Review on various analytical methodologies for Azithromycin

Aakanksha Sinha; S. J. Daharwal*

doi:10.52228/NBW-JAAB.2022-4-1-5

About Journal

editor@newbioworld.org

NewBioWorld

A Journal of Alumni Association of Biotechnology
ISSN: 2583-8105 | Issues per year: Bi-annually

Article in HTML

Review on various analytical methodologies for Azithromycin

Author(s): Aakanksha Sinha¹, S. J. Daharwal*²

Email(s): ¹, ²daharwalresearch@rediffmail.com

Address:

Published In: Volume - 4, Issue - 1, Year - 2022

Keywords:

Cite this article:
Aakanksha Sinha, S. J. Daharwal (2022) Review on various analytical methodologies for Azithromycin. NewBioWorld A Journal of Alumni Association of Biotechnology,4(1):20-27.

View PDF

Please allow Pop-Up for this website to view PDF file.

NewBioWorld A Journal of Alumni Association of Biotechnology (2022) 4(1):20-27

REVIEW ARTICLE

Review on various analytical methodologies for Azithromycin

Aakanksha Sinha and S. J. Daharwal*

University Institute of Pharmacy, Pt. Ravishanker Shukla University, Raipur, Chhattisgarh, India

*Corresponding Author Email- daharwalresearch@rediffmail.com

ARTICLE INFORMATION

ABSTRACT

Article history:

Received

13 April 2022

Received in revised form

18 April 2022

Accepted

28 April 2022

Keywords:

Azithromycin;

UV- Visible Spectrophotometer; HPLC;

HPTLC;

Bioanalytical method

The azithromycin broad-spectrum macrolide antibiotic, is generally used to treat genitourinary, respiratory, and enteric infections. The development of analytical techniques and the different currently employed techniques for estimating azithromycin, both in pharmaceutical dose form or in bulk, are the primary focus of the current study. Analytical techniques are crucial for determining compositions as they allow us to employ state-of-the-art analytical equipment to collect both qualitative and quantitative data. Chromatographic, electrochemical, spectroscopic, and other methods can be used to analyse azithromycin. These methods aid in understanding key process factors and restricting the negative impact they have on accuracy and precision. Analytical technique development is required for maintaining high commercial product quality standards and to comply with rules and regulations. In response to the reference, regulatory bodies in various nations have developed policies and procedures for giving approval, authentication, and registration.

1. Introduction

The antibiotic azithromycin was developed in 1980 by the Yugoslav Pharmaceutical Firm Pliva, and it received medical approval in 1988 under the trade name Sumamed. It obtained a patent in 1981. It was initially authorised by the FDA in 1991 and is listed on the World Health Organization's list of essential medicines (Greenwood et al. 2008). A broad-spectrum antibiotic with a significant amount of tissue penetration and an extended half-life is azithromycin (Pereyre et al. 2016). Macrolide antibiotics specifically bind to the 50S component of the ribosomes in the apicoplast and inhibit protein synthesis (Choemunng et al. 2010). Azithromycin is a 15-membered semi-synthetic antibiotic that belongs to a member of the macrolide family. Its azalactone ring is structurally distinct from erythromycin in that it contains methyl-substituted nitrogen at position C-10, which increases azithromycin's activity by 15 times compared to erythromycin at position C-9 (Mallah et al. 2011). Erythromycin A was converted into azithromycin by treating it with hydroxylamine hydrochloride, a base, and methanol at reflux temperature for 10 hours to produce oxime. The oxime was separated, purified, and then Beckmann's rearrangement was performed on it in aqueous acetone for two hours at 50 degrees in order to produce the intermediary (6,9-iminoether). With the help of sodium borohydride in methanol or by catalytic hydrogenation in the presence of platinum dioxide and acetic acid as solvents, the iminoether was reduced to the secondary amine (Djokić et al. 1986). IUPAC name of azithromycin (2R, 3S, 4R, 5R, 8R, 11R, 13S, 14R)-11-(2S, 3S, 4S, 6R)-4-(dimethyl amino)- 3-hydroxy-6-methyltetrahydro-2H-pyran-2-yl)oxy)-2-ethyl-3,4,10- trihydroxy-13-(2R, 4R, 5S, 6S)-5-hydroxy-4-methoxy-4,6-dimethyltetrahydro-2H-pran-2-yl)oxy)-3, 5, 6, 8, 10, 14- hexamethyl-1-oxa-6- azacyclopentadecan-15-one (PubChem). Patients with acute bronchitis, pneumonia, sinusitis, pharyngitis, tonsillitis, and otitis media have reported success with it. It is extremely efficient against skin and soft tissue infections, sexually transmitted diseases, and upper and lower respiratory tract infections (Chen et al. 2007).

