This document establishes quality standards for prepared, unprocessed vanilla of the species Vanilla fragrans (Salisbury) Ames syn. Vanilla planifolia Andrews in all its forms: beans, bulk, cut, cuts, seeds, and powder.

• Bean: Commercial term for the whole or partial fruit of the vanilla orchid; botanically a capsule.
• Bulk Vanilla: A mixture of vanilla beans and cut vanilla, classified according to Section 5.
• Crosse (or Crochet): The peduncular end of the vanilla bean.
• Cuts: Intentionally cut vanilla beans, with or without splits, of varying lengths, including whole beans less than 10 cm long, free of foreign matter and mold, excluding vanilla from the QCP process.
• Cured Vanilla: Green vanilla that has undergone appropriate treatment to develop its aroma.
• Cut Vanilla Beans: Whole beans, potentially split, with the crosse intentionally removed.
• Fermented or Sour Vanilla: Cured vanilla prepared with excess water, which can give the beans a fermented fruit-like odor.
• Frosted Vanilla: Vanilla bearing crystals of naturally exuded vanillin.
• Moldy Vanilla: Vanilla bearing or having borne mold and exhibiting a characteristic musty odor.
• Oxidized Vanilla: Vanilla with black dots or patches and a characteristic “iron” odor.
• Phenolated Vanilla: Vanilla with a phenolic (medicinal) odor.
• QCP (Quick Curing Process): Controlled and accelerated physical treatment and drying process for cut vanilla beans.
• Rague (or Gall): Raised scar of a different color than the bean itself, excluding the brand mark.
• Re-cured Vanilla: Cured vanilla that has been re-heated to eliminate defects such as mold or oxidation.
• Red Streaks: Fine longitudinal lines of reddish-brown color.
• “Snail-like” Vanilla: Vanilla with depressed scars, from various causes, commercially classified as ragues (or galls).
• Sound Vanilla: Vanilla that has not undergone any alteration due to natural or artificial circumstances, such as mold, insect attacks, injuries, or diseases.
• Split Vanilla: A bean partially open longitudinally from the stem end along the capsule’s dehiscence line.
• Spot: Localized marking with a color or shine different from the normal appearance.
• Stem End: The end of the vanilla bean opposite the crosse.
• Vanilla Powder: Product obtained by grinding dried, sound vanilla beans, without any additions.
• Vanilla Seeds: Seeds extracted from vanilla beans, classified into two types:
  • Type 1: Greasier, more gelatinous, and moist.
  • Type 2: Drier seeds recovered during processing.
    (Note: This does not include seed residue obtained after bean extraction.)
• Woody Vanilla: Severely dried, rigid, and brittle vanilla with often uneven reddish streaks.

Vanilla is classified into six (6) presentations:

• 3.1. Vanilla Beans: Whole beans, potentially split.
• 3.2. Cut Vanilla Beans: Whole beans, potentially split, with the crosse intentionally removed.
• 3.3. Bulk Vanilla: A mixture of vanilla beans and cut vanilla, classified according to Section 5.
• 3.4. Vanilla Cuts: Intentionally cut vanilla beans, with or without splits, of varying lengths, including whole beans less than 10 cm long, free of foreign matter and mold, excluding vanilla from the QCP process.
• 3.5. Vanilla Powder: Product obtained by grinding dried, sound vanilla beans, without any additions.
• 3.6. Vanilla Seeds: See definition in Section 2.

