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Volume 3 Issue 6 November/December 2015

An International, Open Access, Peer reviewed,

Bi-Monthly Journal

Editorial

Editor-in-Chief

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Advisory Board

Dr. S.M. Gopinath, Phd HOD, Dept of Biotechnology, Acharya Institute of Technology, Bangalore, INDIA

Dr. Vedamurthy A.B. Phd

Professor, P.G. Department of Studies in Biotechnology and Microbiology, Karnatak

University, Dharwad, India

Dr. Hari Venkatesh K Rajaraman MD(Ay), PGDHM Manager, R&D, Sri Sri Ayurveda Trust, Bangalore, INDIA

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Editorial Board

Dr. Pushpinder Kaur, Phd Research Associate, CSIR-Institute of Microbial Technology Sector,

Chandigarh, INDIA

Dr. Kavita Sharma, Phd Senior Scientist, Research and Development, Pharmacology Division,

Sigma Test and Research Centre, New Delhi, INDIA

Dr. Kasim Sakran Abass, Phd Associate Professor, College of Nursing,

University of Kirkuk, Kirkuk, IRAQ

Dr. Ashutosh Chaturvedi (BAMS, PEC Diabetes care)

Resident & M.D Scholar, Department of Panchakarma, SDMCAH - Hassan

Index – JOBB, Volume 3, Issue 6 - November/December 2015

Medical Biotechnology

BLEEDING PATTERN OF WOMEN USING HORMONAL CONTRACEPTIVES:

A PHARMACOVIGILANCE STUDY

Huma Dilshad, Rabia Ismail, Mohammad Harris Shoaib, Safila Naveed 319-326

Medical Biotechnology

SIMPLE UV SPECTROPHOTOMETRIC ASSAY OF NORFLOXACIN FORMULATION AND THEIR

COMPARATIVE STUDY

Safila Naveed, Sidra Sajid 327-331

Journal of Biotechnology and Biosafety

Volume 3, Issue 6, November-December 2015, 319-326 ISSN 2322-0406


www.jobb.co.in International, Peer reviewed, Open access, Bimonthly Online Journal

BLEEDING PATTERN OF WOMEN USING HORMONAL CONTRACEPTIVES:

A PHARMACOVIGILANCE STUDY

Research article

_______________________________

Huma Dilshad1, 2*

, Rabia Ismail1,

Mohammad Harris Shoaib1

Safila Naveed1, 2

_____________________________ 1Faculty of Pharmacy, University of

Karachi

2Faculty of Pharmacy, Jinnah

University for Women, Pakistan

*Corresponding Author: Email:

[email protected],

ABSTRACT:

In this study we compared the breakthrough bleeding, due to the use

of monophasic COC contains 0.15mg Levonorgestrol+0.03mg Ethinyl

estradiol (EE/LNG) with an injectable contraceptive Depot medroxy

progesterone (Depo Provera) comprising 150mg/mL of

medroxyprogestrone acetate (DMPA). The study was carried out in

family planning departments of different clinical settings and hospitals

in Karachi. Females maintained on hormonal type contraceptives were

invited and reviewed for their reproductive health and history. The

investigation includes over 12970 women, maintained on different

methods of contraceptives, out of which 1130 women were using

hormonal contraceptives. Total, 847 females were received allocated

treatment Depo-Provera verses Combined Oral Contraceptives n=498

vs n= 349 per group respectively). There are less bleeding/spotting

days recognized by women receiving DMPA than females using COCs.

Data divided into two groups: Group I (n=349) were the users of Drug-

I combined estrogen-progestin oral contraceptives were more likely to

show breakthrough bleeding than Group II (n=498) women maintained

on Drug-II Depot medroxy progesterone acetate (odds ratio [OR],

0.739; 95% confidence interval [CI], (0.56-1.97). The odd ratio of

breakthrough bleeding (BTB) for women in Age –I (19-35 years) odds

ratio [OR], 1.026 at 95% confidence interval [CI], (0.780 -1.350). This

increased risk was more observed in Duration -II, with odds ratio [OR],

1.74; 95% confidence interval [CI], with lower and upper CI limit

(1.55- 1.80). Patient education regarding early warning symptoms of

breakthrough bleeding, and lack of withdrawal bleeding adds an

additional margin of safety and reduces patient questions and

uncertainties. Long-term contraception with hormonal contraceptives

causes the maximum menstrual disturbances.