1.1 Physicochemical Properties:

Figure 1: Chemical structure of Azithromycin

A crystalline powder that is white, tasteless, and odourless and has the chemical formula C₃₈H₇₂N₂O₁₂. Almost insoluble in water, azithromycin is easily soluble in methylene chloride and anhydrous ethanol has an average molecular weight of 748.9845 g/mol and an ionisation constant of 7.34 (Babić et al. 2007, EP 2005).

DOI: 10.52228/NBW-JAAB.2022-4-1-5

1.2 Pharmacodynamic properties:

By preventing protein synthesis and translation, macrolides treat bacterial infections by preventing growth of bacteria. Azithromycin has been used in chronic respiratory inflammatory illnesses for this reason because it also has additional immunomodulatory properties (Marvig et al. 2012).

1.3 Pharmacokinetic properties:

Azithromycin has a 37% bioavailability after oral ingestion. Food has little impact on absorption. P-glycoprotein (ABCB1) efflux transporters are thought to mediate macrolide absorption in the intestines. A primary route of elimination for azithromycin is biliary excretion, primarily as unchanged medication. Over the course of one week, roughly 6% of the prescribed dose gets found in urine as an unmodified medication (Pereyre et al. 2006). Half-life for terminal elimination is 68 hours. In humans, the serum protein binding varies, ranging from 51% at 0.02 g/mL to 7% at 2 g/ml. It causes renal and liver damage (Singlas et al. 1995).

1.4 Approved dosage form of Azithromycin:

The available dosage form in the market along with the brand name, active pharmaceutical ingredients and concentration are presented in Table 1.

2. Need of Analytical method:

Analytical technique advances are incorporated into official test procedures. Therefore, these methods are used by quality control laboratories to assess the functionality, identity, purity, safety, and efficacy of medication products. Regulatory authorities find the analytical methods utilised in production to be of the utmost importance and utility. For regulatory authorities to approve the medication, the applicant must demonstrate control over the entire drug development process using recognised analytical techniques (Ravisankar et al. 2015). The ICH has published analytical guidelines papers for stability testing (Q1), validation of analytical procedures (Q2), contaminants in drug substances and products (Q3), and specifications for new drug substances and products (Q6) (Breaux et al. 2003).

3. Analytical Method Development by UV Spectrophotometer:

UV-visible spectroscopy is the study of interactions between matter and electromagnetic radiation in this range of wavelengths. The ultraviolet's (UV) wavelength ranges from 200 to 400 nanometres (FJ et al. 2004). It is based on the Beer-Lambert rule, which states that the absorbance of a solution and its path length are directly related. It can therefore be used to determine the absorber concentration in a solution for a specific path length. Understanding how quickly absorbance changes with concentration is crucial (Verma et al. 2018). Table 2 includes a few of the examples.

4. Analytical method development by HPLC

High performance liquid chromatography (HPLC) is by far the most widely used separation technique and one of the most well-established analytical processes. It has been used in laboratories all around the world for more than 40 years for a variety of purposes, including pharmaceutical sciences, clinical chemistry, food and environmental assessments, synthetic chemistry, etc. (Zotou et al. 2012). In this method, the stationary phase could be a liquid or a solid. Using a liquid mobile phase, the components of a combination can be separated using HPLC. The term HPLC describes liquid chromatography in which the liquid mobile phase is mechanically pumped through the column while the stationary phase is contained in a column (Yandamuri et al. 2013). The column is the vibrating heart of HPLC systems. A good silica and binding method will result in a repeatable and symmetrical peak, which is necessary for accurate certification. Examples of frequently utilised RP columns include C18 (USP L1), C8 (USP L8), Phenyl (USP L11), and Cyno (USP L18) (Ravisankar et al. 2014). Researchers usually use an HPLC to create the procedure. Table 3 provides examples of some of those.

