Beşeri ilaçlarda klorheksidin glukonat, benzidamin hidroklorür etken maddelerinin ve bozunma ürünlerinin kromatografik tayini
Chromatographic determination of chlorhexidine gluconate, benzydamine hydrochloride active materials and their degradation products in human medicines
Yazar: YASEMİN GÖKDENİZ
Danışman: PROF. DR. AYŞEM ARDA
İstanbul Üniversitesi-Cerrahpaşa / Lisansüstü Eğitim Enstitüsü / Kimya Ana Bilim Dalı / Analitik Kimya Bilim Dalı
Derece: Doktora, 2023
Konu: Kimya = Chemistry
Dizin: HPLC =

Özet:
Bozunma ürünleri, ilaç ürünlerinin üretimi ve depolanması sırasında gelişebilen, farmasötik ürünlerin etkinliğini etkileyebilen istenmeyen kimyasallardır. Ayrıca, hastalarda olumsuz etkilere neden olma potansiyeli nedeniyle, az miktarda bozunma ürünleri bile farmasötik güvenliği etkileyebilir. İlaç ürünlerinin formülasyon gelişimi sırasında kritik değişkenlerin kontrol edilmesi ve bu safsızlıkların bir takip çalışmasının yapılması, ürün kalitesini sağlamak için eşik değerlerden daha yüksek konsantrasyonlarda bulunan bozunma safsızlıklarını önleyebilir. Bir ilaç ürününde etkin maddenin miktarlarının da hesaplanması kritiktir. Bu bağlamda tez kapsamında sprey formundaki ticari ilaçlarda Klorheksidin Glukonat ve Benzidamin Hidroklorür etken maddelerinin miktarının ve olası bozunma ürünlerinin tayin edilmesine olanak verecek duyarlı ve seçimli bir Yüksek Performanslı Sıvı Kromatografisi (HPLC) yöntemi geliştirildi. Ayrıca ürün içerisinde bulunan Etanol miktarı için Gaz Kromatografisi-Alev İyonizasyon Dedektör- (GC-FID) metodu geliştirildi. İlgili etken maddelerin bozunma ürünlerinin bulunduğu bitmiş ürün için HPLC ile bozunma ürünlerinin tayini metodu geliştirilerek, validasyon parametreleri incelendi. Metotta; Phenomenex Gemini C18 kolon, gradient bir sistem kullanıldı. Benzidamin etkin madde 9,4510-5 mg/mL-8,6610-3 mg/mL, Benzidamin Safsızlık A 3,2210-5 mg/mL-7,2410-3 mg/mL, Benzidamin Safsızlık B 6,010-5 mg/mL -7,1310-3 mg/mL, Benzidamin Safsızlık C 3,8210-5 mg/mL7,6410-3 mg/mL, Klorheksidin Diglukonat etkin madde 1,1310-4 mg/mL-6,0410-3 mg/mL, Klorheksidin Safsızlık N 8,2910-5 mg/mL-7,9610-3 mg/mL, Klorheksidin Safsızlık H 1,8910-4 mg/mL-6,0510-3 mg/mL, Klorheksidin p-kloroanilin 8,2310-5 mg/mL-5,6 10-3 mg/mL derişim aralığında tespit edildi.
Geliştirilen ikinci HPLC metodunda; bitmiş ürün için ilgili etken maddelerin HPLC ile miktar tayini metodu geliştirilerek, validasyon parametreleri incelendi. Metotta; Inertsil ODS C18 kolon ile izokritik sistem kullanıldı. Benzidamin etkin madde 3,510-2 mg/mL-8,0110-2 mg/mL, Klorheksidin Diglukonat 2,7310-2 mg/mL- 6,2410-2 mg/mL derişim aralığında tespit edildi.
Klorheksidin Diglukonat, Benzidamin HCl için tanıma metodu olarak Ultraviole  Spektrofotometri (UV) yöntemi geliştirildi. Karakteristik olarak verdikleri UV absorbansları Klorheksidin Diglukonat 255 nm, Benzidamin HCl 306 nm olarak belirlendi.
Sprey formundaki ürün içerisinde bulunan Etanol için GC-FID miktar tayini yöntemi geliştirildi. Etanol 2,10 mg/mL- 3,90 mg/mL derişim aralığında tespit edildi.

