IJPRD, 2013; Vol 5(09): November-2013 (103 014)

International Standard Serial Number 0974 9446

-------------------------------------------------------------------------------------------------------------------------------------------------DEVELOPMENT AND VALIDATION OF STABILITY INDICATING METHOD FOR DETERMINATION OF

LURASIDONE IN BULK DRUG AND PHARMACEUTICAL DOSAGE FORM BY HPLC
Pawanjeet J. Chhabda1*,
M. Balaji2, Srinivasarao .V2, K.M.Ch.Appa Rao2
1

Department of Biochemistry, Ahmednagar College, Ahmednagar, MH,India

Department of Chemistry, Gitam Institute of Science, GITAM University, Visakhapatnam, India.

ABSTRACT
A simple, precise, selective linear and accurate reverse phase HPLC
method was developed and validated for the assay determination
of Lurasidone in bulk drug and dosage form. Isocratic elution at
flow rate 1.0ml/min was employed on waters XBridge c18
(150x4.6) mm 5m at 30c temperature. The mobile phase
consisted of 0.1% perchloricacid: acetonitrile (50:50) (%v/v). The uv
detection wavelength was 230nm and 10l of sample injected. The
retention time for lurasidone was 5.60min and linearity was
observed in the concentration range of 30-225 g/ml with
correlation coefficient of 0.9999. The percentage relative standard
deviation in accuracy and precision studies was found to be less
than 2%. The method was successfully validated as per
International Conference on Harmonization (ICH) guidelines.
Lurasidone undergoes degradation under acidic, basic, oxidation,
dry heat and photolytic conditions, degradation impurities did not
interfere with the retention time of Lurasidone, and assay method
is thus stability indicating. The method was successfully applied for
routine analysis of lurasidone in bulk drug and dosage form.

Correspondence Author
Pawanjeet J. Chhabda
Department of Biochemistry,
Ahmednagar College, Ahmednagar,
MH,India
Email: pawanjeetvps@rediffmail.com

Keywords:- Lurasidone.validation.HPLC.Stability indicating

INTRODUCTION
Diabetes mellitus is regarded as a syndrome, a

Lurasidone is an atypical antipsychotic , It was

approved by the U.S. Food and Drug
Administration (FDA)
for
treatment
of schizophrenia on October 28, 2010 after a
review that found that two of the four Phase
III clinical trials supported efficacy, while one

showed only marginal efficacy and one was not

interpretable because of high drop-out rates. It is
currently pending approval for the treatment
of bipolar disorder in the United States, developed
by Dainippon Sumitomo Pharma(20). Each tablet
contains 20 mg, 40 mg, 80 mg, or 120 mg of
lurasidone hydrochloride market under the brand
name of LATUDA. Lurasidone
is chemically

Available online on www.ijprd.com

103

International Journal of Pharmaceutical Research & Development

(3aR,4S,7R,7aS)-2-[[(1R,2R)-2-[[4-(1,2Benzisothiazol-3-yl)-1-piperazinyl]
methyl]
cyclohexyl]
methyl]hexahydro-4,7-methano-1Hisoindole-1,3(2H)-dione
hydrochloride
with
empirical formula is C28H36N4O2S.HCl and molecular
weight 529.14 (21).
Various methods in the literatures involve
determination of Lurasidone in human, rat plasma
by
LCMS/MS
(1,
2),
pharmacokinetics,
pharmacodynamics (3-13), UV Spectroscopy (14,
15) and HPLC (16, 17). However no method is
available for stability indicating method for assay of
Lurasidone in bulk drug and pharmaceutical dosage
form. In the present work we have developed a
new, simple precise and stability indicating method
for determination of Lurasidone in bulk drug and
pharmaceutical dosage form.

Figure 1: Structure of Lurasidone

Experimental
Chemicals & Reagents
Lurasidone is available as tablets with brand name
LATUDA was purchased from local market,
containing Lurasidone 120mg HPLC grade
acetonitrile, AR grade perchloric acid were
purchased from Merck, Mumbai. High pure water
was prepared by using Millipore Milli-Q plus
purification system.
Chromatographic Conditions
A
Alliance e2695 separation module (Waters
corporation, Milford, MA) equipped with 2998 PDA
detector with empower 2 software used for
analysis. Buffer consisted of 0.1% perchloric acid in
water (1ml of perchloric acid in 1000 ml of water).

