Journal of Pharmaceutical Investigation Online ISSN 2093-6214

DOI 10.1007/s40005-016-0298-0 Print ISSN 2093-5552

ORIGINAL ARTICLE

Preparation and evaluation of oral dissolving ilm containing local

anesthetic agent, lidocaine
Bo-Sik Kim1 · Gyu-Thae Park1 · Min-Ho Park1 · Young G. Shin1 · Cheong-Weon Cho1

Received: 26 October 2016 / Accepted: 12 December 2016

© The Korean Society of Pharmaceutical Sciences and Technology 2016

Abstract A lot of polymeric oral dosage forms have Introduction

been invented for suitable drug delivery and compliance
of patient. Oral dissolving ilm (ODF) ofers a comfort- The majority of registered medicines available on the mar-
able way to enable drug administration. The purpose of this ket are oral solid dosage forms. Their advantages include
study was to develop an ODF using lidocaine, with mod- the dose accuracy, relatively high stability, and possibility
erate mechanical strength and thickness, fast disintegration to modify the drug release proile in order to delay or sus-
time and local anesthetic efect. Hydroxypropyl methyl tain the therapeutic efect, as well as to speed it up. How-
cellulose, sodium alginate, glycerin, sodium lauryl sulfate, ever, the application of solid oral dosage forms in drug
polyvinylpyrrolidone and citric acid were used to build the therapy is still associated with many challenging problems.
polymeric ilm base of the ODF. The optimal composition One of them is diiculty with swallowing, mostly encoun-
was determined by changing the ratio of individual excipi- tered in pediatric or geriatric populations (Nissen et al.
ents. The thickness was insigniicantly varied in the range 2009; Hansen et al. 2008), but also in the case of handi-
of 0.04 ± 0.01 to 0.06 ± 0.01 mm. On the other hand, the capped or bedridden patients. It is estimated that some
tensile strength was 1.23 ± 0.20 to 1.83 ± 0.30 N/cm2. Drug swallowing issues associated with solid dosage forms are
content in the ilms was evaluated and the values were experienced by between 20% and up to 50%, of all patients.
16 mg of lidocaine per each ODF (40 mg/strip). Lidocaine They can involve only a little discomfort when adminis-
ODF formulation showed the release of 91.7% of lido- tering large sized tablets or capsules as well as more seri-
caine within 1 min and 96.3% of lidocaine in 5 min. These ous problems such as a drug sticking to the throat mucosa,
results suggested that lidocaine ODF was well prepared and irritation of the pharyngeal region, coughing or choking
showed the fast release of lidocaine. (Stegemann et al. 2012).
Fast-dissolving drug-delivery systems came into exist-
Keywords Oral dissolving ilm · Lidocaine · Tensile ence in the late 1970s as an alternative to tablets, cap-
strength · Cellular uptake sules and syrups for pediatric and geriatric patients who
experience diiculties in swallowing traditional oral solid
dosage forms (Nagendrakumar et al. 2015). Fast dissolv-
ing oral delivery systems are solid dosage forms, which
dissolve within 1 min when placed in the mouth without
drinking or chewing (Cilurzo et al. 2008). After disintegrat-
ing in mouth, enhanced the clinical efect of drug through
pre-gastric absorption from mouth pharynx and esopha-
* Cheong-Weon Cho gus as the saliva passes down into the stomach (Pathare
chocw@[Link] et al. 2013). Oral ilm includes various ingredients for its
1
College of Pharmacy and Institute of Drug Research
formulation which includes polymers, active pharmaceuti-
and Development, Chungnam National University, 99 cal ingredient, ilm stabilizing agents, sweeteners, lavors,
Daehak-ro, Yuseong-gu, Daejeon 34134, South Korea colors, saliva stimulating agents, preservatives, surfactants

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 B.-S. Kim et al.

