Most pre-Quaternary palynology samples are currently prepared by demineralization of the sediment/sedimentary rock matrix using hydrochloric and hydrofluoric acids (HCl and HF respectively). If a consistently effective alternative to this procedure can be developed, palynological processing will be made significantly less hazardous to both laboratory personnel, and to the wider environment. Furthermore, most non-acid processing methods are normally quicker and cheaper than matrix dissolution using acid. Some authors have previously used hydrogen peroxide (H2O2) to extract palynomorphs by the physico–chemical disaggregation of the clay fraction. However, H2O2 is a powerful oxidizing agent and hence can potentially destroy sedimentary organic material, including palynomorphs. A new method using hot H2O2, where exposure of the sample material to the H2O2 is minimized, has been developed. Crushed sample material in a suitable vessel is placed on a hot plate for one minute, treated with 15–30% H2O2 for 10 minutes, then the residue is diluted with cold distilled water. Disaggregated sample material tends to float, and is decanted into a large vessel containing distilled water to further dilute the H2O2. If any undisaggregated sample remains, the procedure is repeated several times if necessary. Relatively indurated sedimentary lithotypes normally require several treatments. The reason for this stepwise treatment is that the organic material is not exposed to H2O2 for sustained periods, thereby reducing the possibility of palynomorph damage/degradation due to oxidation. When the sample matrix has been fully disaggregated, the residue can be further processed as appropriate.
In this study, eight samples of Carboniferous, Jurassic, Paleogene, and Quaternary age were prepared quantitatively using the new H2O2 method. These were all prepared using 30% H2O2. For comparison, they were also prepared quantitatively using HCl/HF and/or sodium hexametaphosphate [(NaPO3)6]. Quantitative preparations allow the concentration of palynomorphs extracted to be determined, and therefore the effectiveness of the techniques used can be compared objectively. The palynomorph residues derived from these three techniques varied markedly. The H2O2 method does not consistently disaggregate all the sample material, particularly the older and more indurated lithotypes. Some evidence of oxidation effects was observed. Two samples of Mississippian mudstone from the U.S.A. were prepared using H2O2 and (NaPO3)6. Both methods produced abundant miospores, however the H2O2 procedure yielded far higher palynomorph concentrations than the (NaPO3)6 technique. Minor degradation of palynomorphs in the H2O2 preparation was noted. The H2O2 and HCl/HF methods were compared directly on a palynomorph-rich sample of Upper Carboniferous mudstone from offshore Scotland. Both preparations produced abundant miospores. The HCl/HF method had significantly higher recovery levels than the H2O2 procedure. It appears that the H2O2 method simultaneously macerates the matrix, and oxidizes any amorphous organic material (AOM) present. In this sample, the HCl/HF residue was relatively rich in AOM. By contrast, the H2O2 preparation is virtually clear of this phytoclast type, which partially obscures palynomorphs. Two samples of the Middle Jurassic Grantham Formation of eastern England were processed using H2O2 and HCl/HF. The two methods produced abundant palynofloras of similar palynomorph concentrations. Two dinoflagellate cyst acmes within the Danian (Paleogene) part of the López de Bertodano Formation of Seymour Island, Antarctica were also tested using H2O2, (NaPO3)6, and HCl/HF. TheH2O2 preparation completely destroyed the dominant taxon, Palaeoperidinium pyrophorum, in one sample. By contrast, the (NaPO3)6 and HCl/HF preparations produced abundant, fully representative palynofloras. In the other sample, the acme of Spinidinium spp. is completely unaffected by the H2O2 preparation procedure. The final sample of this study is an unconsolidated clay of Late Pleistocene age from offshore Scotland. Both the H2O2 and HCl/HF preparations proved similar in both taxonomic content and overall palynomorph yield.
The new method of preparation using hot H2O2 has proved to be extremely effective. In particular, it appears to be superior to the (NaPO3)6 procedure for indurated lithotypes. However care should be taken because H2O2 can destroy certain dinoflagellate cysts and kerogen macerals which are especially susceptible to oxidation. Further development work, and more comparative testing of the H2O2, (NaPO3)6, and HCl/HF procedures, should be undertaken.