Ir. J. agric. Res.

21:75-83, 1982

DENSITY MEASUREMENTS OF GRASS BY TOLUENE DISPLACEMENT

AND AIR COMPARISON PYCNOMETRY

P. B. McNulty and S. Kennedy

Agricultural Engineering Deportment, University College, Upper Merrion Street, Dublin 2

ABSTRACT
The wet and dry densities of the leaves and stems of Italian ryegrass were determined by toluene
displacement and air-comparison pycnometry at 18" + 2"C. The mean plant dry matter density, defined
as the mass of dry matter per unit volume of dry matter, was 1421 kg/m3 1+ 16 SE). The estimated
compact density, defined as the mass of wet material per unit volume of wet material (i.e., excluding
internal leaf and stem pore space) was 1050 kg./m3 at a mean plant moisture content of 83.490 + 0.590 SE
(wet basis). These data were used to interpret the densities to which grass may be compacted under
various conservation regimes.

INTRODUCTION

Compaction of grasses and other forage crops occurs in such processes as baling,
ensilage and pelleting. Compaction normally involves the expulsion of gases in the
external pore space leading to higher density materials. The extent of compaction is
normally characterised by the bulk or package density of the compacted material.
Bulk density is defined as mass of material per unit volume of storage space which
includes the volume occupied by the dry matter, moisture, internal and all external
pore space (Fig. 1). Package denisty is defined as the mass of material per unit
volume of package space which includes dry matter, moisture, internal pore space
and pore space external to the material but within the package. Typical data for
forage materials (Table l) reveal that the method of forage conservation and
subsequent processing (where applicable) have a major effect on density and
therefore on the space requirements for their storage and transportation.
The objective of this study was to determine the wet and dry densities of grasses
and use the data to interpret the densities to which grass may be compacted under
various conservation regimes. The techniques of liquid displacement and air
comparison pyconometry were used to evaluate grass densities.
l5
76 IRISH JOURNAL OF AGRICULTURAL RESEARCH, VOL.21, NO. I, I982

Liquid displacement
The measurement of density by liquid displacement is based on Archimedes'
Principle. This involves weighing the dry or wet grass first in air and then submerged
in a suitable liquid and calculating the grass volume by the weight of the liquid
displaced. Toluene has been recommended (9) as a suitable liquid for the deter-
mination of densities of biological materials.
Typically, density bottles are used in these evaluations and as such it is
recommended that the bottles containing the material plus liquid should be
completely evacuated to remove all gas from the system. This is essential in
measuring the dry density (q.) because it is defined as the mass of dry matter (or
solid material) per unit volume of dry matter (Fie. 1). This is equivalent to the
specific gravity of the material and as such all external or internal pore space must be
eliminated.
Applying the same technique to wet grass raises difficulties in the sense that what
we wish to measure is the unit density (p,,) which is defined as the mass of wet grass
per unit volume of plant material, i.e. per unit volume of dry matter plus moisture
plus internal pore space, if any (Fig. 1). In this regard, it is reasonable to assume that

(a) (b)
o,
\D
PORE SPACE 96 = m/vo
EXTERNAL TO (bulk density)
PACKAGE
EXTERNAL
PORE
EXTERNAL Po = m/v, SPACE
PORE SPACE (package density)
WITHIN PACKAGE

pu
INTERNAL Pu = m/vu
(unit density) INTERNAL
PORE SPACE
PORE
SPACE

01 = m/v,
WATER (compact density)
WATER

vs

DRY MATTER P, = mr/v, ps

(dry density)
DRY MATTER

Fig. l: Density terminology (a) Packaged mqterial (e.g., bales, pellets): (b) unpack-
oged mqterial (e.g., loose hay, silsge). ms : mass of solid or dry matter; m
: ms + mt : mqss of dry matter plus water (ms, the mass of the gaseous
phase is qssumed to be negligible); V : volume.
 McNIJI,TY AND KENNEDY: GRASS DENSITY

