Interpreting past trophic ecology of a threatened alpine parrot, kea Nestor notabilis, from museum specimens.

When ecosystems are under severe pressure or environments change, trophic position and intraspecific niche width may decrease or narrow, signalling that conservation action is required. In New Zealand, alpine and subalpine ecosystems have been extensively modified through farming since 19th-century European settlement, with consequences for indigenous species such as the kea Nestor notabilis. We investigated feather stable isotope values in the kea and predicted a lower trophic position in modern kea populations, to reflect reduced lowland habitat and a mixed diet with more plant material. We predicted that size and sex would influence trophic values in this sexually dimorphic species, with larger birds more likely to have a high protein diet. We examined potential dietary changes in 68 museum collected kea from 1880s to 2000s, first recording accession details including provenance and sex and measuring culmen length. We used bulk carbon and nitrogen stable isotope analyses (BSIAs) of feathers and a further feather subset using compound-specific stable isotope analyses of amino acids (CSIA-AA) to obtain isotopic values and estimate trophic position. BSIA showed δ15 N values in kea feathers declined through time and could indicate that early century kea were highly omnivorous, with δ15 N values on average higher than in modern kea. Variance in δ15 N values was greater after 1950, driven by a few individuals. Few differences between males and females were evident, although females in the south region had lower δ15 N values. There was a tendency for large male birds to have higher trophic values, perhaps reflecting dominant male bird behaviour noted in historical records. Nonetheless, CSIA-AA performed on a subset of the data suggested that variation in BSIA is likely due to baseline changes rather than relative trophic position which may be more homogenous than these data indicate. Although there was more variability in modern kea, we suggest caution in interpretation. Stable isotope data, particularly CSIA-AA, from museum specimens can reveal potential change in ecological networks as well as sexually dimorphic feeding patterns within species. The data can reveal temporal and regional variation in species trophic position and changes in ecosystem integrity to inform conservation decision-making.

Authenticating bioplastics using carbon and hydrogen stable isotopes - An alternative analytical approach.

RATIONALE: A combination of stable carbon (δ13 C) and hydrogen (δ2 H) isotope ratios and carbon content (% C) was evaluated as a rapid, low-cost analytical approach to authenticate bioplastics, complementing existing radiocarbon (14 C) and Fourier transform infrared (FTIR) analytical methods. METHODS: Petroleum- and bio-based precursor materials and in-market plastics were analysed and their δ13 C, δ2 H and % C values were used to establish isotope criteria to evaluate plastic claims, and the source and biocontent of the samples. 14 C was used to confirm the findings of the isotope approach and FTIR analysis was used to vertify the plastic type of the in-market plastics. RESULTS: Distinctive carbon and hydrogen stable isotope ratios were found for authentic bio-based and petroleum-based precursor plastics, and it was possible to classify in-market plastics according to their source materials (petroleum, C3, C4, and mixed sources). An estimation of C4 biocontent was possible from a C4-petroleum isotope mixing model using δ13 C which was well correlated (R2 = 0.98) to 14 C. It was not possible to establish a C3-petroleum isotope mixing model due to δ13 C isotopic overlap with petroleum plastics; however, the addition of δ2 H and % C was useful to evaluate if petroleum-bioplastic mixes contained C3 bioplastics, and PLS-DA modelling reliably clustered each plastic type. CONCLUSIONS: A combined dual stable isotope and carbon content approach was found to rapidly and accurately identify C3 and C4 bio-based products from their petroleum counterparts, and identify instances of petroleum and bio-based mixes frequently found in mislabelled bioplastics. Out of 37 in-market products labelled as bioplastic, 19 were found to contain varying amounts of petroleum-based plastic and did not meet their bio-based claims.

Geographical traceability of Chinese green tea using stable isotope and multi-element chemometrics.