• 4.1. Vanilla Beans (Whole or Cut):
  • Must be derived from the vanilla species specified in Section 1.
  • Must have undergone appropriate treatment to develop their aroma.
  • Must have a maximum moisture content conforming to their quality category.
  • May be frosted or not and may bear a brand mark.
  • Must not have undergone treatment that could modify their natural vanillin content or other flavor constituents.
  • Must not be infested with mites, moldy, creosoted, oxidized, “poiquées” (damaged crosse), or fermented.
  • Must not present an odor uncharacteristic of vanilla.
  • Must not be re-cured.
• 4.2. Vanilla Cuts:
  • Must be derived from vanilla beans meeting the characteristics mentioned in 4.1.
  • Must be of varying lengths, including those less than 10 cm long.
  • Must be sound and possess a good, characteristic flavor.
  • Must have a dark brown or reddish color.
• 4.3. Bulk Vanilla:
  • Must be derived from vanilla beans meeting the characteristics mentioned in 4.1.
  • Must be sound and possess a good, characteristic flavor.
  • Must have a dark brown or reddish color.
• 4.4. Vanilla Powder:
  • Must have a brown or dark brown color.
  • Must have a pure and pronounced vanilla flavor.
  • Must not contain any food additives or foreign matter.
  • Must not present a musty, creosote, or other foreign odor.
• 4.5. Vanilla Seeds:
  • See definition in Section 2.

The following classifications apply to both split and non-split beans:

CATEGORY	SUB-CATEGORY	Moisture Content (%)	Vanillin Content (%) (On Dry Matter)	Length (cm)
GOURMET	EXTRA BLACK	36-38	>1.8	>16
	BLACK I	34-36	>1.6	>14
	BLACK II (TK)	30-34	>1.5	>14
EXTRACTION	RED I	25<x<29	>1.4	– grade 1: + de 13 cm) – grade 2: 10, 11 et 12 cm)
	RED II	18<x≤25	>1.4
	RED III	<18	>1.4
	CUTS	<18	>1	<10
POWDER		<15	>1
SEEDS	TYPE I	>25	>1
	TYPE II	<25	>0.2

Vanillin content is influenced by various factors, including soil and climate conditions, and bean maturity. Beans with a content exceeding 2.4% should undergo stricter controls due to potential adulteration with synthetic vanillin.

Sampling methods and techniques are detailed in Appendix D of this standard.

Methods for determining vanillin and moisture content are provided in Appendices B and C of this standard.

8.1. Packaging

• 8.1.1. Vanilla Beans:
  • Beans must be bundled in packages of the same length, tied in the middle with a single tie. A second tie is permitted at the split end for split beans.
  • Bundles must be placed in clean, sound, airtight food-grade packaging material that does not affect the product, such as tinplate boxes, gusset bags, or vacuum-sealed bags, in accordance with current regulations.
  • Beans must be stable as defined in Section 5 of this document.
  • Each primary package must be lined with sulfurized or waxed paper, contain a net weight of 5 to 25 kilograms, and be labeled with the vanilla type and net weight.
  • Each primary package must contain vanilla beans of the same variety and category.
• 8.1.2. Cut Vanilla Beans:
  • Cut vanilla beans should be presented in bundles of the same length when long enough and in bulk when bundling is not possible.
  • They must be placed in clean, sound, airtight food-grade packaging material that does not affect the product, such as tinplate boxes, gusset bags, or vacuum-sealed bags, in accordance with current regulations.
  • Each primary package must be lined with sulfurized or waxed paper, contain a net weight of 5 to 25 kilograms, and be labeled with the vanilla type and net weight.
  • Each primary package must contain vanilla beans of the same variety and category.
• 8.1.3. Bulk Vanilla:
  • Bulk vanilla must be placed in clean, sound, airtight food-grade packaging material that does not affect the product, such as tinplate boxes, gusset bags, or vacuum-sealed bags, in accordance with current regulations.
  • Each primary package must be lined with sulfurized or waxed paper, contain a net weight of 5 to 25 kilograms, and be labeled with the vanilla type and net weight.
  • Each primary package must contain vanilla beans of the same variety and category.
• 8.1.4. Vanilla Cuts:
  • Vanilla cuts must be placed in bulk in clean, sound, airtight food-grade packaging material that does not affect the product, such as tinplate boxes, gusset bags, or vacuum-sealed bags, in accordance with current regulations.
  • They must originate from vanilla beans of the same variety.
• 8.1.5. Vanilla Powder:
  • Vanilla powder must be placed in clean, sound, airtight food-grade packaging material that does not affect the product, such as gusset bags or vacuum-sealed bags, in accordance with current regulations.
• 8.1.6. Vanilla Seeds:
  • Vanilla seeds must be placed in clean, sound, airtight food-grade packaging material that does not affect the product, such as gusset bags or vacuum-sealed bags, in accordance with current regulations.

