ISO 23965:2022

Traditional Chinese medicine - Bupleurum chinense, Bupleurum scorzonerifolium and Bupleurum falcatum root

This document specifies minimum requirements and test methods for Bupleurum chinense, Bupleurum scorzonerifolium and Bupleurum falcatum root. This document applies to Bupleurum chinense, Bupleurum scorzonerifolium and Bupleurum falcatum root that are sold and used as natural medicines in international trade, including Chinese materia medica (whole medicinal materials) and decoction pieces.

Médecine traditionnelle chinoise — Racine de Bupleurum chinense, Bupleurum scorzonerifolium et Bupleurum falcatum

General Information

• Status: Published
• Publication Date: 29-Mar-2022
• ICS: 11.120.10 - Medicaments
• Technical Committee: ISO/TC 249 - Traditional Chinese medicine
• Drafting Committee: ISO/TC 249/WG 1 - Quality and safety of raw materials and traditional processing
• Current Stage: 6060 - International Standard published
• Start Date: 30-Mar-2022
• Due Date: 18-Mar-2022
• Completion Date: 30-Mar-2022

Overview

ISO 23965:2022 - Traditional Chinese medicine - Bupleurum chinense, Bupleurum scorzonerifolium and Bupleurum falcatum root defines minimum quality requirements and test methods for Bupleuri Radix used as natural medicines and decoction pieces in international trade. The standard covers macroscopic description, sampling, laboratory testing and requirements for safety-relevant contaminants to support consistent, traceable quality of Bupleurum chinense, B. scorzonerifolium and B. falcatum root in global supply chains.

Key topics and technical requirements

• Scope and description: Identification and morphological characteristics of the three Bupleurum species (root shape, size ranges, surface and fracture features).
• Physical and chemical limits:
  • Moisture: mass fraction ≤ 10.0%
  • Total ash: mass fraction ≤ 9.0%
  • Acid‑insoluble ash: mass fraction ≤ 3.5%
  • Ethanol‑soluble extractives: mass fraction ≥ 11.0%
• Chemical markers and identity testing:
  • TLC identification (thin‑layer chromatography) for marker compounds such as saikosaponin a (matching spot position and colour with reference).
  • Determination of marker compounds including saikosaponin a and d.
• Contaminants and safety testing:
  • Heavy metals: arsenic, mercury, lead, cadmium (determination methods referenced).
  • Pesticide residues: e.g., benzene hexachloride (BHC), DDT, pentachloronitrobenzene.
  • Sulfur dioxide residues determination.
  • Identification of toxic adulterants (e.g., Bupleurum longiradiatum root fingerprinting).
• Test methods and procedures: macroscopic ID, moisture, ash, ethanol‑soluble extractives, TLC, marker compound assay, heavy metals, pesticides, sulfur dioxide; plus sampling, test reporting, packaging, storage and labelling requirements.
• Informative annexes: detailed guidance for test procedures and national/regional value variations.

Practical applications - who uses ISO 23965:2022

• Manufacturers and processors of TCM materia medica and decoction pieces to ensure compliance and product consistency.
• Quality control and analytical laboratories performing identity and contaminant testing.
• Importers/exporters and traders in herbal products to meet international trade requirements and reduce risk of adulteration.
• Regulators and pharmacopoeia committees aligning national requirements with an international baseline.
• Clinical researchers and formulators needing validated reference methods and marker compound data for Bupleuri Radix.

Related standards

ISO 23965 cites several normative references for specific methods and requirements, including:

• ISO 18664 (heavy metals in TCM),
• ISO 21371 (labelling for oral/topical TCM products),
• ISO 22258 (pesticide residues by GC),
• ISO 22590 (sulfur dioxide determination),
• ISO 23723 (general requirements for herbal raw material and materia medica).

Keywords: ISO 23965:2022, Bupleurum chinense, Bupleuri Radix, traditional Chinese medicine, TCM herb testing, saikosaponin, TLC identification, herbal quality control, heavy metals, pesticide residues.

