appendices - cmro.gov.hk

Appendices

Appendix I Sampling

Appendix I: Sampling

The sampling here refers to the method used for selecting CMM sample for examination. As the

representativeness of sampling directly affects the conclusions of the examination, recommended sampling

procedures are detailed in the following paragraphs for reference.

(1) Before sampling, it is necessary to inspect each container or package to see whether the name, source,

specification and packaging of CMM are correct. Examine the intactness, cleanliness and any water

trace of the package. Check any contamination by moulds or of foreign matter, and make detailed notes.

Any abnormal package should be sampled separately.

(2) The following are the general requirements for random CMM sampling in a consignment –

(a) For packages less than five : sample every package

(b) For packages of 5–100 : sample five packages

(c) For packages of 100–1000 : sample 5%

(d) For packages over 1000 : sample 50 packages plus 1% of those in excess of 1000

(e) For precious CMM : sample every package regardless of the number

(3) Take CMM by portions from different parts of the selected packages and mix them into one pooled

sample. For small amount of material, the quantity of the sample taken should not be less than three

times of the amount required for tests; for large amount of material, the quantity of the sample taken is

recommended as follows –

(a) Common CMM : 100–500 g

(b) Powdered CMM : 25 g

(c) Precious CMM : 5–10 g

(4) One third of the samples are used for analysis; one third for verification and the remaining for retention

for at least a year.

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Appendix II General Quality Control Method

Appendix II: General Quality Control Method

General quality control method includes "Description", "Identification", "Tests", "Extractives" and "Assay".

A scheme for the examination of CMM is outlined in the paragraphs below.

(1) Follow the method for sampling of CMM as set out in Appendix I for CMM examinations.

(2) Whenever necessary, use a reference herb that complies with the requirements listed in the Appendix of

the current edition of Pharmacopoeia of the Peoples’ Republic of China to verify the results of the

examinations.

(3) "Description" refers to the macroscopic and organoleptic characteristics including form, size, colour,

texture, fracture, gross internal structures, odour/smell, taste, and other relevant information of CMM

samples. Safety precautions should be taken in handling the samples.

(a) "Form" refers to the shape of dried CMM samples. In general, it is observed without preliminary

treatment while wrinkled herbs, leaves or flowers can be moistened or softened and spread for the

examination. For some fruits and seeds, the pericarp or seed coat can be softened and removed, if

necessary before the examination of the inner characteristics.

(b) "Size" indicates the length, diameter and thickness of CMM samples, measured by a millimeter

ruler. A few variations from the defined values are acceptable. For fine seeds, arrange 10 seeds

closely in a row on a piece of paper with a millimeter scale, measure and calculate the average

value.

(c) The "colour" of CMM samples is observed in daylight. For a description of a combination of two

colours, the latter is the main colour. For example, in ‘yellowish-brown’, the main colour is brown.

(d) The "surface characters", "texture" and "gross internal structure" (including fracture

characteristics) of CMM samples are observed without preliminary treatment. If the striations of

the fracture surface are difficult to observe, it should be re-examined with a smooth cut surface.

(e) The "odour/smell" of CMM samples can be examined by smelling directly, or after fracturing

and rubbing. When necessary, the examination can be carried out after the samples are moistened

with hot water.

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(f) The "flavour" of CMM samples can be examined by tasting a small amount of sample directly or

by tasting its water extract.

(4) "Identification" refers to the verification of CMM samples by means of microscopic examination of

cross sections and powders, physical and chemical tests and chromatographic analysis.

(a) "Microscopic identification" refers to the observation of the characteristics of structural features,

cells and ergastic substances in section, powder, disintegrated tissue or surface slides of

CMM samples. It is usually carried out by making slides in an appropriate way as detailed in

Appendix III.

(b) "Physicochemical identification" refers to the testing of the representative constituents in the

samples by physical or chemical methods.

(c) "Chromatographic identification" refers to the identification of samples by means of TLC,

HPLC or GC.

(5) "Tests" refers to the qualitative and quantitative detection of heavy metals, pesticide residues, mycotoxins

(aflatoxins), foreign matter, ash, water content and other chemical components in the CMM which should

be monitored.

(6) "Extractives" refers to the soluble contents of a CMM as extracted by water, ethanol or other appropriate

solvents.

(7) "Assay" refers to the quantitative determination of the active ingredients or markers of CMM samples.

(8) Chemical reference substances of high purity should be used. For Assay, the purity should not be less

than 97%.

Appendix II General Quality Control Method

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Appendix III: Microscopic Identification

Microscopic identification is a method using a microscope to identify the characters of structural features,

cells and ergastic substances in section, powder, disintegrated tissue or surface slides of CMM samples. Select

a representative CMM sample and conduct the examination in accordance with the methods described below.

(1) Preparation of cross and/or longitudinal sections slides –Select a representative sample for examination.

After softening, cut the material with a razor blade or a sliding microtome to a thickness of 10–20 µm.

Examine the sample under a microscope after treated with glycerol-acetic acid TS, chloral hydrate TS or

other suitable TS. Embed the material in hard paraffin for cutting, when necessary.

(2) Preparation of powder slides – Spread a small quantity of the powder on a slide, treated with glycerol-

acetic acid TS, chloral hydrate TS, or other suitable TS and conduct the examination.

(3) Preparation of surface slides – After moistening and softening, cut the sample apart or tear out the

epidermis, add suitable TS and conduct the examination.

(4) Maceration and preparation of slides – Cut or slice the sample into small pieces of about 2 mm in

thickness for maceration. Depending on the nature of the material, one of the following three methods is

used –

(a) Potassium hydroxide method – Place the sample in a test tube, and add an adequate quantity of

aqueous potassium hydroxide solution (5%, v/v), then heat until the residue can be easily separated

when pressed with a glass rod. Decant the alkaline solution and wash the residue with water,

transfer a small amount of macerated material onto a slide and tease it out with a needle. Mount in

dilute glycerine and examine under a microscope.

(b) Chromic-nitric acid method – Place the sample in a test tube and add an adequate quantity of

chromic-nitric acid TS, then let stand until the material can be easily separated when pressing with

a glass rod. Decant the acidic solution, wash the residue with water, and prepare the slide as

directed above in (4)(a).

(c) Potassium chlorate method – Place the sample in a test tube, add dilute nitric acid (50%, v/v)

and a small quantity of potassium chlorate; warm gently until the effervescence subsides, then add

small amount of potassium chlorate to maintain a slight effervescence. When the tissue shows a

tendency to disintegrate, break the material with a glass rod. Decant the acidic solution, wash the

macerated material with water and prepare the slide as directed above in (4)(a).

Appendix III Microscopic Identification

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For CMM samples with only a few or scattered woody tissues or with parenchyma tissues, use potassium

hydroxide method. Whereas, for hard material mainly with woody tissues or woody tissues grouped into

bundles, use chromic-nitric acid or potassium chlorate method.

(5) Measurements of sizes of cell and its contents – Under a microscope, use an ocular micrometer to

measure the sizes of cell and cell contents of specimens. Place the ocular micrometer scale in the eyepiece

of the microscope, then calibrate with a stage micrometer. For the calibration, turn the eyepiece and

move the stage micrometer to make the divisions on the two scales parallel and their left "O" lines

coincide, then look for another line which coincides to the right.

The value (in micrometer) of one ocular micrometer division can be calculated on the basis of divisions

of the two micrometer scales between the coinciding lines.

To measure the object, multiply the number of object-measuring divisions of ocular micrometer by the

value (in micrometer) of each division. In general, it is carried out under a high power objective, but a

low power objective would be more convenient to measure the lengths of longer fibres and non-glandular

hairs, etc.

Record the maximal and minimal values (in micrometer), values slightly higher or lower than those

specified in the individual monograph are acceptable.

(6) Histochemical detection of cell walls –

(a) Lignified cell wall – Add 1–2 drops of phloroglucinol TS to the specimen on the slide, allow to

stand for a moment, then add 1 drop of hydrochloric acid. Lignified cell walls are stained red or

purplish-red according to the extent of lignification.

(b) Suberized or cuticular cell wall – Add 1–2 drops of Sudan III TS to the specimen on the slide,

allow it to stand for a few minutes or warm gently. Suberized or cuticular cell walls are stained

orange-red or red.

(c) Cellulose cell wall – Add 1–2 drops of zinc chloride-iodine TS and allow to stand for few minutes;

alternatively, add 1–2 drops of iodine TS, allow to stand for a while, and then add dilute sulphuric

acid (66%, v/v). Cellulose cell walls are stained blue or purple.

(d) Siliceous cell wall – Add 1–2 drops of sulphuric acid, no change is observed.


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(7) Histochemical detection of cell contents –

(a) Starch – Add iodine TS, a blue or purple colour is observed.

(b) Aleurone – (i) Add iodine TS, a brown or yellowish-brown colour is observed; (ii) Add mercuric

nitrate TS, a brick red colour is observed. If the material is oily, render it fat-free by immersing in

and washing with ether or petroleum ether before carrying out the test.

(c) Fatty oil, volatile oil or resin – (i) Add Sudan III TS, an orange-red, red or purplish-red colour is

observed; and (ii) Irrigate the material with ethanol (90%), volatile oils are dissolved in the solvent,

while fatty oils are insoluble (except castor oil and croton oil).

(d) Inulin – Add α-naphthol in ethanol (10%, w/v) and then add sulphuric acid, the crystals of inulin

turn purplish-red and dissolve rapidly.

(e) Mucilage – Add ruthenium red TS, a red colour is observed.

(f) Calcium oxalate crystals – (i) Insoluble in dilute acetic acid (6%, v/v), soluble in dilute

hydrochloric acid (9.5–10.5%, v/v) without effervescence; (ii) Dissolve gradually in dilute sulphuric

acid (50%, v/v), needle crystals of calcium sulphate appear after stand for a moment.

