Materials Today: Proceedings xxx (xxxx) xxx

Contents lists available at ScienceDirect

Materials Today: Proceedings

journal homepage: www.elsevier.com/locate/matpr

An investigation on host-guest complexation of Metformin

hydrochloride with hydroxypropyl-a-cyclodextrin for enhanced oral
bioavailability
S. Lizy Roselet a,⇑, J. Prema Kumari b
a
Holy Cross College (Autonomous), Nagercoil, India
b
Scott Christian College(Autonomous), Nagercoil, India

a r t i c l e i n f o a b s t r a c t

Article history: Metformin hydrochloride is an orally administered biguanide which is an oral anti-hyperglycemic agent.
Received 22 May 2019 It shows incomplete absorption from the gastrointestinal tract and the absolute bioavailability is 50–60%
Accepted 16 June 2019 with relatively short plasma half-life of 1.5–4.5 h. In this study the host-guest complexation of
Available online xxxx
hydroxypropyl-a-cyclodextrin with Metformin hydrochloride is discussed. Inclusion complexes of
Metformin hydrochloride with hydroxypropyl-a-cyclodextrin were prepared in solid and liquid state
Keywords: and they are characterized using spectrophotometry, fluorimetry, 1H NMR and phase solubility studies.
Metformin hydrochloride
The subsistence of host-guest interaction makes hydroxypropyl-a-cyclodextrin a suitable candidate for
Anti-hyperglycemic agent
Bioavailability
the enhancement of bioavailability of Metformin hydrochloride.
Hydroxypropyl-a-cyclodextrin Ó 2019 Elsevier Ltd. All rights reserved.
Inclusion complexes Selection and peer-review under responsibility of the scientific committee of the International Confer-
ence on Recent Trends in Nanomaterials for Energy, Environmental and Engineering Applications.

1. Introduction improved chemical orbiological properties compared to the host

Metformin is widely used for the treatment of type-II diabetes.
Chemically, metformin is a hydrophilic base [1]. However, is usu-
ally present in oral dosage forms in its hydrochloride salt form.
Metformin HCl has acid dissociation constant values (pKa) of 2.8
and 11.5 and, therefore, exists very large as the hydrophilic catio-
nic species at physiological pH values (>99.9%) [2]. The lipid solu-
bility of the unionized species is low as shown by its low water-oil
partition coefficient value (log P = 1.43) [3]. This chemical parame-
ter indicates low lipophilicity and, therefore, rapid passive diffu-
sion of metformin trough cell membranes is unlikely [2]. Based
on these properties, metformin HCl is defined as class III (low per-
meability, high solubility) by the Biopharmaceutics Classification
System (BCS). Poor wettability of drug leads to poor dissolution
and hence, shows variation in bioavailability. Metformin has poor
flow properties. Cyclodextrins are used to form host-guest inclu-
sion complexes with various drugs in solution or a solid state have
been recognized as pharmaceutical excipients [4–7]. Inclusion
complexes formed with a host-guest molecule may exhibit
molecule alone. Such inclusion may: (i) improve aqueous solubil-
ity, dissolution, and bioavailability [8]. Inclusion compounds of
cyclodextrins with hydrophobic molecules are able to penetrate
body tissues, these can be used to release biologically active com-
pounds under specific conditions [9,10]. The chemical name of
Metformin hydrochloride is N, N-dimethyl imido dicarbonimidic-
diamide hydrochloride. Metformin hydrochloride is a white to
off-white crystalline compound with the molecular formula
C4H11N5. HCl and molecular weight of about 165.63. Metformin
hydrochloride is freely soluble in water and is practically insoluble
in acetone, ether and chloroform (see Fig. 1.).
This study involves the synthesis of inclusion complexes with
Metformin hydrochloride and hydroxypropyl-a-cyclodextrin and
the host-guest interaction between them so as to opt it as a means
for enhancing the oral bioavailability of Metformin hydrochloride.
2. Materials and methods
2.1. Reagents and materials

⇑ Corresponding author. Tel.: +91 9486011648. Analytical grade of Metformin hydrochloride (MFH),
E-mail address: lizyroselet@holycrossngl.edu.in (S. Lizy Roselet). Hydroxypropyl-a-cyclodextrin (HPa-CD) were purchased from

https://doi.org/10.1016/j.matpr.2019.06.650
2214-7853/Ó 2019 Elsevier Ltd. All rights reserved.
Selection and peer-review under responsibility of the scientific committee of the International Conference on Recent Trends in Nanomaterials for Energy, Environmental and
Engineering Applications.

