Kniga na apstrakti book of abstrcts

Forsirana degradaciona studija izvr{ena 
vrz aktivnata supstanca Lisinopril dihydrate
K. Brzilova, H. Babunovska, V. Dubrova-Koceva, S. Janeva, 
D. Dam~evska, B. Debarlieva, F. Butikoski, T. Kova~evi}-Novakova
A. D. Alkaloid, Oddel za kontrola na kvalitet
Bul. Aleksandar Makedonski br. 12, 1000 Skopje, Makedonija
Aktivnata farmacevtska supstanca Lisinopril dihydrate spa|a vo grupata na inhibitori na angioten-
zin konvertira~kiot enzim. Ovaa aktivna supstanca vleguva vo sostav na Skopryl tabletite, proizvod na
Alkaloid-Skopje, koi se koristat vo lekuvaweto na hipertenzija, kongestivna srceva insuficiencija i
akuten infarkt na miokardot.
Cel na ovaa studija be{e odreduvawe na degradacionite produkti koi se sozdavaat koga aktivna-
ta supstanca Lisinopril dihydrate }e se izlo`i na dejstvoto na pove}e stres vlijanija: toplotna degradacija
(105
0
C); degradacija pod vlijanie na vidlivata svetlina(450-650nm); degradacija pod vlijanie na ultravi-
oletovata svetlina (254nm);hidroliza vo kisela sredina(1N HCl);  hidroliza vo alkalna sredina (1N NaOH)
i oksidacija (2% H
2
O
2
).
Odreduvawe na degradacionite produkti be{e napraveno so HPLC metoda vo koja be{e koristena
LiChrospher 100 RP-8 250mmx4mm, 5µm kolona zagrevana na temperatura od 50
0
C; detekcijata be{e vr{ena na
branova dol`ina od 210nm; eluiraweto be{e vr{eno so mobilna faza A sostavena od 30 volumeni acetoni-
tril i 970 volumeni rastvor od 3.12g/L natrium dihidrogen fosfat doteran do pH=5.0 so rastvor od 50g/L
natrium hidroksid i mobilna faza B sostavena od 200ml acetonitril i 800ml rastvor od natrium dihidro-
gen fosfat doteran do pH=5.0 so rastvor od 50g/L natrium hidroksid; udelot na mobilnite fazi A i B se
menuva{e so linearen gradient; protokot na mobilnata faza be{e 1.0ml/min; bea injektirani 20 µl i 100 µl
od probnite rastvori.
Pri izveduvaweto na ovaa studija be{e utvrdeno deka:
• Koristenata HPLC metoda poka`a dobra selektivnost i zatoa mo`e da se koristi pri odreduvawe-
to na potencijalnite poznati i nepoznati one~istuvawa koi mo`e da se sozdadat pri degradacija na aktiv-
nata supstanca Lisinopril dihydrate;
• Aktivnata supstanca Lisinopril dihydrate podlo`i na degradacija koga }e se izlo`i na vli-
janieto na temperatura od 105
0
C, pri {to se sozdavaat poznatoto one~istuvawe A i nepoznato one~istu-
vawe so RRT 4.08. Ova nepoznato one~istuvawe voedno e i najgolemo detektirano one~istuvawe;
• Aktivnata supstanca Lisinopril dihydrate ne podlo`i na vlijanie na vidlivata svetlina (pri ovie
uslovi ne se sozdavaat degradacioni produkti);
• Aktivnata supstanca Lisinopril dihydrate podlo`i na degradacija koga }e se izlo`i na vlijanieto
na ultravioletova svetlina od 254 nm, pri {to se sozdavaat poznatoto one~istuvawe A, poznatoto
one~istuvawe D i nepoznati one~istuvawa. Poznatoto one~istuvawe A e najgolemoto one~istuvawe koe
se sozdava pri ovoj stres uslov;
• Aktivnata supstanca Lisinopril dihydrate rastvorena vo 1N HCl e stabilna (ne se sozdavaat degrada-
cioni produkti);
• Aktivnata supstanca Lisinopril dihydrate rastvorena vo 1N NaOH e stabilna (ne se sozdavaat degrada-
cioni produkti);
• Aktivnata supstanca Lisinopril dihydrate rastvorena vo 2% rastvor od H
2
O
2
ne e stabilna, pri {to
se sozdavaat poznatoto one~istuvawe A, poznatoto one~istuvawe D i nepoznati one~istuvawa. Poznatoto
one~istuvawe A e najgolemoto one~istuvawe koe se sozdava pri ovoj stres uslov.