(g) Calcium carbonate (stalactile) – Soluble in dilute hydrochloric acid (9.5–10.5%, v/v) with

effervescence.

(h) Silica – Insoluble in sulphuric acid.

(8) Preparation of test solutions (TS) for microscopic analysis –

(a) Chloral hydrate TS – Dissolve 50.0 g chloral hydrate in a mixture of 15 mL of water and 10 mL

of glycerine.

(b) Cuoxam TS – Add a sufficient amount of water to 0.5 g of copper carbonate and grind in a mortar,

then add 10 mL of strong ammonia solution to dissolve.

(c) Ferric chloride TS – Dissolve 1.0 g of ferric chloride in water and make up to 100 mL.


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(d) Fuchsin glycerine gelatin – Dissolve 1.0 g of animal gelatin in 6 mL of water, then add 7 mL of

glycerine and heat with gentle stirring until well mixed; after filter through a piece of gauze into a

Petri dish, add sufficient amount of basic fuchsin solution (dissolve 0.1 g of basic fuchsin in

600 mL of absolute ethanol and 80 mL of camphor oil), then mix well and allow to solidify.

(e) Glycerol-acetic acid TS – Mix well equal volumes of glycerine, glacial acetic acid and water.

(f) Iodine TS – Use 0.1 M iodine solution directly.

(g) Mercuric nitrate TS – Add 3 mL of fuming nitric acid to 4.5 g of mercury, when the reaction is

completed, dilute with an equal volume of water. Preserved in an amber-coloured glass bottle

with a glass stopper and protected from light.

(h) α-Naphthol TS – To 10.5 mL of a solution of α-naphthol in ethanol (15%, w/v), gently add

6.5 mL of sulphuric acid and mix well, then add 40.5 mL of ethanol and 4 mL of water, mix well.

(i) Phloroglucinol TS – Dissolve 1.0 g of phloroglucinol in 100 mL of ethanol (90%) and then filter.

Preserved in an amber-coloured glass bottle and protected from light.

(j) Ruthenium red TS – Add a sufficient quantity of ruthenium red to 1–2 mL of aqueous sodium

acetate solution (10%, w/v) to make a wine red colour. Freshly prepare the solution.

(k) Sudan III TS – Dissolve 0.01 g of Sudan III in 5 mL of ethanol (90%), then add 5 mL of glycerine

and mix well. Preserved in an amber-coloured glass bottle, use within 2 months.

(l) Tissue-disintegrating solution (chromic-nitric acid TS) – (i) Add 10 mL of nitric acid to

100 mL of water and mix well; and (ii) Dissolve 10 g of chromic trioxide in 100 mL of water. Mix

equal volumes of the above solutions (i) and (ii) prior to use.

(m) Zinc chloride-iodine TS – Dissolve 8.0 g of potassium iodide in 8.5 mL of water and add 2.5 g of

anhydrous zinc chloride, the mixture is then saturated with sufficient quantity of iodine. Preserved

in an amber-coloured glass bottle with a glass stopper.


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Appendix IV(A) Chromatography – Thin-Layer Chromatography (TLC)

Appendix IV(A): Chromatography - Thin-Layer Chromatography (TLC)

TLC is a separation technique in which a stationary phase consisting of an appropriate material is spread in a

uniform thin layer on a support (plate) of glass, plastic or aluminum film. Standard and test solutions are

applied separately on the plate and the components are separated by the developing solvent systems. For the

identification of CMM, separated spots obtained from the test solution are compared with the corresponding

spots obtained from the chemical reference substance(s) in the chromatogram.

(1) Apparatus and materials –

(a) TLC plates – The most commonly used coated plates are silica gel G, silica gel F254

, HPTLC

silica gel F254

, silica gel H and silica gel HF254

. Diatomaceous earth, diatomaceous earth G, aluminum

oxide, aluminum oxide G, microcrystalline cellulose and microcrystalline cellulose F254

etc., can

be used as well. Coated plates with the size of 10�5 cm; 10�10 cm; 10�15 cm; 20�10 cm

or 20� 20 cm are commonly used.

(b) Application devices – Micropipettes, micro-syringes, calibrated capillaries or other suitable

application devices can be used for the proper application of solutions to the plates.

(c) Developing chamber – A tank of size suitable for the plates, with a tightly fitting lid and with a

flat bottom or twin trough is usually used.

(d) Spray reagents – Spray reagent for the detection of spots is specified in the individual monograph.

(e) Ultraviolet (UV) light source – An emitting light source in the UV range is used for the examination

of spots in the chromatogram.

(2) Procedure –

(a) Saturation of the developing chamber – Unless otherwise specified, carry out the chromatography

in a saturated chamber. To achieve saturation, pour sufficient amount of the developing solvents

into the developing chamber, replace the lid and allow it to stand for 15-30 min at room temperature.

If necessary, line the inner walls of the developing chamber with filter paper strips, the lower

edges of the filter papers should be immersed in the developing solvents.

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(b) Applying the standard and test solutions – Apply separately the prescribed volumes of the

standard and test solutions in small portions to obtain spots (less than 3 mm in diameter) or bands

on a line parallel to, and 15 mm from, the lower edge of the plate. Pay attention not to apply spots

or bands less than 15 mm from the sides of the plate and no disturbance of each other should

occur.

(c) Developing a chromatogram – Place the plate in the chromatographic tank after the solvent has

evaporated from the applied solutions, ensuring that the sample line are 5 mm above the surface of

the developing solvents. Then cover the chamber tightly with a lid. Remove the plate from the

chamber when the developing solvents have moved over the distance as prescribed in the individual

monograph. Dry the plate and visualize the chromatogram under visible light and/or UV light as

specified in the individual monograph.

(d) Interpretation of the chromatogram – Compare the principal spots or bands observed from the

test solution with the corresponding spots or bands observed from the standard solutions. For

positive identification, the sample must give spots or bands with chromatographic characteristics,

including the colour and the Rf value, similar to those of the chemical reference substances when

observed under visible light and/or UV light as specified in the individual monograph.

The Rf value is defined as the ratio of the distance from the point of application to the centre of the

spot or band to the distance travelled by the solvent front from the point of application:

Rf

=Distance from the point of application to the centre of the spot or band

Distance travelled by the solvent front from the point of application

Appendix IV(A) Chromatography – Thin-Layer Chromatography (TLC)

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Appendix IV(B) Chromatography –High-Performance Liquid Chromatography (HPLC)

Appendix IV (B): Chromatography – High-Performance Liquid Chromatography (HPLC)

HPLC is a separation technique consisting of a solid stationary phase and a liquid mobile phase. The sample

is injected through an injector and carried into the column by the mobile phase, the components are separated

on the stationary phase and pass through the detector in succession, a chromatogram is recorded.

(1) Preparation of test sample – Powder the CMM sample and pass through a No.2 sieve before analysis.

The quantity of the sample to be powdered should be of at least five times as much as those needed for

the analysis.

(2) General requirements for the apparatus – Set up the stationary and mobile phases of the HPLC as

specified in the individual monograph. One of the most commonly used packing material is ODS

chemically bonded to silica. Ion exchange resins are used for ion exchange chromatography and porous

silica or polymers are used for size exclusion chromatography. The column is usually maintained at

room temperature and an UV photometer is used as a detector.

The types of stationary phase, mobile phase and detector as specified in the individual monograph should

not be varied. Other parameters may be varied to fit for the performance of the system suitability test

when necessary.

(3) System suitability test – This is to test the suitability of the instruments according to the requirements

prescribed in the individual monograph. By using specified chemical reference substances, adjust the

following parameters to comply with the requirements specified in the individual monographs, i.e. to

match the n value, the repeatability, the R value and the T value of the column.

(a) Number of theoretical plates of the column (n) – The n value is a measure of the column efficiency.

It should not be less than the value specified in the individual monograph. The n value is calculated

by using the following equation –

Where tR

= the retention time of the marker peak in the standard solution or analyte

peak in the test solution,

Wh / 2

= the peak width at half-height of the marker peak in the standard solution

or analyte peak in the test solution.

n = 5.54t

R

W h / 2

( ) 2

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(b) Repeatability – The repeatability is expressed as an estimated RSD of at least five replicate

injections of the standard solution. The RSD of the peak area and the retention time should comply

with the requirements specified in the individual monograph.

(c) Resolution factor (R) – To ensure the accuracy of quantitative analysis, the R value (Fig. 1) of the

analyte peak with the adjacent peak must be larger than 1.5, unless otherwise specified. The R

value is calculated by using the following equation –

R =2( t

R2 - t

R1 )

W1 + W

2

Where tR1

and tR2

= the retention times of two adjacent peaks 1 and 2, respectively,

W1 and W

2= the widths of two adjacent peaks 1 and 2, respectively.

(d) Tailing factor (T) – It is necessary to inspect the T value (Fig. 2) of the peak, especially when

using the peak height method. It should comply with the requirement specified in the individual

monograph. The T value is calculated by using the following equation –

T =W

0.05h

2d1

Where W0.05h

= the peak width at 0.05 of the peak height,

d1

= the distance between the perpendicular line passing through the peak

maximum and the leading edge of the peak at 0.05 of the peak height.

(4) Quantitative procedure – Set up the HPLC system according to the procedures described in the

manufacturer’s manuals. Under the recommended HPLC conditions, establish the calibration curves by

injecting an appropriate amount of standard solutions of a series of concentrations into the HPLC system

for analysis. Identify the analyte peaks in the chromatogram of the test solution by comparing their

retention times with those of the peaks of the chemical reference substances in the chromatogram of the

standard solution obtained under the same HPLC conditions as specified in the procedure. Alternatively,

spike an appropriate amount of chemical reference substance in one of the analyzing samples to verify

the identified peak.

