## 2 Materials and Methods
### Experimental
#### 2.2.10 Test for Proteinase Action Inhibition  
Proteinases are found in lysosomes and are known to catalyze the hydrolysis of proteins. It has been reported that leukocyte proteinases contribute to tissue damage in inflammatory reactions and proteinase inhibitors offer protection against proteinases-induced tissue damage [19]. Proteinase enzyme inhibitory activity of myrrrh extract was investigated according to our previously published articles [20] using diclofenac sodium (200 \(\upmu\)g/mL) as a reference drug. Various aliquots of myrrrh extract (50, 75, 100, 200, 300, 400, 500, and 600 \(\upmu\)g/mL) were used for this assay. The optical densities of various samples were recorded at a wavelength of 210 nm against blank having Tris HCl buffer only. The negative control setup was made up of 1 mL normal saline (0.9 % NaCl), 1 mL Tris HCl buffer, 0.06 mL trypsin, 1 mL 0.8 percent casein, and 2 mL 70% perchloric acid. \[\text{Inhibition of proteinase action (\%)}=[(\text{Lc}-\text{Ls})/\text{Lc}]\times 100\]  
where, the absorbance of the control, i.e., the test solution without extract or diclofenac, is Lc; the absorbance of the test solution with extract or diclofenac is Ls. 2.2.11. Egg Albumin Denaturation Inhibition for Anti-Arthritic Activity  
An amount of 2.8 mL of fresh phosphate buffer saline (pH 6.4), 0.2 mL of chicken egg albumin, and 2 mL of either diclofenac sodium (200 ug/mL) or myrrh extract (50, 75, 100, 200, 300, 400, 500, and 600 ug/mL) were placed at room temperature for 15 min inside an incubator [17; 21]. Following the incubation, various test solutions were later put at 70 \({}^{\circ}\)C for 5 min. At 660 nm, the absorbance of all solutions was measured against phosphate buffer as a blank. Inhibition of egg albumin denaturation (%) = [(Lc - Ls)/Lc] \(\times\) 100  
where, the absorbance of the control, i.e., the test solution without extract or diclofenac, is Lc; the absorbance of the test solution with extract or diclofenac is Ls. 2.2.12. Screening for Antiglycation Property of Myrrh Extracts  
Incubation of Myrrh extracts with in vitro glycation system  
The Brownlee approach was used to perform the in vitro glycation of BSA [22]. BSA (10 mg/mL) and glucose (500 mM) were put in sterilized tubes together with varied amounts of myrrh extract (50, 75, 100, 200, 300, 400, 500, and 600 g/mL), either with or without a 0.1 M phosphate buffer (pH 7.4). The tubes were kept in the dark at 37 \({}^{\circ}\)C for 15 days. Glass vials that had been autoclaved, closed, and capped were used to ensure sterility. After incubation, the samples underwent overnight phosphate buffer dialysis at room temperature to remove bound and unbound glucose. The concentration of BSA was determined using the molar extinction coefficient method. After that, the samples were kept at \(-20\)\({}^{\circ}\)C until testing could be performed on them. The experiments were all carried out in triplicate. Effect of Myrrh extracts on browning in glycated samples  
Glycation is assumed to play a major role in the development of diabetes and its consequences [2]. The degree of browning in glycated samples is said to be an effective indicator of glycation in many samples [9]. After diluting with distilled water, the browning intensity was evaluated by monitoring the absorbance of various glycated samples at 420 nm [22] that used a 1 cm path length cell. All of the tests were performed in triplicate. The following formula [9] was used to calculate the relative percentage browning intensity. \[\text{Percentage protection}=[(\text{Lc}-\text{Ls})/\text{Lc}]\times 100\]  
where, Lc refers to absorbance of BSA and glucose system (without extract). However, Ls denote absorbance of BSA and the glucose system (with extract).