with a middle incision. The underlying mesenterium and fat were removed, and the sacs were carefully everted with a glass rod (2.5mm diameter). A 7 cm segment, 3-4 cm distant to the ileocaecal junction was excised, and it was removed of the mesenteric attachments carefully without damaging the intestinal architecture. The intestinal segment was transferred to a petri dish containing KRS which continuously bubbled with a gas mixture of 95% O2 and 5% CO2. This segment carefully mounted on the assembly using nylon thread.17

Permeability markers

Phenol red was used as permeability marker as it was reported to have zero permeability. This marker ensures that the intestine is intact and leak proof throughout the experiment.16

Sample preparation

3.01/ 1.5 mM ciprofloxacin solution was prepared by dissolving the drug in KRS solution. Drug solution was placed in the mucosal part of the assembly (Figure 1) along with the 4 drops of phenol red as a permeability marker. Drug transport across everted sac was measured by withdrawing 1 ml sample from serosal side at 5 min intervals using glass syringe followed by replacing the loss using KRS solution. The sample withdrawn diluted suitably with KRS and estimated the drug content using Colorimetry. After each trial, everted tissue carefully dismounted from the assembly, washed with current of potable water, cut open one side to measure the length and breadth for further calculation. The Influence of herbal extracts on the ciprofloxacin absorption was studied by adding the herbal extracts along with the ciprofloxacin solution in the mucosal side of the assembly. The combination of Verapamil (10.9 mM) and ciprofloxacin acts as a positive control.

Estimation of Verapamil hydrochloride by oxidimetry

Aliquots of pure drug solution (1 to 7 ml) were transferred into a series of 10 ml calibrated flasks. To each flask 1ml of 0.2 M H2SO4 acid solution was added followed by 0.5ml of KMnO4 solution. The contents were mixed and the flasks were heated for 10min. These were cooled and 1ml of Rhodamine-B solution was added to each flask, diluted to the mark with water and the absorbance of solution was measured at 557 nm. A standard graph was prepared by plotting the absorbance versus the concentration of drugs and computed from the regression equation derived using Beer's law.18

Estimation of ciprofloxacin by colorimetry

Various aliquots of ciprofloxacin standard solutions ranging from 5 mg to 50 mg prepared in 10 ml volumetric flask. To the solution, 0.5 ml of 0.7% 1,2-naphthaquinone-4- sodium sulphonate reagent solution and 0.2 mL of 0.1 M NaOH aqueous solutions were added. Each solution was made up to the mark with distilled water, and the absorbance of the coloured solutions were measured against a reagent blank at 487 nm. Calibration curve was prepared by plotting absorbance as a function of the drug concentration.19

Permeability determination

The apparent permeability (Papp) value was calculated according to the following equation:

Papp

Where, Papp is apparent permeability co-efficient, dQ/dT the cumulative amount of drug (Q) appearing in the acceptor (serosal) compartment as a function of time, and was obtained from the slope of the linear portion of the amount transported-versus-time plot, A is surface area of the intestine (cm2) Co is the initial concentration of drug in the donor compartment (μg/ml). After each experiment, intestinal segment is cut open and surface area is determined by measuring length and breadth 20.

Histopathology studies

After completion of the experiments, a small piece of the everted intestine was fixed with 10% formalin for 24 h and then dehydrated with a graded alcohol series and fixed by using paraffin wax. It was stained by H and E stain and specimens were observed and photo micro- graphed under a confocal microscope.21

Compatibility studies

FT-IR spectrophotometer (Thermo Nicolet FT-IR system) was used for infra-red analysis of samples to interpret the interactions of drug with other excipients.

Results and Discussion

Phytochemical investigations

Photochemical evaluation and quantitative estimation of herbal extracts are summarized in Table 1 and 2. All ethanolic extracts possess alkaloids in common glycosides were detected in all except turmeric extract. Terpenoids and steroids are present in all except the P. zylanica extract. Tannins are present in all except M. olefera. Quantitative tests indicated that moisture was 10-18%, ash content 4-14% and extractive value 14 to 24% and pH was ranging from 6-7. All the phytochemical investigations are carried out in triplicate.

UV method for estimation of ciprofloxacin and verapamil

Calibration curve of ciprofloxacin in KBS solution using UV absorption spectroscopy was linear with a regression coefficient of 0.998 with a slope of 0.069. Titrimetric analysis of the same drug also indicated good linearity with a slope of 0.0154 and an intercept of 0.0078. As the absorption maxima of Verapamil (276 nm) and Ciprofloxacin (278 nm) are very close, Ciprofloxacin was estimated separately when studied in presence of verapamil. Preliminary experiments indicated that everted sac was viable up to 60-90 min; however, we restricted our experiments to 30 min with 5 min sampling time.