Keywords: Hormonal contraception, Bleeding pattern, COCs (Combine Oral Contraceptives), DMPA

(Depotmedroxyprogestrone acetate).

____________________________________________________________________________________

INTRODUCTION Research proved risks related with hormonal

contraception have driven women’s interest and changed

the approach for using hormone especially during

preceding decade (Andrist LC. 1998; Hunter MS et al.,

1997). The Women’s Health Initiative (WHI) study

outcomes proved the risk of using conventional

hormones, specifically Premarin®and Provera®

(Rossouw JE et al., 2002). Increased risk of coronary

mailto:[email protected]





events, breast cancer, stroke and blood clots are

associated with the use of estrogen combined with

progestin. Moreover females prescribed hormonal

contraception frequently stop it prior to long-term

advantages was identified. Weight gain and undesirable

side effects are the major reasons for discontinuation and

almost one third to two third of females stopping it during

initial two years (Vihtamaki T et al., 1999; Newton KM

et al., 1997; Tonkelaar I et al., 2000; Bjorn N et al.,

1997). Mainly the side effects are associated with the

synthetic progestin portion, and the most frequent

unwanted effects are bloating, irregular bleeding and

breast tenderness (Ettinger B et al., 1999). Other reasons

for discontinuation include risk of cancer and suggestion

by a physician. The most frequent side effect of every

form of hormonal contraceptive is abnormal uterine

bleeding. However this bleeding is not often dangerous

several females take it worrisome and frequently stop

hormonal contraception due to irregular bleeding and

other unwanted side effects (Sangi-Haghpeykar H et al.,

1996; Paul C et al., 1997). Irregular uterine bleeding has

become more frequent when oral contraceptives contain

lower doses of estrogen, which are incapable to maintain

endometrial integrity (Kaunitz AM 1998).

Progestin-induced decidualization and endometrial

atrophy associated with OCPs are the most frequent

reason of breakthrough bleeding. Increased dose and

potency of estrogen and progestin, decreases the

occurrence of breakthrough bleeding [Sulak P et al.,

1999]. Unusual uterine bleeding is associated with the

ratio of progestin to estrogen (Stubblefield PG 1994).

Moreover every woman metabolizes hormone in a

different way (Wallach M et al., 2000). These types of

factors cause more difficulties to elucidate the study

results. The management of irregular uterine bleeding in

females using combined OCPs starts with Psychotherapy

concerning compliance (Hatcher RA et al., 1998). OCs

inhibits ovulation and provide tremendous contraceptive

efficacy (Sitruk-Ware R 2006; Koetsawang et al., 1980;

Hirvonen E et al., 1990; Kivinen S et al., 1996;

Schubert W et al., 1987; Serup J et al., 1979).

In clinical trials, the monophasic and biphasic forms of

oral contraceptives are unacceptable for cycle control

(Sitruk-Ware R 2006; Hirvonen E et al., 1990; Kivinen

S et al., 1996; Schubert W et al., 1987; Serup J et al.,

1979) which may cause elevated rate of early

discontinuation as compare to products contain ethinyl

estradiol. Allied complications have included menstrual

disturbances, breakthrough bleeding/spotting (Kivinen S

et al., 1996; Schubert W et al., 1987) an improved

number of spotting or bleeding days for each cycle

(Hirvonen E et al., 1990) and extended bleeding (Wenzl

R et al., 1993). Depo-Provera (Depot medroxy

progesterone acetate) is an injection containing 150mg

progestin and administered intramuscularly that sustain

proper contraceptive efficacy for about 14 weeks. High

dose of progestin is inhibited ovulation in majority of

women (Hatcher RA. 1998). Merely 0.3% of females get

pregnancy during the first year of using Depo-Provera

injection (Trussell J. 1998). Breakthrough bleeding

means flow of blood at an unpredicted time throughout

the menstrual cycle and it requires to use tampon /sanitary

napkins. Surprising bleeding that does not need any safety

or protection is called as spotting (Thorneycroft IH.

1999).

