Mikrochim.

Acta 108, 107-111 (1992)

Mikrochimica Acta
9 by Springer-Verlag1992 Printed in Austria

Speetrophotometric Method for Phosphine Residue Determination in Coriander

Keralapura K. Padmanabha and Jois R. Rangaswamy* Infestation Control and Protectants Discipline,Central Food TechnologicalResearchInstitute, 570 013 Mysore, India Abstract. Spectrophotometric method for the determination of phosphine (PH 3) residues in coriander has been developed based on the reaction of phosphine with silver nitrate in 2% aqueous isopropanol. The yellow chromophore formed has an absorption maximum at 430 nm and the linear relation between the absorbances at 430 nm and the concentration of PH 3 is obeyed in the range of 0.02 to 0.17 pg. The method is sensitive with a detection limit of 0.008 #g and can be applied for determination of 0.02 pg/g residue in coriander. Recovery of added PH 3 from a closed system ranges from 96 to 101%. Key words: phosphine residues, coriander, spectrophotometric method.

Phosphine from wheat in closed system was flushed exhaustively with dry nitrogen into chilled ethanolic mercuric chloride solution, and the resultant hydrochloric acid was estimated potentiometrically [1]. Desorbed PH3 residues from wheat was flushed with nitrogen into a trap cooled with dry ice, and the residues were quantitated by gas chromatography (GC) [2, 3]. A mixed indicator strip method for determination of phosphine was employed by Kashi and Muthu [4]. Nowicki [-5] has reported a screening method using GC with flame photometric detection for determining phosphine residues in wheat. Rangaswamy reperted sensitive spectrophotometric methods for determination of PH3 residues in wheat [-6], rice types [7], cashew nuts [8] and coffee seeds [9]. Aqueous silver nitrate solution as reagent was employed for PH3 residue determination in wheat, rice and cashew nuts and 2% aqueous methanolic silver nitrate was employed in the case of coffee seeds. PH3 fumigation of spices such as coriander, pepper and cumin is gaining importance as they are attacked in storage by spice beetles. The present study describes a simple and sensitive method for determination of PH 3 residues in 9coriander based on the reaction of microquantities of PH 3 with AgNO3 in aqueous isopropanol.

* To whom correspondenceshould be addressed

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K . K . Padmanabha and J. R. Rangaswamy

Experimental
Preparation of Reagents
Stock solution of AgNO 3: Crystals of AgNO 3 (AR grade, 150 mg) were dissolved in 50 ml glass distilled water. Two ml of this solution was diluted to 100 ml with distilled isopropanol (BDH grade) so that each ml of the solution contains 60 #g AgNO3. Three ml of this solution was used for development of chromophore with PH 3 in each experiment. Standard phosphine was liberated from Celphos pellets and stored in a gas burette over water.

Preparation of the Standard Graph

Standard linear graph was prepared in the range of 0.02 to 0.17 #g PH 3 as reported earlier [6-9].

Recovery Experiments
First procedure Fumigate 10 g coriander with different doses of PH 3 in 50-ml conical flasks with side spout fitted with rubber septum to permit injection of PH 3 with 50- and 100-#1 Hamilton gas-tight syringe. After 48 h fumigation, without disturbing either the flask or the contents, remove glass stopper of the flask for 15 s to drive out free PH 3 in head space. Quickly add 30 ml working standard aqueous isopropanolic AgNO 3 solution to the flask and replace the stopper tightly. Let the extraction of PH 3 and development of chromophore proceed for 20 rain with shaking at intervals. Filter the extract with Whatman No. 1 filter circles of 9 cm dia. Read the absorbance in Spectronic 21 spectrophotometer against crop control filtrate similarly prepared. Also run blank experiments without coriander in pre-calibrated 50-ml conical flasks at every level of PH 3 used, to assess the extent of absorption of added PH 3 by glass, grease and rubber septum. After 48 h withdraw known aliquots of PH 3 borne air with micro gas-tight syringe and analyse (Table 1).

Second procedure Repeat fumigation of 10 g coriander with different doses of PH 3 as under first procedure. Also carry out crop control and blank experiments. After 48 h fumigation, remove stopper of the flask, and quickly transfer the contents to 50-ml tube containing 30 ml aqueous isopropanolic AgNO3 solution. Tightly stopper the tubes and let stand for 20 rain with shaking at intervals. Read the absorbance of the filtrates as under first procedure (Table 1).