RATIONALE: Deliberate and fraudulent origin mislabeling of Chinese green tea motivated by large price differences often brings significant food safety risks and damages consumer trust. Currently, there is no reliable method to verify the origin of green tea produced in China. Stable isotope and multi-element analyses combined with statistical models are widely acknowledged as useful traceability techniques for many agro-products, and could be developed to confirm the geographical origin of Chinese green tea and, more importantly, combat illegal green tea mislabeling and fraud. METHODS: An analytical strategy combining elemental analyzer/isotope ratio mass spectrometry (EA/IRMS) and inductively plasma coupled mass spectrometry (ICP-MS) with chemometrics tools was used to confirm the origin of green tea grown in the main tea production provinces around China. Stable C, N, H, O isotope ratios and twenty elements were measured to build mathematical discriminant models using unsupervised principal component analysis (PCA) and supervised linear discriminant analysis (LDA). Two main problems: (i) tracing the origin of Chinese green tea from different tea growing provinces (Zhejiang, Shandong, and other provinces); (ii) authentication of high-value Westlake Longjing tea from the Westlake region and surrounding areas in Zhejiang province, were investigated and assessed. RESULTS: The results demonstrated that PCA and follow-up LDA based on stable isotope and multi-element signatures can verify the geographical origin of Chinese green tea from different provinces, and even localized zones in the same province could be distinguishable, with discrimination accuracies higher than 92.3% and 87.8%, respectively. CONCLUSIONS: Geochemical fingerprinting techniques coupled with chemometric tools offer an accurate and effective verification method for the geographical origin of Chinese green tea, providing a promising tool to combat fraudulent mislabeling of high-value green tea.

Two new defatted beef reference materials, CAAS-1801 and CAAS-1802, for carbon and nitrogen stable isotope ratio measurements.

RATIONALE: Isotope reference materials are essential to enable reliable and comparable isotope data across multiple laboratories. Although many reference materials already exist, the best reference materials should mimic the unknown samples, so new reference materials continue to evolve with the development of isotope research in new product areas. METHODS: Two defatted beef reference materials, CAAS-1801 and CAAS-1802, with substantially different δ13 C values (due to difference in dietary intake), have been prepared as reference materials for stable C and N isotope analysis of meat tissue. Homogeneity, and short- and long-term stability tests of these reference materials have been performed. The δ13 C and δ15 N values of both materials were measured for two-point isotopic normalization against international reference materials by elemental analyzer/isotope ratio mass spectrometry (EA/IRMS). A total of nine international laboratories were selected for the joint evaluation. Cochran statistical analysis yielded the values reported here. RESULTS: The defatted beef reference material CAAS-1801 from Heilongjiang province has a δ13 C value of -13.58 ± 0.56‰ relative to VPDB and a δ15 N value of 4.23 ± 0.56‰ relative to N2 in air. The defatted beef reference material CAAS-1802 from Sichuan province has a δ13 C value of -25.03 ± 0.45‰ and a δ15 N value of 4.36 ± 0.69‰. CONCLUSIONS: The two defatted beef reference materials were found to be isotopically stable across a range of ambient temperatures, and to have low volatility and toxicity, which enables them to be useful as stable isotope reference materials in the field of authentication and traceability of meat.

Influence of leaf age, species and soil depth on the authenticity and geographical origin assignment of green tea.

