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Project: Customer name: Project objective: A321.02 Skullcap Herb, Organic #SKU-W2O11, W2O12 Trout Lake Farm HPTLC analysis of a Skullcap herb sample using the method from the American Herbal Pharmacopoeia monograph for Skullcap Aerial Parts. November 11, 2011 CAMAG Scientific, Inc., Wilmington, NC Judy Nichols

Date: Laboratory: Analyst:

Final conclusion Based on the resemblance of the chromatogram of the test sample to those of the Skullcap herb reference samples, it may be concluded that the test sample #SKU-W2O11, W2O12 is Skullcap (Scutellaria lateriflora L.) herb. General experimental details Temperature: 22.8C Humidity: 42% Sample and reference identification Sample Sample description number S716 Skullcap herb, organic

Source and batch Trout Lake Farm #SKU-W2O11, W2O12

Reference Reference sample description Source and batch number S228 Scutellaria lateriflora herb BRM Chromadex #30190-2465 S575 Scutellaria lateriflora herb BRM AHP #2585 Reference number R007 R008 R011 Reference standard description Source and batch Hyperoside Rutin Caffeic acid Fluka #1402468 Sigma #67H0165 Sigma #47H0424

Chemicals Name Acetic acid Dichloromethane Diphenylboric acid 2-aminoethyl ester Ethyl acetate Formic acid Methanol

Manufacturer Sigma-Aldrich Fluka Spectrum Sigma-Aldrich Sigma-Aldrich Sigma-Aldrich

Purity or quality 99.7%, ACS ACS, stabilized ACS HPLC, 99.7% 98-100%, puriss HPLC, 99.9%
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Project A321.02 Trout Lake Farm Skullcap SKU-W2O11, W2O12 HPTLC Report

CAMAG LABORATORY
Polyethylene glycol 400 Water Alfa Aesar Spectrum ACS HPLC

Plates Description HPTLC glass 20X10 cm, Silica gel 60 F254 Equipment Description Automatic TLC Sampler 4 Automatic Developing Chamber 2 Chromatogram Immersion Device III TLC Plate Heater III TLC Visualizer Analytical balance BP 211 D Ultrasonic bath 5S6 Filter paper for chamber saturation Standard sieve #45 (355 m)

Manufacturer Merck

Manufacturer CAMAG CAMAG CAMAG CAMAG CAMAG Sartorius Fisher Scientific CAMAG Humboldt

Sample, references and reagent preparation Test sample 0.5 g of the powdered test sample is mixed with 10 mL of methanol, sonicated for 15 min. and filtered. The filtrate is used as the test solution S716-01. Reference samples 0.5 g of the powdered reference samples are mixed with 10 mL of methanol, sonicated for 15 minutes and filtered. The filtrates are used as the reference sample solutions S228-02 and S575-01.

Reference standards - 0.3 mg of hyperoside is dissolved in 1 mL of methanol R007-01. 0.3 mg of rutin is dissolved in 1 mL of methanol R008-01. 0.1 mg of caffeic acid is dissolved in 1 mL of methanol R011-01. Reagents Natural Products (NP) reagent: 1 g of diphenylboric acid aminoethyl ester is dissolved in 200 mL of ethyl acetate. Polyethylene Glycol (PEG) 400 reagent: 10 g of PEG 400 is dissolved in 200 mL of dichloromethane.

Application

Chromatography Plate: HPTLC glass 20x10 cm, Silica gel 60 F254 Chamber: Automatic Developing Chamber 2 with humidity control (MgCl2 39.0% RH) Saturation: 20 minutes with filter paper Developing distance from application position / lower edge: 62 mm / 70 mm Developing solvent: Ethyl acetate, acetic acid, formic acid, water (15:1:1:2 v/v/v/v) Plate drying: In ADC2 for 5 minutes Derivatization NP and PEG 400 Reagents: The HPTLC plate is placed on the CAMAG Plate Heater III set at 100C for 5 minutes and while still warm dipped (time 0, speed 5) into NP reagent using the CAMAG
Project A321.02 Trout Lake Farm Skullcap SKU-W2O11, W2O12 HPTLC Report 2 of 3

CAMAG LABORATORY
Immersion Device III. After 1 minute of drying, the plate is then dipped (time 0, speed 5) into PEG 400 reagent and allowed to dry in the fume hood for 5 minutes.

Results Image of developed plate in UV 254 nm

Image of derivatized plate in UV 366 nm

Conclusions Based on the resemblance of the chromatogram of the test sample to those of the Skullcap herb reference samples, it may be concluded that the test sample #SKU-W2O11, W2O12 is Skullcap (Scutellaria lateriflora L.) herb.

Date: Analyst:

11 November 2011

Date: Approved:

11 November 2011

Judy Nichols Laboratory Manager

Don Oates Operations Manager

Disclaimer Statements and interpretations provided in this report are the opinion of CAMAG Laboratory. They do not represent a declaration of conformity with respect to inspection or product certification. Test results correspond to the listed samples only and may not be generalized. This report or any part of it shall not be copied without written permission of CAMAG Laboratory.

Project A321.02 Trout Lake Farm Skullcap SKU-W2O11, W2O12 HPTLC Report

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