8.2. Labeling

• 8.2.1. Vanilla Beans, Cut Vanilla Beans, Bulk Vanilla, and Vanilla Cuts:
  • The following information must be displayed on each package or label:
    • Product name (corresponding to the botanical species)
    • Classification
    • Presentation
    • Country of origin
    • Country of harvest
    • Primary package serial number
    • Gross weight, tare weight, and net weight
    • Year of harvest
    • Any other information requested by the buyer
• 8.2.2. Vanilla Powder:
  • The information described in 8.2.1 must be displayed on each primary package and shipping package.

8.3. Sealing and Stamping

• Vanilla boxes must be strapped with metal bands at both ends of their longest side.
• Each inspected box must be lead-sealed at both ends. The seal placement should pass through both layers of the metal band, the lid, and the side wall of the box diagonally, intersecting the corner formed by the side wall and lid.

Verification will be conducted at the trader’s request and in public or private warehouses, or preparation facilities.

A.0 Introduction

This appendix describes a simplified method for detecting the aromatic constituents of vanilla, suitable for routine laboratory control. It is important to note that other methods exist for this purpose. This method focuses on identifying key aromatic compounds in vanilla, including aldehydes, alcohols, phenolic acids, and the synthetic flavoring ethyl vanillin and its derivatives.

This method applies to vanilla beans and can also be used for:

• Vanilla-flavored products (after extraction of flavor components)
• Vanilla extracts (by directly analyzing the extract)

A.1 Principle

This method utilizes thin-layer chromatography (TLC) to extract and identify the principal aromatic constituents of vanilla. Aldehydes are revealed using hydrazine sulfate, alcohols and phenolic acids using diazotized p-nitroaniline, and anisyl alcohol using concentrated sulfuric acid.

A.2 Reagents

All reagents used should be of recognized analytical grade. Water should be distilled or of equivalent purity.

A.2.1 Elution Solvents

• A.2.1.1
  • Toluene: 90 volumes
  • Dioxane: 25 volumes
  • Acetic acid: 4 volumes
• A.2.1.2
  • Propanol solution, 1 mL/L: 3 volumes
  • Ammonium hydroxide solution, c(NH4OH) = 8 mol/L: 1 volume
• A.2.1.3
  • Chloroform: 9 volumes
  • Ethyl acetate: 1 volume

A.2.2 Reference Solutions

• A.2.2.1 Reference Solution I
  • Vanillin: 0.1 g
  • p-Hydroxybenzaldehyde: 0.1 g
  • Anisaldehyde: 0.1 g
  • Protocatechualdehyde: 0.1 g
  • Piperonal: 0.1 g
  • Ethyl vanillin: 0.1 g
  • Ethanol, 95% (v/v): to make up to 100 mL
• A.2.2.2 Reference Solution II
  • Vanillic acid: 0.1 g
  • p-Hydroxybenzoic acid: 0.1 g
  • Protocatechuic acid: 0.1 g
  • Ethyl vanillic acid: 0.1 g
  • Ethanol, 95% (v/v): to make up to 100 mL
• A.2.2.3 Reference Solution III
  • Vanillic acid: 0.1 g
  • p-Hydroxybenzyl alcohol: 0.1 g
  • Ethanol, 95% (v/v): to make up to 100 mL
• A.2.2.4 Reference Solution IV
  • Anisyl alcohol: 0.1 g
  • Ethanol, 95% (v/v): to make up to 100 mL

A.2.3 Developers

• A.2.3.1 Dingemans Reagent
  • Saturated aqueous solution of:
    • Hydrazine sulfate: 9 volumes
    • Hydrochloric acid, c(HCl) = 4 mol/L: 1 volume
• A.2.3.2 Diazotized p-Nitroaniline
  • Prepare this reagent immediately before use. Mix:
    • 2 mL of a 0.5% p-nitroaniline solution in hydrochloric acid, c(HCl) = 2 mol/L
    • 3-4 drops of a 5% sodium nitrite solution
    • 8 mL of a 20% sodium acetate solution
• A.2.3.3 Potassium hydroxide, approximately 0.5% ethanolic solution.
• A.2.3.4 Concentrated sulfuric acid.

