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VALIDATION OF TLC – DENSITOMETRIC

METHOD FOR QUALITY CONTROL OF
AMOXICILLIN IN DOSAGE DRUG PRODUCTS

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 WORLD JOURNAL OF PHARMACY AND PHARMACEUTICAL SCIENCES
Valentina et al. World Journal of Pharmacy and Pharmaceutical Sciences

Volume 3, Issue 5, 995-1008. Research Article ISSN 2278 – 4357

VALIDATION OF TLC – DENSITOMETRIC METHOD FOR QUALITY

CONTROL OF AMOXICILLIN IN DOSAGE DRUG PRODUCTS

Valentina Petkova1, Dobrina Doncheva Tsvetkova *1, Danka Petrova Obreshkova1,

Milen Dimitrov1, Stojan Papanov2, Peter Atanasov3
1
Medical University – Sofia, Faculty of Pharmacy, 2-Dunav str., Sofia 1000, BULGARIA
2
Medical University – Plovdiv, Faculty of Pharmacy, Department of Pharmaceutical
Sciences, Section Pharmaceutical Chemistry, Plovdiv, BULGARIA
3
Clinic of THK, UMHATEM, N.I.Pirogov”– Sofia, BULGARIA

Article Received on ABSTRACT

10 March 2014, The aim of current study was the validation of TLC densitometric
Revised on 01 April 2014, method for routine quality control of Amoxicillin in different dosage
Accepted on 24 April 2014 forms. The applied conditions were: TLC aluminium plates (Merck)
precoated with Silicagel G60F254; mobile phase: ethyl acetate: water:
*Correspondence for Author acetic acid = 60 : 20 : 20 v/v. Densitometric analysis was carried out at
Dr. Valentina Petkova λ = 254 nm. The obtained regression equation y = 22001110.x – 8392
Medical University – Sofia, show the proportional accordance 3D = f (C) in linearity range: 5.10-3
Faculty of Pharmacy, 2-Dunav g/ml  1.10-1 g/ml. The calculated correlation coefficient is 0.997. The
str., Sofia 1000, BULGARIA
results of the recovery studies presented in R [%]  RSD (%) suit
respective confidence interval: model mixtures: 1) RС400: 100.25 %  0.5 %; 2) RС500: 99.88
%  0.39 %; 3) RС600: 100.2 %  0.33 % and drug products: Amopen caps. 250 mg: 101.96
%  3.94 %; Duomox talb. 375 mg: 101.58 %  2.32 %; Amopen caps. 500 mg: 100.23 % 
2.49 %; Ospamox filmtabl. 500 mg: 99.91 %  1.54 %. All results for obtained quantity of
Amoxicillin suit respective confidence range: model mixtures: С400: 400.99 mg  1.99 mg;
С500: 499.43 mg  1.95 mg; С600: 601.19 mg  1.98 mg and drug products: Amopen
caps. 250 mg: 254.89 mg  10.06 mg; Duomox talb. 375 mg: 380.91 mg  8.85 mg; Amopen
caps. 500 mg: 501.14 mg  12.49 mg; Ospamox filmtabl. 500 mg: 499.55 mg  7.69 mg;
Ospamox filmtabl. 1000 mg: 999.24 mg  7.67 mg.

Key words: TLC densitometry, Amoxicillin, validation, quality control.

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INTRODUCTION
The term antibiotic was first used in 1942 by Waksman in his journal articles to describe any
substance produced by a microorganism that is antagonistic to the growth of other
microorganisms in high dilution.[1] With the development of pharmaceutical chemistry, most
modern antibacterials are semisynthetic modifications of various natural compounds such as
the beta-lactam antibiotics. β – lactam antibiotics exhibit their bactericidal effects by
inhibiting enzymes involved in cell wall synthesis. [2] Amoxicillin is semi – synthetic, broad –
spectrum, acid stable, orally absorbed β – lactam antibiotic and is applied for the treatment of
common bacterial infections in humans.[3]