Standards Content (Sample)

INTERNATIONAL ISO
STANDARD 23965
First edition
2022-03
Traditional Chinese medicine —
Bupleurum chinense, Bupleurum
scorzonerifolium and Bupleurum
falcatum root
Médecine traditionnelle chinoise — Racine de Bupleurum chinense,
Bupleurum scorzonerifolium et Bupleurum falcatum
Reference number
© ISO 2022
All rights reserved. Unless otherwise specified, or required in the context of its implementation, no part of this publication may
be reproduced or utilized otherwise in any form or by any means, electronic or mechanical, including photocopying, or posting on
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or ISO’s member body in the country of the requester.
ISO copyright office
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Published in Switzerland
ii
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 Macroscopic characteristics . 4
5.2.1 Bupleurum chinense . 4
5.2.2 Bupleurum scorzonerifolium . 4
5.2.3 Bupleurum falcatum . 4
5.3 Moisture . 4
5.4 Total ash . 4
5.5 Acid-insoluble ash . 4
5.6 Thin-layer chromatogram (TLC) identification . 5
5.7 Toxic adulterants of Bupleurum longiradiatum root . 5
5.8 Ethanol-soluble extractives . 5
5.9 Content of marker compounds . 5
5.10 Heavy metals . 5
5.11 Pesticide residues . 5
5.12 Sulfur dioxide residues . . 5
6 Sampling . 5
7 Test methods . 5
7.1 Macroscopic identification . 5
7.2 Determination of moisture content . 5
7.3 Determination of total ash and acid-insoluble ash content . 5
7.4 Determination of ethanol-soluble extractives content . 5
7.5 Thin-layer chromatogram (TLC) identification . 6
7.6 Identification of the toxic adulterants of Bupleurum longiradiatum root . 6
7.7 Determination of marker compound(s). 6
7.8 Determination of heavy metals . 6
7.9 Determination of pesticide residues . 6
7.10 Determination of sulfur dioxide residues . 6
8 Test report . 6
9 Packaging, storage and transportation . 6
10 Marking and labelling . 7
Annex A (informative) Determination of moisture content . 8
Annex B (informative) Determination of total ash and acid-insoluble ash content .9
Annex C (informative) Determination of ethanol-soluble extractives content .10
Annex D (informative) Thin-layer chromatogram (TLC) identification .11
Annex E (informative) Identification of the toxic adulterants of Bupleurum longiradiatum
root .13
Annex F (informative) Determination of marker compounds .16
Annex G (informative) National and regional requirements for Bupleurum chinense,
Bupleurum scorzonerifolium and Bupleurum falcatum root .19
iii
Foreword
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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
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iv
Introduction
Bupleurum chinense, Bupleurum scorzonerifolium and Bupleurum falcatum root, namely Bupleuri Radix,
are well-known traditional Chinese materia medica which have liver-protection, antipyretic and anti-
inflammatory qualities. Bupleuri Radix is sold in many countries as medicinal materials or decoction
pieces. Bupleuri Radix originates from cross-pollinating plants, which can easily lead to a hybrid of
their provenance, in turn resulting in mixed sources of herbs. This can lead to difficulties in herb
authenticity, as well as variations in herb efficacy and safety. Bupleuri Radix is included in several
pharmacopeia, but with different plant origins. Thus, there is a need to standardize the quality to
bring benefits to consumers and the enterprises involved in the processing, management and trade of
Bupleurum chinense, Bupleurum scorzonerifolium and Bupleurum falcatum root.
As national implementation can differ, national standards bodies can modify the values given in 5.3, 5.4,
5.5 and 5.8 in their national standards. Examples of national and regional values are given in Annex G.
v
INTERNATIONAL STANDARD ISO 23965:2022(E)
Traditional Chinese medicine — Bupleurum chinense,
Bupleurum scorzonerifolium and Bupleurum falcatum root
1 Scope
This document specifies minimum requirements and test methods for Bupleurum chinense, Bupleurum
scorzonerifolium and Bupleurum falcatum root.
This document applies to Bupleurum chinense, Bupleurum scorzonerifolium and Bupleurum falcatum root
that are sold and used as natural medicines in international trade, including Chinese materia medica
(whole medicinal materials) and decoction pieces.
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 22258, Traditional Chinese medicine — Determination of pesticide residues in natural products by gas
chromatography
ISO 22590, Traditional Chinese medicine — Determination of sulfur dioxide in natural products by titration
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 terminology 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
root mass
average mass of final samples of root
Note 1 to entry: Root mass is measured in grams.
3.2
root length
largest distance from the bottom of the tap root to the stem scar
Note 1 to entry: Root length is measured in centimetres.
3.3
batch
samples collected from the same particular place at the same time
3.4
sample
representative material taken from a product, or part of a product
[SOURCE: ISO 11596:2021, 3.1.4, modified — Definition revised and note to entry removed.]
3.5
final sample
sample for the test required in this document
[SOURCE: ISO 21316:2019, 3.3, modified — Note to entry removed.]
4 Description
The structures of Bupleurum chinense DC., Bupleurum scorzonerifolium Willd., Bupleurum falcatum L.
and the dried root are shown in Figure 1.
a) Plant of Bupleurum chinense DC. b) Bupleurum chinense root