ISSN: 0974 9446

waters XBridge c18 (150x4.6) mm 5m column and

isocratic mixture of solution A (Buffer) solution B
(Acetonitrile)(50:50)(%v/v) used as stationary and
mobile phase respectively. Methanol used as
diluent. The column oven maintained at 30c with
1.0ml flow rate. An injection volume 10l was
used. The elution compounds were monitored at
230 nm.
Preparation of Stock and standard solutions
Accurately 150mg of Lurasidone standard dissolved
in 100ml diluent to get a concentration of
1500g/ml. Further 10ml of stock solution was
taken in 100ml flask and diluted up to the mark
with diluent to get concentration of 150g/ml.
Preparation of Tablets for assay
The formulation tablets of Latuda were crushed to
give finely powdered material. Powder equivalent
to 150mg of drug was weighed and transferred to
the 100ml flask added 10ml diluent and placed in
an ultrasonicator for 10minites made up to the
volume with diluent, and filtered through a 0.45m
nylon syringe filter. 10ml of this solution was taken
into 100 ml flask and diluted volume with diluent
to get concentration 150g/ml.
Forced Degradation studies
Acid Degradation studies
Acid decomposition was carried out in 0.1N HCL at
concentration of 1500g/ml Lurasidone and after
refluxation for 24hours at 80c, the stressed
sample was cooled, neutralized and diluted as per
requirement with diluents filtered and injected.
The resulting chromatogram is shown in fig.3 (g).
The results are tabulated in table 4.
Alkali Degradation studies
Base decomposition was carried out in 0.1N NaOH
at concentration of 1500g/ml Lurasidone after
refluxation for 24hours at 80c, the stressed
sample was cooled, neutralized and diluted as per
requirement with diluents filtered and injected.
The resulting chromatogram is shown in fig.3(i).
The results are tabulated in table 4.

Available online on www.ijprd.com

104

International Journal of Pharmaceutical Research & Development

Oxidation
Oxidation was conducted by using 4%H2O2
solution at room temperature for 5hours, 10ml of
solution was taken in 100ml flask and diluted up to
the mark with diluent to get concentration of
150g/ml filtered and injected. The resulting
chromatogram is shown in fig.3 (k). The results are
tabulated in table 4.
Temperature Stress studies
1g of Lurasidone sample was taken into a petridish
and kept in oven at 80c for 24hours. 150mg of
sample was taken into 100 ml flask diluted volume
with diluent, further 10ml to 100ml made up with
diluent. The results are tabulated in table 4.
Photo stability
1g of Lurasidone was taken in to a petridish and
kept in photo stability chamber 200 W.hr/m2 in UV
Fluorescent light and 1.2M LUX Fluorescent light.
150mg of sample was taken in 100ml flask,
dissolved in diluent, further 10ml in 100ml flask
diluted volume with diluent. The results are
tabulated in table 4.

ISSN: 0974 9446

RESULTS AND DISCUSSION

HPLC Method Development and Optimization
To develop a rugged and suitable HPLC assay
method for the determination of Lurasidone, the
analytical condition were selected after the
consideration of different parameters such as
diluents, buffer, organic solvent for mobile phase,
column and other chromatographic conditions
(19). Initial trails were performed with different
composition of buffer (acetate and formate) and
organic phase (methanol, teterhydrofuran) with
different column like c8,phenyl,cyno,amino and
basic but Lurasidone peak shape was not good.
Finally 0.1% perchoric acid and acetonitrile with
isocratic and waters XBridge c18 (150x4.6) mm 5
m column was optimized. Different diluents were
tried to dilute sample like water,buffer,
tetrahydrofuran and mixture of water: methanol
and water: teterhydrofuran, buffer:methanol and
buffer:acetonitrile. Lurasidone was not dissolved,
finally methanol was optimized. The detection
wavelength was chosen as 230nm for Lurasidone
because they have better absorption and
sensitivity at this wavelength (fig-2). Hence
selected method was best among the all trails by
many aspects.

