Thérapie 2015 Juillet-Août; 70 (4): 329–335

P HARMACOKINETICS DOI: 10.2515/therapie/2014224


c 2015 Société Française de Pharmacologie et de Thérapeutique

Bioequivalence Study of Atorvastatin Tablets

in Healthy Pakistani Volunteers
Sohail Mohammad1 , Usman Arshad2 , Nasir Abbass3 , Irfan Parvez4 , Ghulam Abbas4 and Wajahat Mahmood4
1 College of Pharmacy, Shifa Tameer-e-Millat University, Islamabad, Pakistan
2 College of Pharmacy, Government College University, Faisalabad, Pakistan
3 Department of Pharmacy, University of the Punjab, Lahore, Pakistan
4 Department of Pharmacy, Comsats Institute of Information Technology, Abbottabad, Pakistan

Text received September 19th , 2013; accepted October 24, 2014

Keywords: Abstract – A two way, randomized cross-over bioequivalence study was conducted to analyse the rate and extent of ab-
bioequivalence; sorption of atorvastatin after a single dose of 80 mg atorvastatin as atorvastatin calcium tablets. The study was carried out
atorvastatin tablets; using healthy male volunteers (N = 24). A high performance liquid chromatography method was employed to determine
hyperlipidemia the level of drug in human plasma. It was concluded that the test and the reference drug exhibited comparable values of
pharmacokinetic parameters. It was also concluded that since there was no significant difference between the rate and ex-
tent of absorption of the drug from the test and the reference formulations: these two formulations could thus be declared
bioequivalent.

Mots clés : Résumé – Étude de bioequivalence des comprimés d’atorvastatine chez des volontaires sains pakistanais. Une étude
bioéquivalence ; de bioéquivalence randomisée en cross-over a été menée pour évaluer l’absorption de l’atorvastatine 80 mg en comparaison
comprimés de l’un de ses génériques. L’étude a été réalisée sur 24 volontaires sains de sexe masculin. Les dosages plasmatiques ont été
d’atorvastatine ; réalisés par chromatographie en phase liquide haute performance. Dans ces conditions, les paramètres pharmacocinétiques
hyperlipidémie mesurés étaient similaires. En particulier, il n’y avait pas de différence d’absorption entre la molécule originale et son
générique l’ensemble permettant de conclure à la bioéquivalence des deux produits.
Abbreviations: see end of article.

1. Introduction price of the alternative brand is only a fraction of that of the

proprietary product. This price disparity and marketing practices
Developing countries represent a major proportion of global
of generic manufacturers makes alternative brands the preferred
coronary heart disease burden. Among these, South Asian coun-
products to prescribe. Currently, bioequivalence is not a prereq-
tries suffer high mortality rates due to large population and lack
uisite for marketing authorisation of generic drugs in Pakistan.
of investment in health-care sector. [1] In Pakistan, hyperlipidemia
Hence the quality of these formulations is not assured beyond the
is one of the most prominent risk factors reported for the devel-
usual compendial in vitro assays for oral solid dosage forms. How-
opment of coronary heart disease. [2–4] Statins are the most widely
ever, benchmarking against the proprietary product is sometimes
used chemical agents among the drugs for the treatment of hy-
carried out to give confidence to the prescriber about the quality
perlipidemia. [5,6] In United States of America (USA), atorvas-
r , Pfizer), a second generation statin was particularly of the generic product.
tatin (Lipitor
favoured by physicians for its superior efficacy. Following a sim- Following an oral administration of 80 mg of two different
ilar trend in Pakistan, proprietary atorvastatin (Lipitor r , Pfizer) atorvastatin preparations in 24 healthy volunteers, this bioequiv-
[7]
showed impressive sale records. Generic manufacturers moved alence study was carried out to compare the efficacy of a generic
quickly to claim their share of these impressive sales figures and atorvastatin calcium formulation from a local manufacturer and
currently there 79 alternative brands are competing for a mar- the proprietary brand Lipitorr . A two period two sequence ran-

ket share of generic atorvastatin tablets. [8] In a number of cases, domised cross-over study design was applied.

