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Printed by: Dang Van Vu Official Date: Official as of 01-Aug-2021 Document Type: DIETARY SUPPLEMENTS @2023 USPC
Do Not Distribute DOI Ref: at8g4 DOI: https://doi.org/10.31003/USPNF_M34988_05_01
1

Change to read: corresponding in RF and color to ginsenoside Rb1 in

Standard solution A, two to three reddish-violet bands
below ginsenoside Rb1, a dark brown band immediately
▲ (USP 1-Aug-2021)Asian
Ginseng ▲Root and
▲
above ginsenoside Rb1, and a brown band above the
Rhizome Dry▲ (USP 1-Aug-2021) Extract dark brown band due to ginsenoside Rc. In the
middle-third section, Standard solution B exhibits four
DEFINITION brown bands, separately due to ginsenosides Rd, Re, Rf,
Change to read: and Rg1 with increasing RF; the latter corresponds in RF
and color to ginsenoside Rg1 in Standard solution A.
▲
▲ (USP 1-Aug-2021)Asian Ginseng ▲Root and Under UV light at 365 nm: Standard solution A exhibits a
Rhizome Dry▲ (USP 1-Aug-2021) Extract is prepared from ▲the reddish-blue band due to ginsenoside Rb1 in the
dried roots and rhizomes of Panax ginseng C.A.Mey by lower-third section and a pinkish-violet band due to
extraction with water or hydroalcoholic ginsenoside Rg1 in the middle-third section. In the
mixtures.▲ (USP 1-Aug-2021) It contains NLT 3.0% ▲of lower-third section, Standard solution B exhibits a band
total▲ (USP 1-Aug-2021) ginsenosides, ▲calculated as the sum of corresponding in RF and color to ginsenoside Rb1 in
ginsenoside Rg1 (C42H72O14), ginsenoside Re (C48H82O18), Standard solution A, two to three reddish-blue bands
ginsenoside Rf (C42H72O14), ginsenoside Rb1 (C54H92O23), below ginsenoside Rb1, a black band immediately above
ginsenoside Rc (C53H90O22), ginsenoside Rb2 (C53H90O22), ginsenoside Rb1, and a blue band above the black band
and ginsenoside Rd (C48H82O18)▲ (USP 1-Aug-2021) on the due to ginsenoside Rc. In the middle-third section,
anhydrous basis; ▲the content of total ginsenosides is NLT Standard solution B exhibits a blue band due to
90% and NMT 110% of the labeled amount.▲ (USP 1-Aug-2021) ginsenoside Rd and three pinkish-violet bands above
ginsenoside Rd, separately due to ginsenosides Re, Rf,

al
It may contain ▲suitable▲ (USP 1-Aug-2021) added substances
▲
as carriers.▲ (USP 1-Aug-2021) and Rg1 with increasing RF; the latter corresponds in RF
and color to ginsenoside Rg1 in Standard solution
IDENTIFICATION A.▲ (USP 1-Aug-2021)
Change to read: Acceptance criteria
▲
Under white light: In the lower-third section, the Sample
• A. ▲HPTLC FOR ARTICLES OF BOTANICAL ORIGIN á203ñ
Standard solution A: 0.5 mg/mL each of USP Ginsenoside
Rg1 RS and USP Ginsenoside Rb1 RS in
ci solution exhibits a band corresponding in RF and color to
ginsenoside Rb1 in Standard solution A and Standard
solution B, two to three reddish-violet bands below
methanol▲ (USP 1-Aug-2021) ginsenoside Rb1, a dark brown band immediately above
Standard solution ▲B: 10 mg/mL of USP Powdered Asian ginsenoside Rb1, and a brown band above the dark brown
ffi
Ginseng Extract RS in methanol. Sonicate for 15 min, band due to ginsenoside Rc (distinction from P.
centrifuge, and use the supernatant.▲ (USP 1-Aug-2021) notoginseng root and rhizome), corresponding in RF and
Sample solution: ▲10 mg/mL of Asian Ginseng Root and color to the same constituents in Standard solution B. In
Rhizome Dry Extract in methanol. Sonicate for 15 min, the middle-third section, the Sample solution exhibits four
centrifuge, and use the supernatant. brown bands corresponding in RF and color to the same
Chromatographic system▲ (USP 1-Aug-2021)
O

