SPDS Aloe Vera Method Book

Date Issued: Supersedes:

Page:

3 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

1.5.1 Chromatograph a series of pullulan standards. Retention time deviation of the standards (M w 805, 48.8, and 6.2 kDa) should be no more than 0.1 minute, and the Relative Standard Deviation (RSD) for calibration standard should be no more than 2.0%. 1.5.2 Obtain standard calibration curve by plotting the logarithm of pullulan molecular weights (Log M) in narrow mode versus retention time (T) of corresponding standards. Combine aloe standards with known M w , M p , M n , and M z into the above calibration curve in broad mode and use a third-order polynomial fit curve.

1.5.3 1.5.4 1.5.5

Run aloe polysaccharide standards.

Run aloe samples

Run a reference check after every six sample injections and at the end of the run.

1.6

Calculations Integrate the chromatograms showing the peaks of interest. Calculate the molecular weight and molecular weight distribution by Empower 3.0 GPC/SEC software according to: Log M = aT 3 + bT 2 + T + c Where: M: molecular weight; T: retention time (min.); a, b, and c: constants determined by software.

2.0

REFERENCES N/A

3.0

FORMS / CHROMATOGRAMS 3.1

Chromatogram 1: Chromatogram of pullulan standards

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