For determination of Amoxicillin alone are described different methods: 1) HPLC with UV –
detection at λ = 229 nm in human plasma on Lichrosorb (RP C18, 250 x 4.6 mm, 10 µm),
mobile phase: 0.01 mol/l phosphate buffer (pH = 4.8) : acetonitrile = 95 : 5 v/v, flow rate :
1.3 ml/min[4]; 2) UV – spectrophotometry at λ = 247 nm[5]; 3) first derivative
spectrophotometry at λ = 255.8 nm; 4) second derivative spectrophotometry at λ = 249.2
nm[5]; 5) spectrophotometry after derivative reaction based on condensation of Amoxicillin
with 4-aminoantipyrine in the presence of potassium persulphate in alkaline medium to red
product measured at λ = 510 nm[6] or condensation with N,N-dimethylaniline in the presence
of potassium hexacyanoferrate (III) in alkaline medium to blue product measured at λ = 660
nm[7]; 6) UV – spectrophotometric method in pharmaceutical preparations, based on
enzymatic reaction in which penicillin acylase selectively cleave off Amoxicillin at D-4-
hydroxyphenylglycine side chain, which after reaction with 2-oxoglutarate by the catalysis of
D-phenylglycine aminotransferase lead to formations of 4-hydroxybenzoylformate measured
at λ = 335 nm.[8]

For determination of Amoxicillin in drug combinations are described the following different
methods: 1) absorbance ratio, compensation method and ratio spectra first order derivative
spectrophotometry for Amoxicillin and Cloxacillin in capsules at λ = 266.2 nm[8]; 2)
spectrophotometry both for Amoxicilin, Ciprofloxacin and Piroxicam after complexation
with tris(o-phenanthroline) iron (II) at λ = 510 nm and with tris(bipyridyl) iron (II) at λ = 522
nm[8]; 2) HPLC with UV – detection for: a) Amoxicillin and Clavulanic acid in human
plasma on Chromolith (RP C18, 100 mm x 4.6 mm) column, isocratic mobile phase: 0.02 M
disodium hydrogen phosphate buffer : methanol = 96 : 4 v/v at λ = 228 nm[9]; b) Amoxicillin
and Cloxacillin on Apollo C18 (150 x 4.6 nm, 5 µm), mobile phase: 0.01 M KH 2 PO 4 :

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Valentina et al. World Journal of Pharmacy and Pharmaceutical Sciences

methanol = 45 : 55 v/v, isocratic flow rate: 1.0 ml/min at λ = 225 nm[9]; c) Amoxicillin and
Flucloxacillin on ZORBAX 300-SCX column, isocratic mobile phase: 0.025 M ammonium
dihydrogen phosphate : acetonitrile = 95 : 5 v/v[12]; d) Amoxicillin and Ambroxol in
tablets[13]; e) Amoxicillin and Metronidazole on C18 column, isocratic buffered mobile phase
(pH 4.0) at λ = 254 nm[14]; 3) HPLC with mas detection in human plasma on Zorbax SB C18
(50 x 4.6 mm, 5 µm) column, mobile phase: acetonitrile : 2 mM aqmmonium acetate = 70 :
30 v/v, flow rate: 0.5 ml/min, column temperature: 30°C, internal standard:
Hydrochlorothiazide[15]; 4) Micellar Electrokinetic Capillary Chromatography for
Amoxicillin, Ampicillin, Sulfamethoxazole and Sulfacetamide[3]; 5) voltammetry in human
urine.[16]

High performance thin layer chromatographic densitometric methods are applied for
simultaneous determination in pharmaceutical dosage forms of: 1) Amoxicillin trihydrate and
Bromhexine hydrochloride on TLC aluminum plates precoated with Silicagel G60F254, mobile
phase: butylacetate : glacial acetic acid : methanol : water = 5 : 2.5 : 2.5 : 1 v/v and detection
at λ = 320 nm[17]; 2) Amoxicillin trihydrate and Ambroxol hydrochloride on TLC Silicagel
G60F254 plates, mobile phase: ethyl acetate : methanol : toluene : water : glacial acetic acid =
6.0 : 3.0 : 2.0: 1.0 : 0.5 v/v and detection at λ = 237 nm[18]; 3) Amoxicillin and Ampicillin on
titanium (IV) silicate precoated TLC plates, mobile phase: 0.1 M K2HPO4 : 0.1 M KH2PO4 =
1 : 1 v/v, spraying the plates with 1 % ninhydrin solution and detection at λ = 546 nm.[19]

From Hancu et al. is reported TLC densitometry for analysis of Amoxicillin, Ampicillin,
Benzylpenicillin and Oxacillin on TLC Silicagel G60F254 plates, mobile phase: ethylacetate :
water : acetic acid 60 : 20 : 20 v/v and detection at λ = 254 nm.[20] The aim of current study
is the validation and application of TLC densitometric method for rutine quality control of
Amoxicillin in different dosage forms combinations.