c) Plant of Bupleurum scorzonerifolium Willd. d) Bupleurum scorzonerifolium root

e) Plant of Bupleurum falcatum L. f) Bupleurum falcatum root
Key
1 flowering branch 5 cross-section of fruit
2 flower 6 stamen
3 inflorescence 7 sepal
4 fruit
Figure 1 — Structures of Bupleurum chinense, Bupleurum scorzonerifolium and
Bupleurum falcatum root
5 Requirements
5.1 General characteristics
The following requirements shall be met before sampling:
a) Bupleurum chinense, Bupleurum scorzonerifolium and Bupleurum falcatum root materials shall be
clean and dry.
b) The presence of living insects, mouldy root and external contaminants which are visible to the
naked eye shall not be permitted.
c) The shape of the root shall be cylindrical or long and conical.
d) The root mass and the number of the residue shall be measured by electronic balance and counting.
e) The outer surface shall be brown with intermittent longitudinal wrinkles.
5.2 Macroscopic characteristics
5.2.1 Bupleurum chinense
The whole root is cylindrical or elongated conical, branched in the lower part, 6 cm to 15 cm in length
and 0,3 cm to 0,8 cm in diameter. The upper part consists of a bulgy root crown, usually composed
of 3 to 15 stem bases as well as the fibrous remnants of the leaf bases. The outer surface is blackish-
brown or light brown, marked with longitudinal wrinkles and showing rootlet scars and lenticel-like
protuberances. The texture is hard and compact, and difficult to break. The fracture shows concentric
fibrous rings in the wood; the bark is thin, light brown or orange-brown, while the wood is whitish-
yellow.
5.2.2 Bupleurum scorzonerifolium
The whole root is thinner than that of Bupleurum chinense, elongated conical, usually unbranched or
very slightly branched in the lower part, up to 15 cm long and 0,3 cm to 0,5 cm in diameter. The root
crown bears numerous fibres from the bases of wilted leaves in a brush-like shape. The outer surface
is blackish-brown or reddish-brown and shows numerous annular striations near the root crown. The
texture is slightly soft and the root breaks easily. The fracture is even and non-fibrous.
5.2.3 Bupleurum falcatum
The whole root is a long conical or column shape, single or branched, 10 cm to 15 cm in length and
0,5 cm to 1,5 cm in diameter. The upper part is thick and the lower part thin. The apex has numerous
hairy fibres from withered leaves. The external surface is pale brown to brown with deep wrinkles.
The texture is easily broken and the fractured surface is somewhat fibrous.
5.3 Moisture
The mass fraction of moisture should not be more than 10,0 %.
5.4 Total ash
The mass fraction of ash should not be more than 9,0 %.
5.5 Acid-insoluble ash
The mass fraction of acid insoluble ash should not be more than 3,5 %.
5.6 Thin-layer chromatogram (TLC) identification
The identification of marker compound, such as saikosaponin a, with TLC shall present the spots or
bands obtained from the test and reference drug solution in the same position with the same colour.
5.7 Toxic adulterants of Bupleurum longiradiatum root
The characteristic fingerprint for the toxic adulterants of Bupleurum longiradiatum root should be
determined.
5.8 Ethanol-soluble extractives
The mass fraction of ethanol-soluble extracts should not be less than 11,0 %.
5.9 Content of marker compounds
The content of marker compounds such as saikosaponin a and d shall be determined.
5.10 Heavy metals
The content of heavy metals such as arsenic, mercury, lead and cadmium shall be determined.
5.11 Pesticide residues
The content of pesticide residues such as benzene hexachloride, DDT and pentachloronitrobenzene
shall be determined.
5.12 Sulfur dioxide residues
The content of sulfur dioxide residues should be determined.
6 Sampling
Sampling of Bupleurum chinense, Bupleurum scorzonerifolium and Bupleurum falcatum root shall be
carried out in accordance with ISO 23723.
7 Test methods
7.1 Macroscopic identification
Samples of not less than 500 g are taken from each batch randomly and observed with the naked eye,
smelled and tasted.
7.2 Determination of moisture content
See Annex A for additional information.
7.3 Determination of total ash and acid-insoluble ash content
See Annex B for additional information.
7.4 Determination of ethanol-soluble extractives content
See Annex C for additional information.
ISO 2
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