Prepare a 5-point calibration curve by plotting the peak areas of the chemical reference substance against

the corresponding concentrations (in milligram per litre) of the standard solutions. Obtain the slope, y-

intercept, the regression equation and the r2 value from the calibration curve. With the calibration curve


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of the corresponding chemical reference substance, calculate the concentration (in milligram per litre) of

the analyte in the test solution by using the following equation –

Concentration of the analyte =A - I

m

Where A = the peak area of the analyte in the test solution,

I = the y-intercept of the 5-point calibration curve,

m = the slope of the 5-point calibration curve.

Calculate the percentage content of the analyte in the sample by using the following equation –

Content (%) of the analyte =C � V � D

10000 W

Where C = the concentration, in mg/L, of the analyte in the test solution,

D = dilution factor, if any,

V = the final make-up volume, in mL, of the test solution,

W = the weight, in g, of the sample used for the preparation of the test solution.


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Figure 2 Parameters for calculation of tailing factor (T)


Figure 1 Parameters for calculation of resolution factor (R)

tR1 t

R2

W1 W

2

Wh / 2

W0.05h

d1

0.05

h

h

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Appendix IV (C): Chromatography – Gas Chromatography (GC)

GC is a separation technique consisting of gaseous mobile phase and a solid or immobilized liquid stationary

phase. The sample is introduced through the sample injection port, heated and vaporized, and carried into the

column by a carrier gas. Components of the test sample are separated in the column and pass through the

detector in succession, a chromatogram is thus recorded.

(1) General requirements of the apparatus – Unless otherwise specified, nitrogen is employed as a carrier

gas. A packed column or a capillary column may be employed for the test. A packed column is made of

stainless steel or glass. The stationary phase of the column consists of active adsorbent, porous polymer

beads or inert solid supports impregnated with liquid phase. A capillary column is made of glass or

quartz with internal diameter of 0.2 or 0.32 mm. The stationary phase may be coated or chemically

bonded to the inner surface of a column or supporting materials. The temperature of the sample injection

port is usually set at 30–50°C higher than that of the column itself. The volume of solution injected is

usually no more than several micro-litres, the smaller the diameter of the column, the smaller the volume

of solution is injected. Flame-ionization detector, electron-capture detector and mass spectrometric detector

can be used to detect the separated components. The temperature of the detector is higher than that of the

column itself, usually set at about 250–350°C, but never below 100°C. This will avoid condensation of

the moisture.

Parameters including the type of the detector, stationary phase and the supporting material of the column

as specified in the individual monograph should not be varied. Other parameters may be varied to fit for

the performance of the system suitability test. These include the internal diameter and the length of the

column; the commercial brand and size of the supporting materials; the concentration of the liquid

stationary phase; the flow rate of the carrier gas; the temperature of the column; the quantity of the

injecting and the sensitivity of the detector, etc.

(2) System suitability test – The criteria for assessing the suitability of the system are the same as those set

out in Appendix IV(B).

(3) Procedure – The procedure is the same as those set out in Appendix IV(B). Pay special attention to the

effect of the change in room temperature and the injection time.

Appendix IV(C) Chromatography – Gas Chromatography (GC)

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Appendix V Determination of Heavy Metals

Appendix V: Determination of Heavy Metals

Heavy metals refer to the heavy metals and their respective compounds arising from external contamination,

and are absorbed and accumulated in CMM. Arsenic (As), cadmium (Cd), lead (Pb) and mercury (Hg) are

those heavy metals with relatively high toxicity to human beings.

Method –

(1) Analysis of heavy metals – The analytical procedures must be validated and satisfy with all of the

following criteria –

(a) the selected method is suitable for the analysis of the targeted heavy metals;

(b) the limits of detection and quantification are determined for each targeted heavy metal;

(c) the limit of quantification for each targeted heavy metal is set at 0.05 mg/kg;

(d) the recovery for each targeted heavy metal is between 75 and 125%;

(e) the repeatability of the method is less than 15% RSD; and

(f) a linear response is obtained from the analytical detector within the calibration range.

(2) Reagents – All reagents used should be of analytical grade or equivalent and free from any contaminant

which may interfere with the analysis.

(3) Apparatus – Before using the laboratory wares which have been in contact with the samples, the standard

and test solutions, clean them with dilute acids and then rinse them with distilled and de-ionized water.

(4) Preparation of test sample – Take a representative CMM sample and cut it into pieces, if necessary,

before grinding. Powder the sample before the analysis. Whenever possible, the quantity of sample to be

powdered should be of at least five times as much as those needed for the analysis.

(5) Procedure – The following procedures are applicable for the quantitative detection of As, Cd, Pb and Hg

contents in CMM samples. It may have to modify the procedures for the analysis of some samples.

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(a) Microwave assisted acid digestion – Weigh accurately 0.5 g of the sample in a PTFE microwave

digestion vessel, add 7.5 mL of nitric acid. Allow the vessel to stand a while until the reaction

ceases, then seal all vessels properly and place the completed vessel modules in the turntable of

the microwave unit. Start the digestion programme, the selection of low-pressure or high-pressure

microwave assisted acid digestion depends on the type of microwave digestion vessel available in

the individual laboratory. Upon the completion of the programme, cool the mixture and vent the

vessel manually. Transfer the digested sample solution to a 50-mL volumetric flask and make up

to the mark with water, then transfer the solution to a centrifuge tube and centrifuge for 5 min.

Pipette 10 mL of this solution into another 50-mL volumetric flask and make up to the mark with

water. This is the test solution for subsequent instrumental analysis.

(b) Quantitative analysis – Quantify the heavy metals by using ICP-MS with indium (In) as an

internal standard. Internal standards other than In can also be used provided that the method is

properly validated.

Use an ICP-MS system that satisfies with all of the following criteria –

• a resolution better than or equal to 0.7 amu at 10% peak height;

• a mass range from at least 6 to 240 amu and a mass accuracy of ±0.05 amu; and

• a data system that allows correction for isobaric interferences and with an application of internal

standard technique.

Prepare at least four standard solutions in dilute nitric acid (3%, v/v) containing all the targeted

heavy metals at concentrations suitable for plotting calibration curves.

Note: The concentration of the internal standard in the test solution should be same as those in

the standard solutions.

The suggested operation parameters are as follows –

Nebulizer Gas Flow : ~ 0.9 L/min

Auxiliary Gas Flow : ~ 1.2 L/min

Plasma Gas Flow : ~ 15 L/min

Integration Time : 1000 ms

ICP RF Power : 1200 W

Detector : Dual mode

Scan Mode : Peak hopping


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Set up and optimize the ICP-MS according to the manufacturer’s recommended procedures.

Calibrate the instrument with the mixed standard solution. Prior to data collection, flush the system

with rinse blank until the signal level returns to the calibration blank level. Where appropriate,

several isotopes of an element may be monitored with appropriate signal correction to counter-

check the presence of spectral interferences. The isotopes recommended for monitoring the heavy

metals are listed in Table 1. The calculation for As, Cd and Hg contents should be based on the

signals of isotopes of 75 m/z, 114 m/z and 202 m/z, respectively. For Pb, the calculation should be

based on the summation of the signal of the isotopes 206 m/z, 207 m/z, and 208 m/z.

Table 1 Recommended isotopes for monitoring the heavy metals

Heavy Metal Isotope for Monitoring (m/z)

Arsenic 75

Cadmium 111, 114

Lead 206, 207, 208

Mercury 200, 202

Limits – The amount of heavy metals in CMM samples should comply with the limits listed in Table 2 below,

unless otherwise specified.

Table 2 The recommended limits of heavy metals in CMM samples

Heavy Metal Limit (Not more than)

Arsenic 2.0 mg/kg

Cadmium 0.3 mg/kg

Lead 5.0 mg/kg

Mercury 0.2 mg/kg


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Appendix VI Determination of Pesticide Residues

Appendix VI: Determination of Pesticide Residues

Pesticide is a synthetic chemical, a natural or biological substance, or a mixture thereof, used for prevention,

termination and/or control of diseases, pests, grass or other living things which are hazardous to agriculture

and forestry; or for regulation of the growth of plants and pests in an intended way.

The targeted pesticides for the analysis of pesticide residues in CMM are listed as follows –

(a) Aldrin and Dieldrin (sum of)

(b) Chlordane (sum of cis-, trans- and oxychlordane)

(c) Dichlorodiphenyltrichloroethane (DDT) [sum of p,p’-DDT, o,p’-DDT,

p,p’-dichlorodiphenyldichloroethylene (p,p’-DDE) and

p,p’-dichloro-2,2-bis(4-chlorophenyl)ethane (p,p’-TDE)]

(d) Endrin

(e) Heptachlor (sum of heptachlor and heptachlor epoxide)

(f) Hexachlorobenzene

(g) Hexachlorocyclohexane isomers (α-, β- and δ-hexachlorocyclohexane)

(h) Lindane (γ-hexachlorocyclohexane)

(i) Quintozene (sum of quintozene, pentachloroaniline and methyl pentachlorophenyl sulphide)

Method –

(1) Analysis of pesticide residues – The analytical procedures must be validated and satisfy with all of the

following criteria –

(a) the selected method is suitable for the analysis of the targeted pesticides;

(b) the limits of detection and quantification are determined for each targeted pesticide;

(c) the limit of quantification for each targeted pesticide is 0.02 mg/kg. Except for cis-chlordane,

trans-chlordane and oxychlordane, each of which is set at 0.01 mg/kg;

(d) the recovery for each targeted pesticide is between 70 and 120%;



A - 18

(2) Reagents – All reagents used should be of analytical grade or equivalent and free from any contaminant

which may interfere with the analysis. Suitable blank tests should be conducted to demonstrate no

occurrence of contamination of the pesticide residues.