Macedonian pharmaceutical bulletin 53 (1,2) 211-212 (2007)
PP - 99
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Determination of impurity profile of morphine hydrochloride 
by HPCL with diode array detector
G. Evgenievska
1
, S. Naumovska
1
, S. Sardzovska
1
, L. Markovska
1
, A. Jovanovic
1
, H. Babunovska
1
, 
B. Sapkareva
1
, J. Bogdanov
2
1
ALKALOID AD - Skopje, Pharmaceutical, Chemical and Cosmetics Company, 
Aleksandar Makedonski 12, 1000 Skopje, Republic of Macedonia
2
Institute of Chemistry, Faculty of Natural Sciences and Mathematics, 
“Sts. Cyril and Methodius University”, P.O. Box 162, 1000 Skopje, Republic of Macedonia
Morphine (1) and its derivatives continue to be useful in medicine both as analgesics and as anesthetics. In
most of the preparations, morphine is supplied as a salt of which the most commonly used is morphine hydrochlo-
ride trihydrate (1*HCl). Since morphine is derived from natural sources it is important to have a fast and reliable
method for qualitative and quantitative determination of related impurities (which can be process-related or can be
degradation products). Impurity profiling is receiving much attention from regulatory authorities (European
Pharmacopeia, British Pharmacopeia, United States Pharmacopeia) which are slowly incorporating limits to allow-
able levels of impurities present in the active substances or formulations. High pressure liquid chromatograph with
diode array detector HPLC-DAD utilizing octylsilyl stationary phase (250 x 4.6mm, 5
µm), sodium octanesulphonate
as ion-pairing reagent and 25/75 acetonitrile/buffer mobile phase has been used to determine the impurity profile of
morphine hydrochloride trihydrate. Under these conditions good resolution between morphine and the impurities
was achieved and retention times (R
t
) and relative retention times (Rel. R
t
) were obtained. From the runs with indi-
vidual primary standards, purity parameters were determined and UV spectra (200 – 367 nm) of each compound
were obtained. The extinction coefficients (
ε) at 244 nm and 283 nm were calculated from area UV spectra and using
the values of 
ε the absorbance ratios A
244nm
/A
283nm
for each compound was determined. For quantification of impu-
rities, from the extinction coefficients at 283 nm the correction factors (for peak areas) with respect to morphine
hydrochloride trihydrate were determined. For identification purposes, one can differentiate between morphine and
related impurities based on retention times and absorbance ratios (A
244nm
/A
283nm
). The HPLC method presented in
this work is fast, reliable, sensitive and robust for detecting of impurities in morphine hydrochloride trihydrate below
limits specified by ICH.
Macedonian pharmaceutical bulletin 53 (1,2) 213-214 (2007)
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Odreduvawe na profil na one~istuvawa 
na morfin hidrorohlorid so HPLC-DAD
G. Evgenievska
1
, S. Naumovska
1
, S. Sarxovska
1
, L. Markovska
1
, A. Jovanovi}
1
, 
H. Babunovska
1
, B. [apkareva
1
i J. Bogdanov
2
1
ALKALOID AD - Skopje, Farmacevtska, Hemiska i Kozmeti~ka kompanija, 
Aleksandar Makedonski 12, 1000 Skopje, Republika Makedonija
2
Institut za Hemija, Prirodno matemati~ki Fakultet, Univerzitet Sveti Kiril i Metodij, P.F.