Screening of absorption enhancement activity

Apparent permeability coefficient (Paa) of ciprofloxacin absorption was estimated in 1.5mM and 3.0 mM concentration. Results indicated that the drug transport was concentration dependent up to 30 min following first order process. Paa values were higher in 3.0 mM than in 1.5 mM concentration. All the extracts found to enhance the Paa when compared to the control alone. Co incubation of Verapamil resulted in a slight increase of Paa of ciprofloxacin (3.0 mM concentration) from 15.245 cm/sec to 16.718cm/sec whereas in 1.5 mM concentration Paa was from 10.124 to 11.403 revealing that ciprofloxacin absorption is slightly influenced by the P-gp transport. Few cell line studies on ciprofloxacin absorption also depict similar results for incidence22-24 Park MS M et al,4 states that the ratio of B to A /A to B was about 10 MDCKI-MDR1 cells indicating ciprofloxacin is P-gp substrate. However in presence of various inhibitors, the ratio of B to A /A to B increased significantly in presence of indomethacin, sulfinpyrozone and probencid rather than verapamil a known P-gp inhibitor. This indicates the absorption is takes various paths.4

Table 1: Observation of phytochemical constituents present in C. longa, P. nigrum, M. oleifera, P. betle, T. ammi, M. koenigii and P. zeylanica.

Test

Plants

 

C. longa

P. nigrum

M. oleifera

P. betle

T. ammi

M. koenigii

P. zeylanica

Carbohydrates

+

+

-

+

+

-

+

Proteins and Amino-acids

+

+

+

-

+

+

-

Alkaloids

+

+

+

+

+

+

+

Glycosides

-

+

+

+

+

+

+

Terpenoids and Steroids

+

+

+

+

+

+

-

Flavonoids

+

+

-

-

+

+

+

Tannins

+

+

-

+

+

+

+

Saponins

-

+

-

-

+

+

+

(+) indicates presence of Phytochemical, (-) indicates absence of Phytochemical constituents.

Table 2: Results of quantities estimation.

Test

Plants

C. longa

P. nigrum

M. oleifera

P. betel

T. ammi

M. oenigii

P. zeylanica

Moisture

10.5

11.0

15.8

18.51

12.72

12.5

14.9

Ash content

13.42

12.31

14.30

7.85

5.87

4.06

12.61

Acid insoluble Ash

0.8

3.2

2.8

1.42

1.08

1.05

1.7

Water soluble Ash

3.9

5.8

5.4

3.22

2.24

1.24

5.8

Extractive value

19.6

24.11

23.71

22.4

14.81

18.3

22.19

pH

6.5

6.7

6.7

6.6

6.8

6.4

6.3

 

Except pH all results are in percentage.

 

Table 3: Table showing the initial concentration, dQ/dt and Permeability co efficient (Paa).

Sl. No

Sample

Co mM

dQ/dt (slope)

Paa (10-6cm/sec)

1.

Cipro

1.5

0.001412

10.124

2.

Cipro

3.0

0.004301

15.245

3.

Cipro+ Verapamil

1.5

0.001653

11.403

4.

Cipro+ Verapamil

3.0

0.005350

16.718

5.

Cipro+ Curcuma longa extract

1.5

0.001363

12.562

6.

Cipro+ Curcuma longa extract

3.0

0.004287

17.863

7.

Cipro + Plumbago zeylanica extract

1.5

0.001582

14.126

8.

Cipro + Plumbago zeylanica extract

3.0

0.004091

16.235

9.

Cipro+ Trachyspermum ammi extract

1.5

0.002010

13.865

10.

Cipro+ Trachyspermum ammi extract

3.0

0.003997

16.115

11.

Cipro+ Piper nigrum extract

1.5

0.002482

14.56

12.

Cipro+ Piper nigrum extract

3.0

0.005944

18.012

13.

Cipro + Murraya koenigii extract

1.5

0.001640

12.565

14.

Cipro+ Murraya koenigii extract

3.0

0.004380

16.78

15.

Cipro+ Moringa oleifera extract

1.5

0.001393

11.233

16.

Cipro+ Moringa oleifera extract

3.0

0.004242

16.253

17.

Cipro + Piper betle extract

1.5

0.001988

13.256

18.