Please cite this article as: S. Lizy Roselet and J. Prema Kumari, An investigation on host-guest complexation of Metformin hydrochloride with hydrox-
ypropyl-a-cyclodextrin for enhanced oral bioavailability, Materials Today: Proceedings, https://doi.org/10.1016/j.matpr.2019.06.650
2 S. Lizy Roselet, J. Prema Kumari / Materials Today: Proceedings xxx (xxxx) xxx
Fig. 1. Chemical structure of Metformin hydrochloride.
Sigma Aldrich. The solvents used were also of analytical grade. Tri-
ply distilled water was used for the preparation of stock solutions.
2.2. Preparation of liquid inclusion complex of MFH :HPa-CD
About 0.0033 g of MFH was accurately weighed and dissolved in
10 ml methanol. About 0.354 g of HPa-CD was dissolved in 30 ml
distilled water in a 250 ml beaker. Inclusion complexes of MFH:
HPa-CD were prepared by varying the concentration of HPa-CD
from 2
103M to 1
102M with MFH.
2.3. Preparation of solid inclusion complex of MFH:HPa-CD
About 0.049 g of MFH was accurately weighed and dissolved in
30 ml methanol. About 0.354 g of HPa-CD was dissolved in 30 ml
distilled water in a 250 ml beaker. Both the solutions were mixed
together in a beaker and put over electromagnetic stirrer to stir
continuously for 48 h at room temperature. The precipitate
obtained after evaporation was dried and used for characterization.
2.4. UV–VIS spectroscopic studies
Inclusion complexes were prepared for Metformin hydrochlo-
ride, with varying HPa-CD and a-CD concentration and absor-
bances were recorded using Systronics Double Beam
Spectrophotometer 2203.
2.5. Fluorescence spectroscopic studies
The fluorescent spectra were recorded for the inclusion com-
plexes of Metformin hydrochloride with HPa-CD using JASCO
Spectrofluorometer FP-8200
2.6. Phase solubility studies
Phase solubility studies were carried out according to the
method reported by Higuchi and Connors [11]. Samples were pre-
pared by adding an excess quantity of Metformin hydrochloride,
into 60 ml of distilled water taken in a beaker. Then the drug solu-
tion is added to 10 ml distilled water containing varying concen-
trations of HPa-CD such as 0, 2, 4, 6, 8, 10 ml in stoppered
bottles. The above samples were shaken for 72 h. After shaking
the solutions were filtered and their absorbances were recorded
at 230 nm for Metformin hydrochloride, The solubility of the Met-
formin hydrochloride in every HPa-CD concentration was calcu-
lated and phase solubility diagram was drawn between the
solubility of the Metformin hydrochloride against different con-
centrations of HPa-CD.
From the diagram, the apparent solubility constant of Met-
formin hydrochloride can be calculated using the equation
slope
Ks ¼
S0 ð1  slopeÞ
where S0 is the aqueous solubility of the Metformin hydrochloride
Please cite this article as: S. Lizy Roselet and J. Prema Kumari, An investigation on host-guest complexation of Metformin hydrochloride with hydrox-
ypropyl-a-cyclodextrin for enhanced oral bioavailability, Materials Today: Proceedings, https://doi.org/10.1016/j.matpr.2019.06.650
2.7. 1H NMR spectroscopic studies
1
H NMR spectroscopic studies were carried out using Bruker
300 MHz FT NMR spectrometer. Chemical shifts were reported in
ppm (d) downfield from tetramethylsilane (TMS) internal reference
0 ppm. Samples were prepared by dissolving in DMSO.
3. Results and discussion
Effect of HPa-CD
3.1. Absorption studies on Hydroxypropyl-a–cyclodextrin inclusion
complexes of Metformin hydrochloride
The inclusion of the guest in the CD cavity might be associated
with a bathochromic or a hypsochromic shift of its maximum
absorbance wavelength. From Fig. 2 and Table 1 it is inferred that
with the addition of HPa-CD to Metformin hydrochloride the
absorption maxima ofMFH:HPa-CD inclusion complexes are
bathochromically shifted from kmax 232.4 nm to 233.2 nm with
increase in intensity. The spectral shifts are indicative of the inclu-
sion complexes formed between Metformin Hydrochloride and
HPa-CD. It is therefore understood that the solubility increases
with increase in concentration of HPa-CD.
Metformin hydrochloride showed an increase in absorbance
with the increase in the concentration of HPa-CD. This implies that
the solubility and hence the stability of the Hydroxypropyl-a-cyclo
dextrin complexes increased upon complexation with HPa-CD.
The Hydroxypropyla-cyclodextrin dependence of Metformin
hydrochloride, can be analysed by the Benesi – Hildebrand plot
as given by
1 1 1
¼ þ   ð2Þ
A  A0 A1  A0 K B A1  A0 ½HPa  CD
where [HPa-CD] represents the concentration of HPa-CD and A0, A
are the absorbances in the absence and presence of HPa-CD respec-
tively and KB is the binding constant. From Fig. 3 a plot of
1
AA0
Vs ½HPa1CD gave a linear relation which indicated the formation
of MFH:HPa-CD inclusion complex with 1:1 stoichiometry. The
binding constant for HPa-CD complex evaluated from absorption
studies was found to be 97.08 M1 for Metformin hydrochloride.
The relatively high value of binding constants indicated the forma-
tion of a stable inclusion complex.
ð1Þ
Fig. 2. Absorption spectra of MFH:HPa-CD inclusion complex.
 S. Lizy Roselet, J. Prema Kumari / Materials Today: Proceedings xxx (xxxx) xxx 3