Article published by EDP Sciences

330 Mohammad et al.

2. Method easy and repeated sampling. Sampling time included one sam-
ple prior to administration of drug (0 hours) and subsequently at
2.1. Study subjects and design 0.15, 0.30, 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5,8, 12, 24, 36, 48 hours af-
2.1.1. Ethical approval ter dosing. Each sample was transferred to a heparinzed tube and
immediately centrifuged at 4500 × g for 5 minutes. Plasma was
This study was conducted at the National Institute of Health, collected and frozen at –80 o C in freezers (Operon, Korea) until
Islamabad, Pakistan in accordance with the requirements of the analysis.
current good clinical practices (approval number PNIH/009/13). There was a washout period of 7 days in between the two
Ethical clearance was sought from the Ethical Committee for Re- periods and the same dosing and blood sampling scheme was re-
search, at National Institute of Health, Islamabad, Pakistan which peated with alternative drug.
independently approved the study protocols. The purpose, nature
and consequence of the study and investigational procedures were
2.3. Calibration curve and method validation
explained to the subjects and only those who wilfully consented to
participate were included. The participants were informed about
Stock solution of atorvastatin (Gudia Pvt. Ltd) and the inter-
the nature of the study and were told that they can withdraw from
nal standard (IS) progesterone were prepared by dissolving the
the study anytime.
compounds in methanol (Sigma-Aldrich) and 50% acetonitrile
(Sigma-Aldrich) respectively. For the construction of calibration
2.1.2. Subjects
curve, blank human plasma was spiked with standard solutions
This study was carried out in 24 healthy adult male Pakistani of atorvastatin and progesterone. Linearity assessment was done
volunteers aged between 22 to 40 years and body mass index in by constructing the calibration curve using eight standard solution
the range of 18 to 25 kg/m2 . The average age, body weight and concentrations in the range of 0.1-40 ng/mL, each sample was run
height of the volunteers was 25.75 years, 64.41 kg and 171.76 cm in triplicate. Lowest concentration on the standard curve was se-
respectively. Each subject was evaluated through a detailed history lected as the lower limit of quantification (LLOQ) for atorvastatin.
for smoking, drug abuse, use of prescription or over-the-counter The precision and accuracy of the plasma assay for atorvas-
drugs, drug hypersensitivity, recent blood donation and alcohol tatin was determined by running quality control (QC) samples
consumption. Physical examination was carried out to check vi- for three days. Quality control samples were run in duplicates
tal signs and laboratory tests included liver function tests, renal as blank plasma spiked with a fixed concentration of the inter-
function tests and haematology. nal standard and a low (0.1 ng/mL), medium (10 ng/mL) and high
(40 ng/mL) concentration of atorvastatin.
2.2. Treatment phase and blood sampling The QC samples in all the three concentrations were also sub-
jected to five freeze thaw cycles to demonstrate analyte stability
2.2.1. Drug administration and sampling under these conditions.

The test preparation (T), Orvastin r (atorvastatin calcium

40 mg film coated tablets), was manufactured by Amson Vaccines 2.4. Method of analysis
& Pharma(PVT) Ltd, Islamabad, Pakistan. Lipitor tablets r (ator-

vastatin calcium 40 mg film coated tablets manufactured by Pfizer, To 500 µL of plasma, 20 µL of progesterone (4 µ g/mL)
Pakistan) were used as reference drug (R). as internal standard, 750 µ L of acetonitrile and 200 µL NaCl
Both the test and reference preparations were administered (Reidel-de-Haen) saturated solution were added. After vortex
to the study subjects as a single dose of two tablets amounting mixing for 30 seconds (Vortex Genie-2, USA), the mixture was
to 80 mg atorvastatin after an overnight 8 hour fast, in the two centrifuged for 15 minutes at 6 000 rpm (Eppendorf 5810R,
study periods forming a cross over design. The drug was taken Germany) and 20 µL of supernatant was injected into the high
with 150 mL of water. Subjects were not allowed to take any food performance liquid chromatography (HPLC) system.
until at least four hours after dosing and any drink including wa- Plasma atorvastatin level was measured by HPLC method. [9]
ter at least one hour after the dosing. All the volunteers were re- Gradient HPLC system (Perkin-Elmer, Inc, USA) with auto sam-
strained from excessive physical activity and monitored for any pler, UV-visible (UV-vis) detector, column Oven and vacuum de-
possible drug related adverse effects. gasser (Series 200) was used. UV-vis spectrophotometer detector
Venous blood samples in a volume of 5 mL were drawn by was operated at wavelength of 247 nm. Chromatographic sep-
performing venepuncture and by putting indwelling cannula for aration was achieved using C18-RT analytical column, Merck