(3) Apparatus – All apparatus to be used should be thoroughly cleaned to ensure that they are free from any

pesticides. Soak the apparatus in a solution of phosphate-free detergent for at least 16 h, then rinse them

with a large quantity of distilled water and wash them with acetone.


before grinding. Powder the sample before the analysis. Whenever possible, the quantity of the sample to

be powdered should be of at least five times as much as those needed for the analysis.

(5) Procedure – The following procedures are applicable for the quantitative detection of pesticide residues

in CMM samples. It may have to modify the procedures for the analysis of some samples. Wherever

possible, it is necessary to use a second capillary column with different polarities and/or MS to confirm

the analytical results.

(a) Extraction – Weigh accurately 10.0 g of the blended sample powder, add about 4.0 g of anhydrous

sodium sulphate and about 100 mL of ethyl acetate. Sonicate in pulse mode by using an ultrasonic

processor for 3 min. Allow the solids to settle and then filter the supernatant solution and collect

the filtrate. Repeat the extraction twice each with 50 mL of ethyl acetate. Combine the filtrates

and the washings and then evaporate to near dryness in a rotary evaporator at about 35°C. Dissolve

the residue in 10 mL of a mixture of dichloromethane and cyclohexane (1:1, v/v) (Solution A).

(b) Clean-up –

(i) Gel permeation chromatography – The chromatographic procedure may be carried out by

using –

• a Bio-beads S-X3 glass column, 60 g in weight and 43 cm in length, or equivalent; and

• a mixture of dichloromethane and cyclohexane (1:1, v/v) as the mobile phase.

Performance of the column – Inject a solution containing corn oil (about 25 mg/mL), bis(2-

ethylhexyl)phthalate (about 1 mg/mL), methoxychlor (about 0.2 mg/mL) and perylene (about

0.02 mg/mL) and proceed with the chromatography. The column is not suitable unless the

resolution of any adjacent peaks is ≥ 0.85. If necessary, calibrate the column using a solution

containing the pesticides [at a suitable concentration and in a mixture of dichloromethane

and cyclohexane (1:1, v/v)] with the lowest molecular weight (for example pentachloroaniline)


A - 19

and that with the highest molecular weight (for example oxychlordane). Determine which

fractions of the eluate contain the target pesticides.

Purification of the test solution – To 10 mL of solution A, add about 1.0 g of anhydrous

sodium sulphate, centrifuge the mixture and get the supernatant layer. Inject an appropriate

volume of the extract and proceed with the chromatography. Collect the fraction as determined

above. Concentrate the solution in a rotary evaporator on a water bath at a temperature below

35°C until the solvent has almost completely evaporated. Then dissolve the residue in 1 mL

of hexane (Solution B).

(ii) Solid phase extraction – The chromatographic procedure may be carried out by using –

• a florisil solid phase extraction column, 75–150 µm in diameter and 1000 mg in weight,

or equivalent; and

• a solution of diethyl ether in hexane (15%, v/v) as the eluting solvent.

If necessary, calibrate the column by using a solution in hexane containing suitable

concentrations of the targeted pesticides. Determine the fractions of the targeted pesticides

from the eluate.

Pack about 10 mm of anhydrous sodium sulphate on the top of the florisil column. Condition

the column with about 5 mL of hexane. Transfer quantitatively solution B onto the florisil

column and proceed with the chromatography. Collect the eluate (Solution C).

(c) Quantitative and qualitative analysis – Examined by GC using 2,4,5,6-tetrachloro-m-xylene as

an internal standard. Another internal standard may be needed if interferences occur.

Use the gas chromatograph that satisfies with all of the following criteria –

• the R value of any analyte peak with the adjacent peak: > 1.5;

• the n value: ≥ 100000 for the peak of α-hexachlorocyclohexane; and

• the RSD of the peak area: ≤ 5%.

Solution (1): Prepare at least five standard solutions in isooctane containing 2,4,5,6-

tetrachloro-m-xylene and all the targeted pesticides at concentrations suitable for plotting

calibration curves.


A - 20

Solution (2): Concentrate solution C in a stream of nitrogen to almost dryness and dilute to

1 mL with isooctane containing 2,4,5,6-tetrachloro-m-xylene as an internal standard [Notes

1 and 2].

Note 1: The concentration of the internal standard in the test solution should be same as

those in the standard solutions.

Note 2: The sulphuric acid treatment in combination with copper powder treatment may

prove useful to remove certain matrix interference arisen from the sample matrix. However,

this treatment will destroy or remove certain targeted pesticides such as aldrin, dieldrin,

endrin, heptachlor epoxide, methyl pentachlorophenyl sulphide and pentachloroaniline.

The chromatographic procedure may be carried out by using –

• a capillary column (0.25 mm � 30 m) of which the internal wall is covered with (14%-

cyanopropylphenyl)-methylpolysiloxan in a layer about 0.25 µm thick;

• a second capillary column of different polarities (0.25 mm � 30 m) of which the internal

wall is covered with (5%-phenyl)-methylpolysiloxane in a layer about 0.25 µm thick;

• nitrogen as the carrier gas;

• an electron-capture detector; and

• a device allowing split/splitless injection. After maintaining the temperature of the column

at 100ºC for 2 min, raise it to 165ºC at a rate of 10ºC/min and maintain at this temperature

for 10 min. Raise the temperature to 230ºC at a rate of 3ºC/min and afterward to 280ºC at

a rate of 15ºC/min, then maintain at this temperature for 10 min. Maintain the temperature

of the injector port at 210ºC and the temperature of the detector at 300ºC.

In the prescribed conditions, inject 1 µL or other appropriate volume of each solution and

record the chromatograms. The reference RRTs of the targeted pesticides obtained are listed

in Table 1. Calculate the content of each targeted pesticide from its peak area and concentration.


A - 21

The results obtained can be confirmed by GC-MS.

The chromatographic procedure may be carried out by using –

• a capillary column (0.25 mm � 30 m) of which the internal wall is covered with (35%-

phenyl)-methylpolysiloxane in a layer about 0.25 µm thick;

• helium as the carrier gas;

• a mass selective detector capable of operating in a scan mode or selective ion mode (m/z

of the monitoring ions for the targeted pesticides are listed in Table 2 for reference); and

• a device allowing split/splitless injection. Maintain the temperature of the column at

100ºC for 2 min, raise to 160ºC at a rate of 15ºC/min and afterward to 270ºC at a rate of

5ºC/min, then maintain at this temperature for 10 min. Maintain the temperature of the

injector port at 250ºC and the temperature of the ion source at 230ºC.

In the prescribed conditions, inject 1 µL or other appropriate volume of each solution and

record the chromatograms. The reference RRTs of the targeted pesticides obtained are listed

in Table 2.


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Table 1 The reference RRTs of the targeted pesticides obtained by GC

RRT

Pesticide [column used: 0.25 mm � 30 m,

(14%-cyanopropylphenyl)-methylpolysiloxane of 0.25-µm thick]

Hexachlorobenzene 1.24

α-Hexachlorocyclohexane 1.55

Quintozene 1.64

Lindane 1.83

Heptachlor 1.94

Pentachloroaniline 2.01

Aldrin 2.09

Methyl pentachlorophenyl sulphide 2.10

β-Hexachlorocyclohexane 2.32

Oxychlordane 2.41

δ-Hexachlorocyclohexane 2.43

Heptachlor epoxide 2.50

trans-Chlordane 2.67

cis-Chlordane 2.71

p,p’-DDE 2.76

Dieldrin 2.82

Endrin 2.92

o,p’-DDT 2.98

p,p’-TDE 3.15

p,p’-DDT 3.21


A - 23

Table 2 The reference RRTs and the monitoring ions of the targeted pesticides obtained by GC-MS

Pesticide RRTPrimary Ion, Secondary Ion,

m/z m/z

Hexachlorobenzene 1.18 284 286, 282

α-Hexachlorocyclohexane 1.22 181 183, 217

Quintozene 1.32 237 249, 214

Lindane 1.35 183 217, 221

β-Hexachlorocyclohexane 1.45 181 183, 217

Heptachlor 1.48 272 274, 270

Pentachloroaniline 1.49 265 267, 263

δ-Hexachlorocyclohexane 1.55 181 183, 217

Aldrin 1.58 263 265, 261

Methyl pentachlorophenyl sulphide 1.63 296 246, 263

Oxychlordane 1.74 185 387, 237

Heptachlor epoxide 1.79 353 355, 351

trans-Chlordane 1.87 373 375, 377, 371

cis-Chlordane 1.91 373 375, 377, 371

p,p’-DDE 2.00 246 316, 248

Dieldrin 2.03 263 261, 265

Endrin 2.14 263 265, 281

o,p’-DDT 2.17 235 237, 165

p,p’-TDE 2.20 235 237, 165

p,p’-DDT 2.30 235 237, 165


A - 24

Limits – The amount of pesticide residues in CMM samples should comply with the limits listed in Table 3

below.

Table 3 The recommended limits of pesticide residues in CMM samples

Pesticide Limit (Not more than)

Aldrin and Dieldrin (sum of) 0.05 mg/kg

Chlordane (sum of cis-, trans- and oxychlordane) 0.05 mg/kg

DDT (sum of p,p’-DDT, o,p’-DDT, p,p’-DDE and p,p’-TDE) 1.0 mg/kg

Endrin 0.05 mg/kg

Heptachlor (sum of heptachlor and heptachlor epoxide) 0.05 mg/kg

Hexachlorobenzene 0.1 mg/kg

Hexachlorocyclohexane isomers (α-, β- and δ- hexachlorocyclohexane) 0.3 mg/kg

Lindane (γ-hexachlorocyclohexane) 0.6 mg/kg

Quintozene (sum of quintozene, pentachloroaniline and methyl1.0 mg/kg

pentachlorophenyl sulphide)


A - 25

Appendix VII Determination of Mycotoxins (Aflatoxins)

Appendix VII: Determination of Mycotoxins (Aflatoxins)

Mycotoxins, including aflatoxins, refer to the toxic metabolites generated by molds and/or fungi. Since there

is concern over the contamination of aflatoxins in CMM, a harmonized method is developed to study the

contents of aflatoxins B1, B

2, G

1 and G

2 in CMM.