5. Analytical Method Development Using HPTLC Method

Both qualitative and quantitative tasks response positively to the powerful analytical method HPTLC (Sharma et al. 2008). Depending on the type of adsorbents used on the plates and the development solvent system, partition, adsorption, or both processes may be to blame for separation. The many components of HPTLC fundamentals include theory, instrumentation, implementation, optimisation, validation, automation, and qualitative and quantitative analysis (Ambati et al. 2021). Table 4 displays the variations between TLC and HPTLC. The method is often developed by researchers using an HPTLC. Examples of a few of those are given in Table 5.

Table 1: Azithromycin formulations with brand Name and Concentrations

S. No.	Drug Formulation	Concentration	Route of administration	Brand Name
1.	Tablet	600 mg	Oral	Act Azithromycin
2.	Tablet	250 mg	Oral	Azithromycin
3.	Solution	10mg/1mL	Ophthalmic	AzaSite
4.	Solution	1 % w/w	Ophthalmic	Azasite
5.	Injection	500 mg/4.8mL	Intravenous	Azithromycin
6.	Powder, for suspension	100 mg / 5 mL	Oral	Azithromycin
7.	Powder, for solution	200 mg / 5 mL	Oral	Azithromycin

Table no: 2 Analytical method development using UV spectrophotometer

S. No.	Sample / Dosage form	Method / Instrument model	Solvent / Solution	Wavelength	References
1.	Tablet	Shimadzu UV-spectrophotometer, model 1800	Distilled water	Area under curve method 219-224 nm and 275-302 nm	(Gulhane et al. 2021)
2.	Bulk powder and laboratory-prepared tablets	UV-VIS spectrometer	Methanol	Direct measurement of absorbance method 482 nm and 224 nm first derivative spectrophotometric method (475–490 nm) and (280–253 nm)	(El-Yazbi et al. 2020)
3.	Tablet	UV-Visible double beam spectrophotometer Shimadzu UV 1800	Methanol	Simultaneous equation method 251 nm and derivative spectroscopic methods Zero order derivatives 251 nm, first order derivative spectroscopic method 245 nm	(Baig et al. 2015)
4.	Tablet	Double-beam Jasco-630 UV- Visible spectrophotometer	Methanol	Simultaneous equation method 254 nm	(Magar et al. 2012)
5.	Bulk Powder	UV/VIS double beam spectrophotometer (model 1800)	0.1N HCl pH 6.8 phosphate buffer	Simultaneous vierodt’s estimation method equation 298 nm	(Malhotra et al. 2021)
6.	Bulk powder	Double-beam UV/Visible spectrophotometer	Bidistilled water	475 nm	(Omara et al. 2014)
7.	Tablet	UV/Vis-1800 double beam UV/Visible spectrophotometer	Methanol	Second order derivative spectra 326.4 nm	(Shah et al. 2012)
8.	Tablet	Shimadzu 1601 UV visible spectrophotometer	Methanol	482 nm	(Haleem et al. 2006)