A.2.4 Diethyl ether.

A.2.5 Ethanol, 95% (v/v).

A.2.6 Ethanol, 50% (v/v).

A.2.7 Anhydrous sodium sulfate.

A.3 Apparatus

Standard laboratory equipment, including:

• A.3.1 Airtight grinder.
• A.3.2 Extraction apparatus.
• A.3.3 Separatory funnel.
• A.3.4 Rotary evaporator.
• A.3.5 Micropipettes, 10 μL capacity, or capillary tubes.
• A.3.6 Silica gel TLC plates without fluorescence indicator.
• A.3.7 Cellulose TLC plates without fluorescence indicator.
• A.3.8 Developing chambers for ascending TLC.

A.4 Procedure

A.4.1 Total Extraction

Extract approximately 5 g of finely chopped or ground vanilla in the extraction apparatus (A.3.2) for 6 hours with 100 mL of ethanol (A.2.6). Evaporate the ethanol using a rotary evaporator (A.3.4) at low temperature to obtain approximately 25 mL of extract.

A.4.2 Resin Removal

Add three times the volume of water and hydrochloric acid to the vanilla extract until the solution reaches a pH of approximately 2. Allow the precipitated resins to settle and centrifuge the solution.

A.4.3 Aromatic Constituent Extraction

Transfer the aqueous solution to a separatory funnel (A.3.3). Extract twice using 25 mL of diethyl ether each time. Combine the ether solutions in a second separatory funnel and wash with a few milliliters of water. Discard the wash solution. Dry the ether solution over anhydrous sodium sulfate (A.2.7). Filter and evaporate at room temperature in a glass dish, preferably non-stick, then in a fume hood.

Redissolve the residue in 95% ethanol (A.2.5) and make up to 10 mL or 50 mL if the product is concentrated.

A.4.4 Aldehyde Identification

Spot 1 μL of the sample solution (or commercial vanilla extract) onto a silica gel TLC plate (A.3.6), 2 cm from the bottom edge, using a micropipette (A.3.5). Frame the sample spots with 2 μL of Reference Solution I (A.2.2.1).

Place the plate in the developing chamber ensuring the base of the plate is submerged in approximately 1 cm of the elution solvent (A.2.1.1).

Develop the plate and remove it when the solvent front reaches approximately 12 cm above the baseline. Allow the plate to air dry.

Spray the plate with Dingemans reagent (A.2.3.1).

Aldehydes appear as yellow spots on a white background. However, if the p-hydroxybenzaldehyde content is low, the spot will only be visible as a green color under ultraviolet light.

Record the results in Table 1.

Table 1 – Approximate Rf Values and Spot Colors for Aldehydes

Compound	Approximate Rf	Color in Daylight	Color in UV Light
Protocatechualdehyde	0.12	Yellow	Yellow
p-Hydroxybenzaldehyde	0.40	Yellow	Green
Vanillin	0.61	Yellow	Yellow
Ethyl vanillin	0.75	Yellow	Yellow
Anisaldehyde (Vanilla tahitensis only)	0.97	Yellow	Light blue
Piperonal (depending on the species)	0.87	Yellow	Yellow

Note: Detecting anisaldehyde is useful for differentiating Vanilla tahitensis from Vanilla fragrans.

A.4.5 Alcohol and Phenolic Acid Identification

Spot 5 μL of the sample solution onto a cellulose TLC plate (A.3.7) using a micropipette (A.3.5). Frame the spots with 2 μL of Reference Solution II (A.2.2.2) and 2 μL of Reference Solution III (A.2.2.3).

Place the plate in the developing chamber containing elution solvent (A.2.1.2).

Develop the plate and remove it when the solvent front reaches approximately 12 cm above the baseline. Allow the plate to air dry and ensure complete evaporation of ammonium hydroxide.