etc, but the irst and far most a very essential ingredient used as plasticizer in the concentration of 2.4% w/w. Cit-
which helps in ilm formation is a polymer (Siddiqui et al. ric acid was used as saliva stimulating agents to increase
2011). The ilm rapidly hydrates and adheres onto the site the rate of production of saliva that would aid in the faster
of application. It then rapidly disintegrates and dissolves disintegration of the rapid dissolving ilm formulations
to release the medication for oromucosal absorption or (Table 1). Aqueous solution was prepared by dissolving
will maintain the quick-dissolving aspects which allow for ilm-forming polymer and plasticizer in speciic propor-
gastrointestinal absorption to be achieved when swallowed tion in distilled water and stirred for 3 h with a magnetic
(Bala et al. 2013). stirrer. Lidocaine was then added in the dispersion. The
Suitable drug candidate for orally dissolving ilms dispersion was casted onto polyethylene ilm after sonica-
(ODF) should possess as below characteristics; no bit- tion to remove all the air bubbles entrapped and dried at
ter taste, good stability in water in saliva, dose should be desiccator for 2 h. Blank ODF without lidocaine was pre-
low (Varun et al. 2011). The dental injection is one of pared with the same procedure as a control.
the causes of patient’s refusal; therefore, in the clinic, the
dentist applies a topical anesthetic to minimize pain and
fear. The local anesthetic most often used in dentistry Determination of mechanical strength
is lidocaine hydrochloride [2-diethylamineoacetate-2′,
6′-xylidide] (Roh et al. 2016). Diferent forms of lidocaine Thickness was calibrated by vernier calipers (Mitutoyo
have been developed for use as topical anesthetics includ- 500-181-20, Mitutoyo, Japan). Thickness was measured
ing ointments, gels and sprays (Gowacka et al. 2009; Pad- at two diferent points in the ilms and mean value was
ula et al. 2003; Stecker et al. 2002). In this study, lidocaine expressed. Tensile strength of ilm specimens was cali-
as local anesthetics was used to prepare ODF for dental brated by using tensile strength tester (IMADA DS2-5N,
applications. IMADA, Japan).
Tensile strength (N∕cm2 ) = Fmax ∕ A
Materials and methods Fmax is the maximum force at break and A is the initial
cross sectional area of the ilm.
Materials

Lidocaine hydrochloride monohydrate was purchased from Drug content

Sigma (Steinheim, Switzerland). Alginic acid sodium salt
and citric acid anhydrous were purchased from Samchun Film (size of 2 × 3 cm2) was taken from diferent areas of
(Pyeongtaek, Korea). Polyvinylpyrrolidone was obtained the ilm and placed in a 10 mL volumetric lask. 10 mL of
from BASF (Ludwigshafen, Germany). Sodium lauryl sul- phosphate bufer pH 6.8 was added and kept aside till the
fate was purchased from Daejung (Cheongwon, Korea). ilm dissolves completely from this solution. 1 mL was
Hydroxypropylmethylcellulose 2910 was received from pipetted out and diluted to 10 mL with phosphate buf-
Richwood trading Co., Ltd (Pyeongtaek, Korea). Glyc- ered saline (PBS) pH 6.8. The solution was analyzed by
erin was purchased from Samjung (Daejeon, Korea). All UV–Visible (UV) spectrophotometer at 254 nm (Achar-
solvents were analytical grade and used without further jya et al. 2010).
puriication.

Cell cultures

Caco-2 cells were purchased from the Korean cell line Table 1 Composition of lidocaine ODF
bank (Seoul, Korea). Caco-2 cells (passage number 46–52) Component (g) F1 F2 F3
were cultured in MEM supplemented with 10% FBS, 1%
HPMC 2910 3.6 3.6 3.6
NEAA, 100 units/mL penicillin and 0.1 mg/mL streptomy-
Sodium alginate 4.8 4.2 5.28
cin in a 5% CO2 atmosphere with 95% humidity in a 37 °C
Citric acid 0.48 1.08 0
incubator.
Glycerin 2.4 2.4 2.4
Sodium lauryl sulfate 0.36 0.36 0.36
Preparation of ODF
PVP 0.36 0.36 0.36
Lidocaine 8 8 8
The two ilm-forming polymers HPMC and sodium algi-
DW 100 100 100
nate were employed to prepare ODF and glycerin was

