ISO/DIS 5228

Traditional Chinese Medicine --Rheum palmatum, Rheum tanguticum, and Rheum officinale root and rhizome

This document specifies the quality and safety of Rheum root and rhizome [the dry root and rhizome of Rheum palmatum L., Rheum tanguticum Maxim.ex Balf. and Rheum officinale Baill.]. This document applies to Rheum root and rhizome which are sold and used as natural medicines in international trade, including Chinese herbal medicines (whole medicinal materials) and decoction pieces derived from these plants.

Titre manque

General Information

• Status: Not Published
• ICS: 11.120.10 - Medicaments
• Technical Committee: ISO/TC 249 - Traditional Chinese medicine
• Drafting Committee: ISO/TC 249 - Traditional Chinese medicine
• Current Stage: 5000 - FDIS registered for formal approval
• Start Date: 26-Jan-2023
• Completion Date: 15-Feb-2023

Standards Content (Sample)

DRAFT INTERNATIONAL STANDARD
ISO/DIS 5228
ISO/TC 249 Secretariat: SAC
Voting begins on: Voting terminates on:
2022-07-14 2022-10-06
Traditional Chinese Medicine — Rheum palmatum, Rheum
tanguticum, and Rheum officinale root and rhizome
ICS: 11.120.10
THIS DOCUMENT IS A DRAFT CIRCULATED
FOR COMMENT AND APPROVAL. IT IS
THEREFORE SUBJECT TO CHANGE AND MAY
This document is circulated as received from the committee secretariat.
NOT BE REFERRED TO AS AN INTERNATIONAL
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ISO/DIS 5228:2022(E)
RECIPIENTS OF THIS DRAFT ARE INVITED
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PROVIDE SUPPORTING DOCUMENTATION. © ISO 2022
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ISO/DIS 5228:2022(E)
DRAFT INTERNATIONAL STANDARD
ISO/DIS 5228
ISO/TC 249 Secretariat: SAC
Voting begins on: Voting terminates on:
Traditional Chinese Medicine — Rheum palmatum, Rheum
tanguticum, and Rheum officinale root and rhizome
ICS: 11.120.10
COPYRIGHT PROTECTED DOCUMENT
THIS DOCUMENT IS A DRAFT CIRCULATED
FOR COMMENT AND APPROVAL. IT IS
© ISO 2022
THEREFORE SUBJECT TO CHANGE AND MAY
This document is circulated as received from the committee secretariat.

All rights reserved. Unless otherwise specified, or required in the context of its implementation, no part of this publication may

NOT BE REFERRED TO AS AN INTERNATIONAL

be reproduced or utilized otherwise in any form or by any means, electronic or mechanical, including photocopying, or posting on STANDARD UNTIL PUBLISHED AS SUCH.

the internet or an intranet, without prior written permission. Permission can be requested from either ISO at the address below

IN ADDITION TO THEIR EVALUATION AS

or ISO’s member body in the country of the requester. BEING ACCEPTABLE FOR INDUSTRIAL,

TECHNOLOGICAL, COMMERCIAL AND
ISO copyright office
USER PURPOSES, DRAFT INTERNATIONAL
CP 401 • Ch. de Blandonnet 8
STANDARDS MAY ON OCCASION HAVE TO
BE CONSIDERED IN THE LIGHT OF THEIR
CH-1214 Vernier, Geneva
POTENTIAL TO BECOME STANDARDS TO
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WHICH REFERENCE MAY BE MADE IN
Reference number
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NATIONAL REGULATIONS.
Website: www.iso.org ISO/DIS 5228:2022(E)
RECIPIENTS OF THIS DRAFT ARE INVITED
Published in Switzerland
TO SUBMIT, WITH THEIR COMMENTS,
NOTIFICATION OF ANY RELEVANT PATENT
RIGHTS OF WHICH THEY ARE AWARE AND TO
© ISO 2022 – All rights reserved
PROVIDE SUPPORTING DOCUMENTATION. © ISO 2022
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ISO/DIS 5228:2022(E)
Contents Page