Table 1
Absorption spectrum of Metformin hydrochloride with HPa-CD anda-CD.

S.No Conc.of HPa-CD (M) kabs Abs (A) log e 1/[HPa-CD]

1 0 232.4 1.025 3.70
2 0.002 232.6 1.189 3.80 500
3 0.004 233.0 1.268 3.82 250
4 0.006 233.2 1.423 3.87 166.6
5 0.008 233.2 1.671 3.94 125
6 0.010 233.3 1.823 3.98 100

rescent intensity upon increase in the concentration of HPa-CD.

The spectral shift confirms the formation of the inclusion complex
between Metformin hydrochloride and HPa-CD.
The Hydroxypropyla-cyclodextrin dependence on the fluores-
cent spectra of Metformin hydrochloride could be analysed using
the Benesi-Hildebrand plot as given by

1 1 1
¼ þ   ð3Þ
I  I0 I1  I0 K B I1  I0 ½HPa  CD

where I0,I are the intensities of fluorescence in the absence and

presence of HPa-CD respectively, [HPa-CD]-concentration of HPa-
CD and KB- Binding constant.
From Fig. 5 a linear relation is obtained in the plot of
1
II0
Vs ½HPa1CD. This indicates that the stoichiometric ratio of the
inclusion complex formed between MFH with HPa-CD is 1:1.
The binding constant was evaluated from fluorescent studies. It
was found that the binding constant of MFH:HPa-CD is 82.1 M1.
Fig. 3. Benesi-Hildebrand plot of MFH:HPa-CD inclusion complex.

3.3. Phase solubility studies

The complexation of Metformin hydrochloride was character-

ized by Phase Solubility Studies. The phase Solubility diagram for
complex formation is shown in Fig. 6. The phase solubility diagram
can be classified as AL type according to Higuchi and Connors in the
case of complexation with HPa-CD. The aqueous solubility
increased linearly as a function of the concentration of HPa-CD.
It was due to the formation of a 1:1 inclusion complex.
The solubility constant KC was calculated from the slope of the
linear plot of the phase solubility diagram according to the equa-
slope
tion K C ¼ S0 ð1slopeÞ
(1)
Where S0 is the solubility of the drug in the absence of HPa-CD
The calculated KC values for HPa-CD complexation was found to be
101.5 M1

Fig. 4. Fluorescent spectra of MFH:HPa-CD inclusion complex. 3.4. 1H NMR spectroscopic studies

Nuclear magnetic resonance (NMR) spectroscopy is a powerful

3.2. Fluorescent spectral characteristics of Metformin hydrochloride
with HPa-CD.
From Fig. 4 and Table 2 it is found that the fluorescent spectra
are bathochromically shifted from kflu 360–365.8 nm for Met-
formin hydrochloride complexes. Also, there is an increase in fluo-
tool and is becoming a routine method for the characterisation of
CD inclusion complexes. It is mainly employed for the elucidation
of the geometric accommodation of the guest inside the CD cavity
[12].NMR provides a direct evidence on the inclusion of the guest
in the CD. This is based on the fact that, if the guest is encapsulated,
then the physical or chemical environment of the guest and cavity

Table 2
Fluorescent spectral data of Metformin hydrochloride with HPa-CD.