Table 3. Analytical method development using HPLC method

S. No.	Sample	Stationary phase/column	Mobile phase	Wavelength (nm)	Flow rate (ml/min)	RT (min)	Reference
1.	Tablet	Octadecylsilyl amorphous organosilica polymer, column 25 cm x 4.6 mm	a. 0.18 per cent w/v solution of anhydrous disodium hydrogen phosphate with the pH adjusted to 8.9 with dilute orthophosphoric acid or with dilute sodium hydroxide solution b. mixture of 25 volumes of methanol and 75 volumes of acetonitrile	210	1	-	(IP 2010)
2.	Tablet	C18 column (100 mm x 4.6 mm, 5 μm particle size)	Methanol: Toluene: 100mMpotassium dihydrogen phosphate buffer (60:30:10, v/v/v)	218	1	5.20	(Afzal et al. 2016)
3.	Bulk	Hypersil GOLD C-18 column packed with deactivated silica (250 mm x 4.6 mm x 5 µm)	Ammonium acetate solution: acetonitrile (18:82, v/v)	210 nm	0.7	-	(Abou et al. 2017)
4.	Bulk and Tablet	Xterra C18 column (150 ×4.6 mm; 5µ)	Acetonitrile and phosphate buffer (pH adjusted to 7.5) in a ratio of 50:50 v/v	215	1	-	(Dewan et al. 2013)
5.	Bulk and Tablet	Kromasil C18	Ortho Phosphoric Acid buffer and Methanol in the ratio of 70:30 v/v	292	1	2.8	(Gundala et al. 2014)
6.	Bulk and Tablet	ODS-3 (250 mm ×4.6 mm x 5 μm)	Methanol: Phosphate buffer (9:1, v/v)	210	1.2	-	(Al-Hakkani et al. 2019)
7.	Tablet	C18 (250 mm × 4.6 mm, 5 μ) column	Phosphate buffer: Methanol (pH adjusted to 5.0 with ortho phosphoric acid)	215	1	-	(Hinge et al. 2015)
8.	Tablet	C18 column (250 mm x 4.6 mm id, 5 µm particle size)	Methanol: potassium dihydrogen phosphate buffer (60:40, v/v	279.6	1	5.08	(Kamble et al.2015)
9.	Bulk and pharmaceutical formulation	supleco C18 (25cm×4.6 mm, 5 µm)	80:20 Na₂HPO₄: Methanol	273	1	2.77	(Nagaraju et al. 2018)
10.	Tablet	Hypersil C18 column (250×4.6 mm, 5μ)	Phosphate buffer(pH-3.5) and acetonitrile in the ratio of 45:55%v/v	240	1	4.472	(Padmavathi et al. 2015)
11.	Tablet	LiChroCART® 125×4.6 mm	Mixture of buffer, acetonitrile and methanol (60:20:20) a	215	1	5	(Zubata et al. 2002)
12.	Tablet	C18 phenomenex Gemini 5m, 250cm x 4.6mm	Acetonitrile and mono basic potassium phosphate buffer of pH 8.5 in the ratio of 65:35 v/v	220	2	6.1	(Raja et al. 2010)
13.	Bulk and tablet dosage	Xterra RP-18 column (250 × 4.6 mm ID, 5 mM)	Water: methanol (63:37% v/v)	210	1.2	-	(Sahoo et al. 2015)
14.	Tablet	Hypersil- keystone C18 (250 X 4.60 mm), 5µm column	Buffer Potassium dihydrogen Phosphate: Acetonitrile (pH 7.5 adjusted with ortho phosphoric acid).	215	1.2	9.761	(Sahu et al. 2015)
15.	Tablet	Xterra RP18 (250 mm × 4.6 mm, 5 µm)	Acetonitrile–dipotassium phosphate (30 mM) (50:50, v/v) (pH 9.0)	215	1.7	11.5	(Shaikh et al. 2008
16.	Tablet	C18 column, 4.6×150mm, 5µm	Ammonium acetate buffer pH 6 ±0.02 pH and methanol (30:70 %v/v)	262	1	4.862	(Vennela et al. 2014)

Table 4. Major Differences between TLC and HPTLC (Gunjal et al. 2022)

Parameters	TLC	HPTLC
Technique	Manual	Instrumental
Efficiency	Less	High
Layer	Lab made	Precoated
Mean particle size	10-12 µm	5-6 µm
Layer thickness	250 µm	100µm
Plate height	30 µm	12 µm
Solid support	Silica gel, Alumina, Kieselguhr	Silica gel-Normal Phase C8 and C18-reverse phase
Spotting of sample	Manual (Capillary/Pipette)	Syringe
Volume of sample	1-5µL	0.1-0.5 µL
Separation	10-15 cm	3-5 cm
Separation time	20-200 min	3-20 min
Analysis time	Slower	Storage migration distance and the analysis time is greatly reduced
Scanning	Not possible	Use of UV/visible/fluorescence scanner

Table 5. Analytical Method Development Using HPTLC Method

S. No.	Sample	Stationary Phase/ Column	Mobile phase	Wavelength (nm)	Reference
1.	Tablet	Aluminium plates precoated with silica gel 60F254	Ethyl acetate: methanol: acetone: toluene: ammonia (1:5:7:0.5:0.5, v/v)	235	(Gawande et al. 2018)
2.	Tablet	Aluminium plates precoated with silica gel 60F254	Chloroform: ethanol :25% ammonia 6:14:0.2 (v/v/v)	483	(Bouklouze et al. 2017)
3.	Tablet	Aluminium plates precoated with silica gel 60F254	Methanol: ethyl acetate: toluene: ammonia (30%) 4.5:3.5:2:0.4%v/v/v/v	450	(Raval et al. 2014)