Foreword ........................................................................................................................................................................................................................................iv

Introduction .................................................................................................................................................................................................................................v

1 Scope ................................................................................................................................................................................................................................. 1

2 Normative references ..................................................................................................................................................................................... 1

3 Terms and definitions .................................................................................................................................................................................... 1

4 Description ........................................................................................................................................... ......................................................................2

5 Requirements .......................................................................................................................................................................................................... 4

5.1 General characteristics ................................................................................................................................................................... 4

5.2 Morphological features ................................................................................................................................................................... 4

5.3 Identification ............................................................................................................................................................................................ 4

5.3.1 Microscopic identification ................................... ....................................................................................................... 4

5.3.2 Thin-layer chromatogram (TLC) identification ..................................................................................... 5

5.4 Rhaponticin ............................................................................................................................................................................................... 5

5.5 Moisture ....................................................................................................................................................................................................... 5

5.6 Total ash ....................................................................................................................................................................................................... 5

5.7 Acid-insoluble ash ................................................................................................................................................................................ 6

5.8 Extractives ................................................................................................................................................................................................. 6

5.9 Heavy metals ............................................................................................................................................................................................ 6

5.10 Pesticide residues ................................................................................................................................................................................ 6

5.11 Marker compounds ............................................................................................................................................................................. 6

6 Sampling ....................................................................................................................................................................................................................... 6

7 Test methods ............................................................................................................................................................................................................ 6

7.1 Macroscopic identification ........................................................................................................................................................... 6

7.2 Thin-layer chromatogram (TLC) identification ........................................................................................................ 6

7.3 Rhaponticin identification ........................................................................................................................................................... 6

7.4 Determination of moisture .......................................................................................................................................................... 6

7.5 Determination of total ash ........................................................................................................................................................... 6

7.6 Determination of acid-insoluble ash ................................................................................................................................... 6

7.7 Determination of extractives .................................................................................................................................................... 7

7.8 Determination of heavy metals ............................................................................................................................................... 7

7.9 Determination of pesticide residues ................................................................................................................................... 7

7.10 Determination of marker compounds ............................................................................................................................... 7

8 Test report .................................................................................................................................................................................................................. 7

9 Packaging, storage and transportation ...................................................................................................................................... 7

10 Marking and labelling .................................................................................................................................................................................... 7

Annex A (informative) Thin-layer chromatogram (TLC) identification ........................................................................ 9

Annex B (informative) Determination of marker compounds by HPLC-UV ............................................................10

Annex C (informative) Reference information of national and regional requirements ............................17

Annex D (informative) Differentiation of the three species ....................................................................................................19

Bibliography .............................................................................................................................................................................................................................22

iii
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ISO/DIS 5228:2022(E)
Foreword

ISO (the International Organization for Standardization) is a worldwide federation of national standards

bodies (ISO member bodies). The work of preparing International Standards is normally carried out

through ISO technical committees. Each member body interested in a subject for which a technical

committee has been established has the right to be represented on that committee. International

organizations, governmental and non-governmental, in liaison with ISO, also take part in the work.

ISO collaborates closely with the International Electrotechnical Commission (IEC) on all matters of

electrotechnical standardization.

The procedures used to develop this document and those intended for its further maintenance are

described in the ISO/IEC Directives, Part 1. In particular, the different approval criteria needed for the

different types of ISO documents should be noted. This document was drafted in accordance with the

editorial rules of the ISO/IEC Directives, Part 2 (see www.iso.org/directives).

Attention is drawn to the possibility that some of the elements of this document may be the subject of

patent rights. ISO shall not be held responsible for identifying any or all such patent rights. Details of

any patent rights identified during the development of the document will be in the Introduction and/or

on the ISO list of patent declarations received (see www.iso.org/patents).

Any trade name used in this document is information given for the convenience of users and does not

constitute an endorsement.