162, 1000 Skopje, Republika Makedonija
Morfinot (1) i negovite derivati prodol`uvaat da bidat korisni vo medicinata kako analgeti-
ci i anestetici. Vo najgolemiot broj na preparati morfinot e vo forma na sol od koja naj~esto koris-
tena e morfin hidrohlorid trihidrat (1*HCl). Morfinot se dobiva od prirodni izvori i e mnogu va`no
da se ima brz i siguren metod za kvalitativno i kvantitativno odreduvawe na srodni one~istotuvawa
(Srodnite one~istuvawa mo`at da poteknuvaat od proizvodstveniot proces ili da bidat degradacioni
produkti). Odreduvaweto na profilot na one~istuvawata dobiva na te`ina kaj regulatornite farma-
cevtski organizacii (Evropska Farmakopea, Britanska Farmakopea, US Farmakopea) koi poleka, gi
inkorporiraat granicite na dozvoleno nivo na one~istuawa prisutni vo aktivnata supstanca ili for-
mulacija. HPLC e najupotrebuvanata metoda za taa cel vo farmacevtskata industrija. Celta na na{eto
istra`uvawe be{e da go odredime profilot na one~istuvawata na morfin hidrohlorid trihidratot so
HPLC-DAD koristej}i oktilsilil stacionarna faza (250 x 4.6mm, 5
µm), natrium oktansulfonat kako jon-
sparuva~ki reagens i 25/75 acetonitril/pufer mobilna faza. Pri ovie uslovi, be{e postignata dobra
rezolucija pome|u morfinot i one~istuvawatate i bea odredeni nivnite retencioni vremiwa (R
t
) i rel-
ativni retencioni vremiwa (Rel. R
t
) . Od ranovite so individualni primarni standardi, parametri na
~istota bea odredeni i snimeni UV spektrite (200 - 367 nm) na sekoe poedine~no soedinenie. Ekstinkcionite
koeficienti (
ε) na 244 nm i 283 nm bea presmetani od UV spektrite i koristej}i gi ε vrednostite, sood-
nosot na absorbancite A
244nm
/A
283nm
bea odredeni. Za kvantifikacija na one~istuvawata,bea odredeni
korekcioni faktori (za povr{ina na pikovite) od ekstinkcionite koeficienti na 283 nm vo odnos na
morfin hidrohlorid trihidrat. Za identifikacioni celi, mo`e da se diferenciraat morfinot i srod-
nite ne~istotii so kombinacija na retencioni vremiwa i soodnos na absorbanci (A
244nm
/A
283nm
). HPLC
metodata prika`ana vo ovoj trud e brza, sigurna, senzitivna i robusna za detektirawe na one~istuvawa
vo morfin hidrohlorid trihidrat pod nivoto specificirano od ICH. 
Macedonian pharmaceutical bulletin 53 (1,2) 213-214 (2007)
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RP- HPLC gradient metod for simultaneous quantification of
Salbutamol sulphate and preservatives in pharmaceutical dosage forms
Katerina Kocova, Lidija Mano Iliev, Irina Batkovska Borozanova, Gordana Trendovska Serafimovska
REPLEKPHARM Company for pharmaceutical-chemical products, Kozle 188, 1000 Skopje, Macedonia
Salbutamol (INN) or albuterol (USAN) is a short -acting 
β
2
– adrenergic receptor agonist used for relief of
bronchospasm in condition such as asthma and chronic obstructive pulmonary disease. Salbutamol sulphate is usu-
aly given by tablets, syrups and sprays for inhalation.
