SUPPLEMENTARY MATERIAL Rapid monitoring of carvacrol in plants and herbal medicines using matrix-solid phase dispersion and gas chromatography flame ionization detector Marzieh Rashidipour 1, Rouhollah Heydari 2,, Alireza Feizbakhsh 3 and Payman Hashemi 4 1 Young Researchers and Elite Club, Khorramabad Branch, Islamic Azad University, Khorramabad, Iran 2 Razi Herbal Medicines Research Center, Lorestan University of Medical Sciences, P.O. Box 68149-89468, Khorramabad, Iran. 3 Department of Chemistry, Islamic Azad University, Central Tehran Branch (IAUCTB), P.O. Box 14676-86831, Tehran, Iran. 4 Department of Chemistry, Lorestan University, Khorramabad, Iran Correspondence to. Tel: +98-661-3204005; fax: +98-661-3204005. E-mail address: rouhollahheydari@yahoo.com Abstract: Matrix-solid phase dispersion (MSPD) method coupled with gas chromatography flame ionization detector (GC-FID) as a quick and easy extraction technique has been developed to extract carvacrol from plants and herbal medicines. Influence of important parameters on MSPD method efficiency, such as the sorbent material, the ratio of sample to sorbent material, elution solvent, and volume of the elution solvent have been evaluated and optimized. Carvacrol was successfully extracted by diatomaceous earth as sorbent with 350 μL of dichloromethane as elution solvent. The calibration curve showed good linearity (r2=0.9965) and precision (RSD<8.16%) in the concentration range of 0.5–100 μg mL-1 for carvacrol. The limit of detection (LOD) and limit of quantification (LOQ) were 0.1 and 0.5 μg mL-1, respectively. The recoveries were in the range of 74.4–80.5% with RSD values ranging from 8.4 to 9.8%. The reported MSPD extraction method revealed to be simpler and faster than conventional methods used to quantify carvacrol from plants and herbal medicines. Keywords: Carvacrol; Matrix-solid phase dispersion; Herbal medicines; GC-FID 3.0 Relative area 2.5 2.0 1.5 1.0 0.5 0.0 Silica gel C18 DE Figure S1. Effect of the nature of the sorbent on the extraction of carvacrol. Extraction conditions: elution solvent, dichloromethane; elution solvent volume, 350 μL; dispersing solvent, 1,4-dioxan; dispersing solvent volume, 100 μL; sample mass to sorbent ratio; 1:2. 3.0 Relative area 2.5 2.0 1.5 1.0 0.5 0.0 Figure S2. Effect of the nature of the elution solvent on the extraction of carvacrol. Extraction conditions: sorbent, diatomaceous earth; elution solvent volume, 350 μL; dispersing solvent, 1,4dioxan; dispersing solvent volume; 100 μL; sample mass to sorbent ratio; 1:2. 4.0 3.5 Relative area 3.0 2.5 2.0 1.5 1.0 0.5 0.0 350 450 550 650 750 Elution Solvent Volume (μL) 850 950 Figure S3. Effect of the elution solvent volume on the extraction of carvacrol. Extraction conditions: elution solvent, dichloromethane; sorbent, diatomaceous earth; dispersing solvent, 1,4-dioxan; dispersing solvent volume; 100 μL; sample mass to sorbent ratio; 1:2. 3.5 Relative area 3.0 2.5 2.0 1.5 1.0 0.5 0.0 No solvent Dioxane Ethanol Ethyl acetate Dispersing Solvent Figure S4. Effect of dispersing solvent on carvacrol extraction efficiency. Extraction conditions: elution solvent, dichloromethane; elution solvent volume, 350 μL; sorbent, diatomaceous earth; dispersing solvent volume; 100 μL; sample mass to sorbent ratio; 1:2. 3.0 Relative area 2.5 2.0 1.5 1.0 0.5 0.0 50 100 150 200 Dispersing Solvent Volume (μL) Figure S5. Effect of dispersing solvent volume on carvacrol extraction efficiency. Extraction conditions: elution solvent, dichloromethane; elution solvent volume, 350 μL; dispersing solvent, 1,4-dioxan; sorbent, diatomaceous earth; sample mass to sorbent ratio; 1:2. 4.0 3.5 Relative area 3.0 2.5 2.0 1.5 1.0 0.5 0.0 1:01 1:02 1:04 1:08 Sample to Sorbent Ratio (%w/w) Figure S6. Effect of sample to sorbent ratio on the extraction of carvacrol. Extraction conditions: elution solvent, dichloromethane; elution solvent volume, 350 μL; dispersing solvent, 1,4dioxan; dispersing solvent volume; 100 μL; sorbent, diatomaceous earth. Figure S7. GC-FID chromatograms of (A) blank extract and (B) carvacrol standard solution (100 μg mL-1) using optimized MSPD method. Figure S8. GC-FID chromatogram of Satureja khuzestanica extract using optimized MSPD method.