МATERIALS AND METHODS

МATERIALS
1. Substances: Amoxicillin trihydrate reference substance batch N : 1000747, sodium
hydrogencarbonate.
2. Drug products: Amopen caps. 250 mg (Antibiotica, N : 2609990902); Duomox talb. 375
mg (Astellas, N : 111413); Amopen caps. 500 mg (Balkanpharma, N : 391103); Ospamox
filmtabl. 500 mg (Sandoz, N : DN 0568);
Ospamox filmtabl. 1000 mg (Sandoz, N : BL 3112).

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3. Reagents: ethylacetate (Sigma Aldrich, SZBC 1280 V UN 1173); glacial acetic acid
(Merck, N : K 22643863 608), distilled water, sodium hydrogencarbonate, distilled water.
4. TLC developing solvent: ethylacetate : glacial acetic acid : distilled water = 60 : 20 : 20
v/v.

Reference substances and reagents used were of analytical grade quality. All solutions were
prepared daily.

МETHOD: TLC – densitometry

I. Instrumentation.
1. Densitometer – VILBER LOURMAT CN-15.LC Serial: 16263
2. Sample applicator – 10 l micropipette (Hamilton, Bonaduz, Switzerland, N: 18005 701)
3. TLC chamber (outer dimensions: 22 cm x 12 cm x 22 cm).
4. TLC aluminum plates precoated with Silicagel G60F254, 20 cm x 20 cm (Sigma Aldrich,
1489896 V).

II. Preparation of standard solutions of Amoxicillin for validation of method for

analytical parameter linearity.
For linearity studies an accurately weighed quantity of reference standard Amoxicillin: 50
mg, 100 mg, 200 mg, 300 mg, 400 mg, 500 mg, 600 mg, 700 mg, 800 mg, 900 mg, 1000
mg was dissolved in separate volumetric flask of 10.0 ml in 4.2 % solution of sodium
–3 –
hydrogencarbonate to obtain solutions with concentration correspondingly: 5.10 g/ml; 1.10
2 –2 –2 –2 –2 –2 –2 –2
g/ml; 2.10 g/ml, 3.10 g/ml, 4.10 g/ml, 5.10 g/ml, 6.10 g/ml, 7.10 g/ml, 8.10 g/ml,
–2 –1
9.10 g/ml, 1.10 g/ml.

III. Preparation of model mixtures with Amoxicillin for validation of method for
analytical parameters accuracy and precision.
3 different model mixtures: I (average weight: AW = 0.5 g), II (AW = 0.6 g), III (AW = 0.7
g) were prepared from supplement starch by separately adding of reference standard
Amoxilcillin respectively equivalent to level of 80 % (400 mg, I), 100 % (500 mg, II) and
120 % (600 mg, III) of theoretical concentration of Amoxilcillin in tablets (500 mg). From
every of model mixtures an accurately weighed quantity equivalent respectively to 400 mg,
500 mg, 600 mg Amoxicillin was dissolved in separate volumetric flask of 10.0 ml in 4.2 %
solution of sodium hydrogencarbonate.

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IV. Preparation of solutions from dosage formulations.

Separately from every drug formulation 20 tablets were weighed accurately and finely
powdered. Procedure was repeated six times for the analysis of homogenous sample from
every drug preparation. From drug formulations: Amopen caps. 250 mg (AW = 0.3923 g);
Duomox talb. 375 mg (AW = 0.5007 g); Amopen caps. 500 mg (AW = 0.6887 g), Ospamox
filmtabl. 500 mg (AW = 0.6739 g); Ospamox filmtabl. 1000 mg (AW = 1.3397 g) an
accurately weighed quantity equivalent respectively to 250 mg, 375 mg, 500 mg, 500 mg,
1000 mg Amoxicillin was weighed and dissolved in 4.2 % solution of sodium
hydrogencarbonate in 10.0 ml volumetric flask. All solutions were filtered.