For an explanation of the voluntary nature of standards, the meaning of ISO specific terms and

expressions related to conformity assessment, as well as information about ISO's adherence to

the World Trade Organization (WTO) principles in the Technical Barriers to Trade (TBT), see

www.iso.org/iso/foreword.html.

This document was prepared by Technical Committee ISO/TC 249, Traditional Chinese medicine.

Any feedback or questions on this document should be directed to the user's national standards body. A

complete listing of these bodies can be found at www.iso.org/members.html.
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ISO/DIS 5228:2022(E)
Introduction

Rheum root and rhizome is the dry root and rhizome of Rheum palmatum L., Rheum tanguticum Maxim.

ex Balf. and Rheum officinale Baill. As one of the four traditional Chinese medicines in China, Rheum

root and rhizome is documented in Shennong material medica (神农本草经). Rheum root and rhizome

has a wide range of clinical applications, high frequency of use, and a long history of medicinal use. It is

recorded in traditional Chinese medicine books such as Yao pin hua yi(药品化义), Yi xue zhong zhong can

xi lu(医学衷中参西录)and Su Wen(素问).

Rheum root and rhizome has a cold property and bitter flavour. Its meridian distribution in the theory

of TCM belongs to the spleen, stomach, large intestine, liver, and pericardium meridians. It can remove

accumulation with purgation, clear heat, purge fire, cool the blood, remove toxin, expel stasis, unblock

the meridians, and drain dampness to abate jaundice, for accumulation, stagnation and constipation

caused by excess heat, hematemesis caused by intestinal abscess, blood-stasis amenorrhea, postpartum

stasis and obstruction, traumatic injuries, dampness-heat dysentery, jaundice, red urine, stranguria,

and edema, topical application for burn and scald. Wine-fried Rheum root and rhizome is good at

clearing heat toxin in the upper energizer blood aspect, which can be applied for red eyes, swollen

throat, and painful swollen gums. The purgation power of prepared Rheum root and rhizome is mild,

and it can purge fire and remove toxins to treat soreness and ulcers caused by fire-toxin. Charred

Rheum root and rhizome can cool the blood, resolve stasis and stop bleeding, which can be applied to

bleeding symptoms caused by blood heat and stasis.

Rheum root and rhizome has a complex chemical composition, including various compounds such as

anthraquinone, anthrone, tannin, and polysaccharide. Modern pharmacological studies have shown

that sennoside and anthraquinone glucosides are the main components of Rheum root and rhizome

that can induce diarrhea; free anthraquinones are antibacterial and antitumor active ingredients of

Rheum root and rhizome; n-butyrophenones have good anti-inflammatory and analgesic effects;

gallic acid glucosides and galloyl proanthocyanidins in the tannins have hypolipidemic effects; and

d-catechin and gallic acid have haemostatic effects. Moreover, Rheum root and rhizome also has the

functions of relieving phlegm, protecting the liver and gallbladder, and protecting cardiovascular and

cerebrovascular diseases.

In global trade, taking China Customs data as an example, Rheum root and rhizome in China is mainly

exported to Japan, South Korea, the United States, Indonesia, Germany, Italy, Singapore, France,

Thailand, Vietnam, Malaysia and so on. From 2012 to 2016, the average annual export trade volume

of Rheum root and rhizome in China was 7.128.400 US dollars, which is one of the main Chinese herbal

medicines exported by China.

There are 152 Chinese patent medicines containing Rheum root and rhizome in the Chinese

Pharmacopoeia (2015 Edition) and 2 preparations in Japanese Pharmacopoeia (17th Edition). Rheum

root and rhizome and its products have applications in a variety of medical fields, and also involve

detoxification and beauty, lipid-lowering, weight-loss health products and food additives. Many

users believe that "Chinese medicine is non-toxic" and fail to take it strictly according to the doctor's

instructions, which leads to excessive and chronic irregular use of Rheum root and rhizome and liver

and kidney damage.