RATIONALE: Stable isotope fractionation occurring during leaf growth provides internal characteristics for identifying the geographical origin, traceability and authentication of tea. Studying the influence of leaf age, species and the relationship with the cultivated soil may reveal previously undocumented stable isotope fractionation mechanisms, and provide a deeper understanding of the physiological isotopic effects on the tractability and authentication accuracy of green tea to combat mislabeling and fraudulent conduct. METHODS: A total of 36 pairs of young (one bud with one leaf) and mature growth (older leaf) samples from two species of Longjing tea (Longjing #43 and Colonial cultivar) and corresponding cultivation soil samples from two different depth layers (0-20 cm and 20-40 cm) were collected in Westlake district, Hangzhou, Zhejiang province, China. Four stable isotope ratios (δ13 C, δ15 N, δ2 H, and δ18 O values) were measured using an elemental analyzer coupled with an isotope ratio spectrometer. Linear correlation and one-way analysis of variance (ANOVA) statistical analyses were performed to investigate isotopic fractionation mechanisms during plant growth, and reflect the dynamic physiological processes from soil to leaf. RESULTS: The carbon and nitrogen isotope ratios (δ13 C and δ15 N values) reflected the absorption, migration and fractionation of carbon dioxide and nitrogenous nutrients during photosynthesis, nutrient uptake, nitrogen fixation and leaf respiration. The water isotope ratios (δ2 H and δ18 O values) reflected the use and fractionation of water by tea plants at different growth stages. CONCLUSIONS: Considerable differences were found for hydrogen and oxygen isotope ratios according to leaf age, revealing complex isotopic fractionation mechanisms and possible interference factors. Leaf maturity effects should be considered, as they will influence the precision and accuracy of models when assigning the geographical origin, traceability and authentication of tea.

Fertilizer Effects on the Nitrogen Isotope Composition of Soil and Different Leaf Locations of Potted Camellia sinensis over a Growing Season.

The nitrogen-stable isotopes of plants can be used to verify the source of fertilizers, but the fertilizer uptake patterns in tea (Camellia sinensis) plants are unclear. In this study, potted tea plants were treated with three types of organic fertilizers (OFs), urea, and a control. The tea leaves were sampled over seven months from the top, middle, and base of the plants and analyzed for the δ15N and nitrogen content, along with the corresponding soil samples. The top tea leaves treated with the rapeseed cake OF had the highest δ15N values (up to 6.6‱), followed by the chicken manure, the cow manure, the control, and the urea fertilizer (6.5‱, 4.1‱, 2.2‱, and 0.6‱, respectively). The soil treated with cow manure had the highest δ15N values (6.0‱), followed by the chicken manure, rapeseed cake, control, and urea fertilizer (4.8‱, 4.0‱, 2.5‱, and 1.9‱, respectively). The tea leaves fertilized with rapeseed cake showed only slight δ15N value changes in autumn but increased significantly in early spring and then decreased in late spring, consistent with the delivery of a slow-release fertilizer. Meanwhile, the δ15N values of the top, middle, and basal leaves from the tea plants treated with the rapeseed cake treatment were consistently higher in early spring and lower in autumn and late spring, respectively. The urea and control samples had lower tea leaf δ15N values than the rapeseed cake-treated tea and showed a generalized decrease in the tea leaf δ15N values over time. The results clarify the temporal nitrogen patterns and isotope compositions of tea leaves treated with different fertilizer types and ensure that the δ15N tea leaf values can be used to authenticate the organic fertilizer methods across different harvest periods and leaf locations. The present results based on a pot experiment require further exploration in open agricultural soils in terms of the various potential fertilizer effects on the different variations of nitrogen isotope ratios in tea plants.

Effects of Water Isotope Composition on Stable Isotope Distribution and Fractionation of Rice and Plant Tissues.

Rice origin authenticity is important for food safety and consumer confidence. The stable isotope composition of rice is believed to be closely related to its water source, which affects its origin characteristics. However, the influence of water availability on the distribution of rice stable isotopes (δ2H and δ18O) is not clear. In this study, three irrigation waters with different isotopic values were used to investigate isotopic water use effects of Indica and Japonica rice, using pot experiments. Under three different water isotope treatments, the δ2H values of Indica polished rice showed significant differences (-65.0 ± 2.3, -60.5 ± 0.8 and -55.8 ± 1.7‰, respectively, p < 0.05) compared to δ13C and δ15N, as did Japonica polished rice. The values of δ2H and δ18O of rice became more positive when applying more enriched (in 2H and 18O) water, and the enrichment effect was higher in rice than in the corresponding plant tissue. In addition, the δ2H and δ18O values of Indica rice leaves decreased at the heading stage, increased at the filling stage, and then decreased at the harvest stage. Japonica rice showed a similar trend. δ2H changes from stem to leaf were more negative, but δ18O changes were more positive, and δ2H and δ18O values from leaf to rice were more positive for both brown and polished rice. The results from this study will clarify different water isotopic composition effects on rice and provide useful information to improve rice origin authenticity using stable isotope-based methods.