TABLE l: Densities and storage space of various forms of grasses (G) and
legumes (L)
Fractional Denth
Bulk Package Storage Storage Data
morsture (m) density den.ity space \Dace
to"t'
content, M (kglm3) tkg'm:) {ml t) 1*''', DM)
Long loose hay (L) NK (-0.2) NR 64 15.6 - 19.5 (1)
Baled hay (L) NK (-0.2) NR 130-220 4.5-7 .74 -s.7-9.6 (1)
Cubed hay (L) 0. l4 NR 400-480 al l< 2.4-2.9 (1)
Chopped hay (L) 0. l5 fop layers 54 18.5 21.8 (2)
0. l5 1.5 74 I 3.5 Is.e (2)
0. l5 3.0 9l 11.0 12.9 (.2)
0. l5 6.0 130 '7.7 e.l (2)
Chopped grass (C) 0.43 l.5b 159 o.J l 1.0 (3)
(zero day) 0.83 l 5b 439 /.) 13.7 (3)
Silaee (G) 0.43 I .5b 234 4. -J 7.s (3)
(day l8) 0.83 l.5b 993 1.0 6. l (3)
Settled silage (G) 0.82 0.91 534 1.9 10.4 (4)
(pit) 2.14 712 1.4 7.8 (1)
Settled silage (G) 0.60 8.4 670 1.5 3.7 (5)
(tower) 9.9 730 1.4 3.4 (5)
Wafers (C) oven dry NR 700- l 000 -2.O-2.8c -2.0-2.8 (6)
Pellets (G) oven dry NR l 140-1300 -1.5-1.8c - 1.5-1.8 (7)
Pellets (L) NK NR up to 1500 down to l.3c (8)

DM = dry matter; NK : not known; NR - not relevant

aFor bales of rectangular cross-section, stacked to exclude pore space external to the package (round bales
of similar package density would require greater storage space).
bAverage densities in experimental silos 2.5 m square and 3.05 m deep (average depth
- 1.5 m).
'Pore space external to the package assumed to be 5090.

internal pore space is present in both the leaves and stems of grasses. For instance,
Nobel (10) indicates that, in general, intercellular air space accounts for about 5090
of the volume of leaves. Unfortunately, no data appear to be available specifically
for grasses. Grass stems have an internode hollow section (ll) and may also have
intercellular air space. The volume occupied by the total interpore space in grass
stems does not appear to be known. However, Hall et al (12) have demonstrated that
the wet cross-sectional area of alfalfa stems was approximately 36Vo of the total
area, i.e., interpore space was about 64Vo of alfalfa stem volume.
As a consequence, if the wet grass plus toluene is completely evacuated in a
density bottle, it will be the compact density, et, r&ther than the unit density, qu,
which will have been measured. The former is defined as the mass of wet grass per
unit volume of dry matter plus moisture (Fig. l). If the material is not completely
evacuated (e.g., Ieaf stomata are closed; non-chopped stems), a density intermediate
between the compact and unit densities will be measured, and this density shall be
termed the apparent wet density.
78 1RISHJOURNALOFACRICULTURALRESEARCH,VOL.2I,NO.I,Igs2

Air comparison pycnometry

An air comparison pycnometer (9, l3) consists basically of two chambers of equal
volume with a piston in each, a valve connecting the two chambers, a differential
pressure indicator, and a digital counter calibrated for volume measurement. When
a sample of unknown volume is placed in the measuring chamber, the position of the
measuring piston will differ from that of the reference piston by a displacement
equivalent to the sample volume when the differential pressure indicator reads zero.
This volume is then read from the digital counter. The standard operating mode
which was used in this study involves pressurising the sample from I to 2 atmos-
pheres (13). As with liquid displacement, it is likely that this technique will measure
the compact rather than unit density provided the internal pore space comes into
equilibrium with the pressurised air in the sample chamber.