As a worldwide drug, Rheum root and rhizome is included in the pharmacopoeia of many countries

and regions, such as China, Japan, Korea, and Europe. At present, the quality control of Rheum root and

rhizome and its preparations is mostly based on the content of anthraquinones. However, the medicinal

ingredients of Rheum root and rhizome are not just anthraquinones. The diversity of ingredients in

traditional Chinese medicine determines that quality control should adopt a multi-index quality

evaluation model.

Furthermore, Rheum root and rhizome is ranked tenth in the ISO/TR 23975: 2019 Traditional Chinese

medicine — Priority list of single herbal medicines for developing standards, which indicates its high

priority. Therefore, it is necessary to establish an international standard of Rheum root and rhizome,

which unifies the quality and safety of Rheum root and rhizome, ensures the safety and effectiveness of

the medication, and regulates trade in the international market. The establishment of an international

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ISO/DIS 5228:2022(E)

standard for Rheum root and rhizome is therefore necessary to guarantee the clinical effectiveness,

safety, and controllability of this valuable medicine in global commerce and trade.

As national implementation may differ, national standards bodies are invited to modify the values given

in 5.5, 5.6, 5.7, 5.8, 5.9, 5.10, and 5.11 in their national standards. Examples of national and regional

values are given in Annex C.
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DRAFT INTERNATIONAL STANDARD ISO/DIS 5228:2022(E)
Traditional Chinese Medicine — Rheum palmatum, Rheum
tanguticum, and Rheum officinale root and rhizome
1 Scope

This document specifies the quality and safety requirements of Rheum root and rhizome [the dried root

and rhizome of Rheum palmatum L., Rheum tanguticum Maxim. ex Balf., and Rheum officinale Baill.].

This document applies to Rheum root and rhizome that is sold and used as natural medicines in

international trade, including Chinese materia medica (whole medicinal materials) and decoction

pieces derived from these plants.

This document does not apply to the processing methods and processed products of Rheum root and

rhizome.
2 Normative references

The following documents are referred to in the text in such a way that some or all of their content

constitutes requirements of this document. For dated references, only the edition cited applies. For

undated references, the latest edition of the referenced document (including any amendments) applies.

ISO 18664, Traditional Chinese Medicine — Determination of heavy metals in herbal medicines used in

Traditional Chinese Medicine

ISO 21371, Traditional Chinese medicine — Labelling requirements of products intended for oral or topical

use

ISO 22217, Traditional Chinese medicine —Storage requirements for raw materials and decoction pieces

ISO 22258, Traditional Chinese medicine — Determination of pesticide residues in natural products by gas

chromatography

ISO 23723, Traditional Chinese medicine — General requirements for herbal raw material and materia

medica
3 Terms and definitions
For the purposes of this document, the following terms and definitions apply.

ISO and IEC maintain terminological databases for use in standardization at the following addresses:

— ISO Online browsing platform: available at https:// www .iso .org/ obp
— IEC Electropedia: available at https:// www .electropedia .org/
3.1
Rheum root and rhizome

dried root and rhizome of Rheum palmatum L., Rheum tanguticum Maxim. ex Balf., and Rheum officinale

Baill.
3.2
batch

samples collected from the same particular place at the same time, no more than 5 000 kg

[SOURCE: ISO 22988:2019, 3.8]
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ISO/DIS 5228:2022(E)
4 Description

Rheum root and rhizome is the dried root and rhizome of Rheum palmatum L., Rheum tanguticum Maxim.