Spray the plate with diazotized p-nitroaniline (A.2.3.2). Allow to air dry in a well-ventilated area, then spray with the potassium hydroxide solution (A.2.3.3).

Record the results in Table 2.

A.4.6 Anisyl Alcohol Identification

Spot 5 μL of the sample solution onto a silica gel TLC plate (A.3.6). Frame the spots with 5 μL of Reference Solution IV (A.2.2.4).

Place the plate in the developing chamber containing elution solvent (A.2.1.3).

Develop the plate and remove it when the solvent front reaches approximately 12 cm above the baseline. Allow the solvent to evaporate.

Reveal the spots with sulfuric acid (A.2.3.4). Anisyl alcohol appears as a red spot on a white background with an Rf value of 0.40.

A.5 Test Report

The test report should include the method used and the results obtained. Additionally, it should mention any procedural details not specified in this standard or considered optional, as well as any incidents that may have influenced the results.

The test report should provide all the information necessary for the complete identification of the sample.

Table 2 – Approximate Rf Values and Spot Colors for Alcohols and Phenolic Acids

Compound	Approximate Rf	Color with Diazotized p-Nitroaniline	Color with Diazotized p-Nitroaniline + Potassium Hydroxide
Protocatechuic acid	0.07	Yellow	Violet
Vanillic acid	0.25	Yellow	Red
p-Hydroxybenzoic acid (more abundant in Vanilla tahitensis)	0.31	Yellow	Violet
Ethyl vanillic acid	0.33	Yellow	Violet
Vanillyl alcohol	0.75	Yellow	Red
p-Hydroxybenzyl alcohol	0.82	Blue-gray

B.0 Introduction

This appendix outlines a method for determining the vanillin content of vanilla. This method is also applicable to powdered vanilla derived from the species described in this International Standard.

B.1 Principle

This method involves extracting vanillin from a test portion using ethanol and then quantifying it using ultraviolet (UV) spectrophotometry.

B.2 Reagents

All reagents used should be of recognized analytical grade. Water should be distilled or of equivalent purity.

• B.2.1 Ethanol, 95% (v/v) solution for UV spectrophotometry.
• B.2.2 Sodium hydroxide solution, c(NaOH) = 1 mol/L.
• B.2.3 Vanillin.

B.3 Apparatus

Standard laboratory equipment, including:

• B.3.1 Airtight grinder.
• B.3.2 One-mark volumetric flasks, 100 mL and 250 mL capacities, complying with ISO standards.
• B.3.3 Pipettes, capable of delivering 10 mL, 20 mL, and 25 mL respectively.
• B.3.4 Desiccator, containing an effective desiccant.
• B.3.5 Extraction apparatus.
• B.3.6 Spectrophotometer, suitable for measurements in the UV range.
• B.3.7 Silica cuvettes for spectrophotometry, with a 1 cm path length.
• B.3.8 Weighing bottle, with airtight lid, 25 mL capacity.

B.4 Procedure

B.4.1 Determination of Specific Absorbance of Vanillin

B.4.1.1 Preparation of Standard Solutions

1. Weigh approximately 30 mg of vanillin (B.2.3), previously dried in a desiccator (B.3.4), into a weighing bottle (B.3.8) to the nearest 0.1 mg.
2. Dissolve in approximately 20 mL of ethanol (B.2.1) and transfer quantitatively to a 250 mL volumetric flask (B.3.2). Rinse the weighing bottle several times with ethanol and add the rinsings to the flask.
3. Make up to the mark with ethanol and mix well (Solution A1).
4. Pipette 25 mL of Solution A1 into a 100 mL volumetric flask (B.3.2). Make up to the mark with ethanol and mix well (Solution B1).
5. Pipette 10 mL of Solution B1 into a 100 mL volumetric flask. Add approximately 60 mL of ethanol and 2 mL of sodium hydroxide solution (B.2.2).
6. Mix well. Make up to the mark with ethanol and mix well (Solution C1).

B.4.1.2 Preparation of Reference Solution

Prepare a reference solution by pipetting 2 mL of sodium hydroxide solution (B.2.2) into a 100 mL volumetric flask and filling to the mark with ethanol. Mix well.