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Preparation and evaluation of oral dissolving ilm containing local anesthetic agent,…

Liquid chromatography–mass spectrometry In vitro release study

The liquid chromatography–mass spectrometry system The formulation F3 of lidocaine ODF was placed in
consisted of two Shimadzu LC-20AD pumps, a Shimadzu 30 mL of distilled water at 37 ± 0.5 °C. At predetermined
CBM-20A HPLC pump controller (Shimadzu Corpora- time intervals (1, 3, 5, 10 min), an aliquot of 3 mL of
tion, Columbia, MD, USA), a CTC HTS PAL autosam- the release media was withdrawn and the concentration
pler (CEAP Technologies, Carrboro, NC, USA) and a of the release media was estimated by UV spectropho-
quadrupole time-of-light (QqTOF) TripleTOF™ 5600 tometer at 254 nm. The medium was replaced with fresh
mass spectrometer (AB Sciex, Foster City, CA, USA). bufer (3 mL) to maintain constant volume.
The analytical column used for this assay was a Phenom-
enex Kinetex XB-C18 column, 2.1 × 50 (2.6 μm). The
mobile phase consisted of: mobile phase A, distilled and Cell viability assay
deionized water containing 0.1% formic acid; and mobile
phase B, acetonitrile containing 0.1% formic acid. The After 24 h incubation of Caco-2 cells (70% conluent)
gradient was as follows: from t = 0 min to t = 0.5 min, 5% with medium solubilized blank ODF or lidocaine ODF,
B; from t = 0.5 min to 1.5 min by a linear gradient from the cytotoxicity was determined by MTT assay accord-
5% B to 95% B; 95% B was maintained for 0.2 min; from ing to the manufacturer’s protocol (Sigma, USA). Briely,
t = 1.7 min to t = 1.8 min by a linear gradient from 95% after incubation, MTT (3-(4,5-dimethylthiazol-2yl)-
B to 5% B, and then 5% B was maintained for 1.2 min. 2,5-diphenyl-2H-tetrazolium bromide, Sigma) was added
The gradient was delivered at a low rate of 0.4 mL/min. to each well and incubated for 2 h at 37 °C. The crys-
The injection volume was 10 μL. TOF-MS/MS scan tals of viable cells were solubilized in isopropanol. The
mass spectra were recorded in the positive ion mode. For absorbance was determined at 570 nm in a microplate
TOF-MS/MS scan, m/z 50–300 with 0.25 s accumulation reader (Sunrise, Tecan, Austria). Cell viability (%) was
time was used. High-purity nitrogen gas was used for the represented with the (OD of samples-treated cells divided
nebulizer/Duospray™ and curtain gases. The source tem- by OD of cells incubated without samples) × 100.
perature was set at 500 °C with a curtain gas low of 30 L/
min. The ion spray voltage was set at 5500 V, decluster-
ing potential was 100 V and the collision energy was Cellular uptake study
21 V. For the quantiication, lidocaine was analyzed using
LC-TOF-MS/MS scan method. [M + H]+ ion for lido- One day before the uptake experiments, Caco-2 cells in
caine (m/z 235.2) was selected and its product ion at m/z 6-well plates were seeded at a density of 5 × 105 cells/
86.0950 was used for quantitative analysis for lidocaine. well. The cells were washed twice with serum-free
MEM, and then exposed to either with serum-free MEM,
200 µg/mL of pure lidocaine solution or medium solubi-
Physicochemical characterization of ODF lized lidocaine ODF for 2 h. Subsequently, the cellular
uptake studies were terminated by aspirating the media
The morphology and surface characteristics of lidocaine and washing the cells three times with phosphate buf-
ODF were examined by scanning electron microscopy ered saline (PBS). The cells were then lysed with 800 μL
(SEM) (JEOL JSM7500 Field Emission Scanning Elec- of 0.2 N NaOH. Aliquots of the cell lysate solution were
tron Microscope, Thermo, USA) operating at an acceler- removed for analysis of lidocaine and protein content,
ating voltage of 20 kV. The samples were mounted onto respectively. After vortexing for 1 min, the mixture was
metal stubs using double sided adhesive tape onto which centrifuged at 15,000 rpm for 10 min. The supernatants
the samples were applied. The stubs were sputter-coated were diluted in water and injected into the HPLC/MS/MS
with gold particles in a sputter coater for 50 s. The lido- system for analysis. The amount of protein in each sam-
caine ODF was characterized at the solid state using Fou- ple was determined by the BCA assay kit (Sigma, Stein-
rier transform infra-red (NICOLET 380 FT-IR, Thermo, heim, Switzerland).
USA). A powder 5 mg with attached ATR prism was
recorded. Lidocaine ODF was scanned in the IR range
from 400 to 4000 cm− 1. The changes of crystallinity were Statistical analysis
observed by XRD (Multilab 2000, Thermo Scientiic,
USA). Data were collected over an angular range com- The Student’s t test was used to compare two difer-
prised between 5° and 70° with a step size of 0.01° ent groups of samples. A p value < 0.05 was considered
signiicant.

