Evaluation of Method Performance and Biological Variations for Oxidative Stress Biomarkers in Plant Extracts: A Method Validation Study

Year 2022, Volume: 25 Issue: 5, 965 - 973, 31.10.2022

Erkan Öner Ergul Belge Kurutas İlter Demirhan Meltem Güngör

https://doi.org/10.18016/ksutarimdoga.vi.997048

Abstract

In this study, it was aimed to determine biological variations and analytical quality specifications to evaluate the acceptability of method performance for oxidative stress biomarkers in Silybum marianum (Milk thistle) and Artemisia absinthium (Wormwood) plant extracts. Silybum marianum and Artemisia absinthium were used in the study. Samples were taken from plants on the zero, 1st, 3rd, 5th, 7th, 15th and 30th days. Index of individuality (II) and reference change value (RCV) were calculated from within-subject (CVI) and between-subject (CVG) variations. Moreover, imprecision, bias and total error were calculated as analytical quality specifications. Oxidative stress biomarkers [catalase (CAT), superoxide dismutase (SOD) and malondialdehyde (MDA)] in plant extracts were measured as spectrophotometric methods. Compared to Artemisia absinthium, higher antioxidant enzyme activities (SOD and CAT) and lower MDA levels in Silybum marianum were found in our study (p<0.05). MDA in plants showed low individuality, and CVI of MDA were larger than CVG. However, CAT and SOD in plants showed strong individuality, but CVI of them were smaller than CVG. CAT and SOD except, RCVs of MDA analyte were relatively higher, because of high CVI, resulting in a higher RCV. Also, the described methodology achieves these goals, with analytical CVs of < 3.5% for all analytes. Goals for bias and total error were 11.2-25.2% and 8.93-27.6%, respectively. The RCV concept for predicting that the plant is diseased by some causative agent resulting in an abnormal physiological process can be a valuable tool in demonstrating laboratory optimization. Moreover, desirable precision targets based on biological variation for measurements of oxidative stress biomarkers in plants can be achieved with current methodologies.

Keywords

Oxidative Stress Biomarkers, Method Performance, Biological Variations, Artemisia absinthium, Silybum marianum, Silybum marianum, Silybum marianum, Silybum marianum, Silybum marianum

Bitki Ekstrelerinde Oksidatif Stres Biyobelirteçleri için Yöntem Performansının Değerlendirilmesi ve Biyolojik Varyasyonlar: Metot Validasyon Çalışması

Abstract

Bu çalışmada, bitki ekstrelerinde oksidatif stres biyobelirteçleri için yöntem performansının kabul edilebilirliğini değerlendirmek için biyolojik varyasyonlar ve analitik kalite spesifikasyonlarının saptanması amaçlandı. Araştırmada Silybum marianum (Deve Dikeni) ve Artemisia absinthium (Pelin Otu) kullanıldı. Bitkilerden 0.,1., 3., 5., 7., 15. ve 30. günlerde örnekler alındı. Bitki ekstrelerinde oksidatif stres biyobelirteçleri [katalaz (CAT) superoksit dismutaz (SOD) ve malondialdehit (MDA)] spektrofotometrik yöntemlerle ölçüldü. Her bir bitkinin kendi-içinde (CVI) ve bitkiler-arası (CVG) varyasyonlarından bireysellik indeksi (Bİ) ve referans değişim değeri (RDD) hesaplandı. Ayrıca analitik kalite spesifikasyonları olarak impresizyon, bias ve toplam hata hesaplandı. Çalışmamızda, Artemisia absinthium’a kıyasla Silybum marianum'da daha yüksek antioksidan enzim aktiviteleri (SOD ve CAT) ve daha düşük MDA düzeyleri tespit edildi (p<0.05). Bitkilerde MDA düşük bireysellik gösterdi ve MDA'nın CVI'sı CVG'den daha büyüktü. Bununla birlikte, CAT ve SOD güçlü bir bireysellik gösterdi, ancak bitkilerdeki CVI'ları CVG'lerden daha küçüktü. CAT ve SOD dışında, MDA analitinin RDD'leri, yüksek CVI nedeniyle nispeten daha yüksekti ve bu da daha yüksek bir RDD ile sonuçlandı. Ayrıca açıklanan metodoloji, tüm analitler için <%3.5 analitik CV ile bu hedeflere ulaşır. Bias ve toplam hata hedefleri sırasıyla %11.2-25.2 ve %8.93-27.6 idi. Bitkinin anormal bir fizyolojik süreçle sonuçlanan bazı nedensel ajanlar tarafından hastalandığını tahmin etmeye yönelik RDD konsepti, laboratuvar optimizasyonunu göstermede değerli bir araç olabilir. Ayrıca, bitkilerde oksidatif stres biyobelirteçlerinin ölçümleri için biyolojik varyasyona dayalı istenen kesinlik hedefleri, mevcut metodolojilerle elde edilebilir.

Keywords

Oksidatif Stres Biyobelirteçleri, Yöntem performansı, Biyolojik Varyasyonlar, Artemisia absinthium;, Silybum marianum

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