V. Chromatographic procedure.
From all solutions 10 l aliquot parts were spotted onto Silicagel G60 F254 plates keeping 10
mm distance between bands. The plate was developed about 1 h min at 25 ± 1oC in ascending
vertical manner in glass chamber, previously presaturated for 1 h with mobile phase :
ethylacetate : water : acetic acid 60 : 20 : 20 v/v. The migration distance of the mobile phase
in all experiments was 15 mm. The developed plates were dried on air. Densitometric
scanning was performed on scanner VILBER LOURMAT CN-15 LC Serial : 16263,
operated in the absorbance mode at λ = 254 nm.

RESULTS AND DISCUSSION

As per ICH guidelines, method validation parameters checked were selectivity, linearity,
accuracy, precision, limit of detection, limit of quantitation, robustness and specificity.[21]

I. Selectivity.
Placebo solution, containing as supplement starch, without the active substance Amoxicillin,
was prepared in the same manner like the standard solution. The selectivity of the applied
method was confirmed by the fact that on chromatogram with placebo preparation did not
exist spot with Rf, corresponding to Rf of the Amoxicillin (0.66). This fact confirm that there
was no interference from the commonly present in the tablets excipient starch.

II. Linearity.
For the investigation of analytical parameter linearity were prepared solutions with increasing
concentration of reference standard Amoxicillin and were analyzed separately by the written
TLC densitometric method. For every concentration (C) in g/ml was measured the respective
value of the 3D volume in relative units. Data for 3D volume were plotted against

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corresponding concentrations and linear regression analysis was performed. The regression
calibration curve was built. The obtained regression equation y = 22001110.x – 8392 showed
the proportional accordance 3D = f (C) in linearity range: 5.10-3 g/ml  1.10-1 g/ml. The
calculated correlation coefficient (R2) was 0.997.

On Fig. 1. is illustrated the calibration curve for linearity.

Fig. 1. Calibration curve for linearity of Amoxicillin.

III. Limit of detection (LOD) and limit of quantitation.

The limit of detection (LOD) and limit of quantification (LOQ) were obtained by calculating
using the standard formula as per the ICH guidelines. LOD is the smallest concentration of
the analyte that gives the measurable response. SD of the response is 41175.

LOD 3.3 x SD of the response

LOD = -------------------------------------- = 5.61.10–3 g/ml
slope of calibration curve

LOQ is the smallest concentration of the analyte, which gives response that can be accurately
quantified.
10 x SD of the response
LOQ = --------------------------------- = 1.87.10–2 g/ml.
slope of calibration curve

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IV. Accuracy and precision (repeatability) for model mixtures with reference standard
Amoxicillin.
For the estimation of analytical parameter accuracy the recovery study was carried out by
applying the method in triplicate to every to 3 different model mixtures containing known
amount of Amoxicillin added separately by the standard addition method respectively at level
of 80 %, 100 % and 120 % of labeled content in tablets. On Table 1. for model mixtures are
summarized data for spot radius (r), spot area (S), 3D volume, Chauvenet’s criterion for 3D
volume (U3D).

Тable 1. 3D volume and Chauvenet’s criterion for 3D volume for model mixtures with
reference standard Amoxicillin.

Model mixture I Model mixture II Model mixture III

N: Amoxicillin 400 mg Amoxicillin 500 mg Amoxicillin 600 mg
r S 3D U3D r S 3D U3D r S 3D U3D
1. 0.45 0.64 869000 1.11 0.58 1.06 1086500 0.91 0.65 1.45 1309800 1.03
2. 0.45 0.64 875000 0.27 0.58 1.06 1089700 0.16 0.7 1.54 1314600 0.07
3. 0.48 0.72 877500 0.84 0.6 1.13 1095000 1.07 0.7 1.54 1318500 0.96
___
X 873833 1090400 1314300
SD 4368 4293 4358
RSD
0.5 0.39 0.33
[%]