AU

203.3
231.4

0.05

315. 3
369.8 384.2

0.00
20 0.00

220.00

240.00

260.00

280.00

300.00

320.00

340.00

360.00

380.00

nm

Fig-2 wavelength spectrum of Lurasidone

Method Validation
Specificity
A study to establish the interference, blank
detection was conducted. Diluent was injected as
per the test method. Solution of standard and
sample were prepared as per test method and
injected into the chromatographic system. The

chromatograms of blank, standard and sample

were shown in the fig a, b, c.
Precision
The precision for assay method was established by
evaluating method precision and intermediate
precision study. Method precision was determined

Available online on www.ijprd.com

105

International Journal of Pharmaceutical Research & Development

by analyzing six independent assays were
performed and calculated the % RSD for replicate
assay determinations. Intermediate precision of
the analytical method was determined by
conducting method precision on another day and
another analyst under same experiment condition.
The result obtained for method precision and
intermediate precision are shown in table 3. The
percentage of RSD was calculated. The %RSD range
was obtained as 0.18 and 0.29 for method
precision and intermediate precision respectively
(Table 3) which is less than 2% indicating that the
method is more precise.
Accuracy
The accuracy of the method was estimated by
determination of recovery for three concentrations
(corresponding to 50,100 and 150% of test solution
concentration) covering the range of the method.
For each concentration three sets were prepared
and injected. The drug concentrations of
Lurasidone were calculated, the results obtained
are shown in table 2.The percentage recovery was
found to be 99.77-99.96% with %RSD 0.03 0.21(<2.0%) indicating that the method is more
accurate (table 2).
LOD and LOQ
The LOD and LOQ were determined at a signal to
noise ratio of 3:1 and 10:1 respectively by injecting
a series of test solutions of known concentrations
within the linearity range. Precision study was also
carried out at the LOQ level by injecting six
pharmaceutical preparations. The LOD and LOQ
were to be 0.07g/ml and 0.23g/ml respectively.
The %RSD value was noticed to be less than 2.0% at
LOQ concentration level.
Linearity
The linearity plot was prepared with six
concentration levels (30, 60,120,150,180 and 225
g/ml of Lurasidone). These concentration levels

ISSN: 0974 9446

were respectively corresponding to 20, 40,

80,100,120 and 150 % of test solution
concentration. The results obtained are shown in
table 1. The peak areas were plotted against the
corresponding concentrations to obtain the
calibration curve (figure 4).
Robustness
Robustness of method was checked by making
slight deliberate changes in chromatographic
conditions like flow rate (0.1 ml/min), %organic
(0.5) and column temperature (5c). In the all
above varied conditions, the components of the
mobile phase were held constant. The results are
tabulated in table 5.
Solution stability and Mobile phase stability
Solution stability checked for stability of standard
and sample solutions. Solution stability checked at
each interval initial 2,4,6,8,12,16,20 and 24 hours.
For standard solution stability and sample solution
stability %assay value calculated at each interval.
%RSD (NMT 2.0%) between initial assay value and
assay value obtained at predetermined time
interval calculated.
Forced Degradation Studies
Stress studies on Lurasidone were carried out
under oxidation, thermal stress, photolysis, acid
and alkali hydrolysis conditions. Significant
degradation was observed in base (fig 3i) and
peroxide of Lurasidone. There was no significant
degradation of Lurasidone upon exposure to dry
heat at 80c for 24hrs, acid and photolysis total
impurity increased to 0.11%, 0.43% and 0.10%
which indicated that the drug was stable against
these stress conditions. The developed method
revealed that there was no interference from the
impurities, degradation products and excipients to
determine the assay of drug substance in pure and
pharmaceutical formulation.