A review of recent compound-specific isotope analysis studies applied to food authentication.

Compound-specific stable isotope analysis (CSIA) of food products is a relatively new and novel technique used to authenticate food and detect adulteration. This paper provides a review of recent on-line and off-line CSIA applications of plant and animal origin foods, essential oils and plant extracts. Different food discrimination techniques, applications, scope, and recent studies are discussed. CSIA δ13C values are widely used to verify geographical origin, organic production, and adulteration. The δ15N values of individual amino acids and nitrate fertilizers have proven effective to authenticate organic foods, while δ2H and δ18O values are useful to link food products with local precipitation for geographical origin verification. Most CSIA techniques focus on fatty acids, amino acids, monosaccharides, disaccharides, organic acids, and volatile compounds enabling more selective and detailed origin and authentication information than bulk isotope analyses.. In conclusion, CSIA has a stronger analytical advantage for the authentication of food compared to bulk stable isotope analysis, especially for honey, beverages, essential oils, and processed foods.

Recent advances in Chinese food authentication and origin verification using isotope ratio mass spectrometry.

Over the last decade, isotope ratio mass spectrometry (IRMS) using up to 5 light stable isotopes (13C/12C, 2H/1H, 15N/14N, 18O/16O, 34S/32S) has become more widely applied for food origin verification as well as food authentication in China. IRMS technology is increasingly used to authenticate a range of food products including organic foods, honey, beverages, tea, animal products, fruits, oils, cereals, spices and condiments that are frequently unique to a specific region of China. Compared to other food authenticity and traceability techniques, IRMS has been successfully used to characterize, classify and identify many Chinese food products, reducing fraud and food safety problems and improving consumer trust and confidence. IRMS techniques also provides scientific support to enhance China's strict government regulatory policies. Isotope testing verifies geographical origin labelling of domestic and imported foods, protects and verifies high value foods that are unique to China, and indicates environmentally friendly farming practices such as 'green' or 'organic' methods. This paper reviews recently published Chinese research to highlight the recent advances of IRMS as a regulatory and verification tool for Chinese food products.

Hydrogen and Oxygen Isotope Fractionation Effects in Different Organ Tissues of Grapes under Drought Conditions.

A study of different grapevine tissues and organs (root, stem, leaf, fruit) water isotope fractionation models from high-quality wine grapes produced in the Helan Mountains, a key wine-producing area in northwestern China, was undertaken. Results showed that δ2H values of local groundwater sources were more negative than rivers and precipitation. Soil water δ2H and δ18O values were significantly higher than those of other environmental water sources. Water from the soil surface layer (0-30 cm, δ2H and δ18O values) was more positive than the deeper layer (30-60 cm), indicating that soil water has undergone a positive fractionation effect. δ2H and δ18O values of tissues and organs from different grape varieties followed a similar pattern but were more negative than the local atmospheric precipitation line (slope between 4.1 to 5.2). The 2H and 18O fractionation relationship in grapevine organs was similar, and 18O has a higher fractionation effect than 2H. δ2H and δ18O values showed a strong fractionation effect during the transportation of water to different grape organs (trend of stem > fruit > leaf). This study showed that 18/16O fractionation in grapes is more likely to occur under drought conditions and provides a theoretical basis to improve traceability accuracy and origin protection of wine production areas.

Effects of Light Shading, Fertilization, and Cultivar Type on the Stable Isotope Distribution of Hybrid Rice.