B.4.1.3 Measurement

Record the spectrum of Solution C1 against the reference solution (B.4.1.2) from 250 nm to 420 nm using the spectrophotometer (B.3.6) and cuvettes (B.3.7).

B.4.1.4 Calculation

The maximum absorbance should be at 350 ± 3 nm, and its value should be between 0.2 and 0.8.

Draw a baseline starting at approximately 270 nm and extending to 380 nm.

Note the absorbance at the maximum (Amax) and at the baseline at the same wavelength as the maximum (Abase).

Calculate the specific absorbance (E1cm1%) of vanillin using the formula:

     E1cm1% = (100 * (Amax – Abase)) / m   

where:

• m is the mass, in grams, of vanillin used to prepare the solution.

B.4.2 Sample Preparation

B.4.2.1 Vanilla Beans

Grind or chop the sample and mix thoroughly.

B.4.2.2 Powdered Vanilla

Mix the sample thoroughly.

B.4.3 Test Portion

Weigh approximately 5 g of the prepared sample to the nearest 0.01 g.

B.4.4 Extraction

Extract the test portion (B.4.3) in the extraction apparatus (B.3.5) with approximately 200 mL of ethanol (B.2.1) for 16 hours.

Transfer the extract quantitatively to a 250 mL volumetric flask (B.3.2). Rinse the extraction flask several times with small quantities of ethanol and add the rinsings to the volumetric flask (B.3.2).

Make up to the mark with ethanol and mix well (Solution A2).

B.4.5 Preparation of Reference Solution

Prepare a reference solution by pipetting 2 mL of sodium hydroxide solution (B.2.2) into a 100 mL volumetric flask (B.3.2) and filling to the mark with ethanol. Mix well.

B.4.6 Determination

1. Pipette 25 mL of Solution A2 into a 100 mL volumetric flask.
2. Make up to the mark with ethanol and mix well (Solution B2).
3. Pipette 20 mL of Solution B2 into a 100 mL volumetric flask. Make up to the mark with ethanol and mix well (Solution C2).
4. Pipette 10 mL of Solution C2 into a 100 mL volumetric flask.
5. Add approximately 60 mL of ethanol and 2 mL of sodium hydroxide solution (B.2.2).
6. Make up to the mark with ethanol and mix well (Solution D2).
7. Record the spectrum of Solution D2 against the reference solution (B.4.5) from 250 nm to 420 nm using the spectrophotometer (B.3.6) and cuvettes (B.3.7).

B.4.7 Expression of Results

The vanillin content, expressed as a percentage by mass of the sample, is equal to:

     (50,000 * (Amax – Abase)) / (E1cm1% * m)

 

where:

• Amax is the absorbance at the maximum.
• Abase is the absorbance at the baseline at the same wavelength.
• E1cm1% is the specific absorbance of vanillin (see B.4.1).
• m is the mass, in grams, of the test portion used for the extraction.

Note: To express the result on a dry matter basis, take into account the moisture content of the product.

B.5 Test Report

The test report should indicate the method used and the results obtained. Additionally, it should mention any procedural details not specified in this International Standard or considered optional, as well as any incidents that may have influenced the results.

The test report should provide all the information necessary for the complete identification of the sample.

0 Introduction

This International Standard applies to most spices. However, due to their number and diversity, modifications to the method or the selection of a more appropriate method may be necessary in specific cases.

These modifications and alternative methods will be indicated in the International Standards specific to the spices in question.

1 Object and Scope of Application

This International Standard specifies a method for determining the moisture content of spices.

2 References

• ISO 948, Spices – Sampling.
• ISO 2825, Spices – Preparation of a ground sample for analysis.

3 Definition

• Moisture content: The amount of water, expressed as a percentage by mass, that is entrained and collected according to the method specified in this International Standard.

4 Principle

This method determines the amount of water entrained by azeotropic distillation using an organic liquid immiscible with water. The entrained water is then collected and measured in a graduated tube.

5 Reagent

• 5.1 Toluene.

Saturate the toluene by shaking it with a small amount of water and distill it. Use the distillate for the moisture content determination.