constituents in Standard solution B, separately due to

Adsorbent: ▲Silica gel with fluorescence indicator F254, ginsenosides Rd, Re, Rf (distinction from P. quinquefolius
precoated plate, for HPTLC▲ (USP 1-Aug-2021) root and rhizome), and Rg1 with increasing RF; the latter
Application volume: ▲3 µL, as 8-mm bands corresponds in RF and color to ginsenoside Rg1 in Standard
Relative humidity: Condition the plate to a relative solution A.
humidity of about 33% using a suitable device. Under UV light at 365 nm: In the lower-third section, the
Temperature: About 25°▲ (USP 1-Aug-2021) Sample solution exhibits a band corresponding in RF and
Developing solvent system: ▲Methylene chloride, color to ginsenoside Rb1 in Standard solution A and
anhydrous ethanol, and water (60: 45: 6.5)
Standard solution B, two to three reddish-blue bands
Derivatization reagent: 10% sulfuric acid in alcohol.
below ginsenoside Rb1, a black band immediately above
[NOTE—Prepare on the day of the test; it can be used for
up to 1 week if stored in a tightly closed container in a ginsenoside Rb1, and a blue band above the black band
cool place. Slowly add sulfuric acid to ice-cold due to ginsenoside Rc (distinction from P. notoginseng
alcohol.]▲ (USP 1-Aug-2021) root and rhizome) corresponding in RF and color to the
Analysis same constituents in Standard solution B. In the
Samples: Standard solution ▲A, Standard middle-third section, the Sample solution exhibits four
solution B,▲ (USP 1-Aug-2021) and Sample solution bands corresponding in RF and color to the same
▲
Apply the Samples as bands and dry in air. Develop in a constituents in Standard solution B, including a blue band
saturated chamber, remove the plate from the chamber, due to ginsenoside Rd and three pinkish-violet bands
and dry in air. Treat the plate with Derivatization above ginsenoside Rd, separately due to ginsenosides Re,
reagent, heat at 105° for 10 min, and examine Rf (distinction from P. quinquefolius root and rhizome),
immediately under white light and UV light at 365 nm. and Rg1 with increasing RF; the latter corresponds in RF and
System suitability color to ginsenoside Rg1 in Standard solution
Samples: Standard solution A and Standard solution B A.▲ (USP 1-Aug-2021)
Suitability requirements
Under white light: Standard solution A exhibits a Change to read:
reddish-violet band due to ginsenoside Rb1 in the
lower-third section, and a brown band due to • B. ▲HPLC
ginsenoside Rg1 in the middle-third section. In the Analysis: Proceed as directed in the test for Content of
lower-third section, Standard solution B exhibits a band Ginsenosides.

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Printed by: Dang Van Vu Official Date: Official as of 01-Aug-2021 Document Type: DIETARY SUPPLEMENTS @2023 USPC
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Acceptance criteria: The Sample solution exhibits peaks Standard solution B: 10 mg/mL of USP Powdered Asian
corresponding to the retention times of ginsenoside Rg1 Ginseng Extract RS in Solvent, sonicate for 10 min. Before
and ginsenoside Rb1 in Standard solution A and Standard injection, pass through a nylon filter of 0.22-µm
solution B. The Sample solution exhibits peaks due to pore size.▲ (USP 1-Aug-2021)
ginsenoside Re, ginsenoside Rf (absent in the roots and Sample solution: ▲Transfer about 100 mg (adjust the
rhizomes of P. quinquefolius and P. notoginseng), amount, if necessary) of Asian Ginseng Root and Rhizome
chikusetsusaponin V (ginsenoside Ro), ginsenoside Rc, Dry Extract, accurately weighed, to a suitable flask, add
ginsenoside Rb2 (distinction from P. notoginseng root and 10.0 mL of Solvent, and close tightly. Weigh the filled flask
rhizome; it does not contain chikusetsusaponin V and accurately, and sonicate for 30 min. Cool to room
ginsenosides Rc and Rb2), and ginsenoside Rd temperature and adjust to the initial weight by adding
corresponding to the retention times of the same Solvent, if necessary. Before injection, pass through a nylon
ginsenosides in Standard solution B. The content ratio of filter of 0.22-µm pore size and discard the first portion of
ginsenoside Rb2 to ginsenoside Rb1 is NLT 0.3 the filtrate.▲ (USP 1-Aug-2021)
(differentiation from P. quinquefolius root and rhizome). Chromatographic system
Malonyl ginsenoside Rb1 peak is between ginsenoside Rb1 (See Chromatography á621ñ, System Suitability.)
Mode: LC
and ginsenoside Rc; one peak between ginsenoside Rc and
Detector: UV 203 nm
ginsenoside Rb2 is due to malonyl ginsenoside Rc; one peak
Column: ▲2.1-mm × 5-cm; 1.7-µm▲ (USP 1-Aug-2021) packing
between ginsenoside Rb2 and ginsenoside Rd is due to
L1
malonyl ginsenoside Rb2 (malonyl ginsenosides are absent Column temperature: ▲40°▲ (USP 1-Aug-2021)
in water extract and steamed roots and rhizomes of P. Flow rate: ▲0.8▲ (USP 1-Aug-2021) mL/min
ginseng); and these peaks correspond to the retention times Injection volume: ▲5▲ (USP 1-Aug-2021) µL
for the same peaks in Standard solution B. The peak intensity