For all of the obtained results for 3D volume is necessary to estimate the Chauvenet’s
criterion (U), because when U for one value is higher than the relevant standard criterion
(USt), the result must be removed as unexpected. The relations: U < 1.68 (Table 1.) show,
that all experimental results for UA are lower, than standard requirement: Umax = 1.68 (n =
3) and it isn’t necessary to remove data for 3D. The content of Amoxicillin in model mixtures
is determined by method of calibration curve using the regression equation. On Table 2. are
indi –cated: C – obtained quantity of Amoxicillin (С400, С500,С600) in model mixtures after
application of densitometric method); R (%) – degree of recovery (RС400, RС500, RС600); UC
– Chauvenet′s criterion for obtained quantity Amoxi – cillin (UС400, UС500, UС600); N –
___
number of the individual measurements (1 ÷ 6); X – arithmetical mean; SD – standard
___
deviation; RSD – relative standard deviation (%); S X – mean quadratic error; P –

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 Valentina et al. World Journal of Pharmacy and Pharmaceutical Sciences

___ ___
confidence possibility (%); t – coefficient of Student; X ± t.S X – confidence interval); E (%)
– relative error.[21]

Tаble 2. Content of Amoxicillin in model mixrutes

Model mixture I Model mixture II Model mixture III

Amoxicillin 400 mg Amoxicillin 500 mg Amoxicillin 600 mg
N:
С400 RС400 UС400 С500 RС500 С600 RС600 UС600
UC500
[mg] [%] [mg] [%] [mg] [%]
1. 398.79 99.7 1.11 497.65 99.53 0.91 599.15 99.86 1.03
2. 401.52 100.38 0.27 499.11 99.82 0.16 601.33 100.22 0.07
3. 402.66 100.67 0.84 501.52 100.3 1.07 603.1 100.52 0.96
___ 400.99  499.43  601.19 
X  SD 1.99 1.95 1.98
__
R [%]  100.25 99.88  100.2 
RSD[%]  0.5 0.39 0.33
SD 1.99 0.5 1.95 0.39 1.98 0.33
RSD [%] 0.5 0.5 0.39 0.39 0.33 0.33
___
S X 1.15 0.29 1.13 0.23 1.14 0.19
P [%] 90.0 90.0 90.0 90.0 90.0 90.0
t 2.92 2.92 2.92 2.92 2.92 2.92
___
t. S X 3.36 0.85 3.3 0.67 3.33 0.55
___
X 397.63  99.4  496.13  99.21  597.86  99.65 
___
404.35 101.1 502.73 100.55 604.52 100.75
t.S X
Е [%] 0.29 0.29 0.23 0.23 0.19 0.19

The relation UC < 1.68 shows, that all experimental results for UC are lower, than standard
requirement: Umax = 1.68 (n = 3).

For the assessment of accuracy and precision is calculated sample standard deviation (SD),
by the applying of the Bessel’s correction, in which the denominator N − 1 (degrees of
freedom) is used instead of N and in this case (S)2 is an unbiased estimator for (SD)2.
Analytical parameter accuracy is presented by the degree recovery R (%) ± RSD (%).[20]
Results from Тable 2. showed that at confidence possibility P = 90.0 % (t = 2.92), all data for
R are included in respective confidantial interval: 1) RС400: 99.4 % 101.1 % (SD = 0.5, RSD
= 0.5 %); 2) RС500: 99.21 %  100.55 % (SD = 0.39, RSD = 0.39 %); 3) RС600: 99.65 % 
100.75 % (SD = 0.33, RSD = 0.33 %). For all model mixtures values for SD and RSD are
lower or equal to 0.5 and mean quadratic error and relarive error are lower than 1.2.

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For the estimation of an analytical parameter precision (repeatability) is used the uncertainty
of the result, which is determined by: SD, RSD and confidence range.[20] From Тable 2. it is
obvious that at confidence possibility 90.0 % (t = 2.92), all results for obtained quantity of
Amoxicillin suit respective confidence range: С400: 397.63 mg  404.35 mg (SD = 1.99, RDS
= 0.5); С500: 496.13 mg  502.73 mg (SD = 1.95, RDS = 0.39); С600: 597.86 mg  604.52 mg
(SD = 1.98, RDS = 0.33).