Management of Irregular Uterine Bleeding

COMBINED ORAL CONTRACEPTIVE PILLS

Abnormal uterine bleeding occurs in 30% women

during first month of using combined oral contraceptives

pills (COCPs), this frequency decreases with the time.

The management of irregular uterine bleeding associated

with COCPs starts with counseling (Wallach M et al.,

2000; Hatcher RA et al., 1998). If bleeding sustain after

three months of pill use so that patient will be treated with

non-steroidal anti inflammatory drugs (NSAID) or

supplemental estrogen (Hatcher RA et al., 1998).

PROGESTIN-ONLY CONTRACEPTIVE

METHODS

Unusual uterine bleeding is commonly associated with

progestin-only contraceptives. Depo-Provera injection

contains higher dose of progestin. During first year of use

the incidents of irregular bleeding occur in 70% females

using injectable contraceptives (Hatcher RA et al.,

1998).

MATERIAL AND METHODS The study was carried out in family planning centers,

established by Government of Pakistan in different

clinical settings of Karachi and was conducted on young

females, maintained on hormonal contraceptives

including oral pills and Injectable (Depo) for at least two

years with the following chemical composition,

Combined oral contraceptives (COC) containing

0.15mg Levonorgestrol + 0.03mg Ethinyl

estradiol.

Depo-Provera (Depo), an injectable consisting of





150mg/mL Medroxyprogestrone Acetate.

Study population A cohort of 12214 women, between 19 to 49 years of

age, between January 2012 and December 2013 was

identified. The selection of the subjects was done on the

basis of following inclusion and exclusion criteria. The

selected subjects maintained on these agents, were

thoroughly evaluated for minor side effects and data was

recorded on predesigned data collection form. The data

collection form was constructed on the basis of such

similar studies conducted by several researchers like

(Talisuna AO et al., 2006; Stergachis A et al., 2010 and

Kuemmerle et al., 2011). Those females who were

qualifying inclusion criteria evaluated for their

reproductive history. Healthy females aged 19–49 years

looking for contraception participated in this study. The

exclusion criteria for this study is included the pregnancy;

incident of b3 menstrual cycles due to childbirth, abortion

/lactation; present use of an intrauterine device; obesity

(body mass index N30 kg/m2); hypersensitivity to any of

the hormonal drug ingredients.

Study assessments

All through study period, participants were requisite to

record tablet intake and bleeding events. Parameters for

bleeding patterns were assessed for every 3-months

reference period (as suggested by the World Health

Organization). Reference Period 1 is 1-90 days of

treatment; Reference Period 2 is next three months (day

91-180) of treatment. Eight Reference Periods were

included in this study. Parameters of bleeding pattern

observed in this study are the number of bleeding/spotting

days and the extent and duration of bleeding/spotting

events.

Statistical analysis:

Statistical analysis was performed by means of age

group, treatment group and duration of treatment. In this

study 457 women aged 19–35 years and 390 women aged

36–49 years per treatment group (total participants = 847)

were selected to estimate of the number of females

required to conduct exact specific comparisons between

treatment groups and between age groups during whole

treatment period for the number of bleeding/spotting days

per reference period.

RESULTS AND DISCUSSION

Table 1: (CROSSTAB AND CHI SQUARE TEST OF COMBINED ORAL CONTRACEPTIVE (COCs)

VERSES DEPO –PROVERA FOR Breakthrough bleeding

EFFECT OF DRUGS

Breakthrough Bleeding TOTAL P-Value

B.BleedingYes B.Bleeding No

COCs(Drug-I) Count 218 179 397

0.031

% B.Bleeding 43.8% 51.3% 46.9%

Depo-

Provera(Drug-II)

Count 280 170 450

% B.Bleeding 56.2% 48.7% 53.1%

Total Count 498 349 847

% B.Bleeding 100% 100% 100%

EFFECT OF AGE


B.Bleeding Yes B.Bleeding No

Age-I ( years)

19-35

Count 270

187

457

0.830

% B.Bleeding 54.2% 53.6% 54.0%

Age-II(years)