Table 1. Recovery of PH 3 from coriander (average of 8 trials Dosed PH 3, #g 7.2 18.0 36.0 144.0 216.0 360.0 First procedure, #g 0.1640 1.3764 3.1527 9.9600 18.0400 20.4400 + 0.1033 0.3097 0.2971 +_ 1.7459 2.6158 1.7328 Second procedure, #g 0.1570 0.1933 0.4945 0.5700 0.6450 2.2050 0.0202 0.0833 0.2968 0.2697 0.2930 0.9085

Spectrophotometric Method for Phosphine Residue Determination Table 2. Recovery of phosphine by third procedure (average of 4 trials + SD) Dosed 720 1440 2160
PH 3

109

#g

Found in empty flask, pg 734.00 + 8.00 (101.9%) 1419.40 __+18.97 (98.6%) 2081.28 _ 18.82 (96.4%)

Found in head space, #g 250.51 + 17.39 517.06 _+ 31.16 645.40 _ 20.27

Absorbed by coriander, #g 483.5 (65.9%) 902.34 (63.6%) 1435.88 (68.9%)

Table 3. Phosphine residue (pg/g) in fumigated and oneday aired coriander (average of 3 fumigations _ SD) Sampling Pre-aired One-day aired 3 tablets/ton 0.0248 _ 0.0216 0.0042 + 0.0007 6 tablets/ton 0.0378 _ 0.0206 0.0181 __+0.0028

Third procedure Expose 80 g coriander to 1.0 to 3.0 ml PH 3 for two weeks in 250 ml stoppered conical flasks with side spout fitted with rubber septum that permits injection and gas sampling. Inject an equal amount of P H 3 into another similar pre-calibrated empty flask to serve as control. At the end of 2 weeks, determine the concentration of PHa in head space over coriander and in the control flask by withdrawing known aliquots of PH3-borne air. Also determine the void volume of each flask after a i r i n g P H 3 by opening the stopper. Compute the amount of P H 3 taken up by 80 g coriander by subtracting head space concentration from that in the control flask (Table 2).

Residue Analysis of Coriander

Two kilograms of coriander in 3 replicates were fumigated at two doses of 3 and 6 tablets/ton by placing Celphos (18 and 36 mg respectively) in a paper pack underneath the coriander in 5-1 flask. The flasks were closed with gas-tight silicon greased glass stoppers. Nearly 6 and 12 mg of P H 3 should have been produced by 18 and 36 mg Celphos pellet respectively. At the end of one week exposure at room temperature (28-30~ P H 3 residue in pre-aired samples was determined by the proposed method. The remaining coriander was aired by spreading as a thin layer in the open for 24 h and the P H 3 residue was again determined by the proposed method (Table 3). P H 3 residue was determined by extracting 10 g coriander with 30 ml of 2% aqueous isoproponolic AgNO3 (60/~g/ml) solution for 20 min. Filtered and the absorbance was read at 430 nm against the corresponding crop control filtrate as blank. Ten g of unfumigated coriander was similarly extracted and used as blank. The same filtrates were also employed for scanning the absorption spectra in the range of 380-600 nm (Fig. 1).

Results and Discussion

W h e n a q u e o u s A g N O 3 s o l u t i o n w a s u s e d for e x t r a c t i o n o f c o r i a n d e r (10 g + 30 ml), the c r o p c o n t r o l g a v e v e r y h i g h a b s o r b a n c e o f 0.29. H e n c e v a r i o u s o r g a n i c s o l v e n t s c o n t a i n i n g 1 - 2 % w a t e r as c a r r i e r o f A g N O 3 w e r e tried. C r o p c o n t r o l e x t r a c t s w i t h

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0.2 1. crop control 2. crop extract 3. PH3-AgNO 3

K.K. Padmanabha and J. R. Rangaswamy

t
lJ till 0 l/I

oN,

0.1 2

<

5oo Wavelength (nrn)

600

Fig. 1. Absorption maximum of crop control extract, PH3-AgNOa chromophore from coriander and PH3-AgNO 3 chromophore in isopropanol.