al
System suitability
of ginsenoside Rd is less than or similar to that of
Samples: ▲Standard solution A and Standard solution
ginsenoside Rb2 (differentiation from aerial parts of P.
B▲ (USP 1-Aug-2021)
ginseng).▲ (USP 1-Aug-2021) Suitability requirements
▲
Resolution: NLT 1.3 between the ginsenoside Rg1 and
Delete the following:
▲
• C. The retention times of the peaks for ginsenosides Rg1,
Re, Rf, Rb1, Rb2, Rc, and Rd in the Sample solution
ci ginsenoside Re peaks; NLT 2.5 between the
ginsenoside Rb1 and chikusetsusaponin V peaks; NLT 1.3
between the peak of ginsenoside Rd and the peak before
it, Standard solution B
chromatogram correspond to those in the Standard
solution, as obtained in the test for Content of Ginsenosides. Tailing factor: NMT 2.0 for ginsenoside Rg1 and
ffi
The ratio of the peak area of Rb2 to the peak area of Rb1 is ginsenoside Rb1 peaks, Standard solution A
NLT 0.4 (differentiation from American Relative standard deviation: NMT 2.0% for
Ginseng).▲ (USP 1-Aug-2021) ginsenoside Rg1 and ginsenoside Rb1 peaks in replicate
injections, Standard solution A▲ (USP 1-Aug-2021)
COMPOSITION Chromatogram similarity: The chromatogram ▲of
Standard solution B▲ (USP 1-Aug-2021) is similar to the
Change to read:
O

reference chromatogram provided with the lot of USP

• CONTENT OF GINSENOSIDES Powdered Asian Ginseng Extract RS being used.
▲
Solution A: ▲0.003% phosphoric acid in▲ (USP 1-Aug-2021) ▲ (USP 1-Aug-2021)
water Analysis
Solution B: Acetonitrile▲▲ (USP 1-Aug-2021) Samples: ▲Standard solution A, Standard
Mobile phase: See Table 1. solution B,▲ (USP 1-Aug-2021) and Sample solution
▲
[NOTE—Protect from light. Standard solution A,
▲
Table 1 Standard solution B, and Sample solution are stable for
Time Solution A Solution B
24 h at room temperature.]
(min) (%) (%) Using the chromatograms of Standard solution A, Standard
solution B, and the reference chromatogram provided
0 83 17 with the lot of USP Powdered Asian Ginseng Extract RS
0.5 83 17 being used, identify the peaks corresponding to
ginsenosides Rg1, Re, Rf, Rb1, Rc, Rb2, and Rd, and the
3.5 81 19
peaks corresponding to chikusetsusaponin V, and malonyl
5.0 77 23 ginsenosides Rb1, Rc, Rb2 if they are available, in the
15.0 70 30
Sample solution.
Separately▲ (USP 1-Aug-2021) calculate the percentages of
16.0 70 30 ▲
ginsenosides Rg1, Re, and Rf against USP Ginsenoside
16.1 5 95 Rg1 RS and the percentages of ginsenosides Rb1, Rc, Rb2,
and Rd against USP Ginsenoside Rb1 RS▲ (USP 1-Aug-2021) in
20.0 5 95
the portion of ▲▲ (USP 1-Aug-2021) Asian Ginseng ▲Root and
20.1 83 17 Rhizome Dry▲ (USP 1-Aug-2021) Extract taken:
25.0 83 17
Result = (rU/rS) × ▲Cs × (V/W) × F × 100▲ (USP 1-Aug-2021)
Solvent: Methanol and water (7:3) rU = peak area ▲of▲ (USP 1-Aug-2021) the relevant
Standard solution A: 0.15 mg/mL each of USP Ginsenoside ginsenoside from the Sample solution
Rg1 RS and USP Ginsenoside Rb1 RS in Solvent