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 B.-S. Kim et al.

Results and discussion Table 2 Mean thickness and tensile strength of lidocaine ODF
F1 F2 F3
Evaluation of ODF
Thickness (mm) 0.04 ± 0.01 0.06 ± 0.01 0.05 ± 0.01
Three kinds of lidocaine ODF was prepared with the prepa- Tensile (N/cm2) 1.63 ± 0.30 1.23 ± 0.20 1.83 ± 0.30
ration process in Fig. 1. Lidocaine ODF was adjusted with
the size of 2 × 3 cm2 and measured with weight of 40 mg/
strip. The thickness of each ilm was measured using ver- uniform quantity of the drug, as per content uniformity
nier caliper (thickness tester) at diferent positions of the studies indicating reproducibility of the technique. It was
ilm and the average was calculated (Table 2). As all the reported that 2% lidocaine was used in dentistry (Rozanski
formulations contained diferent amounts of sodium algi- et al. 1988). Lidocaine ODF might have a potential to elicit
nate, hence the thickness was insigniicantly varied in the the local anesthetic action because drug content in the ilms
range of 0.04 ± 0.01 to 0.06 ± 0.01 mm. On the other hand, was were 16 mg of lidocaine per each ODF (40 mg/strip).
the tensile strength was 1.23 ± 0.20 to 1.83 ± 0.30 N/cm2.
There is no signiicant diference between blank ODF and Physicochemical characterization of ODF
lidocaine ODF in mean thickness and tensile strength. F3
showed the highest tensile strength, suggesting sodium Figure 2 reveals the X-ray pattern of lidocaine and ODF
alginate played an important role in strength of ODF. For in the absence/presence of lidocaine. Lidocaine ODF
buccal administration, strong and lexible ilms are more was appeared that lidocaine was embedded as an amor-
preferable. All the strips were found to contain an almost phous form. Even if blank ODF without lidocaine was

Fig. 1 Lidocaine ODF. a Image (a)

of ODF; b manufacturing pro-
cess of ODF

(b)
Sodium Alginate 5.28g was stirred in 80mL DW at 40°C

HPMC 3.6g, Glycerin 2.4g

Homogenized at 2,000 rpm for 20 min

Lidocaine HCl 8g

(stirred In 20mL DW at 40°C)

Homogenized at 2,000 rpm for 20 min

Cut into single

Degas for 60 min Cast on a PP liner Dry in a desiccator for 3
film sheets (2Ő

3 cm2 )

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Preparation and evaluation of oral dissolving ilm containing local anesthetic agent,…

lidocaine with decreased intensity which could be due to

the dilution of the mixture by the polymer. The results of
FT-IR study showed the absence of new chemical bonds
between lidocaine and the polymer indicating the absence
of interaction between the drug and polymer.

In vitro release study

Lidocaine ODF showed a fast release by in vitro release

test, expecting the rapid absorption and moderate anes-
thetic efect (Table 3). Lidocaine ODF formulation showed
the release of 91.7% of lidocaine within 1 min and 96.3%
of lidocaine in 5 min. There was a report that lupentixol
ODF was prepared using HPMC and carboxymethyl cellu-
lose, which showed more than 95% release of lupentixol in
5 min. Drug release rate was decreased by increasing the
amount of HPMC E5 and CMC in the formulation (Abdel-
bary et al. 2014). The combination of nonionic gelling
polymer such as HPMC and anionic polymer such as car-
boxymethyl cellulose or sodium alginate showed the rapid
release of dug.