EXPERIMENTAL
Grass
Grass was harvested from a 4.5 ha field on the University farm containing Italian
ryegrass, in its second year of growth, as the predominant species. Date of harvest
was 11 July, 1978, I day before the field was cut for a second cut of silage.
Approximately I kg of grass was harvested with a scissors in each of the five random
locations leaving a stubble of 25-50 mm. The average height of the cut grass was
about 600 mm. The grass was sealed in plastic bags, without bending the stems, and
stored at 4"C until required for testing.

Test regime
The grass in each bag was removed from the refrigerator as required and divided
into lots of leaves and stems. These were further divided into two sub-lots which
were then subjected to the following tests (sequential in each sublot using the same
material) at l8o + 2oC.

Sub-lot 1.' Fresh leaves and stems were cut by scissors into approximately 12.7 mm
lengths. The apparent wet density was determined on each sample in duplicate by:
a) Air comparison pycnometry (8 g stem, 6 g leafl using the Beckman Model 930
pycnometer and following the recommended procedure in the standard mode (9, 13).
b) Liquid displacement using toluene and 50 ml density bottles (8 g stem, 5-6 g leaf)
and following the recommended procedure (9).

Sub-lot 2; a) Moisture was determined in duplicate by drying leaves and stems at

103"C for 24 h following standard procedure (14).
b) Dry density was determined in duplicate on the dried and ground material (6-10 g
stem, 5 g leaf) by air-comparison pycnometry as outlined above.
 MCNULTY AND KENNEDY: GRASS DENSITY .79

c) Dry density was determined in duplicate on the dried and ground material (5 g
stem, 3 g leaf) by liquid displacement using toluene and 50 ml density bottles and
following the recommended procedure (9).

RESULTS AND DISCUSSION

Dry density qnd qpparent wet density
Table 2 shows a comparison of the two methods for measuring densities of grass. In
three out of the four cases there was no significant difference between measuring
density by air-comparison pycnometry and by toluene displacement as indicated by
a paired l-test. The mean apparent wet density of the stem, however, was
significantly higher as measured by air comparison pycnometry (1142 + 18 kglm3)
than by toluene displacement (1004 + 42kg/m3).
Table 3 shows that there was no significant difference between the moisture
contents of the leaves and stems of Italian ryegrass. Nor was there any significant

TABLE 2: Comparison of two methods for measuring the mean densities (+ SE)
of the leaves and stems of Italian ryegrass
Fractional Apparent
moisture density Dry density
Methoda Material content (M) (kg,zm3) (kg,um3)

a) TD Stem 0.827 + 0.009 1004 + 42 1381 + 27

b) ACP Stem 0.827 + 0.009 1142 + 18 1419 + 8

r-test a) vs b) df : 9 t:3.22+ 1 - 1.81 NS

c) TD Leaf 0.842 + 0.004 1041 + 39 1460 + 56
d) ACP Leal 0.842 + 0.004 1094 + 19 1424 + t5
l-test c) vs d) dl : 9 I= 1.06 NS 1 : 0.64 NS
ain this and subsequent tables: TD, toluene displacement; ACP, air comparison pycnometry; NS, not
significant; +p(0.05; ++p(0.01.

TABLE 3: Comparison of mean moisture contents and densities (+ SE) of the

leaves and stems of Italian ryegrass
Fractional Apparent
moisture rvet density Dry density
N4aterial Method content (M) (kg/m3) (kg/nr3)

a) Stem ACP 0.827 + 0.009 1142 + l8 1419 + 8

b) Leaf ACP 0.842 + 0.004 109,+ + 19 1421 + t5
/-test a) vs b) df - 9 1.50 NS 1.34** 0.30 NS
c) Stem TD 0.827 + 0.009 t001 + 42 l38l + 27
d) Leaf TD 0.842 + 0.004 1041 + 39 1460 + 56
l-test c) vs d) dl : 9 1.50 NS 0.46 NS 1.03 NS
 IRISH JOURNAL OF AGRICULTURAL RESEARCH, VOL. 21, NO. I, I982