Methods –

(1) Analysis of aflatoxins – The analytical procedures must be validated and satisfy with all of the following

criteria –

(a) the selected method is suitable for the analysis of the targeted aflatoxins and is not susceptible to

the interference from co-extractives;

(b) the limits of detection and quantification are determined for each aflatoxin;

(c) the limit of quantification for each targeted aflatoxin is set at 0.3 µg/kg;

(d) the recovery for each targeted aflatoxin is between 50 and 120%;



(2) Reagents – All reagents used should be of analytical grade or equivalent. Methanol and acetonitrile used

should be at least of HPLC grade.

(3) Apparatus – All apparatus to be used should be thoroughly cleaned to ensure that they are free from any

aflatoxins. Soak the laboratory wares in a solution of household bleach (10%, v/v) for at least 12 h and

then wash them with distilled water.


before grinding. Powder the sample before the analysis. Whenever possible, the quantity of the sample to

be powdered should be of at least five times as much as those needed for the analysis.

(5) Procedure – The analysis of aflatoxins is based on the detection of the characteristic fluorescence emitted

by aflatoxins B1, B

2, G

1, and G

2 after iodine derivatization and UV excitation. It may have to modify the

procedures for the analysis of some samples.

A - 26

(a) Extraction – Weigh accurately 15.0 g of the blended sample, add 3 g of sodium chloride and

75 mL of a mixture of methanol and water (7:3, v/v). Homogenize the mixture for about 2 min and

centrifuge for about 10 min. Check the pH of the extract. Transfer accurately 15 mL of the

supernatant solution of the centrifuged mixture to an amber bottle and reduce to about 5 mL with

a gentle stream of nitrogen at about 60˚C on a water bath [Note 1]. Make up the resultant solution

to 50 mL with a solvent that is compatible with the performance of the immunoaffinity column as

suggested by the manufacturer [Note 2]. Centrifuge the solution for about 10 min and filter through

a glass-fibre filter paper by suction. Collect the filtrate as the test solution.

Note 1: If the sample absorbs solvent significantly (i.e. the volume of the resultant supernatant is

less than 40 mL), repeat the extraction as directed above. However, use 5.0 g of sodium chloride

and 125 mL of a mixture of methanol and water (7:3, v/v) instead, and transfer 25 mL of the

supernatant solution.

Note 2: When handling samples that produce extracts which might affect the normal functioning

of the immunoaffinity column, seek manufacturer’s advice to ensure the column performance. Pay

particular attention to the suggested pH working range of the column and the solvent used for

diluting the sample extract prior to column clean-up.

(b) Clean-up by immunoaffinity column chromatography –

Chromatographic procedure – It may be carried out by using –

• an immunoaffinity column containing antibodies specific for aflatoxins B1, B

2, G

1 and G

2; and

• methanol as the eluting solution.

Performance test of the column – Pass an appropriate volume of the mixed standard solution

containing aflatoxins B1, B

2, G

1 and G

2 through the column, then follow the procedure as described

below ‘Clean-up of the test solution’, and the recovery of the aflatoxins B1, B

2, G

1 and G

2 should

be at least 90%, 80%, 90% and 60% respectively.

Clean-up of the test solution [Note 3] – Condition the column according to the manufacturer’s

instruction, then pass a selected volume of the test solution through the column at a flow rate of

about 3 mL/min. Wash the column with 10 mL of eluent, as recommended by the manufacturer, at

a flow rate of about 3 mL/min, then wash dry the column by passing 10 mL of air through the

column. Elute the column with 1.5 mL of methanol followed by 10 mL of air. Collect all eluates in

a 2-mL volumetric flask and make up to the mark with water.


A - 27

Note 3: It may have to modify the clean-up procedures for different brands of immunoaffinity

column. Please refer to manufacturer’s instruction.

(c) Quantitative analysis – Use a HPLC system that satisfies with all of the following criteria –

• the R value of any analyte peak with the adjacent peak: > 1.5;

• the n value of any analyte peak: ≥ 7000; and

• the RSD of peak area: ≤ 5%.

Individual aflatoxin standard stock solutions – Determine the concentration of individual

aflatoxin stock standard solution (about 10 mg/L) in a mixture of benzene and acetonitrile (98:2,

v/v) by UV spectroscopy according to the following equation –

Concentration of aflatoxin (mg/L) =A

350 � M

w � 1000

ε

Where A350

= the absorbance of the aflatoxin at a wavelength of maximum absorption

close to 350 nm,

Mw

= the molecular weight of the aflatoxin (Table 1),

ε = the molar absorptivity of the aflatoxin in benzene-acetonitrile solution

(Table 1).

Table 1 The molecular weights (Mw) and molar absorptivities (ε) of the aflatoxins

Aflatoxin Molecular Weight ( Mw) Molar Absorptivity ( ε)

B1

312 19800

B2

314 20900

G1

328 17100

G2

330 18200

Mixed aflatoxins standard solutions – Prepare at least five standard solutions in a mixture of

methanol and water (7:3, v/v) containing all the targeted aflatoxins at concentrations suitable for

plotting calibration curves.

Chromatographic procedure – It may be carried out by using –

• a LC column (4.6 � 250 mm) packed with particles of octadecylsilyl groups modified silica

(5 µm in diameter);


A - 28

• a post-column reactor system with the reaction temperature set at 70˚C and 0.5 mM iodine

solution as post-column derivatization reagent;

• a mixture of distilled water, acetonitrile and methanol (3:1:1, v/v) as the mobile phase; and

• a fluorescence detector: λext

= 360 nm and λem

= 450 nm.

Set the flow rate of the mobile phase (LC column) as 1.0 mL/min and the flow rate of the post-

column derivatization reagent as 0.3 mL/min. Under such conditions, aflatoxins are eluted in the

order of G2, G

1, B

2, and B

1 with retention times of about 13, 16, 18, and 23 min, respectively. If

necessary, adjust the retention times by changing the solvent composition. Calculate the content

of each aflatoxin from its peak area and concentration.

Note: Soak all used laboratory wares in a 10% solution of household bleach overnight before

reuse or disposal.

Limits – The amount of aflatoxin B1 and the total amount of aflatoxins (B

1, B

2, G

1 and G

2) in CMM samples

should comply with the limits listed in Table 2 below.

Table 2 The recommended limits of aflatoxins in CMM samples

Aflatoxin Limit (Not more than)

Aflatoxin B1

5 µg/kg

Aflatoxins (sum of B1, B

2, G

1 and G

2 ) 10 µg/kg


A - 29

Appendix VIII Determination of Foreign Matter

Appendix VIII: Determination of Foreign Matter

Foreign matter is material consisting of any of the following –

(1) The biological origin of which is the same as that specified in the monograph concerned but the appearance

or botanical part is different.

(2) The biological origin of which differs from that specified in the monograph concerned.

(3) Foreign mineral matters such as stones, sand, lumps of soil.

Method and Procedure –

(1) Weigh 100–500 g of CMM sample and spread in a thin layer. Sort the foreign matter into groups either

by visual inspection, using a magnifying lens (5–10�), or with the help of a suitable sieve.

(2) Weigh each group of foreign matter separately, and calculate the percentage of foreign matter in the

weight of CMM sample.

Note 1: In case of close resemblance between the foreign matters and the bulk sample in appearance, use

microscopic, physical or chemical methods to identify the foreign matter.

Note 2: For large-sized sample, cut it off when necessary, so as to examine any signs of spoilt or contamination

by insects and moulds.

Limits – The amount of foreign matter in CMM samples should not be more than the percentage specified in

the individual monograph.

A - 30

Appendix IX Determination of Ash

Appendix IX: Determination of Ash


(1) Total ash –

(a) Pulverize CMM sample, pass through a No.2 sieve and mix well. Accurately weigh 2–3 g (3–5 g

for the determination of acid-insoluble ash) of the powdered sample in a tared crucible (to the

nearest 0.01 g). Ignite the sample gently until completely carbonized, keep it from burning, then

gradually increase the temperature to 500–600°C. Continue the ignition until a constant weight of

carbon-free ash is obtained. Calculate the percentage of total ash in the weight of CMM sample.

(b) If a carbon-free ash cannot be obtained in this way, cool the crucible, and moisten the residue with

hot water or 2 mL of aqueous ammonium nitrate solution (10%, v/v), then dry the residue on a

water bath. Ignite the residue again as directed above until a carbon-free ash is obtained.

(2) Acid-insoluble ash –

To a crucible containing the total ash, add 10 mL of dilute hydrochloric acid (10%, v/v), cover with a

watch glass and gently heat for 10 min on a water bath. Rinse the watch glass with 5 mL of hot water and

add the rinsing to the crucible. Transfer the insoluble matter and rinse the remaining residues from the

crucible onto an ashless filter paper, wash with hot water until the filtrate is free of chlorides. Transfer the

ashless filter paper containing the insoluble matter to the original crucible, dry and ignite to constant

weight. Calculate the percentage of acid-insoluble ash in the weight of CMM sample.

Limits – The amount of total ash and acid-insoluble ash in CMM samples should not be more than the

percentages specified in the individual monograph.

A - 31

Appendix X Determination of Water Content

Appendix X: Determination of Water Content

Method – Toluene distillation method is used for the determination of water content in CMM samples.