Sl. No Conc. of HPa-CD (M) kflu Int. (I) 1/[HPa-CD]

1 0 360 21.68
2 0.002 361.1 26.52 500
3 0.004 362.0 33.38 250
4 0.006 363.7 38.45 166.6
5 0.008 364.3 40.21 125
6 0.010 365.8 42.56 100

Please cite this article as: S. Lizy Roselet and J. Prema Kumari, An investigation on host-guest complexation of Metformin hydrochloride with hydrox-
ypropyl-a-cyclodextrin for enhanced oral bioavailability, Materials Today: Proceedings, https://doi.org/10.1016/j.matpr.2019.06.650
4 S. Lizy Roselet, J. Prema Kumari / Materials Today: Proceedings xxx (xxxx) xxx

Table 4
Chemical shifts of MFH and their difference in presence of HPa-CD.

H MFH MFH : HPa-CD Dd

dMFH dMFH: HPa-CD
Ha 2.93 2.91 0.02
Hb,c,d 7.05 6.69 0.36
He,f 7.44 7.14 0.30

Ha – 2 CH3 protons, Hb, Hc, Hd 2 NH and 1 NH2 proton He and Hf 1 NH, 1HCl proton.

Fig. 5. Benesi-Hildebrand plot of MFH:HPa-CD inclusion complex.

Fig. 7. 1H NMR spectra of HPa-CD.

Fig. 6. Phase solubility diagram of MFH:HPa-CD.

hydrogens (H-3 and H-5 of the CD) will be affected leading to a

modification of the NMR spectra [13].1H NMR can provide evi-
dence for the inclusion of guest molecules within the cavity of
the HPa-CD [13–15]. In general, if a guest molecule is located
within the CD cavity there the hydrogen atoms located on the
inner surface of the cavity (H-3 and H-5) will be considerably
shielded by the outer surface (H-1, H-2, H-4) will be unaffected
by the inclusion complex formation. The chemical shift change
(Dd) is defined as the difference in the chemical shift change. A
positive sign means a downfield shift and negative sign means
an upfield shift [16–19].
From the Table 3 and 4 and Figs. 7–9, in MFH:HPa-CD complex,
the H-3 and H-5 protons experience more upfield shift. The H-1
Fig. 8. 1H NMR spectra of Metformin hydrochloride.

Table 3
Chemical shifts (d) of HPa-CD their difference in presence of MFH. and H-2 protons too experienced upfield shift. Further the DdH3 = -
H HPa-CD HPa-CD:MFH Dd DdH5 = 0.09. The H-4 and H-6 signals of HPa-CD disappeared in
dHPa-CD dHPa-CD: MFH the inclusion complex. The same change in chemical shift values
1 4.52 4.78 0.04 of H-3 and H-5 shows that complete inclusion has taken place.
2 3.80 3.72 0.08 Thus the inclusion complex formation of MFH: Hpa-CD is
3 3.35 3.26 0.09 confirmed.
4 3.97 – – When the spectrum of MFH and MFH:HPa-CD are compared, it
5 3.70 3.61 0.09
6 4.20 – –
is found that the Hb, He, Hd, He and Hf protons have experienced
more upfield shift than Ha.