Available online on www.ijprd.com

106

International Journal of Pharmaceutical Research & Development

ISSN: 0974 9446

(a)

(b)

(c)

(d)

Available online on www.ijprd.com

107

International Journal of Pharmaceutical Research & Development

ISSN: 0974 9446

(e)

(f)

(g)

Available online on www.ijprd.com

108

International Journal of Pharmaceutical Research & Development

ISSN: 0974 9446

(h)

(i)

(j)

Available online on www.ijprd.com

109

International Journal of Pharmaceutical Research & Development

ISSN: 0974 9446

(k)
P u rit y
A u t o T h re s h o ld

8 0 .0 0

0 .2 2
0 .2 0
0 .1 8
0 .1 6

7 0 .0 0

6 0 .0 0

5 0 .0 0

AU

0 .1 4
4 0 .0 0

0 .1 2

Degrees

0 .2 4

Lurasidone - 5.578

0 .2 6

0 .1 0
3 0 .0 0

0 .0 8
0 .0 6

2 0 .0 0

0 .0 4
1 0 .0 0

0 .0 2
0 .0 0

0 .0 0

- 0 .0 2
5 .3 0

5 .3 5

5 .4 0

5 .4 5

5 .5 0

5 .5 5

5 .6 0

5 .6 5

5 .7 0

5 .7 5

5 .8 0
M in u t e s

5 .8 5

5 .9 0

5 .9 5

6 .0 0

6 .0 5

6 .1 0

6 .1 5

6 .2 0

6 .2 5

6 .3 0

(l)
P u rit y
A u t o T h re s h o ld

4 .0 0

Lurasidone - 5.567

0 .2 2
0 .2 0
0 .1 8
0 .1 6

3 .5 0

3 .0 0

2 .5 0

AU

0 .1 4
0 .1 2

2 .0 0

0 .1 0

Degrees

0 .2 4

1 .5 0

0 .0 8
0 .0 6

1 .0 0

0 .0 4
0 .5 0

0 .0 2
0 .0 0

0 .0 0
- 0 .0 2
5 .3 5

5 .4 0

5 .4 5

5 .5 0

5.55

5 .6 0

5 .6 5

5 .7 0
M in u t e s

5 .7 5

5 .8 0

5 .8 5

5 .9 0

5.95

6 .0 0

6 .0 5

(m)
P u rit y
A u t o T h re s h o ld

9 0 .0 0

Lurasidone - 5.575

0 .2 2
0 .2 0
0 .1 8
0 .1 6
0 .1 4

8 0 .0 0
7 0 .0 0
6 0 .0 0

AU

5 0 .0 0
0 .1 2
4 0 .0 0

0 .1 0
0 .0 8

Degrees

0 .2 4

3 0 .0 0

0 .0 6
2 0 .0 0

0 .0 4
0 .0 2

1 0 .0 0

0 .0 0
0 .0 0
- 0 .0 2
5 .3 0

5 .4 0

5 .5 0

5 .6 0

5 .7 0

5 .8 0

5 .9 0
M in u t e s

6 .0 0

6 .1 0

6 .2 0

6 .3 0

6 .4 0

(n)
3 Typical chromatograms of (a) Blank (b) Standard (c) Sample (d) precision injections (e) Linearity injections
Fig-3
(f) Acid blank (g) Acid sample (h) Base blank (i) Base sample (j) Peroxide blank (k) Peroxide sample (l)Purity plot
of Acid (m) Purity plot of Base (n) Purity plot of Peroxide

Available online on www.ijprd.com

110

International Journal of Pharmaceutical Research & Development

ISSN: 0974 9446

Fig-4 Linearity of Lurasidone

16000000
y = 92529x - 12579
R = 0.999

14000000

Mean Area

12000000
10000000
8000000
6000000
4000000
2000000
0
0

50

100

150

Concentration(g/mL)

200

Table-1 Results for linearity of Lurasidone

Linearity
%Level
Area
level
1
20
517237
2
40
1064284
3
80
2171507
4
100
2766401
5
120
3313426
6
150
4188742
Correlation co-efficient 0.999933
intercept -62723.4
slope
28231.25

Accuracy
Level

50%

100%

150%

Table-2 Recoveries study for Lurasidone

Accuracy (Recovery) study
Set No Amount Amount Recovery Average
Added
Found
(%)
recovery
(g/ml) (g/ml)
1
75.06
74.84
99.71
2
75.16
75.26
100
99.77
3
75.08
74.78
99.6
1
150.04
149.8
99.84
2
150.16
150
99.89
99.87
3
150.2
150.04
99.89
1
225.06
225.08
100
2
225.2
224.98
99.9
99.96
3
225.14
225.1
99.98

Std
Dev.