36-49

Count 228 162

390

% B.Bleeding 45.8% 46.4% 46.0%


% B.Bleeding 100% 100% 100%





EFFECT OF DURATION


B.BleedingYes B.Bleeding No

Duration-I

6months-1 year

Count 144 123

267

0.041

% B.Bleeding 28.9% 35.2% 31.5%

Duration-II

1year-2 years

Count 354 226

580

% B.Bleeding 71.1% 64.8% 68.5%


% B.Bleeding 100% 100% 100%

Table 2: (LOGISTIC REGRESSION PREDICTING BREAK THROUGH BLEEDING BETWEEN

DIFFERENT DRUGS, AGE GROUP AND DURATION)

Description DRUGS

(Drug-I (COCs)

Drug-II (Depo-Provera)

AGE

Age-I(19-35)

Age-II (36-49)

DURATION (Duration-

I (6months-1 year)

Duration-II (1year-2years)

Number of

reported cases

847

847

847

Omnibus Tests

of Model

Coefficient s

Chi-Square

p-value

Chi-Square p-value

Chi-Square p-value

4.65 0.031 0.033 0.855 3.78 0.042

Variables in the

Equation

Drugs verses Nausea Age verses Nausea Duration verses Nausea

Odds Ratio

(Exp β)

95% CI Odds Ratio

(Exp β)

95% CI Odds Ratio

(Exp β)

95% CI

Lower –

Upper

Lower –

Upper

Lower - Upper

0.739

0.56 – 1.97 1.026 0.780-

1.350

1.74 1.55-1.80

p-Value 0.031 0.855 0.04

DISCUSSION:

When Drug I (n=397) was compared with Drug II

(n=450) and were observed for the presence of

breakthrough bleeding i.e. 218/397 (43.8%) and 280/450

(56.2%) cases were found respectively and statistical

significance was observed by chi-square test (p=0.031)

(Table 1). Omnibus test was also applied which showed

overall significance of the model with the chi-square

value of 4.653. The value of odd ratio (β) obtained was

0.746 showed that the odd of breakthrough bleeding

(BTB) of Drug I was 0.746 times greater than Drug-II

with 95% confidence interval limit 0.56-1.97. Fig 2 shows

comparison of breakthrough bleeding using both drugs.

Similarly when Age -I (n=457) was compared with Age-II

(n=390) were observed for the presence of breakthrough

bleeding 270/457 (54.2%) and 228/390 (45.8%) cases

were observed respectively and statistical insignificance

was observed by chi-square test (P=0.830) (Table 1). By

using Omnibus test overall significance of the model was

obtained with the chi-square value of 0.033. The value of

odd ratio (β) obtained was 1.026 showed non-significant

results between Age and Break through Bleeding, as the

p-value is >0.05 and 95% (CI) confidence interval limit of





1.077- 1.895. Fig 3 shows breakthrough bleeding in

different age group.

When Duration I (n=267) was compared with

Duration-II (n=580) was observed for the presence of

breakthrough bleeding 144/267 (28.9%) and 354/580

(71.1%) were observed respectively and statistical

significance was observed by chi-square test (p=0.041)

(Table 1). We used Omnibus test to analyze the

association between side/adverse effect and duration that

showed overall significance of the model where the chi-

square value of 3.78. However the value of odd ratio (β)

was 1.74 showing that the odd of breakthrough bleeding

of Duration-II was 1.74 times greater than Duration-I with

95% confidence interval limit of 1.55- 1.80. Fig 1 shows

time period for Breakthrough bleeding.

CONCLUSION Abnormal menstrual patterns are most probably

produce discontinuation of hormonal contraception in

females. These disturbances include breakthrough

bleeding/spotting although bleeding due to missed

withdrawal is associated with existing levels of estrogen

and concealed endometrium.

Since breakthrough bleeding recovers with the time

and clinicians must educate women about benefits of

hormonal contraceptives as proper counseling can

discourage the misperceptions to encourage more women

to use contraception during child bearing age. For the

development of efficient treatment it is necessary to have

the knowledge of basic mechanism of bleeding. It is also

essential to review the approaches to the assessment and

treatment of abnormal bleeding associated with the use of

contraceptives.

Fig 1: Time period for Breakthrough bleeding





Fig 2: Comparison of breakthrough bleeding using both drugs

Fig 3: Breakthrough bleeding in different age group





REFERENCES

Andrist LC (1998). The impact of media attention,

familyhistory, politics and maturation on

women’sdecisions regarding hormone replacement

therapy.Health Care Women Int. 19:243-260.

