SPDS Aloe Vera Method Book

Date Issued: Supersedes:

Page:

2 of 4

Procedure Title: Determination of Polysaccharides Molecular Weight in Aloe vera Fresh Leaf and Concentrated Juice Powder by SEC-RI

N/A

Effective Date:

QCL

SOP #:

Revision #:

Prepared By: Yuehong Zhang, Zhichao Bao Department: R&D Applies To: Herbalife International

1.1

100mM NaCl Mobile Phase Preparation 1.1.1

Mobile Phase A: Weigh 5.844mg of sodium chloride (NaCl) into a 1-L volumetric flask and dilute to volume with water. Mix well and filled by the filter kit with 0.22μm Nitrocellulose membrane.

1.2

100mM NaCl Extraction Solution Preparation 1.2.1

Extraction Solution: Weigh 5.844 g of NaCl into a 1-L volumetric flask and dilute to volume with water. Mix well.

1.3

Standard Preparation 1.3.1

Standard Solutions 1.3.1.1 Pullulan Standards Solution: Weigh ~5 mg each of standards into each 5 mL volumetric flask. Add 10 µL of methanol (MeOH). Dissolve and dilute to volume with 100mM NaCl solution. Vortex for 30 seconds and allowed to swell for 20-24 hours at 4 o C. Filter standard solution through a 0.45μm Hydrophilic PTFE filter into a glass HPLC vial. Aloe Standard Solutions 1.3.2.1 Weigh ~5 mg aloe standard into a 5 mL volumetric flask. Add 10 µL of methanol (MeOH). Dissolve and dilute to volume with 100mM NaCl solution. Vortex for 30 seconds and allowed to swell for 20-24 hours at 4 o C. Filter standard solution through a 0.45μm Hydrophilic PTFE filter into a glass HPLC vial. Fresh Aloe Leaf 1.4.1.1 Preparation of fresh leaf samples: for whole leaf sample, cleaned the aloe leaf by water and cut the leaf tip, butt, barbs and edges with a knife. Grind the entire leaves in a high speed blender until the blend was flowing smoothly and no large chunks remained. Squeeze the juice out of the leaf residue and lyophilize the juice to whole leaf juice powder. For inner leaf, filet the cleaned leaf with knife to separate the rind from the inner leaf and lyophilize inner leaf to inner leaf juice powder. 1.4.1.2 Weigh ~100 mg of the leaf juice powder into a 50 mL volumetric flask. Add 100 µL of MeOH. Dissolve and dilute to volume with 100mM NaCl solution. Vortex the sample for 30 seconds and allowed to swell for 20-24 hours at 4 o C. Centrifuge the sample liquid with 4000-5000 rpm for 10 min. Pipette 1-mL supernatant into a glass HPLC vial. Aloe Concentrated Juice Powder 1.4.2.1 Weigh ~250 mg of the concentrated juice powder into a 50 mL volumetric flask. Add 100 µL of MeOH. Dissolve and dilute to volume with 100mM NaCl solution. Vortex the sample for 30 seconds and allowed to swell for 20-24 hours at 4 o C. Centrifuge the sample liquid at 4000-5000 rpm for 10 min. Pipette 1- mL supernatant into a glass HPLC vial.

1.3.2

1.4

Sample Preparation 1.4.1

1.4.2

1.5

System Suitability

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