The effect of fertilizer supply and light intensity on the distribution of elemental contents (%C and %N) and light stable isotopes (C, N, H, and O) in different rice fractions (rice husk, brown rice, and polished rice) of two hybrid rice cultivars (maintainer lines You-1B and Zhong-9B) were investigated. Significant variations were observed for δ13C (-31.3 to -28.3‱), δ15N (2.4 to 2.7‱), δ2H (-125.7 to -84.7‱), and δ18O (15.1‱ to 23.7‱) values in different rice fractions among different cultivars. Fertilizer treatments showed a strong association with %N, δ15N, δ2H, and δ18O values while it did not impart any significant variation for the %C and δ13C values. Light intensity levels also showed a significant influence on the isotopic values of different rice fractions. The δ13C values showed a positive correlation with irradiance. The δ2H and δ15N values decreased with an increase in the irradiance. The light intensity levels did not show any significant change for δ18O values in rice fractions. Multivariate ANOVA showed a significant interaction effect of different factors (light intensity, fertilizer concentration, and rice variety) on the isotopic composition of rice fractions. It is concluded that all environmental and cultivation factors mentioned above significantly influenced the isotopic values and should be considered when addressing the authenticity and origin of rice. Furthermore, care should be taken when selecting rice fractions for traceability and authenticity studies since isotopic signatures vary considerably among different rice fractions.

Towards Verifying the Imported Soybeans of China Using Stable Isotope and Elemental Analysis Coupled with Chemometrics.

Verifying the geographical origin of soybeans (Glycine max [Linn.] Merr.) is a major challenge as there is little available information regarding non-parametric statistical origin approaches for Chinese domestic and imported soybeans. Commercially procured soybean samples from China (n = 33) and soybeans imported from Brazil (n = 90), the United States of America (n = 6), and Argentina (n = 27) were collected to characterize different producing origins using stable isotopes (δ2H, δ18O, δ15N, δ13C, and δ34S), non-metallic element content (% N, % C, and % S), and 23 mineral elements. Chemometric techniques such as principal component analysis (PCA), linear discriminant analysis (LDA), and BP-artificial neural network (BP-ANN) were applied to classify each origin profile. The feasibility of stable isotopes and elemental analysis combined with chemometrics as a discrimination tool to determine the geographical origin of soybeans was evaluated, and origin traceability models were developed. A PCA model indicated that origin discriminant separation was possible between the four soybean origins. Soybean mineral element content was found to be more indicative of origin than stable isotopes or non-metallic element contents. A comparison of two chemometric discriminant models, LDA and BP-ANN, showed both achieved an overall accuracy of 100% for testing and training sets when using a combined isotope and elemental approach. Our findings elucidate the importance of a combined approach in developing a reliable origin labeling method for domestic and imported soybeans in China.

Stable isotope and elemental profiles determine geographical origin of saffron from China and Iran.

Origin verification of high-value saffron is essential for fair trade and to protect consumers' interests and rights. A traceability method using elemental content (% C and % N) and stable isotopes (δ13C, δ2H, δ18O, and δ15N) combined with chemometrics was developed to discriminate saffron from Iran and China and classify major domestic production areas in China. Results showed that Iranian samples had lower % C and % N contents but higher δ13C values than Chinese origin saffron, with δ13C acting as an important variable for origin discrimination. Moreover, δ2H and δ13C isotopes were found to be important variables to classify Chinese regional saffron origin. Two supervised pattern recognition models (PLS-DA) developed to classify Iranian and Chinese saffron, and regional Chinese saffron had a discrimination accuracy of 85.0 % and 80.2 %, respectively. These models provide the basis for a new regulatory inspection procedure to verify saffron origin and label claims, minimizing fraudulent mislabeling and adding value to saffron from specific regions.

Geographical origin classification of peanuts and processed fractions using stable isotopes.