Cell viability assay

ODF should be non-toxic because the ilm rapidly disinte-

grates and dissolves to release the medication for mucosal
absorption or for gastrointestinal absorption to be achieved
when swallowed. Because ODF was applied in mouth, the
cell viability should be performed in relevant oro-mucosal
cells. However, Caco-2 cells were used because it could be
swallowed. In cell viability assay, blank ilm was viable in
Fig. 2 X-ray difraction of lidocaine (a), blank ODF (b) and lido- the concentration according to 0.33 mg/mL of lidocaine
caine ODF (c)
and lidocaine ODF showed the cell viability of 20% at the
same concentration, suggesting ODF ilm was nontoxic
transparent, the crystalline peak was observed. However, (Fig. 4).
the intensity of blank ODF was lower than that for the pure
lidocaine due to the low amount of lidocaine in the ilm. Cellular uptake study
This result was well correlated with the other publication
(Preis et al. 2014). We observed the 96.3% release of lidocaine from lidocaine
The FT-IR studies were carried out to assess any pos- ODF by in vitro release study. Subsequently, the cellular
sible interaction between lidocaine and ODF composition uptake study was carried out whether the released lidocaine
in the solid state. The FT-IR spectrum of lidocaine, blank could be uptake into cells. Based on cytotoxicity results,
ODF, and lidocaine ODF was shown in Fig. 3. The ben- 200 µg/mL of lidocaine was treated into Caco-2 cells. Lido-
zene ring helps produce the signal at about the 3030 cm− 1 caine ODF was treated with the solubilized form into cells.
range. While the C=C produce a strong signal at the Lidocaine solution or lidocaine ODF showed the lidocaine
1450–1600 cm− 1. The H–N–C=O is an amide group which uptake amount of 1.68 ± 0.12 or 0.92 ± 0.03 µg normalized
has a signal appearing in the 3000–3500 cm− 1. The main by protein, respectively (Fig. 5). The percentage of cellu-
absorbance band of HPMC is at 1045 cm− 1 (C–O stretch lar uptake of lidocaine from lidocaine solution or lidocaine
vibration), but several smaller peaks are found between ODF is 0.79 or 0.47%, respectively. This might be due to
1200 and 1500 cm− 1 (Abdelbary et al. 2014). A carbonyl viscosity of HPMC and sodium alginate. This suggested
group is producing a signal in the 1630–1690 cm− 1. Lido- that lidocaine ODF could be used for dental applications
caine ODF presents all the major absorption peaks of instead of lidocaine injection.

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 B.-S. Kim et al.

Fig. 3 FT-IR of lidocaine a, blank ODF b and lidocaine ODF c

Table 3 In vitro release study of lidocaine from lidocaine ODF Conclusion

Time (min) 1 3 5 10
We designed the ODF containing lidocaine by controlling
Absorbance 0.529 ± 0.03 0.547 ± 0.02 0.561 ± 0.02 0.578 ± 0.12 the polymer ratio such as sodium alginate and HPMC. As
(OD) a result, lidocaine ODF had the moderate mechanical prop-
Content 14.67 15.09 15.41 15.80 erty and rapidly released the lidocaine.
(mg)
Released (%) 91.71 94.29 96.30 98.74

Fig. 4 Cytotoxicity of blank

ODF, lidocaine ODF against
Caco-2 cells after 24 h

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Preparation and evaluation of oral dissolving ilm containing local anesthetic agent,…

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Acknowledgements All authors (B.S. Kim, G.T. Park, M.H. Park, Roh JY, Han MR, Kim KN, Kim KM (2016) The in vitro and in vivo
Y.G. Shin, C.W. Cho) declare that they have no conlict of interest. efects of a fast-dissolving mucoadhesive bi-layered strip as topi-
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