c 2015 Société Française de Pharmacologie et de Thérapeutique Thérapie 2015 Juillet-Août; 70 (4)
Bioequivalence Study of Atorvastatin Tablets 331

(150 × 4.6 mm I.D, 5 µm particle size) and guard column (1 cm × 25 Plasma concentrations of test and reference drugs

Drug plasma concentration

4.0 mm I.D., 5 µm particle size). The column oven tempera- 20
ture was set at 62 ◦ C. The mobile phase was 60% acetonitrile
15

ng/uL)
and 40%, 0.05M sodium phosphate (Applichem) buffer adjusted
to pH 5.5 using a pH meter (VWR, SympHony, USA) at flow rate 10

of 1.5 mL/min. [9] The mobile phase was prepared daily and de- 5
gassed by ultrasonication (water bath sonicator, Branson, USA)
0
before use.
0 10 20 30 40 50
Chromatographic data was interpreted and chromatograms Time (hours)
obtained by TotalChrom Navigator (Perkin Elmer Inc.) Ver.
6.3.0.0445. Reference Test

Fig. 1. Overlay comparison of test & reference drugs.

2.5. Data analysis for bioequivalence

3. Results
The drug concentrations obtained from all the volunteers at
both first and the second periods (0 hr-48 hr) were analysed using This study was carried out to analyse the pharmacokinetic
Software for Pharmacokinetic analysis & PK calculations (APO, parameters of a drug formulation prior to its introduction to the
MW PHARM, Ver. 3.60, Mediware, Holland). A one compart- market. The formulation had all the physiochemical parameters
ment model was applied for pharmacokinetic profile analysis of tested and ready for final marketing.
atorvastatine. Peak drug concentration (Cmax ) and time to peak All the 24 subjects in this study were healthy Pakistani men.
drug concentration (Tmax ) were obtained directly from the data. Each one of them had normal values for clinical and laboratory
Area under the plasma concentration time curve (AUC0−t ) was parameters. None of the volunteers reported any adverse event
calculated by trapezoidal method using the last measured plasma due to the drug administration. All the volunteers were admin-
drug concentration. The AUC from time zero to infinity (AUC0−∞ ) istered 80 mg atorvastatin as calcium salt, either as test or the ref-
was calculated as: erence drug. Blood samples were collected at designated intervals
and analysed for the drug concentration in plasma. The profiles of
(AUC0−∞ ) = AUCt + Ct /Ke mean plasma concentration versus time of the test and reference
drug are displayed in figure 1.
where Ct is the drug concentration at time t and Ke is the elimina- Pharmacokinetic profile of the test and reference drug in both
tion rate constant. the first and the second periods is reflected in tables I and II re-
Data obtained from these pharmacokinetic profiles were spectively.
statistically analysed with the help of SPSS Ver. 15.0. [10] The results of ANOVA revealed that in respect of Cmax , ln
and Microsoft Excel with Statistical Data Analysis tool Pack Cmax , Tmax , ln Tmax , AUC(polyexp) , ln AUC(polyexp) , AUC(trap) and ln
Ver. 2007. AUC(trap) , none of the effects subject, period and treatment were
statistically significant.
The pharmacokinetic parameters of the test and reference
2.6. Statistical analysis for bioequivalence
drug are presented in table III. The geometric mean ratio (90%
CI) of these parameters along with the results of bioequivalence
For computation and statistical analysis of bioequivalence,
testing are presented in tables IV and V.
computer software EquivTest Version 1.00 (Statistical Solutions
Ltd.) was used. Analysis of variance (ANOVA) was carried out
using ratios of log transformed values of the pharmacokinetic pa- 4. Discussion
rameters of test drug (T) and the reference drug (R). Effects of
subject, period, sequence and treatment were considered in the The aim of this study was to compare the bioavailability of
analysis. a commercially available formulation of atorvastatin from a local
The 90% confidence intervals (CI) were calculated for these Pakistani pharmaceutical manufacturer to the proprietary product
ratios and used as criteria for the assessment of bioequivalence. Lipitor. As with other bioequivalence studies, the outcome of this
Acceptance criteria for the 90% CI were prospectively defined as study was expected to enable the prescriber to make an informed
80-125% for log transformed AUC and Cmax ratios. decision about generic substation of Lipitor r with the generic