The aim of this study was to develop a simple, fast and accurate, reversed phase HPLC method for simulta-
neous quantification of Salbutamol and accompanyng preservatives (Sorbic acid, Methyl paraben and Propyl paraben)
in pharmaceuticals. These components represent a diverse array of structures, polarities and functional groups. The
wide variation in polarity and hydrophobicity among the analyte components (Salbutamol and Propyl Paraben)
requires application of gradient elution.
Separation was achieved using Lichrospher RP-select B column (125 x 4 mm i.d., 5
µm), emloying simple
mobile phase: 10 mM Potassium Dihydrogen Phosphate, pH=2.6 and Acetonitrile with flow rate of 1.1 ml/min.
Detection was achieved with the photodiode - array detector at 225 nm and the analyze was completed in less than
15 min., including the time for re – equilibration.
Retention times of all components were: 2 minutes for Salbutamol, 7 minutes for Sorbic Acid, 7.7 min. for
Methyl Paraben and 9.5 min. for Propyl Paraben. This good resolution for all 4 components is obtained by applying
a segmented gradient: an initial contentration of 10 % Acetonitrile within 2 minutes is necesary for the retention of
salbumatol and for the pick-shape, and a second concetracation of 45 % acetonitrile within 7 min. to provide reso-
lution of the later eluting components.
The reversed phase HPLC method with gradient elution for simultaneous quantification of Salbutamol and
accompanyng preservatives (Sorbic acid, Methyl paraben and Propyl paraben) is simple, fast and accurate method
which can be applied in sample analysis with wide range of polarity.
Macedonian pharmaceutical bulletin 53 (1,2) 215-216 (2007)
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Reverzno Fazen 
HPLC
-gradient metod za simultana kvantifikacija na
Salbutamol sulfat i konzervansi vo farmacevtski doza`ni formi
Katerina Ko~ova, Lidija Mano Iliev, Irina Batkovska Borozanova, 
Gordana Trendovska Serafimovska
ReplekFarm, Dru{tvo za proizvodstvo na farmacevtsko - hemiski proizvodi,
Kozle 188, 1000 Skopje, Makedonija 
Salbutamol (INN) ili albuterol ( USAN ) e beta
2
-adrenergi~en receptor agonist so kratko dej-
stvo, koj se primenuva za olesnuvawe na bronhospazam kaj pacienti so bronhijalna astma i pri hroni~ni
opstruktivni belodrobni zaboluvawa. Salbutamol sulfatot naj~esto se primenuva vo oblik na sirupi,
tableti i sprej za inhalirawe.
Celta na ovaa studija e da se razvie ednostaven, brz i to~en reverzno-fazen HPLC metod za simul-
tana kvantifikacija na Salbutamol i prisutnite konzervansi (Sorbinska kiselina, Metil i Propil
parabeni) vo farmacevtski formulacii. Ovie komponenti imaat razli~na hemiska struktura, polarnost,
funkcionalni grupi. Golemata razlika vo polarnosta ili hidrofobnosta me|u komponentite vo anali-
tot (salbutamol i propil paraben) uslovuva primena na gradientno eluirawe.
Separacijata be{e izvedena na Lichrospher RP-select B kolona (125 x 4 mm, 5mm) so ednostavna mobil-
na faza sostavena od 10 mM Kalium dihidrogen fosfat so pH 2,6 i acetonitril so protok od 1,1 ml/min.
Detekcijata be{e izvr{ena so “photodiode arra“ detektor na 225 nm i analizata be{e komletno zavr{ena
za pomalku od 15 minuti zaedno so vremeto potrebno za re-ekvilibrirawe na kolonata.
Dobrata rezolucija na site ~etiri komponenti so retencionite vremiwa od 2 minuti za Salbu-
tamol, 7 min za Sorbinskata kiselina i 7,7 min. odnosno 9,5 min. za metil i propil paraben soodvetno, e
postignata so primena na segmentiran gradient: po~etnata koncentracija od 10 % na acetonitril do 2
min. e neophodna za zadr{ka na Salbutamolot kako i za oblikot na pikot, dodeka vo vtora faza-45 % na
Acetonitril vo tek na 7 min. obezbeduva rezolucija na podocna eluira~kite komponenti.