%
RSD

0.21

0.21

0.03

0.03

0.05

0.05

Available online on www.ijprd.com

111

International Journal of Pharmaceutical Research & Development

ISSN: 0974 9446

Table-3 Precision results for Lurasidone

Study

Method precision

Intermediate
precision

Set no

Assay (%)

1
2
3
4
5
6
1
2

100.02
100.15
99.78
99.86
100.1
100.24
99.75
100.22

100.34

4
5
6

99.88
99.9
99.55

Stress condition

Mean
assay(%)

Stdev

RSD%

100.03

0.18

0.18

99.94

0.29

0.29

Table-4 forced degradation results for Lurasidone

Drug recovered (%)
Drug decomposed (%)

Standard drug

100

Acid degradation

99.57

0.43

Alkali degradation

83.81

16.19

Oxidation degradation

89.81

10.19

Thermal degradation

99.89

0.11

Photolytic degradation

99.90

0.10

Table -5 Robustness results for Lurasidone

Robust conditions

Flow

Temperature

%Acetonitrile

variation

Retention
time(min)

USP Tailing

USP Plate
count

0.9ml

6.22

1.37

4235

1.0ml

5.6

1.3

4450

1.1ml

5.01

1.27

4568

25c

5.75

1.34

4325

30c

5.6

1.3

4450

35c

5.25

1.24

4579

45

5.95

1.34

4321

50

5.6

1.3

4450

55

4.98

1.21

4589

Available online on www.ijprd.com

112

International Journal of Pharmaceutical Research & Development

CONCLUSIONS
A validated RP-HPLC method has been developed
for determination of Lurasidone in presence of
degradation impurities . The proposed method was
found to be a new, simple, precise, linear, accurate
and specific. Degradation impurities did not
interfere with the retention time of Lurasidone,
and assay method is thus stability indicating.

6.

7.

ACKNOWLEDGEMENTS
The authors are grateful of M/S GITAM Institute of
Science, GITAM University, Visakhapatnam, India
for providing research facilities.
.
REFERENCES
1. Tae-Sung Koo,Soo-Jin Kim, Jongjoo Lee, Dong-Jin
Ha, Myoungki Baek, Hongsik Moon, Quantification
of lurasidone, an atypical antipsychotic drug, in rat
plasma
with
high-performance
liquid
chromatography with tandem mass spectrometr,
Biomedical Chromatography Volume 25, Issue 12,
pages 1389-1394, December 2011
2. Chae, Yoon-Jee; Koo, Tae-Sung; Lee, Kyeong-Ryoon,
A Sensitive and Selective LC-MS Method for the
Determination of Lurasidone in Rat Plasma, Bile,
and Urine, Chromatographia (2012) 75: 1117-1128 ,
October 01, 2012
3. Takeo
Ishiyama,Kumiko
Tokuda,Tadashi
Ishibashi,Akira Ito, Satoko Toma, Yukihiro Ohno ,
Lurasidone (SM-13496), a novel atypical
antipsychotic drug, reverses MK-801-induced
impairment of learning and memory in the rat
passive-avoidance test, European Journal of
Pharmacology Volume 572, Issues 23, 31 October
2007, Pages 160170.
4. Ishibashi T, Horisawa T, Tokuda K, Ishiyama T, Ogasa
M, Tagashira R, et al. Pharmacological Profile of
Lurasidone, a Novel Antipsychotic Agent with
Potent 5-Hydroxytryptamine 7 (5-HT7) and 5-HT1A
Receptor Activity, J Pharmacol Exp Therapeut 2010;
334:1-11.
5. Nakamura M, Ogasa M, Guarino J, and et al.
Lurasidone in the treatment of acute schizophrenia:

8.

9.

10.

11.

12.