(1) Preparation of test sample – Prepare a suitable quantity of test sample by cutting (or by using other

appropriate means) CMM sample into pieces of less than 3 mm in length or diameter. Flowers, seeds or

fruits of length or diameter less than 3 mm, can be used directly for the examination.

(2) Reagent – Toluene used in this method should be saturated with water and distilled.

(3) Apparatus – The apparatus (Fig. 1) consists of a 500-mL round-bottomed flask (A); a graduated receiving

tube (B); and a reflux condenser (C) of about 40 cm in length. The apparatus should be cleaned and dried

in an oven before being used.

(4) Procedure – Weigh accurately a quantity of the sample which is expected to give 2–4 mL of water,

transfer to the flask and add about 200 mL of toluene into it. When necessary, add a few pieces of glass

beads. After assembly of the apparatus, fill in the narrow part of the receiving tube with toluene through

the condenser, then heat the flask gently by using an electric heater or other appropriate means. When the

toluene begins to boil, adjust the temperature to allow the distillation proceed at a rate of 2 drops per

second until the water has been completely distilled. Rinse the inside of the condenser with toluene.

Continue the distillation for five more minutes, then remove the apparatus away from the heat and allow

it to cool to room temperature. Disconnect the apparatus and dislodge any droplets of water that adhere

to the wall of the receiving tube.

Allow the receiving tube to stand a while until the water and toluene are completely separated [Note].

Record the volume of water and calculate the percentage of water content in the weight of CMM sample.

Note: A small amount of methylene blue may be added to form a bluish aqueous layer to facilitate

observation.

Limits – The water content in CMM samples should not be more than the percentage specified in the individual

monograph.

A - 32

Appendix X Determination of Water Content

Figure 1 Apparatus for the determination of water content in CMM samples

A. Round-bottomed flask

B. Graduated receiving tube

C. Reflux condenser

C

B

A

A - 33

Appendix XI Determination of Extractives

Appendix XI: Determination of Extractives


(1) Determination of water-soluble extractives – Pulverize CMM sample, pass through a No.2 sieve and

mix well.

(a) Cold extraction method – Place 4.0 g of the powdered sample, accurately weighed, in a

250–300 mL conical flask with a stopper. Accurately add 100 mL of water, insert the stopper and

extract for 24 h. Shake frequently during the first 6 h, then allow to stand for 18 h. Filter rapidly

through a dry filter. Accurately transfer 20 mL of the filtrate to an evaporating dish, previously

dried to constant weight, and evaporate to almost dryness on a water bath, then dry at 105°C for

3 h. Cool in a desiccator for 30 min, and then weigh immediately and accurately. Calculate the

percentage of water-soluble extractives with reference to the dried CMM sample.

(b) Hot extraction method – Place 2.0–4.0 g of the powdered sample, accurately weighed, in a 100–

250 mL conical flask with a stopper. Accurately add 50–100 mL of water, insert the stopper and

weigh. Allow to stand for 1 h. Attach a reflux condenser to the flask and boil gently for 1 h, then

cool to room temperature and weigh, readjust to the original weight with water. Shake and filter

through a dry filter. Accurately transfer 25 mL of the filtrate to an evaporating dish, previously

dried to constant weight, and evaporate to dryness on a water bath, then dry at 105°C for 3 h. Cool

in a desiccator for 30 min, and then weigh immediately and accurately. Calculate the percentage

of water-soluble extractives with reference to the dried CMM sample.

(2) Determination of ethanol-soluble extractives –

(a) Cold extraction method – Proceed as in section 1(a) except by using ethanol (70%) in lieu of

water as solvent.

(b) Hot extraction method – Proceed as in section 1(b) except by using ethanol (70%) in lieu of

water as solvent.

Limits – The amount of water-soluble extractives and ethanol-soluble extractives in CMM samples should

not be less than the percentages specified in the individual monograph.

A - 34

Appendix XII Gas Chromatographic and High-Performance LiquidChromatographic Fingerprinting

Appendix XII: Gas Chromatographic and High-Performance Liquid Chromatographic

Fingerprinting

GC and HPLC fingerprinting refers to the identification of CMM samples by the examination of the GC and

HPLC chromatograms of their solvent extracts.

GC and HPLC fingerprinting have the merits of high selectivity, high sensitivity, high separation rate and

requiring only a short analytical time with a small amount of sample. In general, one or more chemical markers

and characteristic peaks can be identified in the chromatographic fingerprinting.

Method – Establishment of a chromatographic fingerprinting

(1) Appropriate extraction method and chromatographic conditions should be selected in accordance with

the nature of the active ingredients or markers contained in CMM samples. The chromatographic

fingerprinting should contain as many well resolved peaks as possible in order to provide adequate

information for the identification. The recommended run time for every GC and HPLC chromatographic

fingerprinting is not more than 60 min.

(2) In the chromatographic fingerprinting established, a well resolved peak corresponding to an available

chemical reference substance can be used as a marker peak for the calculation of the RRTs of other peaks

in the same chromatogram. When necessary, more than one marker peak may be chosen.

(3) The RRT of a characteristic peak is calculated based on the retention time of a chosen marker peak by

using the following equation –

RRT =Retention time of the characteristic peak

Retention time of the marker peak

(4) The reference fingerprint chromatogram, marker peak, characteristic peaks and acceptable ranges of a

sample is drawn up based on the test results of at least 10 batches of representative samples examined in

duplicate (i.e. a total of 20 sets of data).

(5) A fingerprint chromatogram of a sample is then established by using the above procedure. For positive

identification, the sample must give all the characteristic peaks with the RRTs falling within the acceptable

range of the corresponding peaks in the reference fingerprint chromatogram as specified in the individual

monograph.

A - 35

Appendix XIII Detection of Aristolochic Acid I

Appendix XIII: Detection of Aristolochic Acid I

Aristolochic Acid I (AAI) is a known nephrotoxin and potential carcinogen that is commonly present in herbs

derived from plants belonging to the genera of Aristolochia and Asarum of the family Aristolochiaceae. In

view of this, the Department of Health announced that importation and sale of Chinese herbs containing AAI

were prohibited starting from 1st June, 2004. There are unclear factors in local Chinese herbal market that may

lead to inappropriate or misuse of the herbs containing Aristolochic Acid. It is suggested that the trader may

apply the following method to detect AAI in the suspected Chinese herbs.

Method

Carry out the method as directed in Appendix IV(B).

Standard solution

Aristolochic acid I standard stock solution, Std-Stock (50 mg/L)

Weigh accurately 5.0 mg of aristolochic acid I CRS and dissolve in 100 mL of methanol.

Aristolochic acid I standard solution for detection

Measure accurately the volume of aristolochic acid I Std-Stock, dilute with methanol to produce solutions of

0.05 and 5 mg/L for aristolochic acid I.

Test solution

Weigh accurately 0.5 g of the powdered sample and put into a 50-mL centrifugal tube, and add 8 mL of

methanol. Sonicate (490 W) the mixture for 30 min. Centrifuge at about 1800 � g for 10 min. Transfer the

supernatant to a 25-mL volumetric flask. Repeat twice. Combine the extracts and make up to the mark with

methanol. Mix and filter through a 0.45-µm RC filter.

Chromatographic system

The liquid chromatograph is equipped with a detector (396 nm) and a column (4.6 � 250 mm) packed with

ODS bonded silica gel (5 µm particle size). The flow rate is about 1.0 mL/min. The mobile phase is a mixture

of acetonitrile and 1% acetic acid (52:48, v/v). The elution time is about 25 minutes.

System suitability requirements

Perform at least five replicate injections each with 10 µL of aristolochic acid I (standard solution for detection,

5 mg/L). The requirements of the system suitability parameters are as follows: the RSD of the peak area of

aristolochic acid I should not be more than 3.0%; the RSD of the retention time of aristolochic acid I peak

should not be more than 2.0%; the column efficiency determined from aristolochic acid I peak should not be

less than 10000 theoretical plates.

A - 36

Appendix XIII Detection of Aristolochic Acid I

Procedure

Separately inject aristolochic acid I (standard solution for detection, 0.05 mg/L) and the test solution (10 µL

each) into the HPLC system and record the chromatograms. Identify aristolochic acid I peak in the chromato-

gram of the test solution by comparing its retention time with that in the chromatogram of aristolochic acid I

(standard solution for detection). The retention times of aristolochic acid I peaks from the two chromatograms

should not differ by more than 2.0%.

Limit

Aristolochic acid I (C17

H11

NO7) should not be detected in CMM samples, unless otherwise specified.

A - 37

Appendix XIV Detection of Aconitine, Hypaconitine and Mesaconitine

Appendix XIV: Detection of Aconitine, Hypaconitine and Mesaconitine

The Aconitum diester alkaloids mainly include aconitine, mesaconitine and hypaconitine. It is commonly

present in herbs derived from plants belonging to the genera of Aconitum of the family Ranunculaceae. The

major toxicity expresses in cardiotoxicity and neurotoxicity. To reduce the toxicity of the Chinese Materia

Medica, an appropriate processing method should be adopted in order to reduce the total content of Aconitum

diester alkaloids. In view of this, Chinese herbs containing aconitine, mesaconitine and hypaconitine should

be examined and the limits of the above compounds should be established.

Method

Carry out the method as directed in Appendix IV(B).

Standard solutions

Aconitine standard stock solution, Std-Stock (1000 mg/L)

Weigh accurately 5.0 mg of aconitine CRS and dissolve in 5 mL of 0.01 M hydrochloric acid.

Hypaconitine standard stock solution, Std-Stock (2000 mg/L)

Weigh accurately 10.0 mg of hypaconitine CRS and dissolve in 5 mL of 0.01 M hydrochloric acid.

Mesaconitine standard stock solution, Std-Stock (1000 mg/L)

Weigh accurately 5.0 mg of mesaconitine CRS and dissolve in 5 mL of 0.01 M hydrochloric acid.

