INTRODUCTION:

These methods are used to determine the active ingredients in body fluid are seemly most essential for the learning of bio-availability, bio-equivalence (BE), Pharmacokinetics (PK) studies, evaluation of active ingredients, amount of drug and their intermediate substances, advanced drug synthesis, explore in fundamental related to health and pharmaceutical discipline and pharmacological drug monitoring^1,2,15-17, HPLC fewest used practical synthetic method because of its extremely exclusive and higher dependableness, peculiarly in the pharmaceutical, environmental, forensic, surgical, and hospitality division^3,18-21.

Mostly in 2001 by USFDA and freshly the EMEA also projected the guidelines for the bio-analytical system confirmation. Confirmation involves signaling, through the use of particular laboratory examinations, that the execution characteristics of a system are suitable and dependable for the deliberate bio-analytical operations. The adequacy of logical data corresponds to used authorize the system. For important studies that bear non supervisory action for blessing, similar as BE or PK studies, the bioanalytical styles could be completely validated. For advance styles used for the guarantor’s inner decision- timber, lower confirmation may be sufficient when changes are made to a prelude validated system, fresh confirmation may be demanded^4,5,22-24. Frequently undergoes numerous variations; these variations should be validated to confirm suitable performance of the logical system. The organic process alteration need to support specific studies for the different situations of confirmation to prove the validity of system Confirmation of bioanalytical styles carried during.

The responsibility of logical findings is a concern of large significance in rhetorical and medical pharmacology, this is a obligatory for right rendering of pharmacological findings. Fallible reports are non queried in judicature, may track to unwarranted judicial actions for the suspect or to incorrect care of the case.

The amount of medicines insert inner the grocery is adding all time. Those medicines may be either new realities or biased functional revision of the being one. Veritably frequently there's a period pause from the date of preface of a medicine into the grocery to the date of its addition in pharmacopoeias. That will entertain misgivings in the nonstop and wider operation of these medicines, reports of new venom (performing in their pullout from the request), improvement of diligent impedance and preface of better medicines by challengers).

METHOD DEVELOPMENT:

This process modification is the procedure of production a process to unexplored substance or new substance be known and deliberate in a mould. A substance arse rarely be calculated by various performing and the selection of synthetic process relate, synthetic belongings of the substance quantity determination, amounts of the drug, sample cast, price for the investigation process and equipments, velocity and period of the quality determination, exactness and essential instrumentation. It consists of unknown solution distribution formulation of unknown reagents, division of mixtures, discovery and judgement of the solutions.^6,25-28

Analytical methods used for the separation of components are classified as

I. classical:

1. Extraction

2. Precipitation

3. colour

4. odour

5. melting point

II. Instrumental:

1. UV Spectroscopy

2. HPLC

3. Fluorescence

4. Conductivity

5. Chromatography

6. Electrophoresis

Preparation of sample:

Collect various samples from animal source, human body.

HPLC method development:⁷

It is the best development method. Many dosage forms are analysed by HPLC. From this method both soluble internal phases are separated. By using exact mobile phase and stationary phase then we get the optimum range of separation. Solvent selection is also most important.

Accuracy:⁷

It is defined that coincidence values of practical and original values. The practical value is identical to the original value. Typically, quality is depicted and discovered by recuperation survey.

Extraction method:

Liquid-liquid extraction:

It is founded on the generality of dispute miscibility and division balance of measuring particles between water, other non polar liquid. Natural separation of active ingredients mostly pertain that lineage for the chemical between the two liquid phases^8,29-33. Presently solution filtrate displaced with sophisticated procedures are solution low filtrate with membrane filter^9,34.

Solid phase extraction:¹⁰

It is the very essential method utilized in the solution tending for chromatography. This can be performed by both liquid and solid phases.

Protein Precipitation:¹¹

It is related to analyse the body fluids, intra cellular, extra cellular fluids, proteins and other fluids present in the body. Some substances may precipitate proteins due to aggregation of globules or coalescence of globules.

Chromatography:^12,13,35 In which separation of molecules from the mixture of components through the mobile phase travel on the stationary phase both in case of paper and thin layer Chromatography.

Stationary phase: It is a solid phase or liquid molecules attached on the solid surface.

Mobile phase: It is a liquid phase or gaseous phase

Types of chromatography:

Column chromatography

Ion-exchange chromatography

Gel-permeation (molecular sieve) chromatography

Affinity chromatography

Paper chromatography

Thin-layer chromatography

Gas chromatography

Dye-ligand chromatography

Hydrophobic interaction chromatography

Pseudo affinity chromatography

High-pressure liquid chromatography (HPLC)

Column chromatography:

In which separation of components through the column

Fig 1:Column chromatography.

Ion- exchange chromatography:

In which the separation is based on the cation and anion present in the given sample solution

Fig 2:Ion- exchange chromatography.

Gel- permeation (molecular sieve) chromatography: In which separation of molecules from the size of the particles

Fig 3:Gel-permeation (molecular sieve) chromatography.