This study investigates the use of stable isotopes (C, N, H, and O) to characterize the geographical origin of peanuts along with different peanut fractions including whole peanut kernel, peanut shell, delipidized peanuts and peanut oil. Peanut samples were procured in 2017 from three distinctive growing regions (Shandong, Jilin, and Jiangsu) in China. Peanut processing significantly influenced the δ 13C, δ 2H, and δ 18O values of different peanut fractions, whereas δ 15N values were consistent across all fractions and unaffected by peanut processing. Geographical differences of peanut kernels and associated peanut fractions showed a maximum variance for δ 15N and δ 18O values which indicated their strong potential to discriminate origin. Different geographical classification models (SVM, LDA, and k-NN) were tested for peanut kernels and associated peanut fractions. LDA achieved the highest classification percentage, both on the training and validation sets. Delipidized peanuts had the best classification rate compared to the other fractions.

A comparative authentication study of fresh fruit and vegetable juices using whole juice and sugar-specific stable isotopes.

Not-from-concentrate (NFC) juice has better nutrition, flavor and higher price than reconstituted juice. Accordingly, NFC juice is prone to adulteration and is an ongoing industry problem that has not yet been resolved. Undeclared addition of water and sugar are the main forms of NFC juice adulteration. This paper investigates the carbon and oxygen stable isotope ratios (δ13C and δ18O values) of the bulk juice and different juice components from 21 fruit and vegetable juices, and qualitatively and quantitatively analyzes the addition of water and sugar in NFC juices. The results show that the use of fruit pulp can help to qualitatively and quantitatively indicate the presence of C4 plant sugars in NFC juice, and can reliably detect added C4 plant sugars above 7 %. Sugar-specific isotope analysis (SSIA) technology was used to determine the δ13C values of different sugars (sucrose, glucose and fructose) and carbon content to qualitatively infer C3 plant sugar addition. Pulp extracted from juice had a good linear relationship with the juice water δ18O values (R2 >0.90). The addition of water to NFC juice can also be determined by comparing δ18O values of extraneous water, pulp and filtered juice. Stable isotope technology confirmed NFC juice adulteration of in-market samples using the pulp as an internal reference and was found to be a useful tool to detect adulteration of in-market NFC juice.

Determining the geographical origin and cultivation methods of Shanghai special rice using NIR and IRMS.

Songjiang CPB rice (cultivated by a 'combination of planting and breeding') and Chongming NCFP rice (cultivated by using 'no chemical fertilizers and pesticides') are two high quality special rice products developed for Shanghai urban agriculture. Protecting these products from geographical origin mislabeling or cultivation method fraud are very important. To verify the origin of Songjiang and Chongming rice and discriminate their cultivation methods (CPB or N (non) CPB, NCFP or chemical fertilizers and pesticides (CFP)), NIR and IRMS methods combined with chemometrics were developed. Results showed complete separation of Songjiang and Chongming rice from other geographic origins using NIR combined with PLSDA and LVQNN; CPB and NCPB Songjiang rice could be completely identified using NIR or IRMS, while the optimal identification accuracy rate of NCFP from CFP in Chongming rice was 87.5%. It provided a rapid and robust method to identify the origin and cultivation methods of Shanghai special rice.

Chemometric origin classification of Chinese garlic using sulfur-containing compounds, assisted by stable isotopes and bioelements.

Geographical origin discrimination of agro-products is essential to guarantee food safety and fair trade. Garlic samples cultivated in six provinces or major production regions in China were characterized for stable isotopes (δ13C, δ2H, δ18O, δ15N, and δ34S), bioelemental contents (% C, % N and % S), and sulfur-containing compounds (8 organosulfur components and 2 amino acids). Results showed that many of the 18 analyzed garlic variables had significant differences among production regions. Some sulfur-containing compounds found in garlic from different provinces had a strong correlation with sulfur isotopes, suggesting garlic sulfur isotopes were also affected by geographical origin. Two supervised pattern recognition models (PLS-DA and k-NN) were developed using stable isotopes, elemental contents, and sulfur-containing compounds, and had a discrimination accuracy of 93.4 % and 87.8 %, respectively. Chemometric classification models using multi-isotopes, elements and sulfur-containing compounds provides a useful method to authenticate Chinese garlic origins.