Bjorn N, Backstrom T (1997). Compliance to HRT:

thesignificance of negative side effects and

moodsymptoms. Abstract: Eighth Annual Meeting

ofNAMS. Menopause. 4:283.

Den Tonkelaar I, Oddens BJ (2000). Determinants of

longterm hormone replacement therapy and reasons

forearly discontinuation. Obstet Gynecol. 95:507-512.

Ettinger B, Pressman A, Silver P(1999). Effect of ageon

reasons for initiation and discontinuationof hormone

replacement therapy. Menopause. 6:282-289.

Hatcher RA, Guillebaud J. The pill: combined oral

contraceptives. In: Hatcher RA. Contraceptive

technology. 17th ed. rev. New York: Ardent Media,

1998:405–66.

Hunter MS, O’Dea I, Britten N (1997). Decision-

makingand hormone replacement therapy: a

qualitativeanalysis. Soc Sci Med. 45:1541-1548.

Hatcher RA (1998). Depo-Provera, Norplant, and

progestin- only pills. In: Hatcher RA. Contraceptive

technology. 17th ed. rev. New York: Ardent Media. 467-

509.

Hatcher RA, Guillebaud J.(1998). The pill: combined oral

contraceptives. In: Hatcher RA. Contraceptive

technology. 17th ed. rev. New York: Ardent Media. 405-

66.

Hirvonen E, Vartiainen E, Kulmala Y (1990). A

multicenter trial with a new OC using a natural estradiol

and cyproterone acetate for women over 35. Adv

Contracep. 6:248.

Kaunitz AM (1998). Oral contraceptive estrogen dose

considerations. Contraception. 58(3 suppl):S15–21.

Kivinen S, Saure A.(1996). Efficacy and tolerability of a

combined oral contraceptive containing 17 beta-estradiol

and desogestrel. Eur J Contracept Reprod Health

Care.1:183.

Koetsawang S, Mandlekar AV, Krishna UR (1980).A

randomized, double-blind study of two combined oral

contraceptives containing the same progestogen, but

different estrogens. World Health Organization Task

Force on Oral Contraception. Contraception. 21:445-59.

Kuemmerle A, Dodoo AN, Olsson S, Van Erps J, Burri C

(2011) Assessment of global reporting of adverse drug

reactions for anti-malarials, including artemisinin-based

combination therapy, to the WHO Programme for

International Drug Monitoring. Malar J 10: 57.

Paul C, Skegg DC, Williams S (1997). Depot

medroxyprogesterone acetate. Patterns of use and reasons

for discontinuation. Contraception. 56:209–14.

Newton KM, LaCroix AZ, Leveille SG (1997). Women’s

beliefs and decisions about hormonereplacement therapy.

J Womens Health. 6:459-465.

Rossouw JE, Anderson GL, Prentice RL (2002).Risks and

benefits of estrogen plus progestin inhealthy

postmenopausal women: principal resultsFrom the

Women’s Health Initiative randomizedcontrolled trial.

JAMA. 288:321-333.

Sangi-Haghpeykar H, Poindexter A N Bate-man L,

Ditmore JR (1996). Experiences of injectable

contraceptive users in an urban setting. Obstet Gynecol.

88:227–33.

Serup J, Bostofte E, Larsen S, Westergaard J, Lebech PE

(1979). Natural oestrogens for oral contraception. Lancet.

2:471-2.

Schubert W, Cullberg G (1987). Ovulation inhibition with

17 beta-estradiol cyclo-octyl acetate and desogestrel. Acta

Obstet Gynecol Scand. 66:543-7.





Sitruk-Ware R(2006). New progestagens for

contraceptive use. Hum Reprod Update. 12:169-78.

Stubblefield PG (1994). Menstrual impact of

contraception. Am J Obstet Gynecol. 170(5 pt 2):1513–22.

Sulak P, Lippman J, Siu C, Massaro J, Godwin A (1999).

Clinical comparison of triphasic norgestimate/35

micrograms ethinyl estradiol and monophasic

norethindrone acetate/20 micrograms ethinyl estradiol.