Affinity chromatography:

In which the separation is based on the formation of complex between the molecules.

Fig 4: Affinity chromatography.

Paper chromatography:

In which stationary phase is paper and mobile phase is liquid phase.

Thin-layer chromatography:

In which stationary phase is solid adsorbed on the TLC plate and mobile phase is liquid phase.

Gas chromatography:

In which stationary phase is column and mobile phase is gas phase.

Dye- ligand chromatography:

In which separation is between dye and ligand.

Hydrophobic interaction chromatography (HIC):

In which the adsorbents prepared as column material for the ligand binding in affinity chromatography are used.

Pseudo affinity chromatography:

In which ligands can be used are anthraquinone dyes, and azo-dyes because of their attraction especially for dehydrogenases, kinases, transferases, and reductases. The mostly famous type of this kind of chromatography is immobilized metal affinity chromatography¹⁴.

CONCLUSION:

From this article, bio-analytical styles are extensively used to determine the amount of medicines and their intermediate derivatives in tube matrices and the styles should be practical to acquisition in areas of mortal clinical and in human study. Bio-analytical system employed for the quantitative estimation of medicines and their metabolites in natural media and plays an essential portion in computation and rendition of BE, PK, and TK survey. The leading BA part is system improvement, system confirmation, and statistical distribution investigation. Ways similar as HPLC and LC conjugated LCMS- MS are utilized for the BA of medicines in body.

REFERENCES:

1. Thompson M, Ellison SL, Wood R. Harmonized guidelines for single laboratory validation of method of analysis. Pure Appl Chem. 2008; 74(5):835-55.

2. Wood R. How to validate analytical methods. Trends Analyt Chem. 2005; 18:624-32.

3. Chiu ML, Lawi W, Snyder ST, Wong PK, Liao JC, Gau V. Matrix effects: A challenge toward automation of molecular analysis. J Assoc Lab Autom. 2010; 15:233-42.

4. Reid E, Wilson ID. Methodological survey in biochemistry and analysis. Analysis for Drug and Metabolites, Including Anti-Infective Agents. Vol. 20. Cambridge, England. Royal Society of Chemistry.1990. p. 1-57.

5. Surendra B, DeStefano A. Key elements of bioanalytical method validation for small molecules. AAPS J. 2007; 9(1):109-14.

6. Puluido A, Ruusanches I, Boque R, Rius FX. Uncertainty of results in routine qualitative analysis in analytical chemistry. J Pharm Biomed Anal. 2005; 22:647-54.

7. P. Ravisankar*1, 2, S. Gowthami1, G. Devlala Rao3, A review on analytical method development, Indian Journal of Research in Pharmacy and Biotechnology, May-June 2014, 2(3), 1183 -1195.

8. Lang JR, Bolton S. A comprehensive method validation strategy for bioanalytical applications in the pharmaceutical industry--1. Experimental considerations. J Pharm Biomed Anal. 1991; 9(5):357-61.

9. Causon R. Validation of chromatographic methods in biomedical analysis. Viewpoint and discussion. J Chromatogr B Biomed Sci Appl. 1997; 689(1):175-80.

10. Pranay Wal, 1 Brijesh kumar1 Dr. Anil Bhandari1, A. K. Rai, 2 Ankita wal, Bioanalytical Method Development –Determination of Drugs in Biological Fluids, journal of Pharmaceutical Science and Technology. Vol. 2 (10), 2010, 333-347333

11. https://en.wikipedia.org/wiki/Protein_precipitation

12. Cuatrecasas P, Wilchek M, Anfinsen CB. Selective enzyme purification by affinity chromatography. Proc Natl Acad Sci U S A. 1968; 61:636–43. [PMC free article] [PubMed] [Google Scholar]

13. Porath J. From gel filtration to adsorptive size exclusion. J Protein Chem. 1997; 16:463–8. [PubMed] [Google Scholar]

14. Porath J. Immobilized metal ion affinity chromatography. Protein Expr Purif. 1992; 3:263–81.

15. M.Venkataswamy, Viswanadham Manasa, K. Vanitha, P. Prabhakar, Alluri Ramesh “Formulation and Evaluation of Aqueous Enteric Coated Delayed Release Omeprazole Pellets” Research Journal of Pharmaceutical Dosage Forms and Technology. 2016; 8(1): 1-4.

16. M.Venkataswamy, K. Vyshnavi, Madhuri Bhashetty, Namitha Swain, Alluri Ramesh. Evaluation of Prepared Aspirin Modified Release Tablets with Polymer Isolated from Fresh Mosambi Leaves. Research Journal of Pharmaceutical Dosage Forms and Technology. 2017; 9(2): April- June,.

17. M.Venkataswamy, Arul B, Keerthi Sagar A, Dinesh Mohan S, Vanitha K, Ramesh Alluri. Preparation and Evaluation of a Mucoadhesive Polymer from the Extract of Seeds of Annona squamosa Linn. Asian J. Res. Pharm. Sci. 2017; Vol. 7: Issue 3.

