HPLC Determination of Synthetic Pyrethroid Insecticides in Shampoos and Lotions Application

Environmental, Consumer Products

Authors
Coral Barbas and Luis Saavedra Facultad de CC Experimentales y de la Salud Universidad San Pablo-CEU Urbanizacin Monteprncipe, Boadilla del Monte, 28668 Madrid, Spain Andre Dams* Agilent Technologies, Inc. Amstelveen The Netherlands Ronald E. Majors Agilent Technologies, Inc. 2850 Centerville Road Wilmington, DE 19808-1610 USA
*Present address: Andre Dams Analytical Consultancy, Amstelveen The Netherlands

Introduction
Permethrin (Figures 1A and 1B) is a pyrethroid insecticide that is a chlorinated, synthetic form of pyrethrum, a popular natural insecticide derived from chrysanthemum flowers. Permethrin has multiple uses as an insecticide in household insect foggers, tick and flea spray for yards and pets, as a spray for agricultural and livestock products, and for mosquito abatement. When combined with piperonyl butoxide (Figure 1C), a synergist that is used to enhance the affect of a wide variety of insecticides, permethrin formulations are widely used to combat head-lice and are available in the form of a liquid, gel, or shampoo. Recently, the increase of opportunistic infections in HIV-infected patients, like scabies, has lead to an increasing interest in formulations of these substances for therapy. Thus, the analysis of permethrin and it's synergist, piperonyl butoxide (PBO), is important at the formulation level as well as the trace level.

Abstract
The separation of synthetic pyrethroids, cis- and transpermethrin, and the synergist piperonyl butoxide in shampoo and lotion was easily accomplished by dilution, filtration, and injection into a ZORBAX Eclipse XDB-C18 RPC HPLC column. All components could be separated to baseline with symmetrical peaks using a water-methanol isocratic mobile phase.

A
H 3C C H3 O O H H O

Chemical Properties Permethrin has two diastereomers that possess different chemical, physical, and toxicological properties. Both permethrin and PBO are rather hydrophobic and have no polar functional groups. They are very insoluble in water but soluble with water-miscible organic solvents. They can be separated by reversed phase chromatography (RPC) without the use of buffers. Depending on the matrix, however, pH control could be useful to resolve matrix components from the insecticides. Chromatographic Method and Application
O

Cl Cl

cis-permethrin

B
Cl Cl H 3C C H3 H H O O

trans-permethrin

O O

H3C

Piperonyl butoxide

CH3

Figure 1.

Chemical structures for cis-permethrin (A), trans-permethrin (B), and piperonyl butoxide (C)

In the present example, the analysis of these compounds in shampoo and lotion presented no unusual matrix effects, so sample preparation merely involved dilution of the sample with methanol, followed by filtration through a 0.45-m membrane filter, and injection into the HPLC column. Figure 2 shows the RPC separation of the three standards; all three compounds are well separated isocratically on a 4.6 150-mm ZORBAX Eclipse XDB-C18 column with 3.5-m particles, operated at 35 C. Using a water-methanol mobile phase, peaks were quite symmetrical and the separation time was just over 6 minutes. A diode array detector (DAD) was used at 272 nm for detection. Small amounts of unidentified impurities are observed in Figure 2. Experimental conditions are listed in Table 1.

mAU 140 120 100 80 60 40 20 0 0 1 2 3

Piperonyl butoxide

trans-permethrin

cis-permethrin

min

Figure 2.

Chromatogram of permethrin and piperonyl butoxide standards.

Table 1. Experimental Conditions Instrument: Agilent Series 1100 Column: Mobile phase: Flow rate: Temperature: Detector: Injection volume: Standards: Preparation of standards: ZORBAX Eclipse XDB-C18, 4.6- 150-mm, 3.5-m Water-Methanol (10:90 V/V) (G1311A, Quaternary pump) 1 mL/min 35 C (G1316A Column Oven) Series 1100 Diode Array Detector (DAD) (G1315A), 272 nm (10): Ref. 360 nm (100) 20 L Permethrin = 0.064 mg/mL; PBO = 0.24 mg/mL Dissolve 40 mg of each permethrin and 120 mg piperonyl butoxide in 100-mL methanol. Dilute 4 mL to 25 mL with methanol. Dissolve 160 mg of either shampoo or lotion in 25-mL methanol followed by filtration through a 0.45-m membrane filter.

Preparation of samples:

The HPLC chromatogram of a diluted and filtered sample of lotion containing the permethrin cisand trans-diastereomers and the PBO is shown in Figure 3. The chromatogram is rather clean, with only a small additional unknown peak eluting shortly after the void volume (1.4 min). A shampoo extract, shown in Figure 4, revealed the presence of another compound eluting after the PBO, and some enhanced level of lightly retained compounds just after the void volume.

mAU

Piperonyl butoxide

150

125

100

75

50

trans-permethrin
25

cis-permethrin

0 0 1 2 3 4 5 6 7 8 9 min

Figure 3.

Determination of permethrin and piperonyl butoxide in lotion.

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mAU

Piperonyl butoxide
70

60

50

40

30

trans-permethrin

20

10

cis-permethrin

0 0 2 4 6 8 min

Figure 4.

Determination of permethrin and piperonyl butoxide in shampoo.

Conclusions
The separation of synthetic pyrethroids, cis- and trans-permethrin, and the synergist PBO in shampoo and lotion was easily carried out by dilution, filtration, and injection into a C18 RPC HPLC column. All components could be separated to baseline with symmetrical peaks using a water-methanol isocratic mobile phase.

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Agilent shall not be liable for errors contained herein or for incidental or consequential damages in connection with the furnishing, performance, or use of this material. Information, descriptions, and specifications in this publication are subject to change without notice. Agilent Technologies, Inc. 2005 Printed in the USA October 14, 2005 5989-3633EN