Whatman filters paper no 1

The collected plant material was wrapped with plastic sheets during transportation then washed carefully under running tap water to remove dirt and soil then dried at room temperature under shade and reduced to the appropriate size by using mortar and pestles. A total of 2 kg of dried leaves and 900 g of dried roots were extracted by maceration (50 g of dried leaves in 300 mL of 80% methanol and 1 g of dried roots in 8–10mL of 80% methanol) for 72 h. The extraction process was facilitated manually by vigorous shaking and stirring. The mixture was first filtered using a muslin cloth and then with Whatman filters paper No.1. The residue has macerated a total of three times to obtain the maximum yield.
Filtration and collection of the extract were done three times with the whole extraction taking 9 days. After the extraction, methanol was evaporated under an oven at 40°C. The resulting solution was placed in a deep freezer operating at negative 20°C till it forms solid ice and then the remaining solvent (water) was removed using a lyophilizer. After water removal, a dark green gummy residue weighing 250.46g was obtained; giving rise to a percentage yield of 12.49% from the leaf, and a light yellow powder weighing 65g was obtained from the root part, giving rise to a percentage yield of 7.2%. The residue was then stored at 40°C (deep freezer) until use.

Normal isotonic saline (Euro-med Laboratories, Philippines), n-hexane, 2% tween 80, ethyl acetate (Loba chemicals, India), chloroform (Atico, India), citrate dextrose (Deluxe Scientific Surgico, India), Giemsa (Science Lab, USA), ethanol (Nice chemicals, India), Electrical balance, 1 mL syringes, needles, vials, examination glove, permanent marker, Whatman filters paper No.1 (Whatman ^®, England), collecting flask, separatory funnel, hematocrit centrifuge, Micro-Hematocrit Reader, microscope, laboratory glasswares, and lyophilizer (Labfreez, China) were used.

The fresh stem bark was washed so as to remove dead materials and allowed to dry for three weeks under a shade. The dried stem bark was then pulverized by using a grinder. The powdered stem bark (1.52 Kg) was macerated in 80% methanol for 72 hrs in maceration jars, extraction was aided by orbital shaker and intermittent stirring. The extract was first filtered using a muslin cloth and then by Whatman filters paper No.1. For exhaustive extraction of the plant material, the residue was re-macerated for another 72 hrs twice and then filtered. The combined filtrates were dried in an oven dryer at a temperature of 40°C and weighed. Then, the dried extract (crude extract) was kept in a tightly closed amber bottle and stored in a refrigerator at 4°C until further use.

Freshly collected matured seeds of G. lotoides were thoroughly washed with tap water to eliminate any contaminants. After complete drying of the sample sunshade, it was pulverized by an electrical grinder. The dry powdered seeds of G. lotoides were subjected to extraction employing. One kg sample was soaked in a total of 5.7 litter of 80% hydromethanol with continuous shaking employing automatic shaker (Stuart Scientific) at 120 rpm at room temperature for 72 h. Then, the solution was primarily filtered employing muslin cloth and then refiltered with Whatman filters paper No.1. The marc was re-soaked with the same volume of solvents twice. Then subsequent filtrates were dried in to solid mass in a rotary evaporator (YAMATO rotary evaporator RE301, Japan, 40 °C, 60 rpm). The concentrated filtrate was subjected to deep freezer (DN-86W258) and then freeze-dried in a lyophilizer (Labfreeze Instruments Group Co., Ltd., Japan) at −50 °C. Ultimately, the %yield of extract was computed and kept at −4 °C using sealed container for experimentation [26 (link)].
The percentage yield of the rude extract $\text{\%Yield}=\frac{\text{ Seed extract weight obtained}(\text{g})}{\text{ Seed powder employed as stock}}\times 100$