Contraception. 59:161–6.

Stergachis A, Bartlein RJ, Dodoo A, Nwokike J, Kachur

SP (2010). A situational analysis of pharmacovigilance

plans in the Global Fund Malaria and U.S. President's

Malaria Initiative proposals. Malar J 9: 148.

Talisuna AO, Staedke SG, D'Alessandro U (2006)

Pharmacovigilance of antimalarial treatment in Africa: is

it possible? Malar J 5: 50.

Thorneycroft IH (1999). Cycle control with oral

contraceptives:a review of the literature. Am J Obstet

Gynecol. 80(2 pt 2):280-7.

Trussell J (1998). Contraceptive efficacy. In: Hatcher

RA.Contraceptive technology. 17th ed. rev. New York:

Ardent Media, 779-99.

Wenzl R, Bennink HC, van Beek A, Spona J, Huber J

(1993). Ovulation inhibition with a combined oral

contraceptive containing 1 mg micronized 17 beta-

estradiol. Fertil Steril. 60:616-9.

Vihtamaki T, Savilahti R, Tuimala R (1999). Why

dopostmenopausal women discontinue

hormonereplacement therapy? Maturitas. 33:99-105.

Wallach M, Grimes DA, Chaney EJ, Totowa, N.J.: Emron

(2000). Approach to common side effects. In: Modern

oral contraception: updates from The Contraception

Report. 70–6.

Citation of this article: Huma Dilshad, Safila Naveed, Rabia Ismail, Mohammad Harris Shoaib.

Bleeding pattern of women using hormonal contraceptives: A Pharmacovigilance study. Journal

and Biotechnology and Biosafety. 3(6):319-326

Source of Support: Nil Conflict of Interest: None Declared





SIMPLE UV SPECTROPHOTOMETRIC ASSAY OF NORFLOXACIN

FORMULATION AND THEIR COMPARATIVE STUDY

Research article ___________________________________

Safila Naveed1*, Sidra Sajid

2

___________________________________

Faculty of pharmacy, Jinnah University for

women, Karachi.

Corresponding author: [email protected],

[email protected]

ABSTRACT

Norfloxacin belongs to flouoroquinolones class of

antibiotics. Its structure is related to nalidixic acid. The

purpose of following study was to develop an easy, less

time consuming and economically affordable UV

Spectrophotometric method for assay of Norfloxacin in

tablet formulation and comparison of two brands of

Norfloxacin tablets. This assay procedure based on the

absorbance measurements of 2 brands of Norfloxacin

400mg tablet available in market at wavelength of

294nm.These two brands are taken and their solutions of

100ppm, 50ppm, 25ppm, 12.5 and 6.25ppm is prepared.

Results obtained by regression equation and regression

line. Percent assay is calculated by taking absorbance of

sample and standard solutions of 12.5ppm to predict the

availability of drug in both brands i.e. 98.27% and

102.6%. Results indicates that both brands meet USP/BP

limit.

Key words: Norfloxacin, UV spectrophotometer, Assay

_________________________________________________________________________________

INTRODUCTION

Figure 1: Structure of Norfloxacin

Molecular weight: 319.33

Norfloxacin (NFX) l-ethyl-6-fluoro-1,4-di-hydro-4-

oxo-7-(1-piperazinyl)-3-quinoline carboxylic acid, is a

synthetic antifluoroquinolon (Prapaporn B et al.,

2002). It is a synthetic drug was the firstly selected drug

for the treatment of diseases caused by Ecoli,

Salmonella, Campylobacter, V. cholera, Shigella and V.

cholera (Murakawa GJ et al., 2003). The drug is also

prescribed for optical infections as well as gonorrhea

and infection urinary tract (Van Bambeke et al., 2005).

The dose usually recommended is 400mg two times

daily. Half-life of NFX in plasma and serum is

measured3-4 hours and only approx bioavailability of

oral dose is 30–40%. Norfloxacin, aflouroquinolone

antibacterial agent structurally related nalidixic acid,

Merck Index (2001).