CH3 OH, C z H 5OH, ethylacetate showed high absorbance of 0.18 to 0.24. Although C H a C N containing 2~o water and 60 #g AgNO 3 per ml of the solution gave a low absorbance of 0.05 with crop control, the solvent system did not allow interaction of AgNO 3 with PH 3 residue on coriander to form chromophore. Isopropanol containing 2 ~ water and 60 #g AgNO 3 per ml of the solution not only showed a low absorbance of 0.025 with crop control, but also allowed the development of AgNOa-PH 3 residue chromophore. Development ofchromophore was complete at 20 rain both with the standard PH 3 and PH3 residue on coriander. The chromophore was stable for 30 min and thereafter it started to decompose as indicated by the decrease in absorbance. So in all experiments of recovery studies and residue analysis, the absorbance of the extract was read within 30 min. The slightly yellow chromophore formed due to interaction of PH3 with AgNO3 has an absorption maximum at 430 nm (Fig. 1), and the interaction is water dependent as the formation of chromophore did not take place in pure isopropanol. PH 3 residue-AgNO 3 chromophore extracted from coriander has also an absorption maximum at 430 nm, while crop control extract does not have. The linear relationship between absorbance at 430 nm and the concentration of PH 3 is obeyed between 0.01 and 0.17 #g PH 3. Mean values were calculated of 12 replicate determinations and the mean of standard deviations at various points is +0.0045. The minimum weight of PH 3 needed to produce a measurable chromophore is 0.008 #g. Recoveries by the first and second procedures (Table 1) are the amounts of PH3 determined in/on coriander. Unlike with solid or liquid pesticides, complete recovery of fortified fumigant is very difficult to establish due to fugacity of the gas. Some amount of fumigant escapes at every stage of sampling. It is also likely that some portion of PH3 that has settled on coriander might have also escaped when the stopper is removed for 15 s to expel PH3 in the head space (first procedure). The amount of PH3 recovered increases as the dosed PH 3 increases from 7.2 to 360 #g by first and second procedure. If this is taken as an index of the amount of PH3 fortified with coriander, then these values indicate complete recovery of PH 3 fortified suggesting the soundness of the method. As for example, at a dose of 7.2 #g, 0.164 #g PH 3 is retained by the coriander and the same at 360 #g dose is 20.4 #g. Similarly by the second procedure at 7.2 #g dose, 0.157 #g is held back and 360 #g

Spectrophotometric Method for Phosphine Residue Determination

111

dosage the same is 2.20/~g. Comparison of recoveries by these two procedures show that recoveries by the second procedure are lower at any given dose than by the first. This is because of the loss of PH 3 loosely held by coriander seeds as they are disturbed during transfer of coriander to reagent in a Stoppered tube. Within the limits of PH3-holding capacity of coriander, 10 g coriander would hold only a definite amount of PH3 under a given set of conditions including the dosage as shown by the values in Table 1, and this amount has been completely recovered. At all these doses there is 85 to 101% recovery of added PH 3 from empty flasks. The aim of the third procedure (Table 2) is to determine PH3 residue on coriander by computing the head space concentration and the amount of added PH3 remaining after adsorption by glass and grease. Values in Table 2 show 96 to 101% recovery of added PH 3 from empty flasks. By this procedure also the residue on coriander decreases as the dosed PH 3 decreases. The residue on coriander is 250.5 #g at 720 #g dose while the same is 645.4 #g at 2160/tg dose. As seen by the values in parentheses in the last column, about 66% of the available PH3 is taken up by coriander, which represents the PH 3 holding capacity of coriander. PH3 residues on coriander fumigated experimentally at two doses (Table 3) indicate that the method can be applied for determination of PH3 residues as crop extractives from coriander do not interfere with the development of the chromophore. Loss of PH 3 from coriander dosed at 3 Celphos tablets/ton on one-day airing is 83% while the same from that dosed at 6 tablets/ton is 52%.

Acknowledgement. KKP thanks the Director, CFTRI, Mysore, for his interest, CSIR for the award of Junior Research Fellowship and Mr. P. K. Raman for word processing.

References
1-1] [2] [3] [4] 1-5] [6] 1-7] [8] [9] B. Berck, J. Agric. Food Chem. 1968, 16, 415. T. Dumas, J. Assoc. Off. Anal. Chem. 1978, 61, 5. T. Dumas, J. Agric. Food Chem. 1980, 27, 337. K. P. Kashi, M. Muthu, Pestic. Sci. 1975, 6, 511. T. Nowicki, J. Assoc. Off. Anal. Chem. 1978, 61,829. J. R. Rangaswamy, J. Assoc. Off. Anal. Chem. 1984, 67, 117. J. R. Rangaswamy, M. Muthu, J. Assoc. Off. Anal. Chem. 1985, 68, 205". J. R. Rangaswamy, J. Assoc. Off. Anal. Chem. 1988, 71,557. J. R. Rangaswamy, J. Food Sci. Technol. 1990, 27, 33.

Received March 27, 1991. Revision July 11, 1991.