Reverzno fazen HPLC metod so gradientno eluirawe za kvantifikacija na Salbutamol i pridru`-
nite konzervansi e ednostaven, to~en i brz koj mo`e da se primeni za analiza na primerok so {irok rang
na polarnost.
Macedonian pharmaceutical bulletin 53 (1,2) 215-216 (2007)
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Simoultaneous quantitative determination of 2 active components 
and 2 preservatives in liquid dosage form trimosul with HPLC
Piponski M.
1
, Slaveska I., Rusevska T., Mindoseva M., 
Serafimovska-Trendovska G.
1
Quality Control Deparment, Pharmaceutical Company Replekfarm – Skopje, Kozle 188, Skopje, R. Macenodia
We developed and validated simple chromatographic method for quantification of Sulphometoxazol, Tri-
methoprime, Nipagin and Nipasol in liquid pharmaceutical formulation in analysis lasting less than 7 minutes. The
method shown to be selective, accurate, precise, reproducible and rigid, according to statistical calculations for all
four mentioned components.The method is of isocratic mode of run, using analytical column RP Select B 125 x 4
mm, ternary mobile phase composed of methanol, acetonitrile and 20mM potassium phosphate buffer with pH =
2,7. UV absorbance measuring detector set to 242nm wavelength. This method clearly separates components with
satisfactory resolution and good system suitability parameters. The method can be easily rescaled for faster analy-
sis with shorter analytical column for in process control or in more demandable stability study analysis using longer
analytical column with higher resolution needs. All performances of the method, make it very useful for Quality
Control routine laboratory analysis in pharmaceutical companies engaged in large number of analysis per day.
Macedonian pharmaceutical bulletin 53 (1,2) 217-218 (2007)
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Simultano kvantitativno odreduvawe na 2 aktivni komponenti 
i 2 konzervansi vo te~na doza`na formulacija 
na Trimoksazol so HPLC metoda
Piponski M.
1
, Slaveska I., Rusevska T., Mindoseva M., 
Serafimovska-Trendovska G.
Sektor za Kontrola na Kvalitet, Replekfarm - Skopje, Kozle 188, 1000 Skopje, R. Makedonija
Razvien i validiran e ednostaven hromatografski metod za kvantifikacija na Sulphometoxazol,
Trimethoprime, Nipagin i Nipasol, vo te~na farmacevtska formulacija. So analiza koja trae pomalku od 7 mi-
nuti. Metodata poka`uva selektivnost, preciznost, povtorlivost i rigidnost, vo odnos na statisti~kite
kalkulacii za site 4 komponenti. Samata metoda e so izokratski mod na separacija, koja koristi anali-
ti~ka kolona RP Select B 125 x 4 mm, ternarna kompozicija na mobilna faza sostavena od metanol, aceto-
nitril i 20mM kalium fosfaten puffer so kiselost pH=2.7, UV-apsorpcionen detektor doteran na 242
nm branova dol`ina. Metodata jasno gi razdvojuva komponentite so zadovoluva~ka rezolucija i solidni
vrednosti za soodvetnost na sistemot. Istata metoda mo`e lesno da se redizajnira za pobrzi analizi so
pokratka kolona za neposredna proizvodna kontrola ili za poopse`ni analizi pri studiite za stabil-
nost na preparatot so koristewe na podolgi hromatografski koloni, koi baraat pogolema rezolutivnost.
Site performansi na metodata ja ~inat mnogu korisna za rutinski sekojdnevni analizi vo sektorite za
kontrola na Kvalitet pri farmacevstskite kompanii, koi imaat golem broj na analizi vo rabotniot den.