Mixed aconitine, hypaconitine and mesaconitine standard solution for detection

Measure accurately the volume of aconitine, hypaconitine and mesaconitine Std-Stock, mix and dilute with

0.01 M hydrochloric acid to produce a series of solutions of 0.5, 1, 2, 3, 4 mg/L for both aconitine and

mesaconitine, and 5, 10, 20, 30, 40 mg/L for hypaconitine.

Test solution

Weigh accurately 0.5 g of the powdered sample and put into a 10-mL centrifugal tube, then add accurately

5 mL of methanol (50%). Sonicate (490 W) the mixture for 60 min. Centrifuge at about 1800 � g for 5 min.

Filter through a 0.45-µm RC filter.

Chromatographic system

The liquid chromatograph is equipped with a detector (240 nm) and a column (4.6 � 250 mm) packed with

ODS bonded silica gel (5 µm particle size, pH: 1-12). The flow rate is about 1.0 mL/min. Programme the

chromatographic system as follows –

Time Ammonium bicarbonate* AcetonitrileElution

(min) solution (%, v/v) (%, v/v)

0 – 60 70 ➔ 45 30 ➔ 55 linear gradient

A - 38

*Ammonium bicarbonate solution

Dissolve 0.2 g of ammonium bicarbonate in 1 L of water and adjust the pH to 10 with 1 mL of ammonia

solution.

System suitability requirements

Perform at least five replicate injections each with 20 µL of hypaconitine (standard solution for detection,

20 mg/L). The requirements of the system suitability parameters are as follows: the RSD of the peak area of

hypaconitine should not be more than 3.0%; the RSD of the retention time of hypaconitine peak should not be

more than 2.0%; the column efficiency determined from hypaconitine peak should not be less than 30000

theoretical plates.

The R value between hypaconitine peak and the closest peak in the chromatogram of the test solution should

not be less than 1.0.

Calibration curve

Inject a series of the mixed aconitine, hypaconitine and mesaconitine (standard solution for detection, 20 µL

each) into the HPLC system and record the chromatograms. Plot the peak areas of aconitine, hypaconitine and

mesaconitine against the corresponding concentrations of the mixed aconitine, hypaconitine and mesaconitine

(standard solution for detection). Obtain the slopes, y-intercepts and the r2 values from the corresponding 5-

point calibration curves.

Procedure

Inject 20 µL of the test solution into the HPLC system and record the chromatogram. Identify aconitine peak,

hypaconitine peak and mesaconitine peak in the chromatogram of the test solution by comparing their reten-

tion times with those in the chromatogram of the mixed aconitine, hypaconitine and mesaconitine (standard

solution for detection). The retention times of aconitine peaks, hypaconitine peaks and mesaconitine peaks

from the two chromatograms should not differ from their counterparts by more than 2.0%. Measure the peak

areas and calculate the concentrations (in milligram per litre) of aconitine, hypaconitine and mesaconitine in

the test solution, and calculate the percentage contents of aconitine, hypaconitine and mesaconitine in the

sample by using the equations indicated in Appendix IV(B). Calculate the sum of the content.

Limit

The total content of aconitine (C34

H47

NO11

), hypaconitine (C33

H45

NO10

) and mesaconitine (C33

H45

NO11

) in

CMM samples should comply with the limit specified in the individual monograph.

Appendix XIV Detection of Aconitine, Hypaconitine and Mesaconitine

A - 39

Appendix XV Determination of Volatile Oil

Appendix XV: Determination of Volatile Oil

Method – Specified apparatus is used to determine the volatile oil in CMM samples.

(1) Preparation of test sample – Pulverize CMM sample, pass through No.2 or No.3 sieves and mix well,

unless otherwise specified.

(2) Apparatus – The apparatus (Fig. 1) consists of a 1000-mL (500-mL or 2000-mL) round-bottomed flask

(A), a volatile oil determination tube (B) and a reflux condenser (C). All parts are connected via ground

glass joints. The measuring tube of B is graduated in 0.1 mL. The apparatus should be cleaned before use

and all parts of apparatus should be tightly connected to avoid the loss of volatile oil.

Note: The volatile oil determination tube should be set vertically. The connecting point between the side

tube and the graduated tube is at a horizontal level.

(3) Procedure

(a) Method A – This method is used to determine the volatile oils with relative density less than 1.0.

Take a quantity of the powdered sample which is expected to give 0.5 – 1.0 mL of volatile oil,

weigh accurately to the nearest 0.01 g, and put into a round-bottomed flask. Add 300 - 500 mL of

water (or appropriate amount) and a few glass beads, shake and mix well. Connect the round-

bottomed flask to a volatile oil determination tube and then connect the volatile oil determination

tube to a reflux condenser. Add water through the top of reflux condenser until the graduated tube

of volatile oil determination tube is filled and overflows to the round-bottomed flask. Heat the

flask gently until boiling by using an electric heating jacket or other appropriate means. Continue

the gentle boiling for about 5 h until the volume of oil does not increase. Stop heating, allow it to

stand for a while. Open the stopcork at the lower part of volatile oil determination tube and run off

the water layer slowly until the oily layer is 5 mm above the zero mark. Allow to stand for at least

1 h, open the stopcock again, run off the remaining water layer carefully until the oily layer is just

on the zero mark. Record the volume of oil in the graduated tube of volatile oil determination tube

and calculate the percentage of volatile oil in CMM sample.

(b) Method B –This method is used to determine the volatile oils with relative density more than 1.0.

Add 300 mL of water and a few pieces of glass beads into a round-bottomed flask. Connect the

round-bottomed flask to volatile oil determination tube. Add water through the top of volatile oil

A - 40

determination tube until the graduated tube is filled and overflows to the round-bottomed flask.

Add 1 mL of xylene by using a pipette and then connect the reflux condenser to volatile oil

determination tube. Heat the flask until boiling, continue the heating to allow the distillation

proceed at a rate that will keep the middle part of the condenser cold. Stop heating after 30 min,

and allow it to stand for at least 15 min. Record the volume of xylene in the graduated tube of

volatile oil determination tube.

Carry out the procedure as described in Method A beginning at the words "Take a quantity of the

powdered sample". Subtract the volume of xylene previously observed from the volume of oily

layer, the difference in volume is taken to be the content of volatile oil, calculate the percentage of

volatile oil in CMM sample.

Limits – The CMM samples contain not less than the percentage of volatile oil specified in the individual

monograph.


A - 41

C

A

1214

11

811

.5 13.5

0.90.8

7

1.4

Unit : cm

Figure 1 Apparatus for the determination of volatile oil in CMM samples

A. Round-bottomed flask

B. Volatile oil determination tube

C. Reflux condenser

B


A - 42

Appendix XVI Storage

Appendix XVI: Storage

Storage refers to the basic conditions required for storing CMM. Unless otherwise specified, CMM should be

stored in dry and clean containers. Some terms for the general storage conditions are specified as follows –

(1) "Protected from light" refers to the storage of CMM in light resistant containers.

(2) "Well closed container" refers to a container which is able to protect CMM from extraneous matters or

from the loss of its contents.

(3) "Tightly closed container" refers to a container which is able to protect CMM from efflorescence,

deliquescence, volatilization or interference of extraneous matters.

(4) "Tightly sealed container" refers to a container which is tightly sealed with suitable material to protect

CMM against contamination and from permeability of air and moisture.

(5) "Cool and dark place" refers to a dark environment, protected from light, where the temperature is 11–

20°C.

(6) "Cold place" refers to an environment where the temperature is 2–10°C.