ex Balf., and Rheum officinale Baill., as shown in Figure 1. The crude drug is collected in late autumn

when stem and leaves have withered or in spring just before budding. The crude drug is removed

from rootlet and the outer bark, cut into segment or section, and either hung in line for drying or dried

directly.
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ISO/DIS 5228:2022(E)
Key
A rheum palmatum L.
B rheum tanguticum Maxim. ex Balf.
C rheum officinale Baill.
1 inflorescence
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ISO/DIS 5228:2022(E)
2 flower
3 stem
4 root and rhizome
5 leaf
6 xylem, cambium and phloem
7 pith
8 abnormal vascular bundles
Figure 1 — Structure of Rheum root and rhizome
5 Requirements
5.1 General characteristics
The following requirements shall be met before sampling.
a) Rheum root and rhizome shall be clean and free from leave and foreign matter.

b) The presence of living insects, mouldy fruit, and external contaminants which are visible to the

naked eye shall not be permitted.
5.2 Morphological features

The appearance of the medicinal materials of rheum root and rhizome is subcylindrical, conical,

ovoid, or irregular pieces, 3 cm to 17 cm long, 3 cm to 10 cm in diameter. Externally yellowish-brown

to reddish-brown when peeled, sometimes whitish reticulations and scattered star spots (abnormal

vascular bundles) visible, occasionally with brownish-black patches of cork, mainly with a hole through

which the string passed, and coarse wrinkles. The texture is compact and sometimes loose and soft in

the center, facture pale reddish-brown or yellowish-brown, granular. The pith of the rhizome broad,

with star spots arranged in a ring or irregularly scattered. The wood of the root is well developed, lined

radially, cambium ring distinct, without star spots. Odour, delicately aromatic; taste bitter and slightly

astringent, sticky, and gritty on chewing.
NOTE The hole and the string are for hanging Rheum root and rhizome when dry.
5.3 Identification
5.3.1 Microscopic identification

Transverse section, as shown in Figure 2: most cork and phelloderm of root removed. In phloem, sieve

tube groups distinct, parenchyma well developed. Cambium in a ring. Xylem with relatively dense rays,

2 to 4 cells wide containing brown masses; vessels non-lignified, usually single or several grouped,

sparsely arranged. Parenchymatous cells have clusters of calcium oxalate and abundant starch granules.

The pith of the rhizome is broad, usually showing mucilage cavities, containing reddish-brown masses;

abnormal vascular bundles are scattered; cambium is in a ring; xylem is at the inside of cambium, and

phloem is outside. Stellate rays radiate.

Powder: Yellowish-brown. Clusters of calcium oxalate are 20 μm to 160 μm, sometimes up to 190 μm

in diameter. Bordered pitted vessels, reticulated vessels, spiral vessels, and annular vessels are non-

lignified. Starch granules are fairly abundant, single granules are spheroid or polygonal, 3 μm to 45 μm

in diameter, and hilum stellate; compound granules consist of 2 to 8 components.
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ISO/DIS 5228:2022(E)
Key
A rheum palmatum L.
B rheum tanguticum Maxim. ex Balf.
C rheum officinale Baill.
1 cork
2 cortex
3 phloem
4 cambium
5 xylem
6 pith
7 abnormal vascular bundles
a clusters of calcium oxalate
b mucilage cavities
Figure 2 — Transverse section of Rheum root and rhizome
5.3.2 Thin-layer chromatogram (TLC) identification

Spots in the chromatogram obtained with the test solution should correspond in position and colour to

the spots in the chromatogram obtained from the reference drug solution or reference solution.

5.4 Rhaponticin

a) The bright blue fluorescence spot in the chromatogram obtained with the test solution shall not

correspond in position and colour to the spot in the chromatogram obtained with the reference

solution in TLC chromatogram.

b) When HPLC is performed, the chromatographic peak of the test solution shall not correspond

at the retention time to rhaponticin (C H O ) Chemical Reference Standard (CRS) in HPLC

21 24 9
chromatogram.
5.5 Moisture
The content of water in percentage mass should not be more than 15,0 % (w/w).
5.6 Total ash

The content of total ash in percentage mass should not be more than 13,0 % (w/w).

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ISO/DIS 5228:2022(E)
5.7 Acid-insoluble ash

The content of acid-insoluble ash in percentage mass should not be more than 2,0 % (w/w).