Note: Different solvents are used for moisture content determination. Unless otherwise specified in the International Standard specific to the spice in question, toluene should be the solvent used for this determination.

6 Apparatus

• 6.1 Distillation apparatus (a suitable type of apparatus is described in the annex and illustrated in the figure), comprising the following elements connected by ground glass joints:
  • 6.1.1 Flask, short-necked, with a capacity of at least 500 mL.
  • 6.1.2 Reflux condenser.
  • 6.1.3 Receiver, fitted with a graduated tube, placed between the flask and the condenser.
• 6.2 Analytical balance.

7 Sampling

Sample the product according to the method specified in ISO 948.

8 Procedure

8.1 Apparatus Preparation

Clean the entire apparatus with a mixture of potassium dichromate and sulfuric acid to reduce water droplet adherence on the condenser and receiver tube walls. Rinse thoroughly with water and dry completely before use.

8.2 Test Sample Preparation

Prepare the test sample according to the method specified in ISO 2825.

8.3 Test Portion

Weigh approximately 40 g of the test sample (8.2) to the nearest 0.01 g, ensuring that the amount of water collected does not exceed 4.5 mL.

8.4 Determination

1. Quantitatively transfer the test portion (8.3) into the distillation flask (6.1.1) with toluene (5.1). Add enough toluene (approximately 75 mL total) to completely cover the test portion and mix by rotating the flask.
2. Assemble the apparatus and fill the receiver (6.1.3) with toluene by pouring it through the reflux condenser (6.1.2) until it begins to overflow into the distillation flask.
3. If necessary, insert a loose cotton plug at the top of the condenser or attach a small tube containing calcium chloride to prevent atmospheric moisture condensation inside the condenser tube.
4. Wrap the flask and the tube leading to the receiver tube with asbestos to regulate reflux.
5. Heat the flask so that the distillate flow rate is approximately 100 drops per minute.
6. When most of the water has been entrained, increase the distillation rate to approximately 200 drops per minute and continue until no more water collects.
7. Occasionally, during distillation, purge the reflux condenser with 5 mL portions of toluene to rinse any water adhering to the condenser walls.
8. The water in the graduated tube can be separated from the toluene by occasionally passing a copper wire spiral up and down the condenser and graduated tube, causing the water to collect at the bottom of the tube.
9. Continue refluxing until the water level in the tube remains constant for 30 minutes, then stop heating.
10. Wash the condenser with toluene, using the copper wire spiral to remove any adhering water droplets, if necessary.
11. Immerse the tube in water at room temperature for at least 15 minutes or until the toluene layer is clear; then, read the water volume.

9 Expression of Results

The moisture content, expressed as a percentage by mass, is equal to:

     (100 * V) / m

where:

• V is the volume, in milliliters, of water collected.
• m is the mass, in grams, of the test portion.

It is assumed that the density of water is exactly 1 g/mL.

10 Test Report

The test report should indicate the method used and the result obtained. It should also mention any procedural details not specified in this International Standard or considered optional, as well as any incidents that may have influenced the result.

The test report should provide all the information necessary for the complete identification of the sample.

1 Object and Scope of Application

This International Standard specifies a method for sampling spices.

2 Definitions

• 2.1 Consignment: A quantity of spice dispatched or received at one time under a single contract or transport document. It may consist of one or more lots.
• 2.2 Lot: A definite quantity of spice, assumed to have uniform characteristics, formed within the consignment, and from which the quality of the consignment can be estimated.
• 2.3 Increment: A small quantity of spice taken from one point in the lot. A series of increments should be taken from different points within the lot.
• 2.4 Bulk sample: A quantity of spice obtained by combining and mixing all the increments taken from a particular lot.
• 2.5 Laboratory sample: A quantity of spice taken from the bulk sample for analysis or other examination.