Please cite this article as: S. Lizy Roselet and J. Prema Kumari, An investigation on host-guest complexation of Metformin hydrochloride with hydrox-
ypropyl-a-cyclodextrin for enhanced oral bioavailability, Materials Today: Proceedings, https://doi.org/10.1016/j.matpr.2019.06.650
 S. Lizy Roselet, J. Prema Kumari / Materials Today: Proceedings xxx (xxxx) xxx
Fig. 9. H NMR Spectra of MFH:HPa-CD.
This further proves the inclusion of Metformin hydrochloride
into the HPa-CD cavity and this also indicates the complete inclu-
sion of Metformin hydrochloride deeper into the HPa-CD cavity.
4. Conclusion
The spectral shifts confirm the inclusion complexation between
Metformin hydrochloride and Hydroxypropyl-a-cyclodextrin in
both UV–VIS and fluorimetric studies. The Benesi Hildebrand plot
shows the formation of inclusion complexes with 1:I stoichiome-
try. Considerably higher binding constants indicate the inclusion
complexes formed were stable. The phase solubility profiles are
of AL type and the stoichiometry of the complexes formed is 1:1.
Furthermore 1H NMR studies serve as a strong evidence the host-
guest complexation indicating complete inclusion of Metformin
hydrochloride into the HPa-CD cavity. Therefore it could be con-
cluded that Hydroxypropyl-a-cyclodextrin complexation could be
considered as a viable route for the enhancement of bioavailability
of Metformin hydrochloride.
Please cite this article as: S. Lizy Roselet and J. Prema Kumari, An investigation on host-guest complexation of Metformin hydrochloride with hydrox-
ypropyl-a-cyclodextrin for enhanced oral bioavailability, Materials Today: Proceedings, https://doi.org/10.1016/j.matpr.2019.06.650
5
References
[1] G.F. Petrovick, Type 2 diabetes mellitus and metformin hydrochloride usage: a
short review, J. Pharmacol. Ther. Res. 2 (2) (2018) 6–9.
[2] Garry G. Graham, Jeroen Punt, Manit Arora, Richard O. Day, Matthew P.
Doogue, Janna K. Duong, Timothy J. Furlong, Jerry R. Greenfield, Louise C.
Greenup, Carl M. Kirkpatrick, John E. Ray, Peter Timmins, Kenneth M. Williams,
Clinical Pharmacokinetics of Metformin, Clin. Pharmacokinetics 50 (2) (2011)
81–98. http://link.springer.com/10.2165/11534750-000000000-00000,
https://doi.org/10.2165/11534750-000000000-00000.
[3] P.J. Pentikainen, Bioavailability of metformin. Comparison of solution, rapidly
dissolving tablet, and three sustained release products, Int. J. Clin. Pharmacol.
Ther. Toxicol (1986;24(4):) 213–220.
[4] Simone Carneiro Fernanda Costa Duarte Luana Heimfarth Jullyana Siqueira
Quintans Lucindo Quintans-Júnior Valdir Veiga Júnior Ádley Neves de Lima
Cyclodextrin–Drug Inclusion Complexes: In Vivo and In Vitro Approaches IJMS
20(3) 642. https://doi.org/10.3390/ijms20030642 http://www.mdpi.com/
1422-0067/20/3/642
[5] J. Conceição et al., Cyclodextrins as drug carriers in pharmaceutial technology:
the state of the art, Curr. Pharm. Des. 24 (2018) 1–29.
[6] J. Conceição et al., Cyclodextrins as excipients in tablet formulations, Drug
Discov. Today 23 (2018) 1274–1284.
[7] E.M. Davis, E.M. Brewster, Cyclodextrin-based pharmaceutics: Past, present
and future, Nat. Rev. Drug Discov (2004) 1023–1035.
[8] P.J. Salústio et al., Technologies for oral controlled release: cyclodextrins for
oral controlled release, AAPS Pharm. Sci. Tech. 12 (2011) 1276–1292.
[9] K. Uekama, F. Hirayama, T. Irie, Cyclodextrin drug carrier systems, Chem. Rev 9
(5) (1998) 2045–2076.
[10] T. Loftsson, D. Hreinsdóttir, M. Másson, Evaluation of cyclodextrin
solubilization of drugs’, Int. J. Pharm. 302 (1–2) (2005) 18–28.
[11] T. Higuchi, A.K. Connors, Phase-solubility techniques, Adv. Anal. Chem. Inst. 4
(1965) 117–212.
[12] M.K. Foury et al., Characterization of cyclodextrin/volatile inclusion
complexes: a review, Molecules 23 (2018) 1204.
[13] H.J. Schneider et al., NMR studies of cyclodextrins and cyclodextrin complexes,
Chem. Rev. 98 (1998) 1755–1786.
[14] J. Tsao et al., Release of paeonol-b-CD complex from thermo-sensitive poly (N-
isopropylacrylamide) hydrogels, Int. J. Pharm 402 (2010) 123–128.
[15] D.J. Wood, F.E. Hruska, W. Saenger, Proton NMR study of the inclusion of
aromatic molecules in.alpha.-cyclodextrin, J. Am. Chem. Soc. 99 (6) (1977)
1735–1740.
[16] P.V. Demarco, A.L. Thakkar, Cyclohepta-amylose inclusion complexes. A proton
magnetic resonance study, J Chem. Soc. D: ChemComm. 1 (1970) 2–4.
[17] S. KavirajaaPandian et al., Synthesis and characterization of the inclusion
complex of b-cyclodextrin and azomethine, Int. J. Mol. Sci. 14 (2013) 3671–
3682.
[18] F. Djedaïni et al., High-field nuclear magnetic resonance techniques for the
investigation of a b-cyclodextrin:indomethacin inclusion complex, J. Pharm.
79 (7) (1990) 643–646.
[19] C.M. Fernandes et al., Multimodal molecular encapsulation of nicardipine
hydrochloride by b-cyclodextrin, hydroxypropyl-b-cyclodextrin andtriacetyl-
b-cyclodextrin in solution. Structural studies by 1H NMR and ROESY
experiments’, Eur. J. of Pharm. Sci 18 (5) (2003) 285–296.