Abstract:
Degradation products are undesirable chemicals that can develop during the manufacture, storage of pharmaceutical products and affect the efficacy of pharmaceutical products. Furthermore, even small amounts of degradation products may affect pharmaceutical safety due to the potential to cause adverse effects in patients. Consequently, it is crucial to focus on the mechanistic understanding, formulation, storage conditions and packaging to prevent the formation of degradation products that could adversely affect the quality and safety of the drug product. Controlling critical variables during formulation development of pharmaceutical products and performing a follow-up study of these impurities can prevent degradation impurities present at concentrations higher than the threshold values to ensure product quality. It is also critical to calculate the amounts of the active substance in a drug product. In this context, within the scope of the thesis, a sensitive and selective High-performance liquid chromatography (HPLC) method was developed that will allow the determination of the amount of Chlorhexidine Gluconate and Benzydamine Hydrochloride active ingredients and possible degradation products in commercial drugs in spray form. Gas chromatography- Flame Ionization Detector (GC-FID) method was developed to determine the assay of Ethanol in the product. For the finished product containing the degradation products of the relevant active substances, a method for the determination of degradation products by HPLC was developed and the validation parameters were examined. In the method; Phenomenex Gemini C18 column, gradient system was used. Benzydamine active ingredient 9.4510-5 mg/mL-8.6610-3 mg/mL, Benzydamine Impurity A 3.2210-5 mg/mL -7.2410-3 mg/mL, Benzydamine Impurity B 6.010-5 mg/mL -7.1310-3 mg/mL, Benzydamine Impurity C 3.8210-5 mg/mLDegradation products are undesirable chemicals that can develop during the manufacture, storage of pharmaceutical products and affect the efficacy of pharmaceutical products. Furthermore, even small amounts of degradation products may affect pharmaceutical safety due to the potential to cause adverse effects in patients. Consequently, it is crucial to focus on the mechanistic understanding, formulation, storage conditions and packaging to prevent the formation of degradation products that could adversely affect the quality and safety of the drug product. Controlling critical variables during formulation development of pharmaceutical products and performing a follow-up study of these impurities can prevent degradation impurities present at concentrations higher than the threshold values to ensure product quality. It is also critical to calculate the amounts of the active substance in a drug product. In this context, within the scope of the thesis, a sensitive and selective High-performance liquid chromatography (HPLC) method was developed that will allow the determination of the amount of Chlorhexidine Gluconate and Benzydamine Hydrochloride active ingredients and possible degradation products in commercial drugs in spray form. Gas chromatography- Flame Ionization Detector (GC-FID) method was developed to determine the assay of Ethanol in the product. For the finished product containing the degradation products of the relevant active substances, a method for the determination of degradation products by HPLC was developed and the validation parameters were examined. In the method; Phenomenex Gemini C18 column, gradient system was used. Benzydamine active ingredient 9.4510-5 mg/mL-8.6610-3 mg/mL, Benzydamine Impurity A 3.2210-5 mg/mL -7.2410-3 mg/mL, Benzydamine Impurity B 6.010-5 mg/mL -7.1310-3 mg/mL, Benzydamine Impurity C 3.8210-5 mg/mL -7.6410-3 mg/mL, Chlorhexidine Digluconate active ingredient 1.1310-4 mg/mL-6.0410-3 mg/mL, Chlorhexidine Impurity N 8.2910-5 mg/mL-7.9610-3 mg/mL, Chlorhexidine Impurity H 1.8910-4 mg/mL-6.0510-3 mg/mL, p-chloroaniline 8.2310-5 mg/mL-5.6 10-3 mg/mL was detected in the concentration range.
In the developed second HPLC method method; All validation parameters of the method were examined by developing a method for the determination of the relevant active ingredients by HPLC for the finished product. In the method; Inertsil ODS C18 column, isocratic system was used. Benzydamine active substance 3.510-2 mg/mL-8.0110-2 mg/mL and Chlorhexidine Digluconate 2.7310-2 mg/mL- 6.2410-2 mg/mL was determined in the concentration range.
In addition, Ultraviolet-Spectrophotometry (UV) method was developed as a identification method for the active ingredients of Chlorhexidine Digluconate and Benzydamine HCl. Their characteristic UV absorbance was determined. as 255 nm for Chlorhexidine Digluconate and 306 nm for Benzydamine HCl.
The GC-FID method was developed for the ethanol contained in the product in spray form. Ethanol,  2.10 mg/mL-3.90 mg/mL was detected in the concentration range