TABLE 4: Estimation of the fractional phase volumes of Italian ryegrass from

composite leaf-stem data (evenly weighted, + SE) in Table 3. Mean
plant moisture content was 0.834 + 0.005.
Fractional phase volumes
Apparent wet
No. of density Dry density Dry matter Liquid Gaseous
Method runs (kg/nr3) (kg,zm3) Vl Vg

a) TD 20 1023 + 28 1421 + 32 0.119 0.854 0.02'7

b) ACP 20 1118 + 14 1422 + 8 0.1 30 0.934 - 0.064
1{est a) vs b) df =19 2.86* 0.01 NS
Overall mean 40 1421 + 16

difference in three out of the four cases between the mean densities of the leaves and
the stems of Italian ryegrass as indicated by a paired l-test. The mean apparent wet
density of the stem (1142 + l8 kglm3) was, however, significantly higher than the
mean leaf density (1094 + 19 kglm3) as measured by air comparison pycnometry.
The slightly higher stem density may have been due to dry matter compositional
differences between stem and leaf. Typically, the stem is higher in carbohydrate and
lower in protein than the leaf (15) and carbohydrate is a denser material than protein
(16, 17). However, it appears more likely that the air comparison pycnometer has
overestimated both the leaf as well as the stem density as suggested by the data in
Table 4. In these cases leaf and stem data were pooled for each method to provide an
estimate of wet and dry plant densities. These data were then used to evaluate the
fractional volumes of the solid (v,), liquid (v1) and gaseous (vr) phases according to
the following equation which has been derived elsewhere (18):

: I : I
ve
-v,-vr -(1 -M)e,/Q,-MQu/Qr. .... . . . 1

where M : fractional moisture content; Qu : unit density; Qs : dry matter density;

ar : liquid or water density : 1000 kglm3. Values of M and q. are given in Table 4
and the only remaining unknown is the unit density, qu. If the material does not have
a gaseous phase or if, as in this study, the gaseous phase is penetrated by the
measuring fluid, equation I may be written as:

ve:o:l-(1 -M)QtlQr-MQt/Qr ....-2-

In this case complete penetration of the measuring fluid is assumed so that the
fractional gaseous volume (v*) is zero and the unit density (pr) is replaced by the
compact density (9,).

Limitations to methods and sources of error

If we assume that the apparent wet density (p) in Table 4 is the compact density (q,)
we can estimate the fractional phase volumes. Toluene displacement data yielded a
 McNULTY AND KENNEDY: CRASS DENSITY 8t

fractional gas volume (vs) 0.027 rather than zero which may be due to
experimental error or, more likely, to intercellular air space that had not been
penetrated by toluene. Air comparison pycnometry data yielded Vg = - 0.064 rather
than zero, indicating, as previously mentioned, that the technique appears to be
overestimating the compact density. Why this should be so is difficult to understand
particularly since the agreement between the dry densities of both leaf and stem as
determined by toluene displacement and air-comparison pycnometry was very good
(Table 2). Ii the wet material exhibits surface activity or if the water vapour is not
balanced between the chambers, errors may occur in the measurement of sample
volume (13). These problems may be overcome by using helium rather than air as the
measuring fluid, but this procedure was beyond the scope of the present study.
Another source of error may arise if the dry density (e, : t+Zt kglm3) has been
underestimated even though the agreement between the two measurement tech-
niques was excellent (Table 4). This proposal has been prompted by the data of
Hundtoft and Wu (19) who reported a dry density of 1870 kg/m3 for alfalfa. This
vaiue is remarkably high since the dry density of carbohydrates, the major con-
stituent of forage materials, is normally in the range of 1500 to 1600 kg/m3 (16, 17).
Applying Q, : 1870 kglm3 rather than 1421 kg/m3 to the data in Table 4 would yield
fractional gaseous volumes of 0.055 and - 0.033 for toluene displacement and air
comparison pycnometry respectively. In the latter case the error resulting in a
negative gas volume has been reduced but not eliminated. Thus, it is more likely that
the negative gas volume has resuited from an overestimation of the compact density
rather than an underestimation of the dry density. It shouid also be noted that
legumes (such as alfalfa) appear to have a higher dry matter density than grasses.
For instance, Tetlow (8) reported that legumes can be compacted to a package
density of 1500 kg,um3 whereas the corresponding figure for grasses is 1400 kg,/m3.