Stable isotope and photosynthetic response of tea grown under different temperature and light conditions.

The stable isotope and photosynthesis response of tea (Camellia sinensis) is determined under different light and temperature conditions. The results showed that isotopes of young tea leaves were more enriched with increasing light intensity (31 ~ 411 µmol m-2∙s-1). However, the value of δ13C and δ15N seemed depleted, while δ2H and δ18O became enriched as temperature increasing from 15 to 35 °C. Significant isotope differences were found in tea leaves harvested between early growth (0 ~ 10 days) and later growth (10 ~ 21 days) periods (p < 0.05). Pearson's correlation showed a negative correlation between isotopes (δ13C, δ15N and δ2H) and photosynthetic parameters (EVAP and CI) ranging from 0.497 to 0.872, under 25 °C/203 µmol m-2∙s-1. But δ18O had a weak correlation with all photosynthetic parameters under the same conditions. These distinctive correlations between isotopes and photosynthetic parameters provide new insights which could be used to predict tea isotope responses arising from subtle seasonal or climate change conditions.

Predicting isoscapes based on an environmental similarity model for the geographical origin of Chinese rice.

The authentication of geographical origin of food is important using stable isotope analysis. However, the isotopic databank is still short of comprehensive. The isoscapes model based on environmental similarity is used for the first time to predict the geospatial distribution of δ13C, δ2H and δ18O in Chinese rice in 2017 and 2018. 794 rice samples in 2017 were used to build isoscapes model. Independent verification shows that the predicted isotope distribution from this new approach is of high accuracy, with a root mean square error (RMSE) of 0.51 ‰, 7.09 ‰ and 2.06 ‰ for δ13C, δ2H and δ18O values for 2017, respectively. Our results indicate that it is possible to predict the spatial distribution of stable isotopes in rice using an isoscapes model based on environmental similarity. This novel strategy can enrich and complement a stable isotope reference database for rice origin identification at regional scale.

Stable Isotope Effects of Biogas Slurry Applied as an Organic Fertilizer to Rice, Straw, and Soil.

Biogas slurry (BS) is now increasingly used for organic rice production in China. However, the isotopic response and fractionation of different BS application rates to characterize organic rice cultivation have not yet been investigated. In this study, different fertilizer treatments were applied to rice paddy soil including urea, BS with five different application rates and a control with no fertilizer added. Multiproxy analyses (% C, % N, δ13C, δ15N, δ2H, and δ18O) of rice, rice straw, and soil were undertaken using elemental analyzer-isotope ratio mass spectrometry. Rice, straw, and soil showed only minor isotopic and elemental variations across all fertilizer treatments except for δ15N. δ15N values of rice and straw became more positive (+6.1 to +11.2‰ and +6.1 to +12.2‰, respectively) with increasing BS application rates and became more negative with urea fertilization (+2.8 and +3.0‰, respectively). The soil had more positive δ15N values after BS application but showed no significant change with different application rates. No obvious δ15N isotopic differences were found between the control soil and soils fertilized with urea. 15N fractionation was observed between rice, straw, and soil (Δrice-soil -2.0 to +4.3‰, Δstraw-soil -1.9 to +5.3‰) and their isotopic values were strongly correlated to each other (r > 0.94, p < 0.01). Results showed that % C, % N, δ13C, δ2H, and δ18O in rice displayed only minor variations for different fertilizers. However, δ15N values increased in response to BS application, confirming that BS leaves an enriched 15N isotopic marker in soil, straw, and rice, indicating its organically cultivated status. Results from this study will enhance the stable isotope δ15N databank for assessing organic practices using different fertilizer sources.