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Printed by: Dang Van Vu Official Date: Official as of 01-Aug-2021 Document Type: DIETARY SUPPLEMENTS @2023 USPC
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rS = peak area ▲of ginsenoside Rg1 or • MICROBIAL ENUMERATION TESTS á2021ñ: The total aerobic
ginsenoside Rb1▲ (USP 1-Aug-2021) from Standard microbial count does not exceed 300 cfu/g. The total
solution ▲A▲ (USP 1-Aug-2021) combined molds and yeasts count does not exceed
CS = concentration of ▲USP Ginsenoside Rg1 RS or USP 100 cfu/g.
Ginsenoside Rb1 RS▲ (USP 1-Aug-2021) in Standard
solution ▲A (mg/mL) Change to read:
V = volume of Sample solution (mL)
• ▲ABSENCE OF SPECIFIED MICROORGANISMS á2022ñ, Test
W = weight of Asian Ginseng Root and Rhizome Dry
Procedures, Test for Absence of Salmonella Species, and Test
Extract used to prepare the Sample solution
for Absence of Escherichia coli:▲ (USP 1-Aug-2021) Meets the
(mg)
F = conversion factor for the relevant analyte (see requirements▲▲ (USP 1-Aug-2021)
Table 2) SPECIFIC TESTS
Table 2 Change to read:
Conversion Factor • WATER DETERMINATION á921ñ, Method I: NMT
Conversion Factor (F) (F)
Analyte Based on Rg1 Based on Rb1 7.0%▲▲ (USP 1-Aug-2021)
Ginsenoside Rg1 1.00 — Delete the following:
Ginsenoside Re 1.00 — ▲
• ALCOHOL DETERMINATION, Method II á611ñ: NMT
Ginsenoside Rf 0.83 — 0.25%▲ (USP 1-Aug-2021)

al
Ginsenoside Rb1 — 1.00
Add the following:
Ginsenoside Rc — 0.96

Ginsenoside Rb2
▲
• BOTANICAL EXTRACTS á565ñ, Preparations, General
— 0.96
Pharmacopeial Requirements, Residual Solvents: Meets the
Ginsenoside Rd — 0.80▲ (USP 1-Aug-2021) requirements▲ (USP 1-Aug-2021)

Calculate the ▲content of total ginsenosides as the sum of

ginsenosides Rg1, Re, Rf, Rb1, Rc, Rb2, and Rd.
ci ADDITIONAL REQUIREMENTS
Change to read:
Calculate the percentage of the labeled amount of total
ginsenosides in the portion of Asian Ginseng Root and • PACKAGING AND STORAGE: ▲Preserve in tight containers,
ffi
Rhizome Dry Extract taken: protected from light and moisture, and store in a cool
place.▲ (USP 1-Aug-2021)
Result = (P/L) × 100
Change to read:
P = content of total ginsenosides, determined
previously (%) • LABELING: ▲The label states the Latin binomial following the
O

L = labeled amount of total ginsenosides official name of the plant from which the article was
(%)▲ (USP 1-Aug-2021) prepared. The labeling indicates the content of total
ginsenosides as percentage on the anhydrous basis. It
Acceptance criteria meets other labeling▲ (USP 1-Aug-2021) requirements in
▲
Total ginsenosides:▲ (USP 1-Aug-2021) NLT 3.0%,▲and Botanical Extracts á565ñ.
between 90% and 110% of the labeled
amount,▲ (USP 1-Aug-2021) on the anhydrous basis Change to read:

CONTAMINANTS • USP REFERENCE STANDARDS á11ñ

• BOTANICAL EXTRACTS á565ñ, Preparations, General USP Powdered Asian Ginseng Extract RS
Pharmacopeial Requirements, Pesticide Residues: Meets the ▲
USP Ginsenoside Rb1 RS
requirements USP Ginsenoside Rg1 RS▲ (USP 1-Aug-2021)

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