Conclusion

The focus of the study has primarily been on the various analytical techniques used to estimate azithromycin as well as medicines in bulk form. The development of analytical techniques such as UV spectrophotometry, HPLC, HPTLC, and RP-HPLC has been the focus of research. The developed analytical procedures have better levels of automation and sample throughput and are more sensitive, dependable, reproducible, and precise. A literature review is conducted to gather data on various analytical and instrumental analytical techniques. A unique analytical approach could be developed using such data.

Competing Interests

Authors report no conflict of interest concerning this review article.

References

Abou Assi, R., Darwis, Y., Abdulbaqi, I.M. and Asif, S.M., 2017. Development and validation of a stability-indicating RP-HPLC method for the detection and quantification of azithromycin in bulk, and self-emulsifying drug delivery system (SEDDs) formulation. Journal of Applied Pharmaceutical Science, 7(9), pp.020-029.

Afzal, S.J.S., Khan, P.M.A.A. and Baig, M.S., 2016. RP-HPLC analytical method development and validation for azithromycin and cefpodoxime in tablet dosage form.

Al-Hakkani, M.F., 2019. A rapid, developed and validated RP-HPLC method for determination of azithromycin. SN Applied Sciences, 1(3), p.222.

Ambati P, Bala Krishna P, Srinivasa Rao Y, Varaprasada Rao K and Deepthi R. 2021. Instrumentation and future prospectus of HPTLC-A. World Journal of Pharmaceutical Research Vol.10 Issue 6, 650-661

Babić, S., Horvat, A.J., Pavlović, D.M. and Kaštelan-Macan, M., 2007. Determination of pKa values of active pharmaceutical ingredients. TrAC Trends in Analytical Chemistry, 26(11), pp.1043-1061.

Baig, M.S., Anees, M.I., Muntasiruddin, K., Naeem, S. and Khan, J., 2015. Simultaneous estimation of azithromycin and cefpodoxime proxetil from its tablet dosage form by UV Visible spectroscopic methods. World Journal of Pharmacy and Pharmaceutical Sciences, 4, pp.1420-1430.

Bouklouze, A., Kharbach, M., Cherrah, Y. and Vander Heyden, Y., 2017, March. Azithromycin assay in drug formulations: Validation of a HPTLC method with a quadratic polynomial calibration model using the accuracy profile approach. In Annales Pharmaceutiques Francaises (Vol. 75, No. 2, pp. 112-120). Elsevier Masson.

Breaux J, Jones K and Boulas P. Analytical methods development and validation. Pharm. Technol. 2003; 1:6-13.

Chen, L., Qin, F., Ma, Y. and Li, F., 2007. Quantitative determination of azithromycin in human plasma by ultra-performance liquid chromatography–electrospray ionization mass spectrometry and its application in a pharmacokinetic study. Journal of Chromatography B, 855(2), pp.255-261.

Choemunng, A. and Na-Bangchang, K., 2010. An alternative liquid chromatography-mass spectrometric method for the determination of azithromycin in human plasma and its application to pharmacokinetic study. Journal of liquid chromatography and related technologies, 33(16), pp.1516-1528.

Dewan, I., Amin, T., Hossain, M.F., Hasan, M., Chowdhury, S.F., Gazi, M. and Islam, S.A., 2013. Development and validation of a new HPLC method for the estimation of azithromycin in bulk and tablet dosage form. Int J Pharm Sci Res, 4(1), pp.282-286.

Djokić, S., Kobrehel, G., Lazarevski, G., Lopotar, N., Tamburašev, Z., Kamenar, B., Nagl, A. and Vicković, I., 1986. Erythromycin series. Part 11. Ring expansion of erythromycin A oxime by the Beckmann rearrangement. Journal of the Chemical Society, Perkin Transactions 1, pp.1881-1890.

El-Yazbi, A.F., Khamis, E.F., Youssef, R.M., El-Sayed, M.A. and Aboukhalil, F.M., 2020. Green analytical methods for simultaneous determination of compounds having relatively disparate absorbance; application to antibiotic formulation of azithromycin and levofloxacin. Heliyon, 6(9), p.e04819.

