UV kombine RP-HPLC ile İzotonik Serum Fizyolojikte İnsülin Tayini

Abstract

Tip I diabetes mellitus tedavisinde insülin yaygın olarak intravenöz olarak kullanılmaktadır. Bu çalışmanın amacı, % 0.9’luk NaCl solüsyon örneklerinde insülin tayini için yenilikçi, hızlı ve basit bir analiz tekniği geliştirmektir. İnsülin, UV detektörlü HPLC ile analiz edilmiştir. Yöntem, FDA yönergelerine göre onaylanmış ve tatmin edici sonuçlar gözlenmiştir. Örnekler, ACE 5 um C18 100 Å LC kolonu (250 x 4.6 mm) kullanılarak HPLC sistemine manuel olarak enjekte edilmiştir. Mobil faz, bir asetonitril: fosfat tamponu (pH 3.0, 10 mM) (50:50 v / v) karışımıdır. Enjeksiyon hacmi 20 µL’dir. UV saptaması 240 nm’de gerçekleştirilmiştir. Yöntemin, özgün, doğrusal, kesin, doğru ve duyarlı bir yöntem olduğu validasyon çalışmalarıyla ortaya konulmuştur.

Keywords

• Reguler insulin
• HPLC
• izotonik serum fizyolojik
• validasyon
• UV

Determination of Insulin in saline by RP-HPLC combined with UV

Abstract

In the treatment of type I diabetes mellitus, insulin is widely used intravenously. This study aims to develop an innovative, fast and simple analysis technique for insulin determination in 0.9% NaCl solution samples. Insulin was analyzed by HPLC with a UV detector. The method has been validated according to FDA guidelines and satisfactory results have been observed. Samples were manually injected into the HPLC system using ACE 5 µm C18 100 Å LC column (250 x 4.6 mm). The mobile phase was a mixture of acetonitrile: phosphate buffer (pH 3.0, 10 mM) (50:50 v/v). The injection volume is 20 µL. UV detection was carried out at 240 nm. Based on validation studies, the method was found as specific, linear, precise, accurate, and sensitive.

Keywords

• Regular insulin
• HPLC
• saline solution
• validation
• UV

References

1. 1. Mayer JP, Zhang F, and DiMarchi RD: Insulin structure and function. Peptide Science: Original Research on Biomolecules 2007, 88(5): 687-713.
2. 2. Moussa B, Farouk F, and Azzazy H: A validated RP-HPLC method for the determination of recombinant human insulin in bulk and pharmaceutical dosage form. E-Journal of Chemistry 2010, 7(S1): 449-457.
3. 3. Bettelheim FA and Landesberg JM: Laboratory experiments for introduction to general, organic and biochemistry. Cengage Learning; Boston, USA, 2012.
4. 4. Legg KM, Labay LM, Aiken SS, and Logan BK: Validation of a Fully Automated Immunoaffinity Workflow for the Detection and Quantification of Insulin Analogs by LC–MS-MS in Postmortem Vitreous Humor. Journal of analytical toxicology 2019, 43(7): 505-511.
5. 5. Yilmaz B, Kadioglu Y, and Capoglu I: Determination of insulin in humans with insulin-dependent diabetes mellitus patients by HPLC with diode array detection. Journal of chromatographic science 2012, 50(7): 586-590.
6. 6. Yilmaz B and Kadioglu Y: Development and validation of HPLC method for determination of human insulin in pharmaceutical preparation. International Journal of Pharmaceutical Sciences Review and Research 2010, 2(2): 40-43.
7. 7. Salem LI, Bedmar M, Medina M, and Cerezo A: Insulin evaluation in pharmaceuticals: variables in RP-HPLC and method validation. Journal of Liquid Chromatography & Related Technologies 1993, 16(5): 1183-1194.
8. 8. Sarmento B, Ribeiro A, Veiga F, and Ferreira D: Development and validation of a rapid reversed‐phase HPLC method for the determination of insulin from nanoparticulate systems. Biomedical Chromatography 2006, 20(9): 898-903.

9. 9. Lim SC, Roberts MJ, Paech MJ, Peng L, and Jones A: Stability of insulin aspart in normal saline infusion. Journal of Pharmacy Practice and Research 2004, 34(1): 11-13.
10. 10. http://www.fda.gov/downloads/Drugs/GuidanceComplianceRegulatoryInformation/Guidances/UCM368107.pdf (17.03.2021)
11. 11. Hirsch JI, Wood JH, and Thomas RB: Insulin adsorption to polyolefin infusion bottles and polyvinyl chloride administration sets. American journal of hospital pharmacy 1981, 38(7): 995-997.
12. 12. Weber SS and Wood WA: Insulin adsorption controversy. Drug Intelligence & Clinical Pharmacy 1976, 10(4): 232-233.
13. 13. Schildt R, Ahlgren T, Berghem L, and Wendt Y: Adsorption of insulin by infusion materials. Acta Anaesthesiologica Scandinavica 1978, 22(5): 556-562.
14. 14. Twardowski ZJ, Nolph KD, McGary TJ, Moore HL, Collin P, Ausman RK, and Slimack WS: Insulin binding to plastic bags: a methodologic study. American journal of hospital pharmacy 1983, 40(4): 575-579.
15. 15. Bonfilio R, Cazedey ECL, Araújo MBd, and Nunes Salgado HR: Analytical Validation of Quantitative High-Performance Liquid Chromatographic Methods in Pharmaceutical Analysis: A Practical Approach. Critical Reviews in Analytical Chemistry 2012, 42(1): 87-100.
16. 16. Watson DG: Pharmaceutical AnalysisA Textbook for Pharmacy Students And Pharmaceutical Chemists. Churchill Livingstone; Edinburgh, Scotland, 1999.