SPSFAM ERP

Reagents: 1.

Mobile Phase : Concentration varies to achieve good separation of analytes. Range will be from an initial 75 % Acetonitrile to 82 % Acetonitrile at the end of the column’s lifetime. The following recipe is for 75 % Acetonitrile/25 % water. a. Dispense 750 mL HPLC reagent grade acetonitrile into a glass container. b. Dispense 250 mL deionized water into the same glass container. c. Mix contents well and vacuum filter through a 0.45 µm nylon filter membrane. d. Stable for six months stored at room temperature. 2. Strong Needle Wash/Strong Seal Wash: 100 % water a. Dispense 1000 mL deionized water and filter through a 0.45 µm nylon filter membrane. b. Stable for six months stored at room temperature. 3. Weak Wash/Seal Wash/Syringe Wash Solutions: 75 % Acetonitrile/25 % water (or current mobile phase can be used) a. Dispense 750 mL HPLC reagent grade acetonitrile into a glass container. b. Dispense 250 mL deionized water into the same glass container. c. Mix contents well and vacuum filter through a 0.45 µm nylon filter membrane. d. Stable for six months stored at room temperature. 4. Sugar Standard Solutions a. Weigh the sugars as outlined in the chart below into a 250 mL volumetric flask to generate a three tier regression range for fructose and glucose at one linear range, galactose, maltose and lactose at another linear range, and sucrose at a higher linear range to assay samples containing a maximum of 35 %, 14 % and 43 % sugar, respectively (assuming minimum 7.5 g target weight). Sugar Std # mg/mL Glucose & Fructose, g Galactose 1 g Maltose & Lactose 2 g Sucrose g IS Ribose g 1 0 0 0 0 0 0.5 2 0.5/0.5/0.5 0.125 0.125 0.131 0.125 0.5 3 2.5/1/3 0.625 0.625 0.263 0.75 0.5 4 5/2/6 1.25 1.25 0.525 1.50 0.5 5 10/4/12 2.5 0 1.050 3.0 0.5 b. Add 100 mL of deionized water using a class A glass pipette and swirl/sonicate to dissolve the sugars. c. Add 150 mL of acetonitrile using a class A glass pipette (3 x 50 mL). d. Stopper, invert and shake to mix well. Do not dilute to volume. The 10/4/12 standard will need to be at room temperature to completely dissolve, since the addition of acetonitrile lowers the temperature of the solution. 3 1 Due to solubility, the maximum concentration of galactose was achieved with less overall standards. 2 Maltose and lactose are 95% pure because they are monohydrate salts, thus the target weight has been adjusted to account for this.