V. Application of validated densitometric method for analysis of Amoxicillin in drug

preparations.
The validated densitometric method was applied for the identification and determination of
Amoxicillin in Amopen caps. 250 mg, Duomox talb. 375 mg, Amopen caps. 500 mg,
Ospamox filmtabl. 500 mg and Ospamox filmtabl. 1000 mg. The indentity of Amoxicillin in
analysed drug formulations is proved by the correspondence between retardation factors of
Amoxicillin in samples and in reference standard: Rf = 0.66. Representative densitogram for
Amopen caps. 250 mg and Duomox talb. 375 mg is given on Fig. 2.

Fig. 2. Densitogram for Amopen caps. 250 mg and Duomox talb. 375 mg.

On Table 3. for Amopen caps. 250 mg, Duomox talb. 375 mg, Amopen caps. 500 mg,
Ospamox filmtabl. 500 mg and Ospamox filmtabl. 1000 mg are summarized data for spot
radius (r), spot area (S), 3D volume, Chauvenet’s criterion for 3D volume (U3D).

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Тable 3. 3D volume and Chauvenet’s criterion for 3D volume for drug poducts
containing Amoxicillin

Amopen caps. 250 mg Duomox talb. 375 mg Amopen caps. 500 mg

N: 250 mg Amoxicillin 375 mg Amoxicillin 500 mg Amoxicillin
r S 3D U3D r S 3D U3D r S 3D U3D
1. 0.3 0.28 524300 1.27 0.43 0.58 796100 1.72 0.55 0.95 1057500 1.33
2. 0.33 0.34 531200 0.96 0.45 0.64 822100 0.39 0.58 1.06 1072500 0.79
3. 0.33 0.34 546800 0.25 0.45 0.64 830200 0.03 0.58 1.06 1083400 0.39
4. 0.35 0.38 561400 0.41 0.45 0.64 835200 0.28 0.6 1.13 1109500 0.56
5. 0.35 0.38 570700 0.83 0.48 0.72 841200 0.59 0.6 1.13 1112000 0.65
6. 0.35 0.38 580000 1.25 0.48 0.72 853100 1.2 0.63 1.25 1130100 1.31
___
X 552400 829650 1094167
SD 22127 19478 27474
RS
D . 4.01 2.35 2.51
[%]
Ospamox filmtabl. 500 mg Ospamox filmtabl. 1000 mg
500 mg Amoxicillin 1000 mg Amoxicillin
N: r S 3D U3D r S 3D U3D
1. 0.55 0.95 1069200 1.27 1.15 4.15 2169900 1.19
2. 0.58 1.06 1078500 0.72 1.18 4.37 2174300 0.93
3. 0.6 1.13 1086000 0.28 1.18 4.37 2182100 0.47
4. 0.6 1.13 1090100 0.03 1.2 4.52 2198600 0.51
5. 0.6 1.13 1105400 0.87 1.2 4.52 2205000 0.89
6. 0.63 1.25 1114800 1.43 1.2 4.52 2210300 1.2
___
X 1090667 2190033
SD 16914 16876
RS
D 1.55 0.77
[%]

From Table 3. it is obvious that for all of the obtained values of 3D volune for Amoxicillin
the calculated Chauvenet’s criterion (U) is lower than standard Chauvenet’s criterion (Sh =
1.73; N = 6) in analysis of 6 samples.

The content of Amoxicillin in drug preparations is determined by method of calibration curve

using the regression equation. On Table 4. (Amopen caps. 250 mg, Duomox talb. 375 mg,
Amopen caps. 500 mg) and Table 5. Ospamox filmtabl. 500 mg and Ospamox filmtabl. 1000
mg are presented results for: C – obtained quantity of Amoxicillin (С250, С375, С500, С1000) in
drug preparations after application of densitometric method; R (%) – (RС250, RС375, RС500,
RС1000); UC – (UС250, UС375, UС500, UС1000).

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Tаble 4. Content of Amoxicillin in Amopen caps. 250 mg, Duomox talb. 375 mg,
Amopen caps. 500 mg.