Relevance of results to the storage, transport and handling of conserved forages

The average dry matter density (eJ of Italian ryegrass was 1421 -r 16 kg,/m3 and was
independent of the method of measurement (Table 4). This value represents the
maximum extent to which dried grass may be compacted and tends to confirm the
observation of Tetlow (8) that grass pellets of package density 1400 kglm3 were
extremely difficult to produce. If the maximum compaction could be attained and if
the external pore space in a bed of grass pellets were 5070, the minimum srorage
space requirement would be 1.4 m3ltDM. Estimated values based on density data in
Table I range from 1.5 to 1.8 m3 for grass pellets, lrom 2.0 to 2.8 m3/tDM for grass
wafers and from 2.4 to 2.9 m3,/tDM for cubed hay. These values represent small
storage-space requirements compared with conventional hay or silage storage
(Table l).
ln moist crops such as freshly harvested grass the maximum or compact density
attainable (Q,) may be predicted by rewriting equation 2 and using water and dry
matter densities of 1000 and 1421 kg/m3 respectively:
 82 IRISH JOURNAL OF AGRICULTURAL RESEARCH, VOL. 2I, NO. I , I982

Qt: r/ [Orze,l + (l -M)/e,] ....3.

Substituting a fractional moisture content (M : 0.83) (cf Table 1) into equation 3
yields a compact density of 1050 kg/m3. In model silo studies Messer and Hawkins
(3) found that grass density increased dramatically over 18 days as the fermentation
progressed (Table l). The higher moisture crop reached an average bulk density of
993 kg/m3 which is close to the maximum density attainable at that moisture content
of 1050 kg/m3.
In general, bulk densities as high as 993 kglm3 are not obtained under commercial
conditions either in pit (4) or tower (5) silos even though localised high density
regions at depths greater than 6 m have been reported (20). As such the volume
occupied by pore space and water leads to a relatively high storage-space
requirement for silage dry matter (Table 1). In practice this may be desirable for ease
of unloading for feeding purposes even if the quality of the product should suffer
from the relatively large pore space. More information is required before the
optimum density profile within a silo pit can be established in terms of product
quality and ease of unloading.

CONCLUSTONS

Density measurements on the leaves and stems of ltalian ryegrass at 18" + 2oC have
led to the following conclusions:
1. There were no significant differences between the dry matter densities of the
leaves and stemsof Italian ryegrass as measured by either toluene displacement or
air comparison pycnometry. The mean plant dry matter density was 1421 + 16
kg/m3.
2. It was not possible to measure the unit density of freshly harvested leaves or
stems because their internal pore space was penetrated by the measuring fluid in
'\-?, each method.

3. In measuring the apparent wet or compact density, toluene displacement yielded

a plant mean of 1023 + 28 kg/m3, whereas air-comparison pycnometry yielded
1 1 l8 + 14 kg/m3 at a mean plant moisture content of 83.4 + 0.5 7o . Toluene displace-

ment was the more accurate method in that it only slightly underestimated the true
compact density of 1050 kglm3, whereas air-comparison pycnometry significantly
overestimated the true compact density.
4. The maximum extent to which any forage crop may be compacted must lie
between a water density of 1000 kg/m3 and its dry matter density which
for Italian
ryegrass is 1421 + 16 kglm3.
 McNULTY AND KENNEDY: GRASS DENSITY 83

ACKNOWLEDGMENTS

The authors acknowledge Mr. F. Smith for technical assistance and Quigley
Magnesite Division, Pfizer Chemical Corporation, Dungarvan, for the air
comparison pycnometer.

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Received Novetnber 27, 1981.