Professional Documents
Culture Documents
DOI 10.1007/s40005-013-0075-2
NOTES
Received: 23 February 2013 / Accepted: 26 April 2013 / Published online: 10 May 2013
The Korean Society of Pharmaceutical Sciences and Technology 2013
AUCt, Cmax and Tmax were calculated, and computer programs (Equiv Test and K-BE Test 2002) were utilized for
the statistical analysis of the parameters using logarithmically transformed AUCt, Cmax and un-transformed Tmax. The
results showed that the differences between two formulations based on the reference drug, Jeil Berasil tablet, were
2.12, 0.15 and 4 % for AUCt, Cmax, and Tmax, respectively.
There were no sequence effects between two formulations in
these parameters. The 90 % confidence intervals using
logarithmically transformed data were within the acceptance
range of log 0.8log 1.25 (e.g., log 0.9114log 1.0912 and
log 0.8471log 1.1253 for AUCt and Cmax, respectively).
Thus, the criteria of the KFDA bioequivalence guideline
were satisfied, indicating Samchundang Berastolin tablet
was bioequivalent to Jeil Berasil tablet.
Keywords Beraprost sodium Berastolin tablet
Berasil tablet Bioequivalence LC/MS/MS
Introduction
Beraprost sodium (Fig. 1a), a prostacyclin derivative
(PGI2), has marked platelet-aggregation inhibition and
vasodilation activities and relatively week toxicity in animal models. These characteristics make promise therapeutic agent for the treatment of peripheral vascular
diseases. The mechanism of action of beraprost is inhibition of adenylate-cyclase dependent platelet aggregation
(Moriya et al. 2010). Beraprost sodium is an equimolar
mixture of two diastereoisomers. Each of these compounds
has two optical isomers (APS-314 and APS-315). The
inhibitory effects of APS-314d isomer on ADP-induced
platelet aggregation were 18 times more potent than those
of APS-315d isomer on ex vivo human platelets. These
123
252
123
Value
700
250
70
70
10
-4,500
-10
Mode of analysis
Negative
397.2/268.9
379.2/343.0
253
123
254
123
255
110
Table 2 Precision and accuracy for the analysis of beraprost concentration in human serum
100
90
Concentration
(pg/mL)
80
50
40
30
Accuracy
(%, n = 5)
Intra-day CV (%)
(n = 5)
Inter-day CV (%)
(n = 5)
10
8.43
4.67
112
50
7.76
3.13
99.5
200
5.23
3.31
95.5
1,000
3.80
4.50
98.6
70
60
Precision CV (%)
20
10
0
10
15
20
25
30
Time (min)
0
1000
900
800
700
600
500
400
300
200
100
0
0
Time (min)
LC/MS/MS method was successfully applied to a bioequivalence test in which serum concentrations of beraprost in thirty-two healthy male volunteers were
123
256
Table 3 Bioavailability parameters in normal and logarithmic scales for each volunteer obtained after oral administration of Samchundang
Berastolin and Jeil Berasil tablets at the beraprost sodium dose of 60 lg
Subjects
Parameter
AUCt (pg h/mL)
Reference
Value
Cmax (pg/mL)
Test
Log
Value
Tmax (h)
Reference
Test
Reference
Test
Log
Value
Log
Value
Log
Value
Value
0.25
A1
826.25
6.72
979.49
6.89
936
6.84
1,440
7.27
0.25
A2
1,059.83
6.97
976.58
6.88
754
6.63
752
6.62
0.75
0.25
A3
312.75
5.75
236.30
5.47
479
6.17
402
6.00
0.25
0.50
A4
861.63
6.76
1,037.68
6.94
623
6.43
711
6.57
1.00
0.75
A5
923.28
6.83
1,067.28
6.97
357
5.88
428
6.06
2.00
1.75
A6
855.16
6.75
325.65
5.79
1,220
7.11
253
5.53
0.25
0.50
A7
915.98
6.82
843.90
6.74
798
6.68
562
6.33
0.50
0.75
A8
407.54
6.01
382.05
5.95
334
5.81
191
5.25
0.75
1.00
A9
932.70
6.84
1,100.97
7.00
979
6.89
538
6.29
0.25
1.00
A10
574.99
6.35
332.23
5.81
515
6.24
293
5.68
0.25
0.50
A11
622.83
6.43
1,154.58
7.05
404
6.00
1,350
7.21
0.50
0.25
A12
850.93
6.75
864.93
6.76
449
6.11
504
6.22
0.75
1.00
A13
1,154.68
7.05
1,243.90
7.13
492
6.20
940
6.85
0.75
0.25
A14
A15
655.54
616.65
6.49
6.42
484.41
659.48
6.18
6.49
482
369
6.18
5.91
507
416
6.23
6.03
0.75
1.00
0.50
0.75
A16
522.78
6.26
413.94
6.03
331
5.80
359
5.88
1.00
0.25
B1
935.93
6.84
899.65
6.80
956
6.86
844
6.74
0.25
0.25
B2
756.80
6.63
801.00
6.69
473
6.16
677
6.52
0.50
0.50
B3
768.03
6.64
917.08
6.82
454
6.12
552
6.31
0.75
1.00
B4
998.48
6.91
1,555.58
7.35
635
6.45
886
6.79
0.75
0.50
B5
645.53
6.47
522.38
6.26
679
6.52
844
6.74
0.50
0.25
B6
1,313.10
7.18
1,415.28
7.26
859
6.76
992
6.90
0.50
0.75
B7
913.60
6.82
1,129.25
7.03
948
6.85
654
6.48
0.50
1.00
B8
795.51
6.68
647.17
6.47
712
6.57
501
6.22
0.25
0.75
B9
730.05
6.59
839.73
6.73
535
6.28
751
6.62
0.50
0.50
B10
785.05
6.67
936.23
6.84
657
6.49
553
6.32
0.50
0.75
B11
437.15
6.08
487.98
6.19
509
6.23
539
6.28
0.25
0.25
B12
424.06
6.05
701.23
6.55
355
5.87
426
6.05
0.50
0.50
B13
B14
1,009.23
1,190.33
6.92
7.08
838.75
1,240.35
6.73
7.12
670
985
6.51
6.89
460
902
6.13
6.80
0.25
0.50
0.50
0.75
B15
2,379.25
7.77
1,650.35
7.41
1,190
7.08
959
6.87
1.00
0.25
B16
523.88
6.26
581.10
6.36
448
6.10
435
6.08
0.25
0.75
Mean
834.36
6.65
852.08
6.65
643.34
6.39
644.31
6.37
0.59
0.61
SD
369.05
0.39
359.40
0.48
252.79
0.38
290.93
0.46
0.36
0.34
123
257
Table 4 Statistical results of bioequivalence evaluation between two beraprost sodium tablets
Parameters
AUCt
Cmax
Tmax
Difference
2.12 %
0.15 %
4%
FaG
2.3018
3.0368
1.6085
0.9973
0.9764
0.0234
-14.43 % B d B 22.43 %
The AUCt and Cmax values were calculated on the basis of in-transformed data, and the Tmax values on the basis of un-transformed data
a
a = 0.05
Bioequivalence analysis
No significant sequence, subject, formulation or period
effects were detected for any pharmacokinetic parameters.
The point estimates for the mean ratio of the test to the
reference formulation for the AUCt, Cmax were 0.9973,
0.9764, respectively (Table 4). The parametric 90 % confidence intervals were in the range of log 0.9114log
1.0912 and log 0.8471log 1.1253, respectively (Table 4),
which were entirely within the regulatory acceptance limits
for bioequivalence (80125 %) (KFDA 2011). The results
obtained from the two programs (Equiv Test and K-BE
test) were not different. This proved that there was no
significant difference between the bioavailability of reference and test formulations.
Conclusion
This validated LC/MS/MS method was sensitive, reproducible and accurate for the determination of beraprost in
human serum samples collected for bioequivalence studies.
Using this method, the bioequivalence of two different
beraprost sodium tablets was examined at the dose of
60 lg in thirty-two healthy normal male volunteers. No
significant differences in AUCt or Cmax were found
between the test and reference formulations and the calculated 90 % confidence intervals for the ratios of mean
AUCt and Cmax were within the regulatory acceptance
range for bioequivalence (80125 %).
Acknowledgments This study was supported by a contract between
Samchundang Pharm. Co. Ltd. and the Institute of Bioequivalence
and Bridging Study of Chonnam National University. The authors
References
Demolis JL, Robert A, Mouren M, Funck-Brentano C, Patrice Jaillon
(1993) Pharmacokinetics and platelet antiaggregating effects of
beraprost, an oral stable prostacyclin analogue, in healthy
volunteers. J Cardiovasc Pharmacol 22:711716
KFDA (2009) Guideline for Korean good clinical practice 2009-211
KFDA (2011) Guidance for industry, statistical approaches to
establishing bioequivalence 2011-65
KFDA (2012) Guideline for bioequivalence test 2012-103
Lee YJ, Kim YG, Lee MG, Chungv SJ, Lee MH, Shim CK (2000)
Analysis of bioequivalence study using log-transformed model.
Yakhakhoeji 44:308314
Moriya H, Ishioka K, Honda K, Oka M, Maesato K, Ikee R, Hidaka S,
Ohtake T, Kobayashi S (2010) Beraprost sodium, an orally
active prostaglandin I2 analog, improves renal anemia in
hemodialysis patients with peripheral arterial disease. Ther
Apher Dial 14:472476
Nony P, Ffrench P, Girard P, Delair S, Azoulay S, Girre JP,
Dechavanne M, Boissei JP (1996) Platelet-aggregation inhibition
and hemodynamic effects of beraprost sodium, a new oral
prostacyclin derivative: a study in healthy male subjects. Can J
Physiol Pharmacol 74:887893
Pharsight Corporation (19981999) WinNonlinTM Users Guide Ver.
3.0 Statistical Solutions Ltd (2001) Equiv Test 2.0
The Korean Pharmacopoeia IX (KP IX) (2007) Dissolution test
138144
US Department of Health and Human Services, Food and Drug
Administration, Center for Drug Evaluation and Research,
Center for Verternary Medicine (2001) Guidance for industrybioanalytical method validation. http://www.fda.gov/cder/
guidance/index.htm. Accessed 5 March 2011
World Medical Association Declaration of Helsinki: Ethical principles
for medical research involving human subjects. As amended by
the 52nd World Medical Assembly, Edinburgh, Scotland (2000)
http://www.wma.net/en/30publications/10policies/b3/index.html.
Accessed 12 Dec 2011
123