5.8 Extractives

The content of water-soluble extractives in percentage mass should not be less than 25,0 % (w/w).

5.9 Heavy metals

The contents of heavy metals, such as arsenic, mercury, lead, and cadmium, shall be determined.

5.10 Pesticide residues
The contents of pesticide residues shall be determined.
5.11 Marker compounds

The content of marker compounds, such as total anthraquinone, free anthraquinone, and sennoside A,

shall be determined.
6 Sampling

Sampling shall be carried out in accordance with the method described in the Clause 8 of ISO 23723.

7 Test methods
7.1 Macroscopic identification

Samples not less than 500 g are taken from each batch randomly. These samples are examined by naked

eyes observation in sunlight, and smell. The UPLC method specified in Annex D was used to distinguish

the three species of Rheum root and rhizome.
7.2 Thin-layer chromatogram (TLC) identification
See Annex A.1 for additional information.
7.3 Rhaponticin identification

See Annex A.2 and Annex C.4 for additional information. If the results of the two methods are

inconsistent, TLC shall prevail.
7.4 Determination of moisture
The testing method specified in the Clause 7.2.1 of ISO 23723 applies.
7.5 Determination of total ash
The testing method specified in the Clause 7.2.3 of ISO 23723 applies.
7.6 Determination of acid-insoluble ash
The testing method specified in the Clause 7.2.3 of ISO 23723 applies.
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ISO/DIS 5228:2022(E)
7.7 Determination of extractives
The testing method specified in the Clause 7.2.5 of ISO 23723 applies.
7.8 Determination of heavy metals
The testing method specified in ISO 18664 applies.
7.9 Determination of pesticide residues
The testing method specified in ISO 22258 applies.
7.10 Determination of marker compounds
See Annex C for additional information.
8 Test report
For each test method, the test report shall specify the following:
a) all information necessary for the complete identification of the sample;
b) the sampling method used;
c) the test method used, with reference to this international standard;
d) the test result(s) obtained;

e) all operating details not specified in this international standard, or regarded as optional, together

with details of any incidents which may have influenced the test result(s);
f) any unusual features (anomalies) observed during the test;
g) the date of the test.
9 Packaging, storage and transportation

The packaging and transportation shall not transmit any odour or flavour to the product and shall not

contain substances that may damage the product or constitute a health risk. The packaging shall be

strong enough to withstand normal handling and transportation.
The storage condition specified in the Clause 5.2.1 of ISO 22217 shall apply.

The products shall be protected from light, moisture, pollution, and foreign substances entry during

long-distance delivery. Carriers should be well ventilated to keep dry and moisture-proof.

10 Marking and labelling

See the method specified in ISO 21371. The following items shall be marked or labelled on the packages:

a) all quality features indicated in Clause 5, determined in accordance with methods specified in

Clause 7;
b) gross weight and net weight of the package;
c) country of origin and province/state of the products;
d) date of production and expiry date of the products;
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ISO/DIS 5228:2022(E)
e) storage method;
f) any items required by regulatory bodies of the destination country.
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ISO/DIS 5228:2022(E)
Annex A
(informative)
Thin-layer chromatogram (TLC) identification
A.1 TLC identification of Rheum root and rhizome

a) Macerate 0,1 g of the powder in 20 ml of methanol for 1 hour, and gravity filter. Evaporate 5 ml of

the filtrate to dryness, dissolve the residue in 10 ml of water, add 1 ml of hydrochloric acid, heat

under reflux on a water bath for 30 minutes and cool immediately. Extract by shaking with 20 ml

of ether twice, combine the ether extracts, evaporate to dryness and dissolve the residue in 1 ml of

chloroform as the test solution;

b) Prepare a solution of Rheum root and rhizome reference drug in the same manner as the reference

drug solution;

c) Dissolve rhein CRS in methanol to produce a solution containing 1 mg per ml as the reference

solution;
d) Use the upper layer of a mixture
...