ISSN: 0974 9446

a double-blind, placebo-controlled trial, J Cli

Psychiat 2009; 70 (6): 2936.
Enomoto T, Ishibashi T, Tokuda K, Ishiyama T, Toma
S, Ito A .Lurasidone reverses MK-801-induced
impairment of learning and memory in the Morris
water maze and radial-arm maze tests in rats.
Behavioural Brain Research 2008; 186 (2): 197207.
Tadashi Ishibashi, Tomoko Horisawa, Kumiko
Tokuda, Takeo Ishiyama, Masaaki Ogasa,Rie
Tagashira, Kenji Matsumoto, Hiroyuki Nishikawa,
Yoko Ueda, Satoko Toma,Hitomi Oki, Norihiko
Tanno,
Ikutaro
Saji,
Akira
Ito,Yukihiro
Ohno,andMitsutaka Nakamura, Pharmacological
Profile of Lurasidone, a Novel Antipsychotic Agent
with Potent 5-Hydroxytryptamine 7 (5-HT7) and 5HT1AReceptor Activity, the journal of pharmacology
and experimental therapeutics Vol. 334,
167346/3600152.
Hussar DA, Shah A,New drugs. Azilsartan
medoxomil,
belimumab,
and
lurasidone
hydrochloride, J Am Pharm Assoc (2003) 2011 MayJun; 51(3):444-7.
Yasui-Furukori, Norio, Update on the development
of lurasidone as a treatment for patients with acute
schizophrenia , Drug Design, Development and
Therapy 2012;6:107-115.
Takeshi Enomoto, Tadashi Ishibashi, Kumiko
Tokuda, Takeo Ishiyama, Satoko Toma, Akira Ito,
Lurasidone reverses MK-801-induced impairment of
learning and memory in the Morris water maze and
radial-arm maze tests in rats, Behav Brain Res. 2008
Jan 25;186 (2):197-207.
Norio Yasui-Furukori, Update on the development
of lurasidone as a treatment for patients with acute
schizophrenia, Drug Des Devel Ther. 2012 ;6 :107-15
.
Fabio Fumagalli, Francesca Calabrese, Alessia Luoni,
Francesca Bolis, Giorgio Racagni, Marco A Riva,
Modulation of BDNF expression by repeated
treatment with the novel antipsychotic lurasidone
under basal condition and in response to acute
stress, Int J Neuropsychopharmacol. 2011 Feb 24;112.

Available online on www.ijprd.com

113

International Journal of Pharmaceutical Research & Development

13. Jonathan M Meyer, Antony D Loebel, Edward
Schweizer, Lurasidone: a new drug in development
for schizophrenia, Expert Opin Investig Drugs. 2009
Sep 28.
14. Muvvala S Sudhir and Ratnakaram V Nadh, Simple
and Validated Ultraviolet Spectrophotometric
Method for the Estimation of Lurasidone in Bulk
Form , Research Journal of Pharmaceutical,
Biological and Chemical Sciences, January March2013Volume 4 Issue 1 Page No. 609
15. Mali Nikita, Patel Jignesh, Patel Mandev, Validated
Spectrophotometric Methods For The Estimation of
Lurasidone
Hydrochloride
in
Bulk
And
Pharmaceutical Dosage Forms, IJRPS 2012,2(2),4450
16. Nirav K. Joshi, Nehal J. Shah, Development and
validation of RP-HPLC method for estimation of

17.

18.

19.
20.
21.

ISSN: 0974 9446

lurasidone hydrochloride: A novel antipsychotic

drug in bulkdrug and pharmaceutical, pharma
science monitor, Vol-3, Issue-4, Suppl-2, Nov 2012 .
Katasani Damodar Srinu Bhogineni, Bala
Ramanjaneyulu, RP-HPLC Method Development and
Validation for the Analyisis of Lurasidone in
Pharmaceutical Dosage Forms, Drug Invention
Today Year: 2012 Vol: 3 Issue: 12.
ICH Q2 (R1), Validation of analytical procedures:
Text and Methodology, Fed. Reg (19 May 1997)
62:27463
Snyder LR, Kirkland JJ, Glajch JI. PracticalHPLC
Method Development.2nd ed.; 1997.p. 2-21
www.wikipedia.org/wiki/ Lurasidone
www.chemblink.com/products/367514-88-.

*****

Available online on www.ijprd.com

114