Goldenseal
DEFINITION
Goldenseal consists of the dried roots and rhizomes of Hydrastis canadensis L. (Fam. Ranunculaceae). It contains NLT 2.0% of hydrastine (C21H21NO6) and NLT 2.5% of berberine (C20H18NO4), calculated on the dried basis.
IDENTIFICATION
• A. Thin-Layer Chromatographic Identification Test
Standard solution:
0.5 mg/mL each of USP Berberine Chloride RS and USP Hydrastine RS in methanol
Sample solution:
Finely powder the rhizome and the root. Transfer 0.5 g of the powder to a suitable glass vial, and add 0.5 mL of 10% sodium carbonate. Add 5 mL of methanol, and heat for 10 min in a water bath at 60
. Cool to room temperature, filter, and dry under a stream of nitrogen. Add 0.5 mL of methanol to dissolve the residue.
. Cool to room temperature, filter, and dry under a stream of nitrogen. Add 0.5 mL of methanol to dissolve the residue.
Adsorbent:
0.25-mm layer of chromatographic silica gel mixture, typically 20 cm long (TLC plates)
Application volume:
10–20 µL, as bands
Developing solvent system:
Ethyl acetate, butyl alcohol, formic acid, and water (5:3:1:1)
Analysis
Samples:
Standard solution and Sample solution
Develop the chromatograms until the solvent front has moved about three-fourths of the length of the plate, in a saturated chamber. Remove the plate, air-dry, and examine under UV light at 365 nm.
Acceptance criteria:
The chromatograms show zones having a lemon-yellow fluorescence due to berberine at an RF value of about 0.53 and a blue-white fluorescence due to hydrastine at an RF value of about 0.42.
COMPOSITION
• Content of Berberine and Hydrastine and Limit of Palmatine
Mobile phase:
Dissolve 9.93 g of monobasic potassium phosphate in 730 mL of water, add 270 mL of acetonitrile, mix, filter, and degas.
Solvent:
A mixture of 0.1 M monobasic potassium phosphate and acetonitrile (60:40)
Standard solution:
0.05 mg/mL each of USP Berberine Chloride RS and USP Hydrastine RS in a mixture of methanol and water (1:1)
System suitability solution:
Prepare a solution of palmatine in a mixture of water and methanol (1:1) having a known concentration of about 0.05 mg/mL. Mix equal volumes of this solution and the Standard solution.
Sample solution:
Finely powder a quantity of Goldenseal, and transfer about 0.12 g, accurately weighed, to a 50-mL volumetric flask. Add 40 mL of Solvent, sonicate for 5 min, and shake for 10 min. Dilute with Solvent to volume, mix, and filter.
Chromatographic system
(See Chromatography 
621
, System Suitability.)
621, System Suitability.)
Mode:
LC
Detector:
UV 235 nm
Column:
4.6-mm × 150-mm; packing L1
Flow rate:
1.8 mL/min
Injection size:
10 µL
System suitability
Samples:
Standard solution and System suitability solution
Suitability requirements
Resolution:
NLT 1.5 between the berberine and palmatine peaks, and NLT 1.5 between the hydrastine and palmatine peaks, System suitability solution
Capacity factor:
NLT 3.0, determined from the hydrastine and berberine peaks, Standard solution
Column efficiency:
NLT 5000 theoretical plates determined from the hydrastine and berberine peaks, Standard solution
Tailing factor:
NMT 2.0 determined from the hydrastine and berberine peaks, Standard solution
Relative standard deviation:
NMT 2.5% determined from the hydrastine and berberine peaks in repeated injections, Standard solution
Analysis
Samples:
Standard solution and Sample solution
Calculate the percentages of berberine and hydrastine in the portion of Goldenseal taken:
Result = (rU/rS) × CS × (V/W) × 100
| rU | = | = peak area of berberine or hydrastine from the Sample solution |
| rS | = | = peak area of berberine or hydrastine from the Standard solution |
| CS | = | = concentration of berberine or hydrastine in the Standard solution (mg/mL) |
| V | = | = volume of the Sample solution (mL) |
| W | = | = weight of powdered Goldenseal used to prepare the Sample solution (mg) |
Using the values obtained from the chromatogram of the Sample solution, divide the peak area of berberine by the peak area of any peak at the locus for palmatine (if present).
Acceptance criteria:
NLT 2.0% of hydrastine (C21H21NO6) and NLT 2.5% of berberine (C20H18NO4), on the dried basis. The ratio of the berberine peak area to any peak area at the locus for palmatine is more than 50:1.
CONTAMINANTS
• Heavy Metals, Method III 231:
NMT 20 ppm
231:
NMT 20 ppm
• Articles of Botanical Origin, General Method for Pesticide Residues Analysis 561:
Meets the requirements
561:
Meets the requirements
SPECIFIC TESTS
• Botanic Characteristics
Macroscopic:
The rhizome is knotty, subcylindrical, and occasionally has an aerial stem. It is 1–5 cm in length and 2–10 mm in diameter. Externally, the rhizome is brown to dusky yellowish orange, deeply furrowed, and marked by numerous stem and bud scale scars. Numerous brittle roots arise roughly from the same side of the main axis. Fractures are short and resinous, with a dark yellow to yellowish-brown bark, greenish-yellow margins, and a yellowish-orange center that is waxy in appearance. An interrupted circle of small, radially elongated fibrovascular bundles are also present. The roots are filiform, up to 35 cm in length and 1 mm in diameter, and are either curved or twisted, tangled together, or broken. Fractures are short and brittle, and show an internal color of yellowish orange to greenish yellow.
Histology
Transverse section of rhizome and root:
The rhizome has polygonal, yellowish-brown, thin- to slightly thick-walled cork cells. Wedge-shaped vascular bundles are separated by wide medullary rays. Tracheary elements are lignified and have slit-shaped pits. A few large vessels with reticulate thickenings are also present. The parenchyma tissue is composed of polygonal cells with abundant simple or compound starch grains up to 8 µm in diameter. A few irregularly shaped resin cells are present in the cortex and the pith. Masses of granular, orange-brown matter are also present in the parenchyma tissues. The roots have a single layer of irregularly elongated cork cells. The tracheary elements are associated with lignified fibers. Fragments of the epidermis are sometimes present near the base of the rhizome and are composed of cells with thick, lignified, beaded walls.
• Loss on Drying 731:
Dry 2 g of finely powdered Goldenseal at 100 for 5 h: it loses NMT 12.0% of its weight.
731:
Dry 2 g of finely powdered Goldenseal at 100 for 5 h: it loses NMT 12.0% of its weight.
ADDITIONAL REQUIREMENTS
• Packaging and Storage:
Preserve in tight containers, and store protected from light, moisture, and heat.
• Labeling:
The label states the Latin binomial and, following the official name, the parts of the plant contained in the article.
Auxiliary Information—
Please check for your question in the FAQs before contacting USP.
| Topic/Question | Contact | Expert Committee |
|---|---|---|
| Monograph | Maged H. Sharaf, Ph.D.
Principal Scientific Liaison 1-301-816-8318 |
(DS2010) Monographs - Dietary Supplements |
| Reference Standards | RS Technical Services 1-301-816-8129 rstech@usp.org |
USP35–NF30 Page 1343
Pharmacopeial Forum: Volume No. 30(3) Page 952