Following are few developed methods for assay of

Norfloxacin. The UV spectrophotometer was used for

mailto:[email protected]





the determination of the Norfloxacin in tablets at

wavelength 337nm using 0.05M NaOH as solvent

(Stankov, M et al., 1993). A reversed-phase high-

performance liquid chromatographic assay for

Norfloxacin (NFLX) has been developed. The method

gave good linearity (r2≥0.999) in the range 1–20μg/mL

using a Lichrosorb-RP-8 column (10μm, 20cm×g

4.6mm) at wavelength (278nm) at normal temperature.

Above method is also useful for the analysis of photo

degraded NFLX samples, and was useful to study the

photo-stability of NFLX tablets under different

conditions i.e., UV light, sunlight light or fluorescent

light (Córdoba-Borrego et al., 1999). To measure the

quantity or the content of Norfloxacin in its tablet by

UV spectrophotometer, the assay was carried out at the

wavelength of maximum absorption in hydrochloric

acid solution 0.1mol/L (Samanidou, V. F et al., 2003).

A simple titration method is also used for

Norfloxacin analysis as weight and crushes tablet equal

to 250mg of Norfloxacin. Dissolve in 80mL of

anhydrous acetic acid and titrate with 0.1N per chloric

acid, crystal violet used as indicator.

MATERIALS AND METHODS

UV visible 1601 Shimadzu double beam

spectrophotometer was used to measurement of Spectra.

The solvents which were used for the assay was

distilled water.

Wavelength Selection

About 100 to 6.25ppm of Norfloxacin was accurately

prepared in water. These solutions were scanned in the

200-400nm UV regions. The wavelength kept was

294nm and this wavelength was adopted for absorbance

measurement.

Standard Stock solution

Accurately weighed 10mg of Norfloxacin standard was

transferred to a volumetric flask and add sufficient

methanol to produce 100mL. This was sonicated 5 min

to dissolve it. Make up volume up to 100mL with water.

Transfer 6.25mL from stock solution to 50mL

volumetric flask and make up volume with water.

Sample preparation

Two different brands of Norfloxacin from the

marketed sample were weighed and crushed uniformly

with the help of a mortar and pestle. By calculating the

average weighed sample powder equivalent to 10mg of

Norfloxacin was transferred into a volumetric flask

containing 10mL water. The solutions were sonicated

for about 5 min and then make up volume up to 100ml

with water. Now transfer 6.25mL from stock solution

into 50mL volumetric flask and make up volume with

water (Safila N, 2014).

PROCEDURE After preparation of standard and tablet solutions,

strength of solution 12.5ppm in 50mL Absorbance of

the sample preparation and standard preparation in 1cm

cell at the wavelength of maximum. Absorbance at

about 294nm using a spectrophotometer, using the

blank solution, calculate the quantity in mg, of

Norfloxacin per tablet.

RESULTS AND DISCUSSIONS

Using spectrophotometer on both brands of

Norfloxacin tablets during the study carried out

pharmaceutical assay. Table-1.1 shows brand as A and

B identification and % assay of both brands i.e. 98.27%

and 102.6% respectively. Table 1.2 and 1.3 shows

absorbance of brand A and brand B respectively.

Graphical representation of linearity of equation and

regression value for both brands given in figure 2 and 3.

These results indications that Absorbance is directly

proportional to concentration thus, it obeys to Beers

lambert law. Our research group work on different

assay method those were useful for pharmacy

professionals (Safila et al., 2014,15).





TABLE 1.1: SHOWS % ASSAY OF TWO BRANDS

Brand ID ABS at 294nm % Assay

Brand A 0.342 98.27%

Brand B 0.358 102.6%

Standard (Norfloxacin) 0.348 --------

TABLE 1.2: SHOWS ABS OF BRAND A AT DIFFERENT CONC.

Conc ppm Abs

100 2.42

50 1.25

25 0.718

12.5 0.342

6.25 0.17

TABLE1.3: SHOWS ABS OF BRAND B AT DIFFERENT CONC.

Conc ppm Abs

100 2.59

50 1.28

25 0.625

12.5 0.358

6.25 0.19

FIGURE 2: LINEARITY OF EQUATION FOR BRAND A

y = 0.0237x + 0.0606 R² = 0.9982

0

0.5

1

1.5

2

2.5

3

0 20 40 60 80 100 120

Series1

Linear (Series1)





FIGURE 3: LINEARITY OF EQUATION FOR BRAND B

CONCLUSION

It is concluded that brand B is more potent than brand A while both brands shows good linearity of

equation and regression values. The proposed method for the assay of commercially available Norfloxacin

tablet formulation is very simple, accurate, least time consuming and rapid. It can be easily used for routine

quality control for monitoring the assay, in process samples and tablet formulation.