A - 43

INDEXES

m~ÖÉ=kçK

fK lÑÑáÅá~ä=k~ãÉë fJN

ffK `ÜáåÉëÉ=k~ãÉë fJO

fffK `ÜáåÉëÉ=mÜçåÉíáÅ=k~ãÉë fJP

fsK pÅáÉåíáÑáÅ=k~ãÉë =fJQ

sK `ÜÉãáÅ~ä=oÉÑÉêÉåÅÉ=pìÄëí~åÅÉë fJR

Indexes

Index I Official Names

lÑÑáÅá~ä=k~ãÉë m~ÖÉ=kçK

`~ìäáë=`äÉã~íáÇáë=^êã~åÇáá NN

`çêíÉñ=j~Öåçäá~É=lÑÑáÅáå~äáë ON

cäçë=j~Öåçäá~É PR

eÉêÄ~=aÉëãçÇáá=píóê~ÅáÑçäáá QR

eÉêÄ~=béÜÉÇê~É RT

o~Çáñ=^ÅÜóê~åíÜáë=_áÇÉåí~í~É SV

o~Çáñ=^Åçåáíá=mê~Éé~ê~í~ TV

o~Çáñ=^åÖÉäáÅ~É=mìÄÉëÅÉåíáë UV

o~Çáñ=^ìÅâä~åÇá~É NMN

o~Çáñ=_ìéäÉìêá NNP

o~Çáñ=`çÇçåçéëáë NOR

o~Çáñ=Éí=oÜáòçã~=dÉåíá~å~É NQN

o~Çáñ=Éí=oÜáòçã~=däóÅóêêÜáò~É NRR

o~Çáñ=Éí=oÜáòçã~=oÜÉá NSV

o~Çáñ=m~Éçåá~É=^äÄ~ NUT

o~Çáñ=m~Éçåá~É=oìÄê~ NVT

o~Çáñ=mä~íóÅçÇá ONN

o~Çáñ=mçäóÖçåá=jìäíáÑäçêá OOP

o~Çáñ=p~éçëÜåáâçîá~É OPR

oÜáòçã~=`Üì~åñáçåÖ OQT

oÜáòçã~=`áãáÅáÑìÖ~É ORT

oÜáòçã~=`çéíáÇáë OST

oÜáòçã~=`ìêÅìã~É OUN

oÜáòçã~=Éí=o~Çáñ=kçíçéíÉêóÖáá OVT

I - 1

Index II Chinese Names

`ÜáåÉëÉ=k~ãÉë m~ÖÉ=kçK

�� NSV

�� NN

�� OQT

�� ORT

�� NMN

�� SV

�� NRR

�� NUT

�� OOP

�� NVT

�� PR

�� OPR

�� OVT

�� ON

�� ONN

�� NNP

�� RT

�� OUN

�� OST

�� TV

�� ! QR

�� UV

�� NQN

�� NOR

I - 2

`ÜáåÉëÉ=mÜçåÉíáÅ=k~ãÉë m~ÖÉ=kçK

_~áëÜ~ç NUT

`Ü~áÜì NNP

`ÜáëÜ~ç NVT

`Üì~åãìíçåÖ NN

`Üì~åñáçåÖ OQT

a~Üì~åÖ NSV

a~åÖëÜÉå NOR

aìÜìç UV

bòÜì OUN

c~åÖÑÉåÖ OPR

d~åÅ~ç NRR

dì~åÖàáåèá~åÅ~ç QR

eÉëÜçìïì OOP

eçìéç ON

eì~åÖäá~å OST

gáÉÖÉåÖ ONN

içåÖÇ~å NQN

j~Üì~åÖ RT

jìñá~åÖ NMN

káìñá SV

ná~åÖÜìç OVT

pÜÉåÖã~ ORT

uáåóá PR

wÜáÅÜì~åïì TV

Index III Chinese Phonetic Names

I - 3

pÅáÉåíáÑáÅ=k~ãÉë m~ÖÉ=kçK

^ÅÜóê~åíÜÉë=ÄáÇÉåí~í~=_äK SV

^Åçåáíìã=Å~êãáÅÜ~Éäá=aÉÄñK TV

^åÖÉäáÅ~=éìÄÉëÅÉåë=j~ñáãK=ÑK=ÄáëÉêê~í~=pÜ~å=Éí=vì~å UV

^ìÅâä~åÇá~=ä~éé~=aÉÅåÉK NMN

_ìéäÉìêìã=ÅÜáåÉåëÉ=a`K NNP

`áãáÅáÑìÖ~=ÜÉê~ÅäÉáÑçäá~=hçãK ORT

`äÉã~íáë=~êã~åÇáá=cê~åÅÜK NN

`çÇçåçéëáë=éáäçëìä~=Ecê~åÅÜKF=k~ååÑK NOR

`çÇçåçéëáë=éáäçëìä~=k~ååÑK=î~êK=ãçÇÉëí~=Ek~ååÑKF=iK=qK=pÜÉå NOR

`çÇçåçéëáë=í~åÖëÜÉå=läáîK NOR

`çéíáë=ÅÜáåÉåëáë=cê~åÅÜK OST

`çéíáë=ÇÉäíçáÇÉ~=`KvK`ÜÉåÖ=Éí=eëá~ç OST

`ìêÅìã~=âï~åÖëáÉåëáë=pK=dK=iÉÉ=Éí=`K=cK=iá~åÖ OUN

`ìêÅìã~=éÜ~ÉçÅ~ìäáë=s~äK OUN

`ìêÅìã~=ïÉåóìàáå=vK=eK=`ÜÉå=Éí=`K=iáåÖ OUN

aÉëãçÇáìã=ëíóê~ÅáÑçäáìã=ElëÄKF=jÉêêK QR

béÜÉÇê~=ëáåáÅ~=pí~éÑ RT

dÉåíá~å~=êáÖÉëÅÉåë=cê~åÅÜK NQN

dÉåíá~å~=ëÅ~Äê~=_ÖÉK NQN

däóÅóêêÜáò~=áåÑä~í~=_~íK NRR

däóÅóêêÜáò~=ìê~äÉåëáë=cáëÅÜK NRR

iáÖìëíáÅìã=ÅÜì~åñáçåÖ=eçêíK OQT

j~Öåçäá~=ÄáçåÇáá=m~ãéK PR

j~Öåçäá~=çÑÑáÅáå~äáë=oÉÜÇK=Éí=táäëK ON

j~Öåçäá~=çÑÑáÅáå~äáë=oÉÜÇK=Éí=táäëK=î~êK=ÄáäçÄ~=oÉÜÇK=Éí=táäëK ON

kçíçéíÉêóÖáìã=áåÅáëìã=qáåÖ=Éñ=eK=qK=`Ü~åÖ OVT

m~Éçåá~=ä~ÅíáÑäçê~=m~ääK NUTI=NVT

m~Éçåá~=îÉáíÅÜáá=ióåÅÜ NVT

mä~íóÅçÇçå=Öê~åÇáÑäçêìã=Eg~ÅèKF=^K=a`K ONN

mçäóÖçåìã=ãìäíáÑäçêìã=qÜìåÄK OOP

oÜÉìã=çÑÑáÅáå~äÉ=_~áääK NSV

oÜÉìã=é~äã~íìã=iK NSV

oÜÉìã=í~åÖìíáÅìã=j~ñáãK=Éñ=_~äÑK NSV

p~éçëÜåáâçîá~=Çáî~êáÅ~í~=EqìêÅòKF=pÅÜáëÅÜâK OPR

Index IV Scientific Names

I - 4

`ÜÉãáÅ~ä=oÉÑÉêÉåÅÉ=pìÄëí~åÅÉë m~ÖÉ=kçK

OI=PI=RI=Q’ J=íÉíê~ÜóÇêçñóëíáäÄÉåÉJOJlJβJaJÖäìÅçëáÇÉ OORI=OOUI=OOVI=OPMI=OPOI=OPP

Q’lJβJaJÖäìÅçëóäJRJlJãÉíÜóäîáë~ããáåçä OPTI=OQMJOQR

^ÅçåáíáåÉ ^JPUI=^JPVI=US

^äçÉJÉãçÇáå NTUI=NTVI=NUNI=NUPJNUR

^êáëíçäçÅÜáÅ=~ÅáÇ=f ^JPSI=^JPTI=NV

_ÉåòçóäãÉë~ÅçåáåÉ UNI=UPJUT

_ÉêÄÉêáåÉ=ÅÜäçêáÇÉ OSVI=OTQJOTSI=OTUI=OTV

`ÜêóëçéÜ~åçä NTUI=NTVI=NUNI=NUPJNUR

`çäìãÄá~åÉíáå=~ÅÉí~íÉ VNI=VQI=VS

`çëíìåçäáÇÉ NMPI=NMSJNNN

aÉÜóÇêçÅçëíìë=ä~ÅíçåÉ NMPI=NMSI=NMUJNNN

bÅÇóëíÉêçåÉ TQJTS

bJÑÉêìäáÅ=~ÅáÇ OQVI=OROI=ORQ

bãçÇáå NTUJNUNI=NUPJNURI=ORRI=OOUJOPM

béÜÉÇêáåÉ=ÜóÇêçÅÜäçêáÇÉ SOJSQI=SSI=ST

c~êÖÉëáå PTI=QMI=QO

dÉåíáçéáÅêáå NQUJNRMI=NROJNRQ

dÉêã~ÅêçåÉ OVMJOVOI=OVQJOVS

däóÅóêêÜáòáÅ=~ÅáÇ NRTI=NSOJNSQI=NSSJNSU

eÉëéÉêáÇáå NSJNU

eçåçâáçä OQI=OVI=PNI=PPI=PQ

eóé~ÅçåáíáåÉ ^JPUI=^JPVI=US

fëçÑÉêìäáÅ=~ÅáÇ ORVI=OSOI=OSPI=OSRI=OSS

fëçáãéÉê~íçêáå OVVI=PMOJPMS

fëçîáíÉñáå QTI=ROJRS

iáèìáêáíáå NRTI=NSOJNSQI=NSSJNSU

içÄÉíóçäáå NPQJNPSI=NPUJNQM

j~Öåçäáå PTI=QMJQQ

j~Öåçäçä OQI=OVJPNI=PPI=PQ

jÉë~ÅçåáíáåÉ ^JPUI=^JPVI=US

kçíçéíÉêçä OVVI=PMOI=PMQ

läÉ~åçäáÅ=~ÅáÇ NPI=NSI=TNI=TQI=TTI=TU

lëíÜçäÉ VNI=VQJVSI=VUI=VV

m~ÉçåáÑäçêáå NUVI=NVOJNVSI=NVVI=OMQJOMSI=OMUI=OMV

Index V Chemical Reference Substances

I - 5

Index V Chemical Reference Substances

`ÜÉãáÅ~ä=oÉÑÉêÉåÅÉ=pìÄëí~åÅÉë=EÅçåíáåìÉÇF m~ÖÉ=kçK

m~äã~íáåÉ=ÅÜäçêáÇÉ OSVI=OTQI=OTSI=OTUI=OTV

mÜóëÅáçå NTVI=NUNI=NUPJNURI=OORI=OOUJOPM

mä~íóÅçëáÇÉ=b ONPI=ONSJOON

mêáãJlJÖäìÅçëóäÅáãáÑìÖáå OPTI=OQMI=OQOJOQR

mëÉìÇçÉéÜÉÇêáåÉ=ÜóÇêçÅÜäçêáÇÉ SPI=SQI=SSI=ST

oÜÉáå NTUI=NTVI=NUNI=NUPJNUR

p~áâçë~éçåáå=^ NNRI=NNUI=NOMI=NOOI=NOP

p~áâçë~éçåáå=` NNRI=NNU

p~áâçë~éçåáå=a NNRI=NNUI=NNVI=NOM

wJäáÖìëíáäáÇÉ OQVI=OROJORS

I - 6

appendices - cmro.gov.hk

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