Macedonian pharmaceutical bulletin 53 (1,2) 217-218 (2007)
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Development of HPLC method for simoultaneus quantification 
of dexchlorfeniramine, paracetamol, pseudoephedrine 
and dextrometorphan in solid pharmaceutical dosage forms
Piponski M.
1
, Rusevska T., Slaveska I., Hristova O., Serafimovska-Trendovska G.
1
Quality Control Deparment, Pharmaceutical Company Replekfarm – Skopje, Kozle 188, Skopje, R. Macedonia
There are many pharmaceutical preparations on the Macedonian market which are composed different com-
binations of active substances like DEXCHLORFENIRAMINE, PARACETAMOL, PSEUDOEPHEDRINE and
DEXTROMETORPHAN. In this work we describe HPLC method for simultaneous determination of all 4 analytes,
even they do not exist together in any pharmaceutical product on the market. The method is competent for their quan-
tification regardless of the used combination of these analytes are not simple for separation because of their chemi-
cal structure and especially because of the large differences in quantities incorporated in pharmaceutical dosage
forms, and their molar UV absorption coefficients. We tested many different HPLC columns and mobile phase com-
positions, and the best results were gain with Supelco LC-8-DB and RP Select B chromatographic matrixes using
binary composition of mobile phase with buffer and acetonitrile. The mixture of mentioned components were sep-
arated in less than 5 minutes run with satisfying resolution and system suitability parameters. Both columns were
used in methods which shown to be simple, fast, accurate, precise, reproducible, the features especially essential for
routine analyses in Quality Control Laboratories in pharmaceutical companies.
Macedonian pharmaceutical bulletin 53 (1,2) 219-220 (2007)
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Razvoj na izokratska HPCL metoda za simultana kvantifikacija na
Dexchlorfeniramine, Paracetamol, Pseudoephedrine i Dextrometorphan
vo cvrsti farmacevtski dozazni formi
Piponski M.
1
, Rusevska T., Slaveska I., Hristova O., Serafimovska-Trendovska G.
1
Sektor za Kontrola na Kvalitet, Farmacevtska Kompanija Replekfarm- Skopje, 
Kozle 188, 1000 Skopje, R. Makedonija
Postojat poveke farmacevtski preparati na makedonskiot pazar koi vo svojot sostav imaat inkor-
porirano razlicni kombinacii na aktivni supstancii kakvi {to se DEXCHLORFENIRAMINE, PARACETA-
MOL, PSEUDOEPHEDRINE i DEXTROMETORPHAN. Vo ovoj trud e opi{an HPLC metod za simultano deter-
minirawe na site 4 analiti, iako tie ne postojat zaedno vo nitu eden od preparatite od farmacevtskite
produkti na pazarot. Metodata e kompetentna za kvantifikacija na navedenite komponenti nezavisno od
kombinacijata na istite vo gotovite preparati. Ovie analiti ne se lesni za separacija poradi nivnata
hemiska struktura i posebno poradi golemite razliki so koi istite se zastapeni vo farmacevtskite doza-
zni formi, kako i nivnite molarni eksticioni koeficienti. Nie testiravme poveke razlicni HPLC
koloni i sostavi na mobilni fazi, i najdobri rezultati bea dobieni so Supelco LC*-DB i LiChrospher RP
Select B hromatografski matriksi pri koristewe na binaren sostav na mobilna faza so pufer i acetoni-
tril. Miksturata od spomenatite komponenti bese separirana za pomalku od 5 minuti anliza, so zadovo-
litelni parametri na hromatografska soodvetnost na sistemot. Dvete koloni bea koristeni vo metodi
koi se pokazaa deka se ednostavni, brzi, to~ni, precizni i reproducibilni, osobini koi se posebno bitni
za rutinski analizi vo laboratoriite za Kontrola na kvalitet vo farmacevtskata industrija.
Macedonian pharmaceutical bulletin 53 (1,2) 219-220 (2007)
PP - 103
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