REFRENCES

Boonme P., Phadoongsombut N., Ingkatawornwong S.,

& Faroongsarng D (2002). The formulation

development and stability study of norfloxacin

suspension. Thammasat Int. J. Sc. Tech. 7(1):1-4.

Córdoba-Borrego M., Córdoba-Dıaz M., & Córdoba-

Dıaz D (1999). Validation of a high-performance liquid

chromatographic method for the determination of

norfloxacin and its application to stability studies

(photo-stability study of norfloxacin). Journal of

pharmaceutical and biomedical analysis. 18(6):919-926.

Naveed, Safila. (2014). Simple UV spectrophotometric

assay of Atorvastatin API formulation and their

comparative study. Global Journal of Medical Research

14.2

Sable, D., & Murakawa, G. J. (2003). Quinolones in

dermatology. Clinics in dermatology. 21(1):56-63.

Safila Naveed, Lailoona Jaweed (2014). UV

spectrophotometric assay of Ketoconazole oral

formulations American Journal of Biology and Life

Sciences. 2(5):108

http://www.openscienceonline.com/journal/archive?jour

nalId=704

Safila Naveed, Hina Qamar, Wardha Jawaid, Urooj

Bokhari (2014). Simple UV Spectrophotometric Assay

of Amlodipine. American Journal of Chemistry and

Application. 1(4):66-69.

http://www.aascit.org/journal/archive?journalId=905

Safila Naveed,Farya Zafar (2014). UV

Spectrophotometric Assay of Famotidine Formulations.

y = 0.0256x + 0.0161 R² = 0.9995

0

0.5

1

1.5

2

2.5

3

0 20 40 60 80 100 120

Series1

Linear (Series1)

http://www.openscienceonline.com/journal/archive?journalId=704







American Journal of Pharmacy and Pharmacology.

1(3):28-31.


Safila Naveed and Fatima Qamar (2014) Simple UV

spectrophotometric assay of Metronidazole Open

Access Library Journal. 1: e615:1-4.

http://dx.doi.org/10.4236/oalib.1100615

Safila Naveed, Hina Qamar, Wardha Jawaid, Urooj

Bokhari (2014) Simple UV spectrophotometric assay of

Glimepride Journal of Clinical Medicine. 2(4): 94

http://www.openscienceonline.com/journal/archive?jour

nalId=717

Safila Naveed, Hina Rehman, Fatima Qamar, Syeda

Zainab (2015). Method development of Perindopril

using UV spectrophotometer. International journal of

pharmaceutical Quality Assurance. 6(1)

http://ijpqa.com/#

Safila Naveed, Hina Rehman, Fatima Qamar, Syeda

Zainab (2015) Development of a spectrophotometric

method for the assay of Diclofenac potassium JIPBS. 2

(1): 7-11.

Samanidou V. F., Demetriou C. E., Papadoyannis I. N.

(2003). Direct determination of four fluoroquinolones,

enoxacin, norfloxacin, ofloxacin, and ciprofloxacin, in

pharmaceuticals and blood serum by HPLC. Analytical

and bioanalytical chemistry. 375(5): 623-629.

Stankov M., Stankov D., Milićević Z., Veselinović D.,

Djurdjević P (1993). Fluorometric and derivative

spectrophotometric determination of norfloxacin.

Spectroscopy letters. 26(9): 1709-1714.

Van Bambeke F., Michot J. M., Van Eldere J., &

Tulkens P. M (2005). Quinolones in 2005: an update.

Clinical Microbiology and infection. 11(4): 256-280.

Citation of this article: Safila Naveed, Sidra Sajid (2015). SIMPLE UV

SPECTROPHOTOMETRIC ASSAY OF NORFLOXACIN FORMULATION AND THEIR

COMPARATIVE STUDY. Journal and Biotechnology and Biosafety. 3(6):327-331

Source of Support: Nil Conflict of Interest: None Declared




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