Amopen caps. 250 mg Duomox talb. 375 mg Amopen caps. 500 mg

250 mg Amoxicillin 375 mg Amoxicillin 500 mg Amoxicillin
N:
С250 R С250 С375 R С375 С500 R С500
U С250 U С375 U С500
[mg] [%] [mg] [%] [mg] [%]
1. 242.12 96.85 1.27 365.66 97.51 1.72 484.47 96.89 1.33
2. 245.26 98.10 0.96 377.48 100.66 0.39 491.29 98.26 0.79
3. 252.35 100.94 0.25 381.16 101.64 0.03 496.24 99.25 0.39
4. 258.98 103.59 0.41 383.43 102.25 0.28 508.11 101.62 0.56
5. 263.21 105.28 0.83 386.16 102.98 0.59 509.24 101.85 0.65
6. 267.44 106.98 1.25 391.57 104.42 1.2 517.47 103.49 1.31
380.91 501.14
___ 254.89 
X  SD  
10.06
8.85 12.49
__
R [%]  101.96 101.58 100.23
RSD[%]  3.94  2.32  2.49
SD 10.06 4.02 8.85 2.36 12.49 2.50
RSD [%] 3.95 3.94 2.32 2.32 2.49 2.49
___
S X 4.11 1.64 3.61 0.96 5.10 1.02
P [%] 98.0 98.0 99.9 99.9 98.0 98.0
T 3.37 3.37 6.86 6.86 3.37 3.37
___
t. S X 13.85 5.53 24.76 6.59 17.19 3.44
356.15 483.95
___ ___ 241.04  96.43  94.99  96.79 
X  t.S X  
268.74 107.49 108.17 103.67
405.67 518.33
Е [%] 1.61 1.61 0.95 0.95 1.02 1.02

Tаble 5. Content of Amoxicillin in Ospamox tabl. 500 mg and Ospamox tabl. 1000 mg.
Ospamox tabl. 500 mg Ospamox tabl. 1000 mg
500 mg Amoxicillin 1000 mg Amoxicillin
N:
U
С500 [mg] R С500 [%] С1000 [mg] R С1000 [%] U С1000
С500
1. 489.79 97.96 1.27 990.08 99.01 1.19
2. 494.02 98.80 0.72 992.08 99.21 0.93
3. 497.43 99.49 0.28 995.63 99.56 0.47
4. 499.29 99.86 0.03 1003.13 100.31 0.51
5. 506.24 101.25 0.87 1006.04 100.60 0.89
6. 510.52 102.10 1.43 1008.45 100.85 1.2
___
X  SD 499.55  7.69 999.24  7.67 1.2
__
R [%]  RSD[%] 99.91  1.54 99.92  0.77
SD 7.69 1.54 7.67 0.77
RSD [%] 1.54 1.54 0.77 0.77

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 Valentina et al. World Journal of Pharmacy and Pharmaceutical Sciences

___
S X 3.14 0.63 3.13 0.31
P [%] 99.0 99.0 98.0 98.0
T 4.03 4.03 3.37 3.37
___
t. S X 12.65 2.54 10.55 1.04
___ ___ 97.37  98.88 
X  t.S X 486.90  512.20 988.69 1009.79
102.45 100.96
Е [%] 0.63 0.63 0.31 0.31
From Table 4. and Table 5. it is obvious that for all of the obtained results for content
Amoxicillin the calculated Chauvenet’s criterion (U) is lower than standard Chauvenet’s
criterion (Sh = 1.73; N = 6) in analysis of 6 samples.

CONCLUSION
The proposed validated TLC densitometric method is appropriate for routine quality control
of Amoxicillin in commercially available tablets and capsules. The results of the recovery
studies are: Amopen caps. 250 mg: 96.43 %  107.49 %; Duomox talb. 375 mg: 94.99 %
108.17 %; Amopen caps. 500 mg: 96.79 %  103.67 %; Ospamox filmtabl. 500 mg: 97.37
% 102.45 %; Ospamox filmtabl. 1000 mg: 98.88 % 100.96 %. All data for obtained
content of Amoxicillin in tablets correspond to pharmacopoeial requirements: (– 7.5 % +10
%): Amopen caps. 250 mg: 231.25 mg  275 mg; Duomox talb. 375 mg: 346.87 mg  412.5
mg; Amopen caps. 500 mg and Ospamox filmtabl. 500 mg: 462.5 mg  550 mg; Ospamox
filmtabl.1000 mg: 925 mg  1100 mg.

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