3 General

• 3.1 Sampling should be carried out by a person authorized by agreement between the buyer and the seller and, if desired by either party, in the presence of the buyer (or their representative) and the seller (or their representative).
• 3.2 During sampling, preparation, storage, and handling of samples, care should be taken to avoid altering the product characteristics. The following precautions and guidelines should be observed:
  • 3.2.1 Samples should be taken in a sheltered place, not exposed to humid air, dust, or soot.
  • 3.2.2 Sampling equipment should be clean and dry.
  • 3.2.3 Precautions should be taken to protect the samples, the product being sampled, and the sample containers from any possible contamination.

4 Apparatus

The apparatus required is of the following types:

• 4.1 Sampling from bags
  • Specially designed spear probes for bags.
•  
• 4.2 Mixing and reducing
  • Hand scoops and sample dividers.

5 Lot Formation

All packages within a consignment of spice belonging to the same variety, year of production, and grade should constitute a lot.

If the consignment is declared or found to comprise different varieties, grades, or years of production, or if it appears that the lot is heterogeneous, packages containing products with similar characteristics should be grouped, and each group thus formed should constitute a separate lot.

6 Method for Taking Increments

• 6.1 The number (n) of packages to be selected from a lot of stacked packages depends on the size of the lot and should be in accordance with the following table.

Table – Number of Packages to be Selected for Sampling

Lot Size (N)	Number of Packages to be Selected (n)
1 to 5 packages	All packages
6 to 49 packages	5 packages
50 to 100 packages	10% of packages
Over 100 packages	Square root of the number of packages rounded up to the nearest whole number

The packages should, as far as possible, be taken at random from the lot. For this purpose, a table of random numbers, agreed upon by the buyer and the seller, should be used. If such a table is not available, the following procedure should be adopted:

Starting with a randomly selected package, count the packages in the lot 1, 2, 3 … to r, and so on. Select for sampling every rth package thus counted; the value of r is equal to:

     r = N / n

where:

• N is the total number of packages in the lot.
• n is the number of packages to be selected (see table).

If r is a fractional number, its value should be taken as the whole number portion.

• 6.2 When the product is in motion, samples can be taken during loading or unloading of the packages. For this purpose, the number of packages to be selected should also be in accordance with the table. The value of r should be calculated as stated above, and every rth package, counted during loading or unloading, should be selected for sampling.
• 6.3 Take the increments using a suitable sampling instrument from different parts of each selected package.

7 Bulk Sample

Thoroughly mix all the increments taken as described above to obtain the bulk sample. The size of the bulk sample should be at least three times the quantity of the product required to carry out all the tests prescribed in the individual spice specification.

8 Laboratory Sample

Divide the bulk sample into three or more equal parts, depending on the desired number of laboratory samples. Each part thus obtained constitutes a laboratory sample; one of these samples is intended for the buyer and another for the seller. The third sample, bearing the seals of the buyer and the seller (or their representatives) if they are present at the time of sampling, or of the person who sampled the lot, is intended for arbitration in case of a dispute between the buyer and the seller; it should be stored in a place agreed upon by both parties.

9 Packaging and Marking of Samples

• 9.1 Sample Packaging

Laboratory samples should be placed in clean, sound, airtight glass containers or other suitable packaging that does not affect the product. Sample containers should be of such a capacity that they are almost completely filled by the sample. After filling, each sample container should be made airtight by means of a lid or other suitable closure and then sealed so that it is possible to detect if the container has been opened and resealed.

• 9.2 Sample Marking

Laboratory samples should be labeled, and the label should state the following information concerning the sample and full details of sampling, that is:

• Date of sampling.
• Name and address of the person who took the sample.
• Name of the product.
• Grade.
• Variety.
• Year of production.

If any infestation is detected at the time of sampling, this should be noted in the details of sampling on the container holding the sample.

10 Storage and Dispatch of Samples

Laboratory samples should be stored in such a way that the product temperature does not differ greatly from normal atmospheric temperature. Samples that are to be kept for any length of time should be stored in a cool, dark place.

Laboratory samples for which an analysis is required should be dispatched to the laboratory as soon as possible.

11 Sampling Report

If a sampling report is drawn up, it should state, in addition to the usual particulars, the condition of the spice being sampled, the technique employed if different from that described in this International Standard, and any circumstances likely to have influenced the sampling.