18. M. Venkataswamy, Chiluveri Sanjuna, Pindrathi Pravalika, Chikoti Sneha Priya, Alluri Ramesh. The Insidious Disease from Insects: Lyme disease. Research Journal of Pharmaceutical Dosage Forms and Technology. 2018; 10(1): 17-22.

19. M.Venkataswamy, K. Prathyusha, Jaggareddy Gari Manasa Reddy, Alluri Ramesh. Pico technology: Instruments used and Applications in pharmaceutical field” Research Journal of Pharmaceutical Dosage Forms and Technology. 2018; 10(1): 34-41.

20. M. Venkataswamy, Jaggareddy Gari Manasa Reddy, K. Prathyusha, Alluri Ramesh. Spreading of Swine flu disease: Past and Present. Research Journal of Pharmaceutical Dosage Forms and Technology. 2018; 10(2).

21. M.Venkataswamy, K. Vanitha, Sanam Niharika, Alluri Ramesh. Formulation Development and Evaluation of Mebeverine extended release Pellets. Asian J. Pharm. Tech. 2018; 8 (2):71-77.

22. M. Venkataswamy, M. Santhoshini, J. P. Priyanka, K. Prathyusha, Jaggareddy Gari Manasareddy and Ramesh Alluri. Preparation and evaluation of biphasic bilayered buccal tablet containing ketorolac immediate release layer and domperidone maleate sustained release layer. World Journal of Pharmaceutical Research. Volume 7, Issue 11, 905-949.

23. M. Venkataswamy, S. Naveen Kumar, B. Ramesh, G. Anantha Ramulu, M. Bharath, K. Sridivya Goud. Preparation And Evaluation of Mucilage from Fresh Leaves of Psidium Guajava. Research Journal of Pharmaceutical Dosage Forms and Technology. 2018; 10(3): July- September.

24. M.Venkataswamy, Neha Thahaseen, Md Kareemuddin, JP Priyanka, Jaggareddy Gari Manasa Reddy. Formulation And Evaluation of sustained release matrix tablets using drug Cytisine isolated from Fava Beans to quit Smoking. Research Journal of Pharmaceutical Dosage Forms and Technology. 2018; 10(3): July- September.

25. M. Venkataswamy, Ifath Jabeen, Jawariya Sadaf, Maistry Nishitha Reddy, A.V. Naga Mallika, Mamidi Sushmitha. Drug abuse, addiction, its causes and treatment. Research Journal of Pharmaceutical Dosage Forms and Technology. 2018; 10(3): July- September.

26. M. Venkataswamy, Sandhya Mandadi. Formulation And Evaluation Of Metformin Transcutaneous Patches. 2018; WJPR, Vol 7, Issue 18.

27. M. Venkataswamy, Ch. Sreerekha, Y. Manasa, Boppana Rohini, Kadannagari Surekha, Kilari Sri Kavya. Generic Drugs a Benison to Mankind. Research Journal of Pharmaceutical Dosage Forms and Technology. 2019; 11(2): 121-125.

28. M. Venkataswamy, V. Nimitha. Medicinal Plants used for the Treatment of Parkinson's Disease. Research Journal of Pharmaceutical Dosage Forms and Technology. 2019; Vol 11(2), Apr- June.

29. M. Venkataswamy, Ch. Sanjuna. An Arthropod Virus: Chikungunya its Pathophysiology and Herbal Treatment. Research Journal of Pharmaceutical Dosage Forms and Technology. Vol 11(2), Apr- June 2019.

30. M. Venkataswamy, Sana. Preparation and Evaluation of Ginger Mouth Gargle. Research Journal of Pharmaceutical Dosage Forms and Technology. 2019; 11(2): Apr- June.

31. M. Venkataswamy, K. Prathyusha. Thalassemia -A Blood Disorder, its Cause, Prevention and Management. Research Journal of Pharmaceutical Dosage Forms and Technology. 2019; 11(3): Aug- Oct.

32. M. Venkataswamy, Ch. Sneha. Medicinal Plants used for the Treatment of Bacterial Meningitis. Research Journal of Pharmaceutical Dosage Forms and Technology. 2019; 11(3): Aug- Oct.

33. M. Venkataswamy, Sandhya M. Formulation and Evaluation of Herbal Anti-bacterial gel containing Ethanolic extracts of Rosa Indica and Tagetes Minuta. Research Journal of Pharmaceutical Dosage Forms and Technology. 2020; 12(2): Mar- May.

34. M. Venkataswamy, V. Harshitha. Nanoemulgel: A Process Promising in Drug Delivery System. Research Journal of Pharmaceutical Dosage Forms and Technology. 2020; 12(2): Mar- May.

35. M. Venkataswamy, Ramanamurthy Venkata Kolapalli. Formulation and evaluation of microspheres loaded with methotrexate drug. Drug Discovery. 2020; 14(34).

Received on 28.05.2022 Modified on 22.06.2022

Asian J. Pharm. Ana. 2023; 13(2):89-92.

DOI: 10.52711/2231-5675.2023.00015