c 2015 Société Française de Pharmacologie et de Thérapeutique Thérapie 2015 Juillet-Août; 70 (4)

332

Table I. Pharmacokinetic profile of test drug in 1st period.

Pharmacokinetic Subject identification

parameters 1 2 3 4 5 6 7 8 9 10 11 12 13 14 I5 16 17 I8 19 20 21 22 23 24
Area under curve 219.50 504.70 221.60 355.10 254.60 96.26 348.60 201.50 228.50 194.00 131.60 287.63 141.20 104.30 289.90 255.70 282.90 162.20 195.90 131.30 129.90 198.70 168.10 221.10
(ţg.hr/L)
AUC trapezoidal 212.10 454.30 209.10 324.80 268.20 93.73 322.30 211.60 231.50 182.10 115.10 272.20 158.60 122.20 264.80 232.80 254.00 146.60 180.80 143.30 120.30 233.60 139.40 194.50
rule (ţg.hr/L)

c 2015 Société Française de Pharmacologie et de Thérapeutique

AUC polyexponential 217.10 358.70 219.50 349.70 254.40 93.85 267.60 201.40 224.20 190.10 131.50 281.00 140.80 104.30 275.90 255.70 271.80 160.10 191.90 130.20 128.30 198.40 155.20 208.00
(t = 1/2) [ţg.hr/L]
AUC trapezoidal 219.95 368.18 204.85 336.90 269.40 95.69 266.03 212.64 230.16 190.05 122.33 271.43 163.48 134.85 263.88 236.45 223.93 152.23 184.08 154.46 126.01 245.53 144.85 193.11
Rule (t = 1/2) [ţg.hr/l]
CL (L/h) 364.10 158.50 361.00 225.30 314.20 831.10 229.50 396.90 350.10 412.30 608.10 278.20 566.00 767.00 217.00 312.80 282.70 491.20 408.40 609.40 615.90 402.70 476.00 361.80
VD (L) 2850.00 3015.00 3675.00 1915.00 1582.00 5317.00 5533.00 1888.00 3129.00 3766.00 2977.00 2634.00 2277.00 1389.00 3248.00 1262.00 4180.00 4145.00 3889.00 3008.00 4981.00 2109.00 4536.20 4562.00
Elimination rate 0.13 0.05 0.10 0.12 0.20 0.16 0.04 0.21 0.11 0.11 0.20 0.11 0.25 0.55 0.08 0.25 0.07 0.12 0.10 0.20 0.12 0.19 0.10 0.08
constant (L/h)
Eimination half life 5.12 13.19 7.06 5.89 3.49 4.43 16.71 3.30 6.20 6.33 3.39 6.56 2.79 1.26 8.16 2.80 10.25 5.84 6.62 3.42 5.61 3.63 6.35 8.74
(t = 1/2) [L/h]
MRT (h) 7.82 19.02 10.18 8.50 5.03 6.40 24.11 4.76 8.94 9.13 4.90 9.47 4.02 1.81 11.77 4.03 14.79 8.43 9.54 4.94 8.09 5.24 9.16 12.61
MIT (h) 4.09 2.99 3.08 3.83 6.01 1.82 1.37 5.25 285.00 3.24 5.00 3.61 4.44 6.77 2.33 8.11 3.23 2.13 3.16 5.41 2.11 5.66 4.53 2.41
Absorpion rate 0.31 0.39 0.38 0.29 0.20 0.70 1.77 0.21 0.36 0.35 0.21 0.30 0.25 0.19 0.51 0.13 0.33 0.73 0.37 0.20 0.62 0.19 0.24 0.50
constant (Ka) [L/h]
Absorption half life 2.21 1.79 1.80 2.36 3.49 0.99 0.39 3.30 1.92 2.01 3.30 2.32 2.77 3.68 1.37 5.39 2.08 0.95 1.88 3.42 1.12 3.63 2.82 1.37
(t=1/2_a) [h]
Lag time t0 [h] 0.91 0.40 0.48 0.42 0.97 0.40 0.80 0.49 0.08 0.34 0.23 0.26 0.44 1.46 0.35 0.33 0.24 0.76 0.45 0.48 0.49 0.42 0.45 0.43
Tmax (h) 3.50 3.00 3.00 3.00 3.00 3.00 3.00 3.00 3.50 3.50 3.00 3.50 3.00 4.00 3.50 3.50 3.00 4.00 3.00 3.50 3.50 4.50 3.00 3.50
Cmax (ţg/L) 23.50 21.90 29.30 29.90 29.90 23.80 28.00 15.50 30.80 38.70 22.20 31.50 26.30 14.80 34.30 34.40 21.60 30.00 20.50 34.70 22.90 32.80 14.40 32.80