European Pharmacopoeia, edition vol. 16. 2005,1649.

FJ, SDH and Nieman DA, Introduction to UV Spectroscopy in Principle of Instrumental Analysis. 2004, 5th ed., Thomson book/Cole, 156.

Gawande, V.T., Bothara, K.G. and Satija, C.O., 2018. Validated stability-indicating HPTLC method for cefixime and azithromycin with preparative isolation, identification, and characterization of degradation products. Acta Chromatographica, 30(4), pp.212-218.

Greenwood, D., 2008. Antimicrobial Drugs: Chronicle of a Twentieth Century Medical Triumph. OUP Oxford.

Gulhane, C.A., Motule, A.S., Manwar, J.V., Sawarkar, H.S., Ajmire, P.V. and Bakal, R.L., 2021. UV-Visible Spectrophotometric estimation of azithromycin and cefixime from tablet formulation by area under curve method. World Journal of Pharmaceutical Sciences, pp.163-168.

Gundala, U., Bonagiri, C. and Nayakanti, D., 2014. Simultaneous estimation of azithromycin and cefixime in bulk and pharmaceutical formulation by Reverse Phase High Performance Liquid Chromatography (RP-HPLC), World Journal of Pharmaceutical Research Vol.3.

Gunjal Sanket, B and Dighe, PR, Analysis of herbal drugs by HPTLC: A review. Asian Journal of Pharmaceutical Research. 2022;10(2):125-8.

Haleem, D., Shireen, E., Haleem, M., Kaye, W., Bailer, U., Frank, G., Wagner, A. and Henry, S., 2006. Degradation studies of azithromycin and its spectrophotometric determination in pharmaceutical dosage forms. Pak. J. Pharm. Sci, 19(2), pp.98-103.

Hinge, M.A., Desai, N.A., Mahida, R.J. and Dalvadi, H.P., 2015. Spectrophotometric and High-Performance Liquid Chromatographic Determination of Ofloxacin and Azithromycin in Pharmaceutical Tablets. Pharmaceutical Methods, 6(1), p.26.

https://pubchem.ncbi.nlm.nih.gov/#query=azithromycin.

Kamble, R.N., Kumar, A.P. and Mehta, P.P., 2015. RP-HPLC Analytical Method Development and Validation for Azithromycin and Levofloxacin in Tablet Dosage Form. Int. J. Pharm. Sci. Rev. Res, 31, pp.162-165.

Magar, S.D., Tupe, A.P., Pawar, P.Y. and Mane, B.Y., 2012. Simultaneous spectrophotometric estimation of cefixime and azithromycin in tablet dosage form. Journal of Current Pharma Research, 2(3), p.535.

Malhotra, D., KALIA, A. and Tomar, S., 2011. Validated UV spectroscopic method for simultaneous estimation of Azithromycin and Prednisolone. International Journal of Pharmacy and Pharmaceutical Sciences, 3, pp.299-302.

Mallah, M.A., Sherazi, S.T.H., Mahesar, S.A. and Rauf, A., 2011. Assessment of azithromycin in pharmaceutical formulation by Fourier-transform infrared (FT-IR) transmission spectroscopy. Pakistan Journal of Analytical and Environmental Chemistry, 12(1 and 2), p.7.

Marvig, R.L., Søndergaard, M.S., Damkiær, S., Høiby, N., Johansen, H.K., Molin, S. and Jelsbak, L., 2012. Mutations in 23S rRNA confer resistance against azithromycin in Pseudomonas aeruginosa. Antimicrobial Agents and Chemotherapy, 56(8), pp.4519-4521.

Nagaraju, K. and Chowdary, Y.A., 2018. Analytical Method Development and Validation for The Simultaneous Estimation of Azithromycin and Cefixime by RP-HPLC Method in Bulk and Pharmaceutical Formulations. International Journal of Research in Science and Technology, Vol. 4.

Omara, H.A., Ahmed, H.A., El-Mahdy, A.A. and Musbah, S.A., 2014. New spectrophotometric determination of azithromycin in pure and dosage forms using N-bromosuccinimide and potassium permanganate as oxidants. World J. of Pharmacy and Pharmaceutical Sciences, 3(4), pp.100-112.