Cipro+ Piper betle extract

3.0

0.005279

17.596

Table 4: Results of histological examinations

Sl. no

Sample

Cellular infiltration

Villi damage

Intercellular junction

Lumen dilation

Sub-mucosal gland destruction

 

Control

+

-

-

-

-

 

Cipro

+

-

+

-

+

 

Cipro+ Verapamil

+

-

+

+

-

 

Cipro+ Trachyspermum ammi

+

+

+

+

+

 

Cipro + Plumbago zeylanica

+

+

+

+

+

 

Cipro+ Piper nigrum

+

+

+

+

+

 

Cipro+ Murraya koenigii

+

-

+

+

+

 

Cipro+ Moringa oleifera

+

+

+

+

-

 

Cipro+ Piper betle

+

-

+

+

+

Ethanolic plant extracts showed dose dependent increased Paa for Ciprofloxacin indicating more in vitro absorption. Paa (3.0 mM and 1.5 mM) was found in the following order ranging from highest to lowest. P. nigrum >C. longa >P. betle >M. koenigii >M. oleifera >P. zeylanica >T. ammi. Ethanolic extract of P. nigram and turmeric showed absorption 18.24% and 17.2% higher than pure drug respectively indicating the bioenhancing activity. Many studies on cell lines revealed that both, P. nigrum and turmeric showed absorption enhancing action by down regulating p-gp transport and the key contents of P. nigrum is piperin; turmeric are curcumin, demethoxycurcumin and bisdemethoxycurcumin may be responsible for this action.25-29 Further the presence of vital phytochemical constituents like alkaloids, flavonoids, terpenoids and steroids can modulate the main ABC transporters responsible for P-glycoprotein and multidrug resistance-associated protein.30 Especially, flavonoids with high inhibitory activity could modify pharmacokinetics and drug levels of Ciprofloxacin drug and are capable of binding with both the ATP-binding site and the vicinal steroid-binding hydrophobic region (SBHR) within the cytosolic domain of P-gp.30,31 Several reports have also shown that alkaloids and saponins present in these plant extracts are capable of inhibit P-gp function probably by substrate competition activity.32 This could be the reason of greater Paa values of Ciprofloxacin in combination with other extracts of T. ammi, M. koenigii and P. zeylanica. T. ammi is quite popular in India as an appetizer, carminative.33 The essential oil and many phenolic components are present in the seed may be responsible for increased Paa of ciprofloxacin.

These results indicated lucidly that all the extracts are bioenhancers for ciprofloxacin. It is unclear to predict the exact mechanism of action. However, co incubation of these dietary products definitely improve the bioavailability of ciprofloxacin.

FT-IR studies

FTIR studies were conducted in order to explore the possibility of drug extract interaction. As shown in Figure 2, the FT-IR analysis of Ciprofloxacin showed characteristic peaks at 3531.004 m-1 due to hydroxyl group O-H stretching vibration, intermolecular H-bonded, 2930.54 cm-1 due to Aromatic, cyclicenes, 1701.62 cm-1 due to CO group of acid peak assignment C=O stretching vibration, at 1621.06 indicate quinolones because δN-H bending vibration, at 1492.42 indicate carbonyl group because υC-O, at 1307.19 peak assignment by δO-H bending vibration and at 1023.33 indicate fluorine group due to C-F stretching. Moreover, we can observed that FT-IR spectra of Turmeric and P. nigrum showed characteristic peaks (Figure 3 and 4) at the same as Ciprofloxacin with slight variation but within range was observed for the mixture of drug with different herbal extract turmeric and Piper nigram . Hence, it was found that all the herbal extract and animal product used in formulations were compatible with Ciprofloxacin.

Figure 2: FT-IR spectra of Ciprofloxacin.

Figure 3: FT-IR spectra of Ciprofloxacin-Verapamil hydrochloride combination.

 

Figure 4: FT-IR spectra of Ciprofloxacin-turmeric extract combination.

Figure 5: FT-IR spectra of Ciprofloxacin-piper nigram extract combination.

Figure 6: FT-IR spectra of Ciprofloxacin-Drum stick extract combination.

(a) Ciprofloxacin treated.

(b) Ciprofloxacin+ Trachyspermum ammi treated.

(c) Ciprofloxacin +Verapamil treated.

(d) Ciprofloxacin +Turmeric treated.

(e)Ciprofloxacin + Moringa oleifera treated

(f) Ciprofloxacin +Piper nigrum treated

(g) Ciprofloxacin +Betel leaf treated

(h) Ciprofloxacin +Murraya koenigii leaf treated

Figure 7: Histopathological examination of the everted rat intestine with various tested groups.

Histological examination

Histological examination of rat intestine (obtained by everted sac technique) treated with Ciprofloxacin, ciprofloxacin in combination with ethanolic extract of plants, were examined and compared with healthy intestine. Results of histological examinations are shown in Table 4.

Conclusions

In this study, we found that ethanolic extract of plants enhanced the intestinal permeability of Ciprofloxacin. Therefore the permeability of ciprofloxacin can be enhanced by consuming these herbs along with the diet or these herbal extracts can be recommended for better therapeutic efficacy. As no significant interaction was detected in amongst the extracts and the drugs by FTIR, safety can be predicted. As a futuristic approach, Physician can advise the patient to consume the above herbs/ extracts when treated with ciprofloxacin.

Funding: No funding sources

Conflict of interest: None declared

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