AUC: area under curve; C: elimination rate constant; CL: clearance; Cmax : peak concentration; MIT: mean input time; MRT: mean residence time; Tmax : time to peak
concentration.
Mohammad et al.

Thérapie 2015 Juillet-Août; 70 (4)


Table II. Pharmacokinetic profile of reference drug in 2nd period.

Pharmacokinetic Subject identification

parameters 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24
AUC (ţg.hr/L) 166.80 122.40 125.30 93.67 198.40 200.10 145.00 233.30 237.10 41.30 176.70 207.10 242.60 242.60 205.80 246.20 202.30 40.34 177.30 158.00 269.00 141.50 267.50 200.80
Bioequivalence Study of Atorvastatin Tablets

AUC trapezoidal rule 171.90 112.10 116.00 133.90 174.70 185.60 135.90 212.30 224.00 39.00 168.70 194.90 223.90 212.80 186.80 228.80 188.20 44.80 202.90 147.30 266.00 114.60 240.80 264.90
(ţg.hr/L)

c 2015 Société Française de Pharmacologie et de Thérapeutique

AUC polyexponential 158.80 114.60 124.20 93.57 188.20 197.30 142.40 232.10 228.40 40.84 176.30 197.80 230.40 224.70 150.80 243.70 201.10 39.70 176.70 156.60 268.70 127.60 267.40 200.70
(t = 1/2) [ţg.hr/L]
AUC trapezoidal rule 167.63 110.95 116.99 135.05 192.53 196.18 140.00 216.78 194.55 39.33 173.83 192.40 22.95 205.83 181.35 228.30 189.85 45.50 206.70 153.69 268.33 120.10 240.09 266.21
(t=1/2) [ţg.hr/l]
CL (L/h) 479.50 653.70 638.50 854.00 403.10 399.70 155.70 342.90 337.40 1937.00 452.80 386.30 329.80 329.70 388.70 324.90 395.50 1983.00 451.20 506.05 297.10 565.30 299.10 398.30
VD (L) 5625.00 8190.00 4813.00 2873.00 3131.00 3290.00 4841.00 2352.00 486.00 15110.00 3479.00 4432.00 3920.00 4530.00 3268.00 3315.00 2012.00 16420.20 1891.00 3805.00 2270.00 8539.00 1822.00 2035.00
Elimination rate 0.09 0.08 0.01 0.03 0.13 0.12 0.11 0.15 0.07 0.13 0.13 0.09 0.08 0.07 0.12 0.10 0.20 0.12 0.24 0.13 0.13 0.07 0.16 0.20
constant (c) [L/h]
Eimination half lfe 8.13 8.68 5.23 2.33 5.38 5.70 6.08 4.76 10.00 5.41 5.33 7.95 8.24 9.25 56.83 7.07 3.53 5.74 2.91 5.21 5.30 10.47 4.22 3.54
(t=1/2) [L/h]
MRT (h) 11.73 12.53 7.54 3.36 7.77 8.23 8.78 6.86 14.43 7.80 7.68 11.47 11.89 13.74 8.41 10.20 5.09 8.28 4.19 7.51 7.64 15.11 6.09 5.11
MIT (h) 2.29 1.71 1.40 3.00 2.23 2.82 4.07 7.90 1.71 1.30 3.55 5.10 3.11 2.01 2.55 2.38 5.52 1.50 4.51 2.62 6.42 1.74 6.30 5.23
Absorpion rate constant 0.54 3.52 1.01 0.50 0.73 0.53 0.31 0.15 0.76 3.03 0.33 0.21 0.37 0.55 0.61 0.70 0.20 1.76 0.24 0.43 0.17 1.35 0.16 0.20
(Ka) [L/h]
Absorption half life 1.27 0.20 0.68 1.39 0.95 1.31 2.27 4.76 0.92 0.23 2.13 3.24 1.86 1.26 1.14 0.99 3.52 0.39 2.91 1.62 4.17 0.52 4.22 3.54
(t=1/2_a) [h]
Lag time t0 (h) 0.45 1.43 0.42 0.99 0.96 0.94 0.71 0.23 0.39 0.97 0.47 0.43 0.42 0.26 0.90 0.96 0.45 0.93 0.32 0.31 0.40 0.99 0.21 0.12
Tmax [h] 3.00 3.00 3.00 3.50 3.50 3.50 3.50 3.00 3.00 3.00 3.50 3.50 3.50 3.50 3.00 3.50 3.00 3.50 3.00 3.50 3.50 3.50 3.00 3.50
Cmax (ţg/L) 18.10 22.50 38.70 26.00 28.70 28.70 16.70 16.50 19.20 16.10 31.40 13.00 20.70 28.20 23.20 37.20 24.00 19.80 20.80 22.70 17.60 16.20 31.60 29.90

AUC: area under curve; C: elimination rate constant; CL: clearance; Cmax : peak concentration; MIT: mean input time; MRT: mean residence time; Tmax : time to peak
concentration.
333

Thérapie 2015 Juillet-Août; 70 (4)

334 Mohammad et al.

Table III. Statistical analysis of pharmacokinetic profile for test and reference drug in 1st and 2nd periods.

Statistical analysis
Pharmacokinetic profile Test Drug Reference Drug
Mean SD SE Min Max Mean SD SE Min Max
AUC (hr.ng/mL) 221.88 92.89 18.97 96.26 504.7 180.88 63.53 12.97 40.34 269
AUC trapezoidal (hr.ng/mL) 212.05 82.17 16.78 93.73 454.3 174.97 60.26 12.3 39 266
Clearence (L/hr.) 420.87 170.64 34.84 158.5 831.1 571.07 447.72 91.41 297.1 1983
Volume of distribution (L) 3244.84 1237.36 252.63 1262 5533 4867.92 3785.33 772.83 1822 16420
Elimination rate constant (L/hr) 0.15 0.1 0.02 0.04 0.55 0.13 0.06 0.01 0.07 0.30
Elimination half life (h) 6.14 3.47 0.71 1.26 16.71 6.11 2.21 0.45 2.33 10.47
Mean residence time (h) 8.86 5.01 1.02 1.81 24.11 8.81 3.2 0.65 3.36 15.11
Mean input time (h) 3.89 1.69 0.34 1.37 8.11 3.34 1.76 0.36 1.30 7.09
Absorption rate constant (L/h) 0.41 0.33 0.07 0.13 1.77 0.76 0.87 0.18 0.15 3.52
Absorption half life (h) 2.35 1.14 0.23 0.39 5.39 1.89 1.37 0.28 0.20 4.76
Lag time (h) 0.5 0.29 0.06 0.08 1.46 0.61 0.35 0.07 0.12 1.43
Time to peak (h) 3.33 0.41 0.08 3 4.5 3.29 0.25 0.05 3 3.5
Cmax (µ g/L) 26.69 6.67 1.36 14.6 38.7 23.97 7 1.43 13 38.70
AUC: area under curve; Cmax : peak concentration.

Table IV. Pharmacokinetic parameters of test and reference drugs. formulations of atorvastatin, the drug appeared in plasma between
Parameter Test Reference half to one hour and remained detectable up to 36 hours in all the
In AUC (poly exp) [h.µ g/L] 5.28 ± 0.07 5.07 ± 0.10
subjects in the post-dose period. The pharmacokinetic parameters
In AUC (trapezoidal) [h.µ g/L] 5.29 ± 0.07 5.08 ± 0.10 of atorvastatin in the study subjects were found to be in agree-
In Cmax (µ g/L) 3.25 ± 0.06 3.14 ± 0.06 ment with other studies conducted internationally in foreign pop-
In Tmax 1.20 ± 0.02 1.19 ± 0.02 ulations. [12,13]

AUC: area under curve; Cmax : peak concentration; Tmax : time to peak
concentration. 5. Conclusion
product. As a secondary aim, this study was also expected to re- The results of ANOVA revealed that in respect of Cmax , ln
veal any substantial differences in pharmacokinetic parameters of Cmax , Tmax , ln Tmax , AUC(polyexp) , ln AUC(polyexp) , AUC(trap) and ln
atorvastatin that might be present in indigenous population as a AUC(trap) , none of the effects subject, period and treatment were
result of unique genetic makeup or environmental factors. statistically significant. When confidence interval (90%) was cal-
Twenty four healthy subjects were recruited for the purpose culated for ln transformed parameters, all the values were within
of this study. None of the subjects reported any adverse effects the prescribed limits of bio-equivalence (80%-125%). The test and
due to the administered drug. As per the guidelines of United the reference drugs can thus be declared bioequivalent and used
States Food and Drug Administration (FDA), [11] the sampling pe- interchangeably.
riod was extended well beyond three half lives of atorvastatin. The
chosen dose for this study was within the range of clinically ad- Acknowledgements. The authors wish to thank the laboratory staff at the
ministered doses and the study was conducted under fasting con- National Institute of Health, Islamabad for their technical support to the
ditions to avoid the interference of food with absorption kinetics. project.
For the purpose of this study, AUC and Cmax were defined
Conflicts of interest. The authors declare no conflicts of interest.
as the main parameters for the declaration of bioequivalence. As
per US FDA criteria, [11] two different formulations of the same Abbreviations. AUC: area under curve; CI: confidence in-
drug can be declared bioequivalent if the 90% CI of geometric terval; Cmax : peak drug concentration; FDA: Food and Drug
mean ratio for AUC and Cmax of the test and reference drugs Administration; HPLC: high performance liquid chromatography;
lie in the range of 80% to 125%. The results of present study IS: internal standard; LLOQ: lower limite of quantification; QC:
showed that after oral administration of both the test and reference quality control; R: reference drug T: test preparation; USA: United


c 2015 Société Française de Pharmacologie et de Thérapeutique Thérapie 2015 Juillet-Août; 70 (4)
Bioequivalence Study of Atorvastatin Tablets 335

Table V. Log transformed values of pharmacokinetic parameters of test and reference drugs.

Standard Limits Observed Within equivalence limits

Parameters
Lower Upper Lower Upper Lower Upper
In AUC (polyexponential) 80% 125% 0.996 1.09 Yes Yes
In AUC (trapezoidal) 80% 125% 0.999 1.08 Yes Yes
In Cmax 80% 125% 0.992 1.08 Yes Yes
In Tmax 80% 125% 0.974 1.04 Yes Yes
AUC: area under curves; Cmax : peak concentration; Tmax : time to peak concentration.

States of America; Tmax : time to peak drug concentration; UV-vis: 8. Drug information system. Lipitor (Pfizer)
UV-visible. http://druginfosys.com/AlterBrandResult.aspx?code=3211&
packing=5189
Accessed October 27th , 2014
9. Zarghi A, Shafaati A, Foroutan SM, et al. A simple and rapid HPLC method
References for the determination of atorvastatin in human plasma with UV detec-
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360 (9338): 10158 10. SPSS 15.0 for Windows, 2006, SPSS Inc Chicago, IL
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