Padmavathi, K. and Rao, M.S., 2015. A new validated RP-HPLC method for the simultaneous determination of ambroxol and azithromycin in combined dosage form. Bulletin of Pharmaceutical and Medical Sciences (BOPAMS), 3(4).

Pereyre, S., Renaudin, H., Charron, A., Bébéar, C. and Bébéar, C.M., 2006. Emergence of a 23S rRNA mutation in Mycoplasma hominis associated with a loss of the intrinsic resistance to erythromycin and azithromycin. Journal of Antimicrobial Chemotherapy, 57(4), pp.753-756.

Pharmacopeia, Indian. Vol-I and II Indian Pharmacopeia Commission. Ghaziabad, Govt of India Ministry of Health and Family Welfare. 2010: 861-62.

Raja, M.S., Shan, S.H., Moorthy, S. and Perumal, P., 2010. RP-HPLC method development and validation for the simultaneous estimation of azithromycin and ambroxol hydrochloride in tablets. International Journal of Pharm Tech Research, 2(1), pp.36-9.

Raval, P.L., Mehta, F.A., Ahir, K.B. and Bhatt, K.K., 2014. Simultaneous estimation of azithromycin dihydrate and cefixime trihydrate in pharmaceutical formulation by HPTLC method. Journal of Liquid Chromatography and Related Technologies, 37(13), pp.1805-1818.

Ravisankar P, Gowthami S and Rao GD. A review on analytical method development. Indian Journal of Research in Pharmacy and Biotechnology. 2014;2(3):1183.

Ravisankar P, Navya CN, Pravallika D and Sri DN. A review on step-by-step analytical method validation. IOSR J Pharm. 2015;5(10):7-19.

Sahoo, D.K. and Sahu, P.K., 2015. Chemometric approach for RP-HPLC determination of azithromycin, secnidazole, and fluconazole using response surface methodology. Journal of Liquid Chromatography and Related Technologies, 38(6), pp.750-758.

Sahu, A., Jatavb, M.P., Vinodec, R. and Yadavd, K., 2015. Development and validation of a reversedphase HPLC method for simultaneous estimation of azithromycin in tablet dosage form. International Journal Pharmaceutical Chemistry, 5, p.09.

Shah, V. and Raj, H., 2012. Development and validation of derivative spectroscopic method for simultaneous estimation of cefixime trihydrate and azithromycin dihydrate in combined dosage form. International Journal of Pharmaceutical Sciences and Research, 3(6), p.1753.

Shaikh, K.A., Patil, S.D. and Devkhile, A.B., 2008. Development and validation of a reversed-phase HPLC method for simultaneous estimation of ambroxol hydrochloride and azithromycin in tablet dosage form. Journal of Pharmaceutical and Biomedical Analysis, 48(5), pp.1481-1484.

Sharma A, Shanker C, Tyagi LK, Singh M and Rao CV 2008. Herbal medicine for market potential in India: an overview. Academy Journal of Plant Sci.;1(2):26-36.

Singlas, E., 1995. Clinical Pharmacokinetics of Azithromycin. Pathologie-Biologie, 43(6), pp.505-511.

Vennela, K., Reddy, M.M. and Subramanian, S., 2014. A New RP-HPLC Method for the Simultaneous Estimation of Azithromycin and Levofloxacin in it’s Pure and Pharmaceutical Dosage Form as per ICH Guidelines. International Journal of Pharma Research and Health Sciences Volume 2 (6), Page-507-513.

Verma G and Mishra M. Development and optimization of UV-VIS spectroscopy-a review. World Journal of Pharmaceutical Research. 2018;7(11):1170-1180.

Yandamuri N, Srinivas Nagabattula KR, Kurra SS, Batthula S, Nainesha Allada LPS and Bandam P. Comparative study of new trends in HPLC: a review. International Journal of Pharmaceutical Sciences Review and Research.2013; 23:52-57.

Zotou A. An overview of recent advances in HPLC Instrumentation. Open Chemistry. 2012;10(3):554-569.

Zubata, P., Ceresole, R., Rosasco, M.A. and Pizzorno, M.T., 2002. A new HPLC method for azithromycin quantitation. Journal of Pharmaceutical and Biomedical Analysis, 27(5), pp.833-836.

Related Images: